Three-Dimensional Study of the Terminal Portion in Sprague-Dawley Rat Ejaculatory Ducts.

In mammals, a pair of ejaculatory ducts exists in the urethra at the seminal colliculus. The detailed anatomical structures of the distal end of the ejaculatory ducts of Sprague-Dawley rats were investigated by the computer-assisted three-dimensional reconstruction analysis using light-microscopic serial sections. A three-dimensional reconstruction revealed that in adult rats, the ejaculatory sinus pair consists of two parts: the cranial section - a compartment region composed of a fusion of the ampullary gland duct and the seminal vesicle duct, and the caudal section - a grooved region composed of a long slitlike ejaculatory ostium that extends into the urethra on both sides of the seminal colliculus. But the sphincter structure was not observed. The long axis of the compartment region was approximately 58 µm in length, and that of the groove region was approximately 495 µm. Although many epithelial glands ducts were distributed throughout the ejaculatory sinuses, the prostate and coagulation gland ducts did not open in these sinuses. The urethra was composed of transitional epithelium, while the ejaculatory sinuses were composed of single to stratified cuboidal epithelium. The ejaculatory ducts continued to the ejaculatory ostium in male adult Sprague-Dawley rat were composed of the seminal vesicle ducts received the ampullary gland ducts.

Postoperative early hemolytic anemia due to inverted teflon felt strip after emergency repair for type A dissection.

A 39-year-old man underwent emergency surgery for type A acute aortic dissection complicated by paraplegia. However, hemolytic anemia increased significantly due to severe stenosis of the proximal anastomosis one month after surgery. He finally underwent a redo procedure 4 months after the initial operation whereupon it was verified that half of the inner felt strip used for proximal stump fixation had turned up and was protruding into the inner lumen. We report here on a rare case of survival of postoperative early hemolytic anemia due to severe graft stenosis caused by an inverted inner Teflon felt strip without any extra vascular compression.

Apical voltage-driven urate efflux transporter NPT4 in renal proximal tubule.

Uric acid (urate) is the end product of purine metabolism in humans. Human kidneys reabsorb a large proportion of filtered urate. This extensive renal reabsorption, together with the fact that humans do not possess uricase that catalyzes the biotransformation of urate into allantoin, results in a higher plasma urate concentration in humans compared to other mammals. A major determinant of plasma urate concentration is renal excretion as a function of the balance between reabsorption and secretion. We previously identified that renal urate absorption in proximal tubular epithelial cells occurs mainly via apical urate/anion exchanger, URAT1/SLC22A12, and by facilitated diffusion along the trans-membrane potential gradient by the basolateral voltage-driven urate efflux transporter, URATv1/SLC2A9/GLUT9. In contrast, the molecular mechanism by which renal urate secretion occurs remains elusive. Recently, we reported a newly characterized human voltage-driven drug efflux transporter, hNPT4/SLC17A3, which functions as a urate exit pathway located at the apical side of renal proximal tubules. This transporter protein has been hypothesized to play an important role with regard to net urate efflux. An in vivo role of hNPT4 is supported by the fact that missense mutations in SLC17A3 present in hyperuricemia patients with urate underexcretion abolished urate efflux capacity in vitro. Herein, we report data demonstrating that loop diuretics and thiazide diuretics substantially interact with hNPT4. These data provide molecular evidence for loop and thiazide-diuretics-induced hyperuricemia. Thus, we propose that hNPT4 is an important transepithelial proximal tubular transporter that transports diuretic drugs and operates functionally with basolateral organic anion transporters 1/3 (OAT1/OAT3).

Human uric acid transporter 1 (hURAT1): an inhibitor structure-activity relationship (SAR) study.

The current study describes the chemical synthesis of a series of (2-ethylbenzofuran-3-yl)(substituted-phenyl)methanone compounds and their subsequent in vitro testing via oocytes expressing hURAT1. The experimental data support the notion that a potent hURAT1 inhibitor requires an anion (i.e., a formal negative charge) to interact with the positively charged hURAT1 binding pocket. An anion appears to be a primary requirement in order to be a hURAT1 substrate (i.e., urate) or inhibitor. We discuss the inhibitor structure-activity relationship and how electronically donating or withdrawing groups attached to the B-ring can decrease or increase inhibitory potency, respectively.

