Search

Original Article, [Purpose] This study investigated the multifaceted factors of inpatients and community-dwelling women with equivalent exercise capacity. [Participants and Methods] The subjects were 56 women (age 80.9 +- 7.0 years) discharged from the convalescent rehabilitation ward and 74 community-dwelling women (age 74.7 +- 5.3 years). We conducted a two-step test to examine subjects with regard to Locomo risk level 1 for age, grip strength, SMI, walking speed, Alb value, FFM, Kihon checklist, number of medications, presence or absence of low hone mineral density, and cognitive decline. [Results] There were significant differences in Alb value, walking speed, and low bone mineral density between the hospitalized group and the community-dwelling group. [Conclusion] It was suggested that the hospitalized group had undernutrition and a high fracture risk despite the similarity in exercise capacity.〔目的〕本研究の目的は女性で同等の運動能力の入院患者と地域在住者の多面的要因を調査することである。〔対象と方法〕対象は回復期リハビリテーション病棟を退院となった女性56名(平均年齢80.9±7.0歳)と地域在住女性74名(平均年齢74.7±5.3歳)とした。 2ステップテストを用いてロコモ度1に該当した者に対して、 年齢、 握力、 SMI、 歩行速度、 Alb値、 FFM、 基本チェックリスト、 服薬数、 骨密度低下の有無、 認知機能低下の有無の検討を行った。 〔結果〕入院群と地域在住群では、 Alb値、 歩行速度、 骨密度の低下に有意差を認めた。 〔結語〕運動能力が同等であっても入院群は栄養状態が低く、 骨折リスクが高いことが示唆された。

The residue generated by the black soldier fly (Hermetia illucens, BSF) during the processing of organic waste is considered a suitable crop fertilizer. However, no detailed studies have investigated the fertilizer value of the residue obtained from processing household organic waste. In this study, experimental household organic waste (EHOW) was processed by BSF at 200 mg of EHOW per head for 15 days at 27 &deg, C. To evaluate the fertilizer value of the obtained BSF larvae production residue (BSFR), the chemical composition and microbiota were analyzed, and Komatsuna (Brassica rapa var. perviridis) cultivation tests were conducted. BSFR results demonstrated higher ammonium nitrogen and lower nitrate nitrogen, and the highest above-ground dry matter weight of Komatsuna. Although the relative abundance of Escherichia was low, the relative abundance of Xanthomonadaceae, which contains a genus that causes disease in plants, was high. Therefore, the presence of plant pathogens in the BSFR microbiota should be considered. Finally, the effects of BSFR on the external environment requires more detailed investigation.

The grasscutter (also known as the greater cane rat, Thryonomys swinderianus) is a large rodent native to West Africa that is currently under domestication process for meat production. However, little is known about the physiology of this species. In the present study, aiming to provide information about gut microbiota of the grasscutter and better understand its physiology, we investigated the intestinal microbiota of grasscutters and compared it with that of other livestock (cattle, goat, rabbit, and sheep) using 16S rRNA metagenomics analysis. Similar to the other herbivorous animals, bacteria classified as Bacteroidales, Clostridiales, Ruminococcaceae, and Lachnospiraceae were abundant in the microbiome of grasscutters. However, Prevotella and Treponema bacteria, which have fiber fermentation ability, were especially abundant in grasscutters, where the relative abundance of these genera was higher than that in the other animals. The presence of these genera might confer grasscutters the ability to easily breakdown dietary fibers. Diets for grasscutters should be made from ingredients not consumed by humans to avoid competition for resources and the ability to digest fibers may allow the use of fiber-rich feed materials not used by humans. Our findings serve as reference and support future studies on changes in the gut microbiota of the grasscutter as domestication progresses in order to establish appropriate feeding methods and captivity conditions.

