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Objectives: To evaluate the efficacy of tegafur–uracil (UFT), a prodrug of 5-fluorouracil, plus cisplatin and dexamethasone in patients with docetaxel-refractory prostate cancers. Methods: Twenty-five patients with docetaxel-refractory prostate cancer were administered oral UFT plus intravenous cisplatin (UFT-P therapy) and dexamethasone. Treatment responses were assessed monthly via prostate-specific antigen (PSA) level measurements. Treatment-related adverse events and overall survival were also assessed. Results: UFT-P therapy resulted in decreased PSA levels in 14 (56%) patients and increased PSA levels in 11 (44%). In patients with increased PSA levels, 7 (64%) of the 11 patients displayed decreased PSA doubling times. The UFT-P therapy response rate was 84% (21/25 patients). Imaging studies revealed that tumor shrinkage during UFT-P therapy occurred in 1 patient in whom bilateral hydronephrosis caused by lymph node metastasis improved. The median survival time from docetaxel initiation was 36 months. In UFT-P-treated patients, the median PSA progression and overall survival times were 6 and 14 months, respectively. UFT-P treatment-related adverse events were mild diarrhea, general fatigue, and anorexia. Treatment was not discontinued for any of the patients. UFT-P therapy did not cause serious hepatic or renal dysfunction or pancytopenia. Conclusions: UFT-P therapy is a safe and effective treatment for patients with docetaxel-refractory prostate cancer, although large-scale, multicenter, prospective studies are needed to validate these findings.

Tragulidae is a primitive ungulate family within the order Cetartiodactyla, suborder Ruminantia. Domestic ruminants such as cattle, sheep, and goat have two types of Peyer's patches (PP): jejunal and ileal PP, in which there are morphological and functional differences. In this study, lesser mouse deer (Tragulus javanicus) PP was studied by gross anatomical and histological procedures. At the fetal stage, both types of PP were formed in the small intestine. Ileal PP was observed as a single continuous aggregation of lymphatic follicles extending cranially from the ileo-caecal junction. However, jejunal PP was observed as multiple and discrete accumulations of lymphatic follicles. This study showed that the lesser mouse deer has two types of PP in the small intestine. In addition, the anatomical and histological characteristics of jejunal and ileal PP are quite similar to those of other ruminants' jejunal and ileal PP. Further studies are needed to analyze immune function of both PP in lesser mouse deer in order to determine the evolutionary process of Cetartiodactyla.

This study demonstrated the immunohistochemical localization of cytoskeletal proteins, cytokeratin, desmin, vimentin, α-tubulin and α-smooth muscle actin (α-SMA) in the testes of adult and immature lesser mouse deer (Tragulus javanicus). Desmin was found in the sub-epithelial myoid cells of epididymal ducts. The immunoreactivity for vimentin was shown in the peritubular myoid cells and around the nuclei of the Sertoli cells of which the positions indicated various arrangements. In the Sertoli cells, vimentin filaments between the basement membrane and the nucleus were elongated with the migration of the nucleus toward the upper part. Furthermore, vimentin was detected in the Leydig cells and the sub-epithelial myoid cells of epididymal ducts. Alpha-tubulin was apparently extended to the tip of each Sertoli cell cytoplasm, and α-SMA was found in the peritubular myoid cells and the sub-epithelial myoid cells of epididymal ducts. On the other hand, cytokeratin and desmin were not detected in the Se...

1. Inter-breed morphological comparisons were made among 11 breeds of Japanese native chickens (Gifujidori, Hinaidori, Shokoku, Totenko, Tomaru, Satsumadori, Shamo, Koshamo, Koeyoshi, Chabo and Nagoya), White Leghorn, broiler chickens (Chunky) and red junglefowl collected in the Philippines, based on results of direct measurements and analysis by computer vision techniques of the skull. 2. Analysis of direct measurements identified two groups of chicken: a small type that included the Chabo, Koshamo, red junglefowl, Gifujidori and Shokoku and a large type that included the remaining breeds studied. These groupings were made based on size determined both in the first (PC1) and second principal component (PC2). The greatest length of the cranium and condylobasal length greatly contributed to the morphological differences between these two groups. 3. Analysis by computer vision techniques, however, identified three groups: the Bantam group (which includes red junglefowl), Shokoku group and Shamo group. White Leghorn clustered within the Shokoku group while the broiler chicken belonged to the Shamo group. The region around the junction of the neural cranium and the visceral cranium contributed greatly to the morphological differences among breeds, both in the PC1 and PC2.

We examined unique erythrocyte pits of the peripheral blood and bone marrow in the lesser mouse deer, Tragulus javanicus, using scanning electron microscope (SEM) and transmission electron microscope (TEM). Under the SEM observation, the pit was observed as a hole on both mature erythrocytes of the peripheral blood and immature erythrocytes of the bone marrow. By the TEM, the mature erythrocytes had a vacuole, which showed complicated shape and occupied considerable space within the cytoplasm. The vacuole was communicated extracellularly by perforation, which corresponded to the hole on the cell surface. In the bone marrow, erythroblast and reticulocytes have a cytoplasmic vacuole. This abnormal feature of the erythrocytes is peculiar to the mouse deer, and not found in other tropical ruminants. Despite the disadvantage of volume loss from the small erythrocytes, the mouse deer were healthy and showed no signs of anaemia.