Expression of non-muscle type myosin heavy polypeptide 9 (MYH9) in mammalian cells.

Myosin is a functional protein associated with cellular movement, cell division, muscle contraction and other functions. Members of the myosin super-family are distinguished from the myosin heavy chains that play crucial roles in cellular processes. Although there are many studies of myosin heavy chains in this family, there are fewer on non-muscle myosin heavy chains than of muscle myosin heavy chains. Myosin is classified as type I (myosin I) or type II (myosin II). Myosin I, called unconventional myosin or mini-myosin, has one head, while myosin II, called conventional myosin, has two heads. We transfected myosin heavy polypeptide 9 (MYH9) into HeLa cells as a fusion protein with enhanced green fluorescent protein (EGFP) and analyzed the localization and distribution of MYH9 in parallel with those of actin and tubulin. The results indicate that MYH9 colocalizes with actin stress fibers. Since it has recently been shown by genetic analysis that autosomal dominant giant platelet syndromes are MYH9-related disorders, our development of transfected EGFP-MYH9 might be useful for predicting the associations between the function of actin polymerization, the MYH9 motor domain, and these disorders.

Relation of the stages of change for exercise behaviors, self-efficacy, decisional-balance, and diet-related psycho-behavioral factors in young Japanese women.

BACKGROUND: A large proportion of young Japanese women is inactive. Exercise has important health benefits, however, abnormal weight/eating concerns and excessive dieting practices among physically active young women also have been reported in many cross-sectional studies. The purpose of this study was to examine the relationships between stages of change for exercise behaviors and exercise/dieting related psycho-behavioral factors using the Transtheoretical Model of behavior change as a theoretical framework. METHODS: A cross-sectional study included 450 young Japanese women aged 18 to 21 (18.4+/-0.67 years). Subjects in precontemplation (n=111, 24.7%), contemplation (n=120, 26.7%), preparation (n=177, 39.3%), action (n=17, 3.8%), and maintenance (n=25, 5.6%) were compared on physique, body composition, current exercise practices, exercise self-efficacy, decisional balance (benefits and costs exercise), as well as dieting behaviors and weight/eating concerns. RESULTS: Stages of change for exercise behaviors were significantly related to exercise self-efficacy and perceived benefits as well as to dieting behaviors and weight/eating concerns. Subjects in the higher stages had higher self-efficacy, perceived benefits of exercise, and healthy dieting behaviors; however, some of them also had unhealthier dietary practices, higher phobia of obesity and obsession with eating than those in lower stages. CONCLUSIONS: These findings provide support for applying the transtheoretical model of exercise behavioral change to Japanese young women. Additionally, it is also important to pay attention to stage specific psycho-behavioral factors related to their dieting.

Atypical membranoproliferative glomerulonephritis in a cat.

Membranoproliferative glomerulonephritis was observed in a 2-year-old male Japanese domestic cat with clinical renal failure. In the glomeruli, moderate mesangial hypercellularity with an increased mesangial matrix and thickening of the capillary walls were prominent. In addition, frequent duplication of the capillary walls, splitting, and spike formation were observed in the glomerular basement membrane. Granular cat IgG and complement component deposition were detected globally along the glomerular capillary walls and in the mesangium. Transmission electron microscopy revealed dense deposits in the subendothelial and subepithelial regions and the mesangium. Mesangial interposition was also observed. These glomerular lesions are also found in humans with membranoproliferative glomerulonephritis type III, which has not been reported in animals.

Prognostic status of p53 gene mutation in canine mammary carcinoma.