The 〈001〉β-fiber texture, which is a highly desirable orientation for β-Ti alloys with low Young's modulus, was formed successfully in cold-groove-rolled Ti-5.5Mo-8Al-6Zr (mol.%) alloy wires. The effect of a reduction in the cross-sectional area (CSA) on the evolution of the rolling and recrystallization texture, as well as the Young's modulus, was systematically investigated. The formation of α″-martensite was induced by the cold-groove-rolling process, with a 〈010〉α″-fiber texture developing in specimens with CSA reductions of 93% or more. This texture corresponds to the 〈110〉β-fiber texture considering the lattice correspondence of the martensitic transformation between the β (bcc) - and α″ (orthorhombic) -phases. The 〈001〉β-fiber recrystallization texture was formed in solution-treated specimens with CSA reductions of 80% or more and developed dramatically with a reduction of 98%. The development of the 〈001〉β-fiber texture as the dominant component has not been reported previously in conventional β-Ti alloys, although the 〈110〉β-fiber texture is the same as the deformation texture of conventional bcc alloys. The Young's modulus of the solution-treated specimens along the rolling direction decreased with an increase in the CSA reduction rate. This was attributed to the development of the 〈001〉β-fiber texture.

Simple Summary Black soldier fly (BSF) larvae and pre-pupae were raised on nutritionally resembling household organic waste. Next, whole (non-defatted) BSF larvae and pre-pupae were dried and added to the diets of laying hens as a replacement of soybean meal and oil contents. Eggshell thickness and microbiota diversity in the cecum of hens supplemented with BSF pre-pupae showed higher values than those of hens fed with the control diet. It is suggested that chitin, an indigestible substance found in BSF, as well as BSF fat, possibly increased eggshell thickness and microbiota diversity values. Further investigation of the effect of BSF fat added to poultry feed is recommended. Abstract Black soldier fly (BSF) larvae and pre-pupae could be satisfactorily raised on household organic waste and used as poultry feed, offering a potential sustainable way to recycle untapped resources of waste. The present study was conducted to determine if whole (non-defatted) BSF larvae and pre-pupae raised on experimental household waste could substitute soybean meal and oil as ingredients for laying hen diets. While no significant differences in feed intake and the egg-laying rate of hens were observed throughout the experiment, egg weight and eggshell thickness were greater in the pre-pupae-fed group than in the other groups. Moreover, although diversity of the cecal microbiota was significantly higher in the pre-pupae-fed than in the control group, no significant differences in bacterial genera known to cause food poisoning were observed when comparing the treatment groups. Nonetheless, Lactobacillus and Bifidobacterium populations were significantly lower in the treatment than in the control group. Fat content in BSF was possibly related with the changes in the cecal microbiota. Hence, since BSF fat was deficient in essential fatty acids, special attention should be paid to the fat content and its fatty acid composition in the case of regular inclusion of BSF larvae and pre-pupae oil as an ingredient in poultry diets.

The standard for rotational alignment of the tibial component in total knee arthroplasty (TKA) remains unclear. Cases often require positioning of the tibial component, prioritizing adequate coverage of resected bone surface rather than alignment with the tibial rotational axis. We investigated tibial component position in TKA, prioritizing maximum coverage of resected bone surface, and evaluated the correlation with the tibial anteroposterior (AP) axis. We analyzed preoperative computed tomography images for primary TKA in 106 cases and 157 knees, using three-dimensional planning software. Tibial component position prioritizing maximum coverage of resected bone surface was simulated, and results were compared with the AP axis. Rotational alignment angle was defined as that between a line perpendicular to the tibial AP axis and a line connecting the posterior edge of the tibial component. The simulated tibial component was more externally rotated by a mean 4.5° ± 4.2°. The alignment angle showed normal distribution, but variability was large, ranging from 5.1° internal rotation to 16.2° external rotation. In 138 of 157 (87.9 %) knees, the tibial component was positioned in the externally rotated position with respect to the AP axis. The tibial component was aligned within the medial one-third of the patellar tendon in 122 of 157 (77.7 %) knees. The tibial component aligned using coverage prioritizing was externally rotated, although large variability was observed. Rotational alignment was optimal in 79 % of cases when the tibial component was aligned with coverage prioritizing, but hyperexternal rotation was observed in patients with severe knee deformation.