The Tragulidae are the living relics of the basal ruminant stock. They have a diffuse placenta, with no aggregations of the placental villi into localised placentomes characteristic of all other ruminants. Despite this difference, this ultrastructural and immunocytochemical investigation demonstrates that in Tragulus the trophoblast binucleate cell (BNC) plays the same central role in development and structure as in all other ruminants. It shows an identical development and ultrastructure, produces granules reactive with bovine placental lactogen and pregnancy associated glycoprotein antibodies, and migrates when mature through the trophoblast tight junction to fuse into a mosaic of syncytial plaques from which the granules are released to the mother and which have replaced the uterine epithelium. Unlike the persistent plaques in the sheep and goat placenta, in Tragulus they are transient, dying by apoptosis with the fragments phagocytosed by the trophoblast. This brings the trophoblast into direct endotheliochorial apposition to maternal tissue until BNC migration and fusion replace the dead plaque. This intimate fetomaternal confrontation has not been shown in any other ruminant, and could be a relic of the evolutionary development of the synepitheliochorial from the original basic eutherian endo- or hemo-chorial placenta.

The geographical variations of the skull in the lesser false vampire (Megaderma spasma) were osteometrically compared between the traditionally-accepted subspecies from Philippines, Borneo, Java, North and South Thailand-Peninsular Malaysia. In female the population from Philippines was larger than any other population in skull size, whereas in male, the size tendency was not clearly found. The population of North Thailand was smaller than the four other populations at least in mean value of measurements in both sexes. The canonical discriminant analysis essentially separated the populations in the scattergrams of discriminant scores. However, the plots of Java, North Thailand and South Thailand-Peninsular Malaysia populations were intermingled in male, and Borneo population could not be obviously separated from the Java and South Thailand-Peninsular Malaysia populations in female.

Hair length in mammals is generally regulated by the hair cycle, and its disruption leads to abnormal hair morphogenesis in several species. FGF5, one of the hair cycle regulators, has a role in inducing catagen, and that mutation causes abnormal hair length in both sexes in humans, mice, dogs, and cats. Male-dominant long-haired coat (MALC) is an inbred strain of Syrian hamster exhibiting spontaneous long hair in males. After castration, MALC exhibited significantly shorter hair than the control individuals, but testosterone administration to castrated MALC showed reversion to the original phenotype. Moreover, flutamide administration led to MALC phenotype repression. Histological analysis revealed that hair follicle regression was shown in the wild-type 4 weeks after depilation, but that of MALC remained in the anagen phase. We detected a c.546delG of Fgf5 in MALC (Fgf5malc) that might lead to truncation resulting from a frame shift in FGF5 (p.Arg184GlyfsX6). Additionally, homozygous Fgf5malc was only detected in long-haired (Slc:Syrian×MALC)F2 and (J-2-Nn×MALC)F2 progenies, and all homozygous wild and heterozygous Fgf5malc individuals showed normal hair length. Thus, Fgf5malc leads to male-dominant long hair via a prolonged anagen phase which is affected by testosterone in hamsters. To our knowledge, this report is the first to present the sexual dimorphism of hair length caused by the Fgf5 mutation.

Summary The considerable phylogenetical differences between mouse deer and other ruminants have been established by means of DNA sequence analysis and anatomical observations. To clarify the physiological role of the uteroplacenta of the mouse deer, immunohistochemical observation was attempted by using GRP, which has been suggested as a novel regulatory peptide in the female reproductive tract, as an indicator to compare with other ruminants. Strong positive reactions for the GRP were detected in the uterine glands of the pregnant animals, but not in the non-pregnant ones. Although the placenta of the mouse deer is categorized as a diffuse placenta that is different from other ruminants’ polycotyledonary placenta, in terms of GRP immunoreactivity, the mouse deer placenta can be classified as a synepithecholial placenta like the other ruminants’. The secretion of GRP from the uterine glands is of some importance to the fetus in the mouse deer.

Summary The transition of fetal haematopoietic sites in mice was examined histologically from 12.5-day embryos (E12.5) until 10 days of age (D10), and the expression of the adhesive molecules VCAM-1 and fibronectin was examined immunohistologically. Erythropoiesis occurred in the liver (E12.5–18.5), spleen (E18.5–D4) and bone marrow (D6–10), in that sequence. Even at D10, some erythropoiesis occurred in the liver, and more so in the spleen, although the active haematopoietic site was the bone marrow. Similarly, granulopoiesis of neutrophils occurred in the liver, spleen and bone marrow in turn. Granulopoiesis still occurred in the spleen at D10, but no neutrophils were found in the liver after D4. VCAM-1 appeared in the liver, spleen and bone marrow in parallel with active erythropoiesis and granulopoiesis. The co-expression of VCAM-1 and fibronectin was recognized in the endothelial cells of the sinus at the onset of haematopoiesis. This study showed that haematopoiesis in the liver, spleen and bone marrow overlapped peri-natally, although it shifted sequentially. VCAM-1 appears to be closely associated with erythropoiesis and granulopoiesis, and the co-expression of VCAM-1 and fibronectin plays a role in inducing haematopoietic stem cells to move to the tissues.