BACKGROUND: The p53 gene mutations have been associated with the development of human breast and canine mammary neoplasms; breast carcinoma patients with alterations of p53 gene are considered to have a poor prognosis. Mammary carcinoma represents the most common malignant tumor in female dogs. However, the prognostic significance of p53 gene mutation in the dog has been unclear. STUDY DESIGN: The alteration in exons 5-8 of p53 gene in 69 canine mammary carcinomas were investigated by PCR-SSCP with direct sequence analysis and statistically analyzed to compare with other clinicopathological parameters including age, neuter, tumor size, stage, histology, p53 expression, recurrence and death from carcinoma. RESULTS: 12 out of 69 (17%) carcinomas showed p53 gene mutations. After a follow-up period of 30 months, multivariate regression analysis revealed that p53 gene mutation was only an independent risk factor for increased risk of the recurrence and death from mammary carcinoma. CONCLUSION: The p53 gene alterations might contribute to the prognostic status in canine mammary carcinomas, in a way comparable to that of human tumors.

Hepatic fatty acid-binding proteins of a teleost, Lateolabrax japonicus. The primary structures and location of a disulfide bond.

Two fatty acid-binding proteins (FABP), FABP-1 and FABP-2, were purified from the liver cytosol of the teleost, Lateolabrax japonicus (Japan sea bass), and characterized. The complete primary structure of FABP-2 was determined by protein analysis to be the following: MDFSGTWQVY AQENYEEFLR AMELPADVIK MAKDIKPITE IKQSGNDFVV TSKTPGKTVT NSFTIGKEAD ITTMDGKKIR CVVNLEGGKL VCNTGKFCHI QELRGGEMVE TLTMGSTTLI RKSKKM. Partial peptide sequences of FABP-1 were also determined. Phylogenetic analysis indicates that FABP-2 is a homologue of mammalian hepatic FABP, whereas FABP-1 is most similar to the members of mammalian cardiac FABP subfamily. L. japonicus FABP-2 contains three cysteine residues, and a disulfide bond is identified between Cys-81 and Cys-92. A theoretical model of FABP-2 generated by a homology modeling method indicates close proximity of the two cysteine residues in the three-dimensional structure. This is a rather rare case of cytosolic protein having a disulfide bond under the normally reducing conditions of the cytosol, though the presence or absence of disulfide bonds does not seem to affect the ligand-binding ability.

p53 gene mutations occurring in spontaneous benign and malignant mammary tumors of the dog.

Sixty-three cases of benign and malignant canine mammary tumors were analyzed to define the alteration of exons 5-8 for the p53 tumor suppressor gene using polymerase chain reaction direct sequence analysis with paraffin-embedded tissues. Four missense mutations were found in 38 benign mammary tumors (11%), and five missense (one tumor had two missense mutations) and one nonsense mutations were found in 25 mammary carcinomas (20%). These data suggest that the p53 gene alterations might be initiated at an early stage of canine mammary carcinogenesis and p53 mutations might be associated with malignancy. However, there was no evidence of any relationship between the p53 alterations and the histologic types of tumors or breeds of dogs.

Expression of vascular endothelial growth factor in N-butyl-N-(4-hydroxybutyl)nitrosamine-induced rat bladder carcinogenesis.

Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are proteins implicated in tumor-associated microvascular angiogenesis. Expressions of VEGF and bFGF in various stages of chemical-induced rat bladder carcinogenesis were immunohistochemically investigated. Thirty-two male 6-week-old Wistar rats were given drinking water containing 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) for 20 weeks. VEGF and bFGF were not detected in the normal bladder epithelium. In simple hyperplasia, intensive expression of VEGF was observed in a few epithelial cells, and the expression of epithelial VEGF became more pronounced in papillary or nodular (PN) hyperplasia and papilloma. In carcinoma, heterogeneous expression of VEGF was observed in focal tumor cells, intensely expressed in the invading tumor cells. Ultrastructurally, carcinoma cells showed VEGF immunoreactivity in the cytoplasmic matrix and some rough endoplasmic reticulum, and VEGF-positive and -negative carcinoma cells were also clearly defined. High levels of VEGF mRNA were observed in the carcinoma. However, bFGF was not detected in the epithelium throughout the carcinogenesis. Increased microvessel counts appeared at simple hyperplasia and became more pronounced in PN hyperplasia, papilloma, and carcinoma (F-test; P < 0.05). In the carcinoma, the microvessel counts of the VEGF-expressing tumor areas were significantly higher than that of the non-VEGF-expressing tumor areas (U-test; P < 0.05). The present study suggests that upregulation of epithelial VEGF may begin at a quite early stage in BBN-induced rat bladder carcinogenesis, but bFGF may not be involved.