Texture formation in Ti-7Mo-8Al-6Zr cold-groove-rolled alloy wires was investigated and compared to that of Ti-5.5Mo-8Al-6Zr. Irregularly textured -fiber was formed in Ti-5.5Mo-8Al-6Zr. The Ti-7Mo-8Al-6Zr cold-groove-rolled wire mainly composed of the β phase though α″ martensite was induced by cold-groove-rolling in the Ti-5.5Mo-8Al-6Zr alloy. From the TEM observations of the severely deformed cold-rolled sheet, α″ martensite was assumed to be induced during cold-groove-rolling and trace α″ was assumed to be retained after cold-groove-rolling. This texture is common in bcc metals and corresponds to the α″ -fiber which is formed in the Ti-5.5Mo-8Al-6Zr alloy when the lattice correspondence between β and α″ is considered. On the other hand, the -fiber was strongly developed in the cold-groove-rolled wire with a reduced cross sectional area of 98% following solution treatment, while this texture was not observed in the specimen with a 60% reduced cross sectional area. The texture formed in solution-treated Ti-7Mo-8Al-6Zr alloy wire was similar to that of the Ti-5.5Mo-8Al-6Zr alloy, although the dominant phase in the cold-groove-rolled specimen differed.

The present study aimed to examine the relationship between the structure and composition of dentin and odontoblasts and the role of melatonin during the calcification process. The expression of MT1 and MT2 melatonin receptor was confirmed in the odontoblasts of the control group. In addition, the expression of MT1 was stronger than that of MT2. A strong expression of MT1 was detected in the odontoblasts in the melatonin-treated groups. MALDI-TOF MS analysis revealed that two peaks of 795 m/z and 818 m/z were found in dentin. These peaks increased commensurately with the amount of melatonin. The number and size of calcospherites in predentin increased in proportion to the concentration of melatonin. The degree of mineralization increased slightly in the melatonin-treated group using CMR analysis. Two peaks could be clearly detected in the melatonin-treated group at nighttime using X-ray diffraction analysis. Melatonin may participate in the dentin composition and the calcification mechanism of dentin.

Calcium (Ca) storage in bone has a relationship with eggshell production. Forced molting by feeding restriction for older hens improves eggshell quality but leads to a decline in the bone quality. Dietary minerals improve the eggshell and bone quality; however, the effect on bone quality post-molting has not been clarified. This study evaluated the effects of dietary Ca and minerals on the eggshell and bone quality during both pre- and post-molting periods. The bone quality was evaluated by measurement of bone density and Fourier transform infrared spectroscopy (FT-IR) analysis. The eggshell quality was evaluated by morphological observation of the mammillary cores by scanning electron microscope and FT-IR analysis. The high Ca concentration feed group showed a low bone density post-molting and a high carbonate/phosphate ratio pre-molting. In high mineral concentration feed group, the eggshell strength, the thickness, and the proportion of large mammillary core areas were significantly higher (p < 0.05) than control group post-molting. These results suggest that the eggshell strength increases as the proportion of mammillary core areas increases. Furthermore, the high carbonate/phosphate ratio promoted a decrease in bone density. Therefore, the concentration of dietary mineral is strongly related with the maintenance of eggshells and bone quality post-molting.

Collagen in an eggshell membrane is important for egg preservation, medical burn treatment and manufacturing of cosmetics. Because collagen in the membrane is little, it is a need to improve the accumulation in the membrane to develop these applications. Wood charcoal powder with vinegar (WCV) is a natural substance that improves poultry production. In hen fed with WCV, total collagen in the eggshell membrane increased with an increase in dietary WCV and significantly increased in the 1.0% WCV group (p < 0.05). Scanning and light microscopic images revealed that this group had thicker eggshell membranes and a fine mesh structure composed of finer and more densely distributed fibres than in the control. Eggs from WCV group showed slow Haugh unit decrease during egg storage and the decrease correlated with total collagen in eggshell membrane. In intact chicken, type I and type III collagens were found in different specific locations in the oviduct but not in the membrane. The finding that collagen accumulates in the eggshell membrane under WCV feeding suggests that feeding chicken with WCV will permit long-term storage of eggs in poultry production, and the increased volume of total collagen will facilitate its application in medicine and cosmetics.