Summary Fibre connections of the chick nucleus geniculatus lateralis ventralis (GLv) were investigated using the axonal tracing method with wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). After an injection of WGA-HRP into the GLv, many labelled neurons were observed in layer i of the stratum griseum et fibrosum superficiale (SGFS) in the ipsilateral tectum opticum (TO) and in the nucleus lentiformis mesencephali (LM). In the TO-GLv projection, cells of origin were located in the deeper part of layer i of the TO and were topographically distributed along the direction from the rostrodorsal part to the caudoventral part of the TO relating to a rostrocaudal axis of the GLv. In the LM-GLv connection, the dorsal and ventral parts of the LM connected reciprocally with the rostral and caudal halves of the GLv, respectively. In contrast, in the GLv efferent connection, labelled axon terminals spread widely in the ipsilateral area pretectalis without any clear topographical arrangement.

Mouse and vole embryos were allogeneically and xenogeneically transferred into pseudopregnant CD-1 and immunodeficient (scid) female mice, and we investigated the distribution of immunocompetent cells, uterine natural killer (uNK) cells, mast cells and macrophages, in the implantation sites on days 6, 7 and 8 of gestation. The survival rate of the vole embryos decreased gradually with increased gestation, but the rate was higher in the scid uteri than in the CD-1 mice. The number of uNK cells increased markedly at the mesometrial triangle and the outer decidual area in the CD-1 uteri containing vole embryos; by contrast, scid uteri having vole embryos showed almost the same number as those having mouse embryos. Mast cells were present in large numbers at the myometrium, but rarely in the decidua in all types of pregnant uteri. Cells at the myometrium were more numerous in xenogeneic than in allogeneic transfer. Many mast cells appeared in the inner decidua where xenogeneically transferred vole embryos were dead and aborted. Macrophages were present in the outer decidua and myometria in all types of pregnant uteri, and their distribution pattern did not change even in aborted uterine sites. These results suggest: (1) the response of macrophages to dead embryos is completely inhibited, (2) uNK cells and mast cells increase near dead and aborted embryos, and (3) the increment in uNK cells responding to xenogeneic embryos is suppressed in scid mice, and the suppression may contribute partly to survival of the embryos.

We examined the geographical variation of the skull size and shape of the lesser mouse deer (Tragulus javanicus) from Laos, Thailand, Peninsular Malaysia, Sumatra, Java, Borneo, Langkawi and some Islands of Tenasserim in Myanmar. Although the influence of the climatic condition on skull size was not confirmed in the mainland populations, the skull became rostro-caudally longer in the populations of Tenasserim and Sumatra because of island isolation effect. The skull size was classified into the following three clusters of localities from the matrix of Q-mode correlation coefficients: 1) Langkawi and Tenasserim, 2) Laos and Thailand, 3) Sumatra and Borneo. The skulls in the population of Java belong to the cluster of Langkawi and Tenasserim in male, however were morphologically similar to those in the cluster of Borneo and Sumatra. The canonical discriminant analysis pointed out that the Laos and Tenasserim populations were separated from the other ones and that the populations of Sumatra, Java and Borneo were intermingled each other.

The skull adaptation was functional-morphologically examined in 14 species of the tree shrews. From the data of the proportion indices, the similarities were confirmed between T. minor and T. gracilis, T. tana and T. dorsalis, and T. longipes and T. glis. We demonstrated that the splanchnocranium was elongated in terrestrial T. tana and T. dorsalis and shortened in arboreal T. minor and T. gracilis from the proportion data. In both dendrogram from the matrix of the Q-mode correlation coefficients and scattergram from the canonical discriminant analysis, the morphological similarities in the skull shape suggested the terrestrial-insectivorous adaptation of T. tana and T. dorsalis, and the arboreal adaptation of T. minor and T. gracilis. Since the osteometrical skull similarities were indicated among the three species of Tupaia by cluster and canonical discriminant analyses, the arbo-terrestrial behavior and its functional-morphological adaptation may be commonly established in T. montana, T. longipes and T. glis.

An experimental model to study interspecific pregnancy using voles, Microtus arvalis, and green fluorescent protein gene-induced transgenic mice is presented. Xenogeneic blastocysts from the vole were transferred into the uteri of pseudopregnant mice along with allogeneic blastocysts from green fluorescent protein gene-induced transgenic mice. The uteri containing xeno-allo combined transfers were examined from day 6 to 13 of gestation. Although the vole embryos implanted, the uteri containing vole embryos were smaller compared with those having allogeneic mouse embryos. On day 8, the uteri containing vole embryos hemorrhaged internally and no vole embryo was found in the pregnant uterus after day 11. Allogeneic mouse embryos developed normally despite the presence and abortion of the vole embryos. In uteri implanted with vole embryos, decidua were formed and numerous blood vessels were distributed around the embryo. Maternal blood cells infiltrated into the celomic cavity of the vole embryo through the discontinuous region of trophoblast. Periodic acid-Schiff-positive granulated metrial gland cells were remarkably increased in the decidual sites. These findings suggest that a disorder of embryo–maternal interaction might induce the appearance of numerous granulated metrial gland cells and rejection of the embryos.