Mutation, allelotyping, and transcription analyses of the p73 gene in prostatic carcinoma.

A novel gene, p73, encoding a protein with significant homology to p53, was recently identified at 1p36. To investigate penetrance of p73 in prostatic carcinogenesis, mutation, allelotyping, and transcription analyses of p73 were performed in prostatic carcinoma. No types of mutation causing amino acid substitutions or frameshifts were found in 106 cases examined. Loss of heterozygosity in the gene was found in 2 of 38 cases (5.3%). Various expression levels of p73 alpha variant were observed in tumor compared with those in normal tissue. These data suggest that the p73 gene is not playing an essential role, but expression of p73 may associate with tumor growth in prostatic carcinogenesis.

Testicular efferent ductule cyst of a dog.

A 9-year-old male Shetland Sheepdog had a small mass in the left testis. Grossly, the round to oval cyst was present at the upper pole of the testicular parenchyma near the head of the epididymis. Histologically, the cyst was lined by a single layer of nonciliated and ciliated epithelial cells. Immunohistochemically, the epithelial cells of the cyst showed expression of the low- and high-molecular-weight cytokeratins, vimentin, and desmin similar to that of normal efferent ductules in the dog. The testicular cystic dysplasia was thought to originate from the efferent ductules.

p16 (CDKN2/MTS1) gene deletions are rare in prostatic carcinomas in the United States and Japan.

PURPOSE: The incidence of clinically apparent prostatic carcinoma is much higher in the United States than in Japan. Alterations in the p16 tumor suppressor gene have been identified in various tumor types, including cultured prostatic carcinoma cell lines. We studied the possible deletions of either exon 2 or 3 of this gene in primary clinical prostatic carcinomas from Japan and the United States. MATERIALS AND METHODS: Genomic DNA was extracted from 36 formalin-fixed, paraffin-embedded clinical prostatic carcinomas from Japan and 27 carcinomas from the United States. Exons 2 and 3 of the p16 gene were amplified using comparative multiplex polymerase chain reactions (PCR) and then analyzed for possible deletions of either exon. RESULTS: Two out of 36 (5.6%) carcinomas from Japan clearly demonstrated deletion of p16 exon 2, but this deletion was not detected in any of the 27 carcinomas from the United States. CONCLUSIONS: Although slightly higher in Japan than in the United States, the frequency of p16 exon deletions in clinical prostatic carcinomas is very low, and probably is not important in the development of this neoplasm.

Immunohistochemical localization of the epidermal growth factor-receptor in rhesus-monkey prostate.

Epidermal growth-factor receptor (EGF-r), a membrane-bound glycoprotein activated by EGF, is important in maintaining the integrity and function of the prostate. To investigate EGF-r presence in the prostate of the rhesus monkey, monoclonal-antibody immunohistochemical examination was performed. The monkey prostate consisted of the cranial and caudal lobes, and the prostatic epithelial cells were composed of the secretory and basal cells. The distribution patterns of EGF-r in the prostatic epithelial cells were quite different between the cranial and caudal lobes. In the caudal lobe, EGF-r was seen in both secretory and basal cells, whereas, in the cranial lobe, the EGF-r was seen exclusively in the basal cells. The stromal cells of both lobes did not show EGF-r. This study revealed that each prostatic lobe contains specific binding sites for EGF, indicating a biological difference between the two lobes of the prostate of the rhesus monkey.

Lectin histochemical evaluation of glycoconjugates in dog efferent ductules.