Hypothalamic terminals were investigated in the rat posterior pituitary (PP). Injection of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) and co-injection of WGA-HRP with Rab3A-siRNA were made into the hypothalamus, respectively. Additional injection of WGA-HRP was made into the hypothalamus in the animals exposed to ethanol. These injections resulted in heavy labeling of fibers exclusively confined to the PP. Ultrastructural observations showed terminals, fibers, pituicytes, capillaries and vascular spaces in the PP. Although the majority of terminals were observed to contain large dense core vesicles (LDCVs) and HRP-reaction products (HRP-RPs), exocytosis of LDCVs in close proximity to cell membrane was not found. Interestingly, a few terminals showed alteration of cell membrane called "apocrine-like structure" containing LDCV and RP. The narrow neck portion of the structure gave the appearance that it may have been in some stage of separating from terminals. Other remarkable feature was that terminals occasionally reveal the structure of "leakage" of RP discharged into vascular spaces crossing cell membrane. Such hormone-releasing mechanism might be involved in one of "diacrine-like secretion". In the present study secretion-related structures of hypothalamic terminals in the PP are quite different from normal vesicular exocytosis.

This letter presents the experimental investigation to increase the sensitivity of an optical fiber refractive index sensor with a multimode interference (MMI) structure. It is confirmed that the interference wavelength can be set in the long-wavelength region by adjusting the sensing-part length. Moreover, it is shown that the fineness of the cores in the input and output fibers is essential to obtain a sharp interference signal. We demonstrate the high sensitivity of an MMI-structured optical fiber sensor by applying it to refractive index measurements of ethanol/water solutions. This refractive index sensor reveals a resolution as fine as 1.8 × 10-5, which is the highest value so far reported in this type and has one order higher sensitivity than a commercially available Abbe refractometer (∼ 10-4).

Hypoxia promotes neural stem cell proliferation, the mechanism of which is poorly understood. Here, we have identified the nuclear orphan receptor TLX as a mediator for proliferation and pluripotency of neural progenitors upon hypoxia. We found an enhanced early protein expression of TLX under hypoxia potentiating sustained proliferation of neural progenitors. Moreover, TLX induction upon hypoxia in differentiating conditions leads to proliferation and a stem cell-like phenotype, along with coexpression of neural stem cell markers. Following hypoxia, TLX is recruited to the Oct-3/4 proximal promoter, augmenting the gene transcription and promoting progenitor proliferation and pluripotency. Knockdown of Oct-3/4 significantly reduced TLX-mediated proliferation, highlighting their interdependence in regulating the progenitor pool. Additionally, TLX synergizes with basic FGF to sustain cell viability upon hypoxia, since the knockdown of TLX along with the withdrawal of growth factor results in cell death. This can be attributed to the activation of Akt signaling pathway by TLX, the depletion of which results in reduced proliferation of progenitor cells. Cumulatively, the data presented here demonstrate a new role for TLX in neural stem cell proliferation and pluripotency upon hypoxia.

原発性骨粗鬆症の患者に非外傷性に発症した胸骨骨折の1症例を経験したので報告する．症例：80歳，女性．多発性胸椎腰椎圧迫骨折の既往あり．2010年1月，外傷の既往なく，前胸部の激しい疼痛を自覚したため当院を受診した．初診時，胸骨に強い圧痛を認め，胸骨単純X線像にて胸骨体の上1/3の部位に骨折を認めた．また病的骨折を疑わせる所見は見られなかった．躯幹骨二重X線吸収（DXA）法による骨量測定値は腰椎にて若年成人平均値(YAM)の58％，大腿骨にてYAMの38％とWHOの骨折リスクはHigh Riskであった．経過：非外傷性胸骨骨折と診断し，保存療法が可能と考えバストバンド固定のうえ入院加療を行った．その後疼痛は漸減した．考察：本症例の発症機序として原発性骨粗鬆症に起因する後弯変形を原因に胸骨に持続的なストレスがかかったことによって非外傷性に骨折が生じたものと考えられた．