We describe an unusual case of metastasis of the skull base from prostate cancer. A 64-year-old man was referred for progressive cranial nerve palsies. Brain magnetic resonance imaging showed a metastatic tumor in the right petrous bone. He was presenting Garcin syndrome indicating involvement of the right III, IV, V, VI, VII, VIII, IX, X, and XII cranial nerves. The serum PSA level was 5118 ng/mL (standard threshold 4 ng/mL). A transrectal needle biopsy was performed, and poorly differentiated adenocarcinoma was detected in the prostate. Therefore, he was finally diagnosed with skull base metastasis of the prostate cancer. Androgen deprivation therapy and external beam radiation therapy to the skull base resulted in complete resolution of the cranial nerve palsies for more than 30 months. This is the first reported case of metastatic prostate cancer presenting as Garcin syndrome.

INTRODUCTION AND OBJECTIVES: Lysine-specific demethylase 1 (LSD1), the first discovered histone demethylase, is a novel target for prostate cancer therapy. LSD1 expression positively correlates with prostate tumor malignancy. NCL1, a selective cell-active inhibitor of LSD1, was discovered at our university. Here, we verified the mechanism of prostate cancer growth and examined the anti-cancer effect of NCL1 using an animal model. METHODS: We used the prostate cancer cell lines LNCaP, PCai1, and PC3. LSD1 expression was examined by western blotting in LNCaP, PCai1, and PC3 cells. Cell viability was assessed by a WST-1 assay in LNCaP, PCai1, and PC3 cells treated for 48 h with vehicle or 1e100 mM NCL1. LNCaP cells were incubated with or without 30 mM NCL1 for 210 min. Chromatin was collected, and ChIP and real-time polymerase chain reaction were used to confirm the methylation status of the promoter regions of certain genes. Nude mice were subcutaneously injected with 3 106 PCai1 cells. Four days later, mice were injected intraperitoneally with NCL1 at doses of 0.5 mg/kg or 1.0 mg/kg. Mice were treated twice a week for 8 weeks. RESULTS: LSD1 was expressed in all cell lines. The number of viable cells were reduced after NCL1 treatment in all cell lines in a dose-dependent manner. On ChIP analysis, NCL1 treatment induced H3K9me2 accumulation at the androgen response element 1/2 (ARE1/2) and at ELK4 and KLK2 promoter regions. Tumor volume was significantly lower in mice treated with 1.0 mg/kg NCL1 than in controls. Mice that received 1.0 mg/kg NCL1 showed vacuolar degeneration, low Ki67 positivity, and high TUNEL intensity. There were no adverse effects in terms of body weight, relative organ weight, histological findings, and blood biochemical examination results. CONCLUSIONS: LSD1 is involved in prostate cancer cell proliferation via H3K9me2 demethylation at ARE1/2 and ELK4 and KLK2 promoter regions. NCL1 is safe and effective in vivo and induces an anti-cancer effect through apoptosis induction. NCL1 is promising for the development of therapeutics for prostate cancer.

Distribution pattern of the vagal preganglionic neurons in the dorsal motor nucleus of the vagus nerve (DMN) and nucleus ambiguus (NA) projecting into the stomach of the Japanese field voles (Microtus montebelli) was examined using horseradish peroxidase (HRP). The HRP was injected into ten different stomach regions covering the forestomach, glandular region and pyloric stomach. The labeled neurons in the DMN were in every injected case, while labeled neurons in the NA were in the HRP injected into the forestomach and pyloric stomach. After a separate injection of HRP into the ventral and dorsal stomachs, the labeled neurons were predominant in the left and in the right DMNs, respectively. Medio-lateral and rostro-caudal distribution of HRP-labeled neurons showed that many of them were in the medial part and near the level of the obex in the DMN. Especially innervating neurons of glandular region were remarkably restricted in the medial part and at the level of the obex in the DMN. The present investigation showed that majority of neurons supplying each stomach region were in the DMN with site specific and topographic arrangement, and a considerable number of labeled neurons were distributed symmetrically in the NA.

The distribution in the chicken of motoneurons innervating the hyolingual muscles, i.e. the M. branchiomandibularis (BM), M. ceratoglossus (CG), M. interceratobranchialis (CB), M. serpihyoideus (PH), M. stylohyoideus (YH) and one of the mandibular muscles, M. depressor mandibulae (DM), was examined by retrograde transport of horseradish peroxidase conjugated with wheatgerm agglutinin. Labelled motoneurons are found in the three subnuclei of the facial (VII) nucleus, as well as hypoglossal (XII) nucleus. The distribution of motoneurons projecting to the DM are observed in the three subnuclei, those of the BM, PH and YH in the intermediate and ventral subnuclei and those of the CB and CG in the intermediate subnucleus. Motoneurons projecting to the PH, YH, CB and CG are also distributed in the XII nucleus. The ratio of labelled motoneurons of the VII to XII nuclei decreases the PH, YH, CB and CG in that order, and the ratio of labelled ones of the ventral to intermediate subnuclei decreases the BM, PH and YH in that order. The topographical and functional aspects of the subdivision of the motor nucleus are discussed.