Glycoconjugates in the epithelial cells of the efferent ductules in the dog were investigated using lectin histochemistry. These ductules connect the extratesticular rete with the epididymis. The epithelium of the ductules consisted both of ciliated and nonciliated cells. Whereas the apical zone of ciliated cells showed selective binding with WGA, SWGA, SNA, MAA and neuraminidase-PNA, that of nonciliated cells bound to all lectins used in the present study: WGA, SWGA, SNA, MAA, PNA, neuraminidase-PNA, RCA1, DBA and SBA. The nonciliated cells were divided into 3 types: type A cells which lacked both specific granules and vacuoles, type B cells which were characterised by a few specific apical vacuoles and many large specific granules, and type C cells which were characterised by some specific apical vacuoles and small basal granules. The specific granules and vacuoles of type B cells showed binding with WGA, SWGA and MAA. The specific granules of type C cells showed binding with WGA, SWGA, SNA, MAA, PNA and neuraminidase-PNA, while their specific vacuoles showed binding with WGA, SWGA, SNA and MAA. The Golgi zone both of ciliated and type A cells did not bind with any lectins used in this study, while type B and C cells showed similar lectin binding patterns between the Golgi zone and their specific granules. Specific lectin binding patterns revealed a different carbohydrate composition of each type of cell, indicating a biological difference between the ciliated cells and the 3 types of nonciliated cells in dog efferent ductules.

Prevalence of androgen receptor gene mutations in latent prostatic carcinomas from Japanese men.

The incidence rate of clinically apparent prostatic carcinoma is 8-fold higher in the United States than in Japan, while the prevalence of latent prostatic carcinoma, a presumed precursor to clinical carcinoma, is similar in the two countries. The purpose of this study was to investigate the hypothesis that this profound difference in incidence rates of clinical carcinoma reflects distinct profiles of molecular genetic alterations in the latent precursor lesions that occur in the two countries. A significant fraction of latent carcinomas from Japanese men were found to contain inactivating mutations of the androgen receptor gene, while no such mutations were found in latent carcinomas from American men. No mutations were found in clinical carcinomas from either country. These data offer a potential molecular genetic explanation that may partially account for the distinct prostatic carcinoma incidence rates in these two populations.

Nuclear bodies appearance rate in canine testicular Sertoli cell tumor.

A number of nuclear bodies (NB) were observed in the canine testicular Sertoli cell tumors (SCT). We statistically examined nineteen cases of canine SCT concerning the NB appearance rate (NBAR), and also examined the NBAR in four cases of the normal testicular Sertoli cells. The mean value of the total number of the NBAR of SCT was significantly higher than that of normal Sertoli cells. The SCT were classified into three groups according to the Nielsen and Lein's histological classification (1974): intratubular SCT without invasion, intratubular SCT with invasion, and diffuse type SCT. The mean value of NBAR of the diffuse type SCT was significantly higher than that of the intratubular SCT with and without invasion, and there was no significant difference between the mean values of NBAR of the latter two groups. The distribution of NBAR of the diffuse type SCT was significantly different from that of the intratubular SCT with and without invasion. On the other hand, the individual differences of NBAR of the diffuse type SCT and the intratubular SCT with invasion was significantly higher than that of the intratubular SCT without invasion. The present study suggests that the increase of NBAR in canine SCT might be correlated with the tumor invasive progression.

Tumour angiogenesis in latent prostatic carcinoma.

Unrestrained growth of various malignant tumours has been shown to depend upon a critical number of tumour cells which have switched to the angiogenic phenotype. Angiogenic phenotypes were noted in the early stage of prostatic carcinoma (PCa). We investigated 65 cases of latent PCa to define the correlation between tumour angiogenesis and tumour volume. Tumour angiogenesis was determined by the blood capillary density ratio (BCDR) evaluated by a colour image analysis system. Using experimental regression analysis, the correlation between the BCDR and PCa volume was divisible into two distinct stages. When the PCa showed a volume of more than 83 mm3, there was a significant positive correlation between the BCDR and PCa volume (rS-test P < 0.001). However, when the PCa showed a volume of less than 83 mm3, the BCDR remained at a low level which did not change until larger volumes were present (rS-test, NS; ANOVA, NS). The present study suggested that latent PCa showing a volume of less than 83 mm3 would be 'early' indolent carcinoma which, on undergoing additional events concerning tumour angiogenesis, would assume more aggressive growth.