The hypothesis that apoptotic factors play some roles in the denucleation of erythroblasts has been confirmed by the immunohistological detection of both phosphatidylserine and thrombospondin as phagocytosis-inducing factors in general apoptotic events. Both phosphatidylserine and thrombospondin were detected on the surface of cell membrane of mature erythroblasts, while thrombospondin was also detected in more immature erythroblasts. The intensities of their immune reactions increased as the erythroids matured. During denucleation, the positivities of both phosphatidylserine and thrombospondin were restricted on the surface of the cell membrane surrounding the protruding nuclei. Thus, the apoptotic process involves denucleation of erythroblasts and phosphatidylserine, and thrombospondin acts as phagocytosis-inducing factors in the denucleation event.

It has been hypothesized that Rab3A, a small GTPase, may be closely involved in the process of dense core vesicle exocytosis in various cell types. This possibility was investigated by disrupting the expression levels of Rab3A-mRNA using a small interfering RNA of the Rab3A GTPase (Rab3A-siRNA) and examining the effect of this on transcytosis of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP). Rab3A-siRNA and WGA-HRP were injected into the right vagus nerves of adult rats which were killed 12, 24 or 48 hours later. In some animals, portions of the brain stem containing the nucleus of solitary tract (NST) were prepared for electron microscopy. In other animals, the nodose ganglion of the vagus nerve was used to determine the levels of expression of Rab3A-mRNA using RT-PCR techniques. It was found that the expression of Rab3A-mRNA was markedly depressed in animals at 12 h after the Rab3A-siRNA injection. In the NST, there was an accumulation of HRP-reaction product (RP), recognized as electron dense lysosomal-like structures, in both axons and terminals in the NST 12 h after injection. Some HRP-RP was found in membrane bound vesicles in close proximity to cell membranes and appeared to be in the process of transcytosis. This neuronal transcytosis of HRP-RP appeared to occur at random locations over the axodendritic membranes. These findings indicate that inhibiting the expression of Rab3A-mRNA using Rab3A-siRNA can modulate the level of transcytosis of proteins across neuronal membranes confirming the potentially important role of this GTPase in the process of transcytosis.

The orphan nuclear receptor TLX has been proposed to act as a repressor of cell cycle inhibitors to maintain the neural stem cells in an undifferentiated state, and prevents commitment into astrocyte lineages. However, little is known about the mechanism of TLX in neuronal lineage commitment and differentiation. A majority of adult rat hippocampus-derived progenitors (AHPs) cultured in the presence of FGF express a high level of TLX and a fraction of these cells also express the proneural gene MASH1. Upon FGF withdrawal, TLX rapidly decreased, while MASH1 was intensely expressed within 1h, decreasing gradually to disappear at 24h. Adenoviral transduction of TLX in AHP cells in the absence of FGF transiently increased cell proliferation, however, later resulted in neuronal differentiation by inducing MASH1, Neurogenin1, DCX, and MAP2ab. Furthermore, TLX directly targets and activates the MASH1 promoter through interaction with Sp1, recruiting co-activators whereas dismissing the co-repressor HDAC4. Conversely, silencing of TLX in AHPs decreased beta-III tubulin and DCX expression and promoted glial differentiation. Our results thus suggest that TLX not only acts as a repressor of cell cycle and glial differentiation but also activates neuronal lineage commitment in AHPs.

XY females are rare individuals who carry a Y chromosome but are phenotypically female. In approximately 80-90% of these cases, there are no mutations in the SRY gene, a testis-determining gene on the short arm of the Y chromosome, and the pathophysiology of XY females without SRY mutation remains unclear. In the present study, we used a molecular data mining technique to analyze the pathophysiology of an XY female with functional SRY and pericentric inversion of the Y chromosome, and compared the results with those of a normal male. Interestingly, upregulations of numerous genes included in the development category of the Biological Process ontology, including genes associated with sex determination and organ morphogenesis, were seen in the patient. Additionally, the transforming growth factor-beta (TGF-beta) signaling pathway and Wnt signaling pathway, in which most cell-cell interactions during embryonic development are involved, were altered. Alterations in the expression of numerous genes at the developmental stage, including alterations at both the gene and pathway levels, may persist as a vestige of anomalies of sex differentiation that presumably began in the fetal period. The present study indicates that a data mining technique using bioinformatics contributes to identification of not only genes responsible for birth defects, but also disorders of sex development (DSD)-specific pathways, and that this kind of analysis is an important tool for clarifying the pathophysiology of human idiopathic XY gonadal dysgenesis. Our findings could serve as one of the basic datasets which will be used for future follow-up investigations.