The semidominant gene Dh (Dominant hemimelia) induces skeletal and visceral abnormalities of various degrees and failure of the spleen in mice. The homozygous individual (Dh/Dh) seems to be lethal. The present experiment was designed to investigate the ability Dh cells to form a spleen and the genesis of the hind limb malformations by Dh/Dh and Dh/+ cells in chimeric mice. The Dh/Dh and Dh/+ embryos were produced in the F2 progeny of a cross between inbred strains of Dh/+ and DDD mice. They were aggregated with C3H/He or C57BL/6 embryos to make chimeras. Identification of Dh/Dh or Dh/+ embryos was carried out by Pep-3, and chimerism was analyzed by Gpi-1. Of 25 chimeras carrying the Dh gene, four mice formed a small spleen, two mice had a vestigial spleen, and the others no spleen. The tissues of the incompletely developed spleens were normal histologically and Dh cells were involved in the tissues of the spleen. In the chimeric mice, hindlimb malformation by the Dh gene was reduced in severity and the lethality of the homozygote (Dh/Dh) was rescued.

Retinal projections in the Japanese field vole (Microtus montebelli) were determined by anterograde transport of horseradish peroxidase (HRP). Injection of HRP into the unilateral vitreous body demonstrated that the terminal labeling of the optic projections was seen bilaterally in the suprachiasmatic nucleus (SCH), the ventral (GLv) and dorsal (GLd) lateral geniculate nuclei, the intergeniculate leaflet (IGL), the medial pretectal nucleus (NTOM) of the pretectum (PT) and the superficial layer of the superior colliculus (CS), with contralateral predominance, and only contralaterally labeled terminals were found in the lateroposterior thalamic nucleus (LP), the lateral pretectal nucleus (NTOL) of the PT, the dorsal (DTN) and medial (MTN) terminal nuclei of the accessory optic system (AOS). The distribution area of the retinofugal terminals was divided into a three laminar arrangement in the GLd, i.e., layers 1 and 3 and layer 2, received the retinal input from contralateral and ipsilateral eye, respectively, as in arboreal squirrels. The contralateral CS received retinal fibers in the superficial layer, while ipsilateral optic fibers projected sparsely to the stratum opticum of the colliculi. Retinal connections to the DTN and MTN of the AOS were clearly discerned but no lateral terminal nucleus with retinal afferents was found. In addition, the AOS had no inferior fasciculus. These findings indicate that the vole has a contradictory features of a well- and a less-developed sense of vision. Namely, the image forming visual system such as the retino-GLd was as well-developed as in a squirrel, on the other hand, the non-image forming visual system such as the retino-AOS was less-developed as in an insectivore's brain.

Tetraploid mouse embryos usually cease to develop early after implantation, though they can develop to blastocysts. To characterize the failure of development in detail, tetraploid mouse embryos at the preimplantation period were examined as to both their morphology and number of cells. The tetraploid embryos were produced by 12 hr treatment with cytochalasin B (CB) at the 2-cell stage of backcross of (C57BL/6 x C3H/He) F1 x C3H/He. The tetraploid embryos in the preimplantation period exhibited compaction at 72 hr after hCG injection and blastocyst formation at 96 hr, as well as diploid embryos, but the number of cells composing the embryos was significantly smaller than that in the diploid embryos. At the term 60-96 hr after hCG injection, mean cell cycles were 14.03 hr in the tetraploid embryos, but 12.02 hr in the diploid. When tetraploid embryos were transferred into the oviducts of pseudopregnant recipients immediately after CB treatment, the number of cells in tetraploid blastocysts was increased compared with the embryos cultured in vitro, though the number did not reach that of diploid embryos. These results suggested that compaction and blastocyst formation in preimplantation development of tetraploid embryos depended on the time after hCG injection, irrespective of the number of cells or the length of the cell cycle. The lengthening of the cell cycle in tetraploid embryos may be one of the causes of failure in postimplantation development.

Mouse and vole embryos were transferred into pseudopregnant CD-1 and scid female mice. Cellular changes involved in the formation of decidua in the pregnant mouse uterus up to day 8 of pregnancy were examined by histological, electron microscopic, and histochemical techniques. On day 6 of pregnancy, the vole embryos were laid in interstitium of antimesometrial side of the uterus, as well as in intraspecific pregnancy. Compared with intraspesific pregnant mouse, blood vessels were numerous in the decidua around the vole embryos in interspecific pregnancy. Both distribution and dilation of the blood vessels were increased on day 8. A part of cells in the inner cell mass had not nuclei, suggesting damaged vole embryos on day 8. At the implantation site, the uterine decidua was invaded by extravillous trophoblast (EVT) cells whose function is to destroy the walls of the uterine spiral. Moreover, this decidua was infiltrated by a population of natural killer (NK) cells and macrophage. These cells were particularly numerous in the decidua basalis at the implantation site where they come into close contact with invading EVT cells. These results suggest that interaction between NK, macrophage, and EVT provides the controlling relationship of embryo-maternal in intraspecific and interspecific pregnancy.

Protrusion of hyperplastic upper molar teeth (macrodonts) into the cranial cavity was observed in the Japanese field vole. Protrusions of the upper molars occurred on both sides, but were large on the left than on the right. The teeth invaded the cranial cavity and brain parenchyma. Large concavities on the ventral side of the brain were interposed between the lateral olfactory tract and the optic tract. Invation by one macrodont reached the basal nuclei, hypothalamic and thalamic nuclei. The brain was deformed bilaterally, and nearly the entire olfactory bulb, globus pallidus, internal capsule, and lateral preoptic nucleus, and a portion of the hypothalamus, thalamus and anterior commissure were absent on the left side.

Adrenocortical zonation in the laboratory Japanese field vole (Microtus montebelli) was investigated histologically with special attention to the sex and species specific differences. The whole cortex and zona reticularis showed clear sex differences. They were thicker in females than in males. In this respect the vole was similar to the mouse and rat, but different from the Syrian hamster and mastomys. Male cortices were characterized by thick z. fasciculata and meagre z. reticularis, while both zones were thick in females. The z. fasciculata in both sexes and z. reticularis in females seemed to be divided into two sublayers by the cellular arrangement. No distinctive additional zones were identified. The histological observations indicated that the Japanese field vole might have interesting endocrinological characteristics and serve for studying the mechanism of the zone formation in the mammalian adrenal cortex.

OBJECTIVES To establish a novel and simple method of preventing post-retropubic prostatectomy (RRP) inguinal hernia. Inguinal hernias occur in 8%-22% of men within 1-2 years of RRP. Although manipulation during RRP might weaken the normal fascia structure at the internal inguinal ring with the vas deferens, the exact mechanism of post-RRP inguinal hernia remains unknown. METHODS Several surgeons performed RRP at our hospital on 271 patients between April 2004 and September 2009. Among these patients, post-RRP measures to prevent inguinal hernia were applied to 101 patients (group A) and not applied to 170 patients (group B). We released the bilateral spermatic cord from the peritoneum before suturing the wound, which should prevent the intestinal tract coated with the peritoneum from pushing through the internal inguinal tract. We compared the incidence of postoperative inguinal hernia between the 2 groups. RESULTS The patients were followed up for an average of 11.6 (range: 2-22 months) and 23.9 (range: 23-24 months) months in groups A and B, respectively. Inguinal hernia developed in no patients in group A and in 20 (11.8%) in group B. The hernia-free rate was significantly lower in group B than group A. All postoperative inguinal hernias were indirect. The median interval between surgery and hernia diagnosis was 10.6 months (range, 2-24), and 16 patients (80%) were diagnosed within 12 months. CONCLUSIONS We developed a simple method of preventing inguinal hernia after RRP. Our technique is simple enough to complete within a few minutes, and the outcome is excellent.

The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d-galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N-acetyl-d-galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N-acetyl-D-glucosamine and sialic acid (wheat germ agglutinin WGA, s-WGA), D-mannose and d-glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), L-fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA-E) sugar residues. In Golgi-, cap-, and acrosome-phase spermatids, lectin-bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s-WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA-E). s-WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA-E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA-E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA-E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin-bindings noted in the testis of lesser mouse deer included the limited distribution of s-WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications.

The skin and lymphoid organs of Mexican hairless dogs and their hairless offspring were examined histologically. The hairless dogs lacked most hairs except for sparse hairs on the head, tail and feet. The skin of newborn pups consisted of a thick epidermis with epidermal ingrowths forming the rudiments of hair follicles. In older dogs more than 2 months of age, however, the epidermis was thin and the ingrowths were few. Neither hair follicles nor skin glands were present. The hairy skin of the head and tail had hair follicles with sebaceous glands. Regarding the lymphoid organs, the newborn pups possessed a thymus like haired pups. But in the older dogs more than 2 months of age, the thymus was atrophied and the lymphocyte population was too sparse to demarcate the cortex and the medulla. Lymphocyte accumulation in older dogs was also poor in the spleen and mesenteric lymph nodes. The present findings indicate that the hairlessness of the Mexican hairless dogs and their descendants is accompanied by early atrophy of the thymus after birth, and is followed by poor accumulation of lymphocytes in the thymus-dependent area of the spleen and the mesenteric lymph nodes. The defect of the thymus in the hairless dog seems to be different from that in athymic nude mice and rats. Further studies are needed to elucidate the immunological response and function in hairless dogs.

We examined the expression of VCAM-1 and MAdCAM-1 after bone marrow transplantation (BMT). We also examined the influence of alpha(4)beta(7) integrin blockade on the homing of cells to the bone marrow and spleen. The expression of VCAM-1 and MAdCAM-1 by endothelial cells in the spleen and bone marrow was examined by immunoelectron microscopy using colloidal gold and was analyzed semiquantitatively. To examine the role of alpha(4)beta(7) integrin in donor cells, a homing assay was conducted following alpha(4)beta(7) integrin blockade in bone marrow-derived hematopoietic cells or spleen colony cells. Immediately after BMT, the expression of VCAM-1 and MAdCAM1 markedly decreased, but expression recovered significantly between 12 and 24 h after BMT. VCAM-1 recovered more acutely than MAdCAM-1 from 12 h onward. In the group transplanted with anti-alpha(4)beta(7) integrin antibody-treated bone marrow cells, the numbers of homing cells in the spleen and bone marrow were significantly decreased in an antibody dose-dependent manner. However, the number of homing cells was not different in either the spleen or bone marrow between anti-alpha(4)beta(7) integrin antibody treated and untreated spleen colony cells. It has been reported that alpha(4)beta(1) integrin and its receptor VCAM-1 play major roles in the homing of hematopoietic cells to bone marrow. Our study indicates the importance of MAdCAM-1 and its ligand, alpha(4)beta(7) integrin, in the homing of bone marrow-derived hematopoietic cells, but not spleen colony-derived cells, to both the spleen and bone marrow.