In normal ontogenetic development, the expression of the sex-determining region of the Y chromosome (SRY) gene, involved in the first step of male sex differentiation, is spatiotemporally regulated in an elaborate fashion. SRY is expressed in germ cells and Sertoli cells in adult testes. However, only few reports have focused on the expressions of SRY and the other sex-determining genes in both the classical organ developing through these genes (gonad) and the peripheral tissue (skin) of adult XY females. In this study, we examined the gonadal tissue and fibroblasts of a 17-year-old woman suspected of having disorders of sexual differentiation by cytogenetic, histological, and molecular analyses. The patient was found to have the 46,X,inv(Y)(p11.2q11.2) karyotype and streak gonads with abnormally prolonged SRY expression. The sex-determining gene expressions in the patient-derived fibroblasts were significantly changed relative to those from a normal male. Further, the acetylated histone H3 levels in the SRY region were significantly high relative to those of the normal male. As SRY is epistatic in the sex-determination pathway, the prolonged SRY expression possibly induced a destabilizing effect on the expressions of the downstream sex-determining genes. Collectively, alterations in the sex-determining gene expressions persisted in association with disorders of sexual differentiation not only in the streak gonads but also in the skin of the patient. The findings suggest that correct regulation of SRY expression is crucial for normal male sex differentiation, even if SRY is translated normally.

Alcohol ingestion affects both neuropsychological and motor functions. We hypothesized that one of the key factors involved in such functions are neurotrophins and their receptors. We have therefore examined the effects of short-term ethanol exposure on the mRNA expression and protein levels of neurotrophin ligands and receptors in the cerebellum using real-time RT-PCR and Western blotting techniques. Male BALB/C mice were fed a liquid diet containing 5% (v/v) ethanol. The pair-fed control mice were fed an identical liquid diet except that sucrose was substituted isocalorically for ethanol. The cerebellum of mice exhibiting intoxication signs of stage 1 or 2 were used in the present study. We found that exposure to ethanol resulted in elevated levels of nerve growth factor (NGF) and TrkA mRNA expression but a decreased level of brain-derived neurotrophic factor (BDNF) mRNA expression. The expression of TrkB and p73 mRNA was unchanged. Changes in the level of these proteins were found to mirror these mRNA expression levels. We conclude that exposure to ethanol for a short period can cause a differential responsive in the various neurotrophin ligand/receptor systems. The functional consequences of these changes are unknown at present.

【はじめに】癌患者において脊椎転移のため疼痛や神経麻痺を生じADLが著しく障害されることをしばしば見受ける．今回当科において癌の脊椎転移に対し手術治療を施行した症例のADL障害を評価し報告する．【対象と方法】対象は2005年5月から2007年11月に癌の脊椎転移に対し手術を行った10症例．適応は徳橋の術前予後判定法を参考に決定．手術方法は徳橋のスコアリングに従い決定し，術前，術後のFrankel分類，Performance statusを評価した．【結果】術後，全症例においてFrankel分類，Performance statusによる評価で改善を認めた．生存中の歩行可能期間は8症例が術後生存期間の7割以上の期間歩行可能であった．【考察と結語】多くの報告と同様に手術を行った全症例で麻痺の改善，ADLの向上を速やかに認めた．10症例中8症例が自宅療養となり価値の大きい治療であると考えられた．