Short-term dextran sodium sulfate (DSS) treatment has been shown to notably accelerate colorectal tumor development in rats initiated with 1,2-dimethylhydrazine (DMH). In the present study, to clarify mechanisms underlying the DSS influence, time-course studies of histopathological and immunohistochemical characteristics and beta-catenin gene mutations in colorectal mucosa in early stages of this model were conducted. F344 males were given three subcutaneous injections of DMH (40 mg/kg body wt) within a week, followed by free access to drinking water containing 1% DSS for a week. At weeks 1, 4, 6 and 8 after the DSS treatment, rats were euthanized and colorectal samples were collected. At week 1, the colorectal mucosa demonstrated extensive erosion along with significant inflammatory cell infiltration and neighboring reactive hyperplasia. By week 4, the mucosal damage was repaired and regenerative mucosa, partly characterized by Paneth cell metaplasia and altered subcellular localization of beta-catenin, was apparent. Areas with Paneth cells/beta-catenin accumulation were significantly more likely to be accompanied by interstitial inflammation and 17 of 24 dysplastic foci were found in regenerative mucosa with Paneth cells. Furthermore, adenomas/carcinomas frequently featured various degrees of Paneth cell differentiation. Point mutations mainly in codons 34 and 41 of beta-catenin gene were detected in 6 of 27 samples of regenerative mucosa with Paneth cells and four of nine dysplastic foci/adenomas/carcinomas. These findings indicate that inflammation-associated regenerative mucosa with Paneth cell metaplasia and alteration in the APC/beta-catenin/Tcf signal transduction pathway are possibly involved in the acceleration of colorectal carcinogenesis in this DMH-DSS rat model.

Some visual information is sent to the nucleus geniculatus lateralis ventralis (GLv) via the cells in layer I (I cells) of the tectum in birds and is used for color vision, papillary reflex, and kineoptic functions. To reveal the morphological features of 'I cells' projecting to the GLv, they were retrogradely labeled with DiI (1,1'-dioctadecyl-3, 3, 3', 3'-tetramethylindo-carbocyanine perchlorate) in chicks. Two different types of neurons, 'spear dendritic I cells' and 'forked dendritic 1 cells' were identified. The former had small spindle-like soma and an apical dendrite extending to the tectal surface, and the latter had somewhat larger triangular or polygonal soma and plural ascending dendrites. Most of the labeled dendritic endings bifurcated horizontally in layer F, and showed the ending patterns similar to the terminals of optic nerve fibers.

The Tragulidae may be a type that represents the earliest ruminant families to evolve. The female reproductive organs including ovary, oviduct, uterus and placenta were anatomically and histologically investigated. The structures of the ovary and oviduct were very similar to other ruminants. However, the gross structure of the placenta was diffuse and thus noticeably different from other ruminants which are polycotyledonary. Histologically, the placenta of Tragulidae appears to be epitheliochorial and therefore similar to other ruminants. Numerous trophoblastic binucleated cells which are characteristic of all other ruminants were observed. These results suggest that the placenta of Tragulidae is a transitional type between diffuse epitheliochorial and polycotyledonary synepitheliochorial categories.

Summary Organization of the fibre connections in the chick nucleus rotundus (Rt) was investigated by an axonal tracing method using wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). After an injection of WGA-HRP into the Rt, labelled neurones were observed in the striatum griseum centrale (SGC) in both sides of the tectum (TO) and in the ipsilateral nucleus subpretectalis/nucleus interstito-pretecto-subpretectalis (SP/IPS). Labelled fibres and terminals were also found in the ipsilateral ectostriatum (Ect). These fibre connections were topographically organized rostrocaudally. In the TO-Rt projection, the rostral and the dorsocaudal parts of the Rt received afferents from the superficial part of the SGC, the middle part of the Rt received afferents from the intermediate part of the SGC, and the ventrocaudal part of the Rt received mainly fibres from the deep part of the SGC. These topographic projections were accompanied by a considerable number of diffuse projections to the thalamic regions surrounding the Rt. In addition, the rostral and middle caudal parts of the Rt received afferents from the lateral and medial parts of the SP/IPS, respectively, and respective parts of the Rt sent efferents to the lateral and medial parts of the Ect.

The Sertoli cell of the lesser mouse deer, Tragulus javanicus, was examined using light and transmission electron microscopy. Similar to other ruminants, a multivesicular nuclear body (MNB) and laminated smooth endoplasmic reticulum (sER) were observed in the lesser mouse deer Sertoli cell. The MNB was present within the Sertoli cell nucleus, and consisted of vesicles, irregular tubules and ribosome-like structures. It was infrequent in the lesser mouse deer, which differs from domestic ruminants. Vesicles and irregular tubules seem to contain some materials with low and/or middle electron density, and be surrounded by electron dense materials. The diameter of vesicles was between 30 nm and 180 nm. Since the MNB, though less developed compared to that of bulls and goats, was present even in the Sertoli cell nucleus of the primitive ruminant-lesser mouse deer, it should be a common structure of ruminant Sertoli cells.

The mastication muscles were examined in the lesser (Tragulus javanicus) and greater mouse deer (Tragulus napu) to clarify the form of the mastication muscles in these primitive artiodactyls. The M. masseter was well-developed in both species, however the attachment area of its origin was not confirmed in the rostral facial part. The masseter bundles were not observed on the lateral side of the maxilla bone, and their origin was restricted to the zygomatic arch area. This suggests that the M. masseter may not act as a motor raising the mandible rostro-dorsally, but pull the insertion vertically unlike the highly derived grazer of Bovidae. The Crista temporalis was weak and the M. temporalis was thin in the mouse deer, and this indicates that the M. temporalis may not be important in the mastication in the primitive artiodactyls. These findings suggest that the browser such as mouse deer has been adapted for the feeding on soft leaves, and functional-morphologically different in mastication strategy from the grazer such as developed Bovidae species. The architecture of the mastication muscles was not different between the two species. However, in the muscle weight ratios per body weight, the M. temporalis and the M. digastricus were significantly smaller in greater mouse deer than in lesser mouse deer.

Morphological characterization of the skeletal malformations caused by the Dh gene was carried out. Skeletal preparations from newborn mice of Dh/Dh, Dh/+ and +/+ genotypes, and adult Dh/+ mice were examined. The Dh gene induced skeletal deformities at the axial regions from the thorax to the tail. Attachment of the ribs to the sternum was sometimes irregular, and normal segmentation of the sternum was also disrupted. Abnormal fusion of the ventral part of the rib was observed in some cases. Lumbar vertebrae were fewer in number and abnormal in shape. Although it was rare, a curled tail was also observed due to distortion of the caudal vertebrae.

Angiotensin (AT) II, the bioactive octapeptide in the renin-angiotensin system that plays a key role in cardiovascular homeostasis, exerts its multiple effects through the different types of AT receptors, AT1a, AT1b, and AT2. Previously, we showed chronic hypotension in angiotensinogen (the precursor of AT)-deficient mice and a dramatic increase in renin mRNA levels in its kidney, but it remains unclear which types of AT receptors regulate the blood pressure and renin gene expression. In order to elucidate the physiological roles of AT1a receptor, we generated mutant mice with a targeted replacement of the AT1a receptor loci by the lacZ gene. In the heterozygous mutant mice, the strong lacZ staining was found in the glomerulus and juxtaglomerular apparatus of the renal cortex, which coincided with that of the signals detected by in situ hybridization. Chronic hypotension was observed in the heterozygous and homozygous mutant mice, with 10 and 22 mm Hg lower systolic blood pressure, respectively, than that of wild-type littermates. Both levels of renin mRNA in the kidney and plasma renin activity were markedly increased only in the homozygous mutant mice. These results demonstrated that an AT1a-mediated signal transduction pathway is, at least in part, involved in the regulation of blood pressure and renin gene expression.

Ultrastructural changes of the cells forming germinal centers were examined in the chick spleen after intravenous injection of sheep red blood cells. The germinal center was formed by a cluster of large lymphoid cells and a septum of flattened reticular cells. The large lymphoid cell was characterized by numerous polysomes and a large euchromatic nucleus with a prominent nucleolus. In the developing germinal center the large lymphoid cells proliferated and became smaller in size. After the peak of development of the center they were gradually atrophied and finally became degenerative to exhibit pyknotic nuclei, condensed cytoplasm, some vacuoles and many cytoplasmic processes. From the formation of the germinal center through its degeneration, the transformation of lymphoid cells into plasma cells did not occur within the center. Plasma cells were appeared in the red pulp prior to the germinal center formation, and then they proliferated to aggregate around a periarterial lymphatic sheath.

The primary formation of germinal centers was induced in the chick spleen by intravenous injection of sheep red blood cells (SRBC) at 21 days of age. In the spleen, SRBC were trapped in the ellipsoids soon after injection. The number of cells of periellipsoidal lymphoid tissues (PELT) was gradually depleted. Large pyroninophilic cells appeared in the periphery of periarterial lymphatic sheaths (PALS) 4 hrs after injection. Then they increased in number and migrated centralward in the PALS. On the 4th day they were accumulated in the vicinity of the central arteries to make nodular structures. Morphologically mature germinal centers encapsulated by connective tissue were formed for the first time on the 6th day after injection. They were most numerous on the 8th day and then decreased gradually. It was presumed that the primary germinal centers in the chick spleen might have been formed by the cells of the PELT which were originally derived from the bursa of Fabricius. The process of proliferation of germinal centers in the secondary immune response was discussed.

The splenic microcirculation of the White Leghorn chicken was studied by means of the scanning and transmission electron microscopes. Corrosion casts of the splenic vascular network and tissues of the perfused spleen were examined. The corrosion cast presented the passageway of blood in the spleen, in which arterial vessels became veins as continuous canals. The intervening portions connecting both vessels varied in diameter and exhibited tortuous contours. In the perfused spleen, no direct continuity with a continuous endothelium was recognized between the arterial termination and the primordial vein. There was a spongework consisting of reticular cells in the pulp cord. The arterial terminal cepillary opened into the interstice of the spongework. Blood cells remaining in it drained into a venous vessel through the preformed apertures of this vessel.