Herein, we show that both exogenously transfected and endogenously activated p53 repress promoter activity and expression of PDGFRB. p53 binds the proximal promoter containing the CCAAT motif as examined by EMSA and chromatin immunoprecipitation. However, gradual induction of p53 in tet-onSAOS2 cells resulted in a transient increase of the PDGFRB-promoter activity and its expression. As binding of p53 to the promoter increased, previously bound p73, DeltaNp73, c-Myc, HDAC1 and HDAC4 were dismissed from the repressed promoter, and p300 was recruited. The transient increase of the promoter activity was therefore induced by the release of the p73, Myc and HDACs, previously shown to act as repressors to this promoter. Along with further increase of p53, p300 was replaced by HDAC1 and HDAC4, resulting in decreased PDGFRB expression. For the repression, acetylation of the C-terminal lysines of p53 is important, and both acetyl-K373p53 and methyl-K370p53 became bound to the promoter. The acetyl-K373p53 was accumulated in the nucleus and colocalized with promyelocytic leukemia protein. Mitomycin treatment of MEF induced similar epigenetic modification of p53 and its binding to the promoter chromatin. Addition of a PDGFR tyrosine-kinase inhibitor to p53-inducing tet-onSAOS2 increased the number of apoptotic cells. These results suggest that p53 represses the PDGFRB promoter, facilitating the p53-induced apoptosis, whereas tumor cells with p53 mutation or a high level of DeltaNp73 or Myc could become refractory to the regulation.

Maternal alcohol ingestion during pregnancy adversely affects the developing fetus, often leading to fetal alcohol syndrome (FAS). One of the most severe consequences of FAS is brain damage that is manifested as cognitive, learning, and behavioral deficits. The hippocampus plays a crucial role in such abilities; it is also known as one of the brain regions most vulnerable to ethanol-induced neurotoxicity. Our recent studies using morphometric techniques have further shown that ethanol neurotoxicity appears to affect the development of the dentate gyrus in a region-specific manner; it was found that early postnatal ethanol exposure causes a transitory deficit in the hilus volume of the dentate gyrus. It is strongly speculated that such structural modifications, even transitory ones, appear to result in developmental abnormalities in the brain circuitry and lead to the learning disabilities observed in FAS children. Based on reports on possible factors deciding ethanol neurotoxicity to the brain, we review developmental neurotoxicity to the dentate gyrus of the hippocampal formation.

We report a considerable number of cells in the ventricular and the subventricular zones (SVZ) of newborn mice to stain positive for the PDGF beta-receptor (PDGFRB). Many of them also stained for nestin and/or GFAP but less frequently for the neuroblast marker doublecortin and for the mitotic marker Ki-67. The SVZ of mice with nestin-Cre conditional deletion of PDGFRB expressed the receptor only on blood vessels and was devoid of any morphological abnormality. PDGFRB(-/-) neurospheres showed a higher rate of apoptosis without any significant decrease in proliferation. They demonstrated reduced capacities of migration and neuronal differentiation in response to not only PDGF-BB but also bFGF. Furthermore, the PDGFR kinase inhibitor STI571 blocked the effects of bFGF in control neurosphere cultures. bFGF increased the activity of the PDGFRB promoter as well as the expression and phosphorylation of PDGFRB. These results suggest the presence of the signaling convergence between PDGF and FGF. PDGFRB is needed for survival, and the effects of bFGF in migration and neural differentiation of the cells may be potentiated by induction of PDGFRB.

We aimed to elucidate the mechanism of action of estrogenic endocrine disruptors and the rescue of reproductive function, particularly the responsiveness of testes to eCG and/or activin A (ACT) after establishing reproductive disorders. Newborn male mice (n = 29) were randomly divided into an untreated group and three treatment groups that received diethylstilbestrol (DES; 100 mug per animal) subcutaneously on Postnatal Day 3 to establish reproductive disorders and daily treatment with PBS (controls: DES + PBS), eCG (eCG group: DES + eCG), or eCG + ACT (eCG + ACT group: DES + eCG + ACT) at 6-8 wk of age prior to mating. After treatment, the controls showed diminished Leydig cells in the testes and thin germ cell layers containing pyknotic germ cells and multinucleated cells. In the eCG and eCG + ACT groups, spermatids and Leydig cells increased markedly. The immunoexpression of androgen receptors in the eCG group and steroidogenic acute regulatory (STAR) protein in the eCG and eCG + ACT groups recovered to approximately the levels in the untreated group; plasma LH and testosterone levels also increased relative to those in the controls. In addition, the cell proliferation index, which is estimated from 5-bromo-2'-deoxyuridine immunoexpression in spermatogonia, increased significantly under eCG treatment, and even more with eCG + ACT. However, the numbers of germ and Leydig cells decreased at 12 wk of age. Thus, ACT and eCG help the testes to recover from the dysfunction induced by neonatal DES administration. Furthermore, the permanent male reproductive disorder induced by neonatal exposure to estrogenic agents may be more likely to result from dysfunction of the hypothalamic-pituitary axis than from dysfunction of the lower reproductive organs.

The expression of calcium binding proteins (CaBPs) has been linked to protection of neurons. The present study investigated the effects of monocular enucleation on the distribution of parvalbumin immunoreactive (PV-IR) neurons and glial fibrillary acidic protein immunoreactive (GFAP-IR) astrocytes in both the dorsal (dLGN) and ventral (vLGN) regions of the lateral geniculate nucleus (LGN). Our results demonstrated an increase in PV-IR neuronal density on the contralateral vLGN at 1-week post-enucleation (PE), which was maintained without significant change for 12 weeks. By contrast, PV-IR neurons in dLGN decreased significantly at all time point examined. The number of GFAP-IR astrocytes showed an initial increase from 1 to 4 weeks PE and then gradually decreased until 48 weeks in both regions of the LGN with contralateral side predominance. The present results suggest that monocular enucleation results in variable expression of PV-IR neurons and GFAP-IR astrocytes in the LGN complex, which may play an important role in neuronal degeneration and neuroplasticity of the rat visual system.

The present study was undertaken to evaluate the effects of monocular enucleation on the calbindin-D 28k (CB) and c-Fos immunoreactive (IR) neurons in the lateral geniculate nucleus (LGN) complex of adult rats. The enucleation resulted in neuronal degeneration and decrease of neurons in the LGN complex. Our study demonstrated a decrease of CB-IR neuronal density on the contralateral side of the ventral (vLGN) and dorsal LGN (dLGN) until 12 weeks post-enucleation (PE). On the ipsilateral side, CB-IR neuronal density in the dLGN and vLGN showed significant and continuous decrease until 48 and 12 weeks PE, respectively. In an additional experiment, c-Fos-IR neurons were increased at 2 days PE in the vLGN with ipsilateral predominance. At 7 days, c-Fos-IR neurons on the ipsilateral vLGN were still higher than those of pre-enucleated rats. The present results suggest that monocular enucleation affects the expression of the CB and c-Fos in the LGN complex. It is indicated that these may play an important role in the neuronal degeneration and neuroplasticity of the subvisual system in rats.

The effects of maternal deprivation (MD) during early postnatal life on the brain-derived neurotrophic factor (BDNF) level were investigated in the present study. Wistar rats were assigned to either maternal deprivation or mother-reared control (MRC) groups. MD manipulation was achieved by separating rat pups from their mothers for 3 h a day during postnatal days (PND) 10-15. At 16, 20, 30, and 60 days of age, the level of BDNF mRNA in the hippocampal formation of each group was determined using real-time PCR analysis. Early postnatal maternal deprivation of rat pups resulted in a significant increase in body weight at 60 days of age. The expression of BDNF mRNA in the hippocampus was significantly decreased at 16 days of age, and increased at 30 and 60 days of age. These data indicate that even a brief period of maternal deprivation during early postnatal life can affect hippocampal BDNF expression.

We investigated mechanisms of the p73alpha-mediated repression of the platelet-derived growth factor beta-receptor (PDGFRB) promoter caused by its interaction with NF-Y. Treatment of cells with the histone deacetylase (HDAC) inhibitor, Trichostatin A, increases PDGFRB promoter activity through the CCAAT motif and counteracts the repression caused by p73alpha. Activation of the PDGFRB promoter by the co-activator p300 also occurs through the CCAAT motif. Expression of p73alpha counteracts both p300- and P/CAF-mediated activation of the PDGFRB promoter, and expression of p300 or P/CAF attenuates the p73alpha-mediated repression of the promoter activity. In concordance, p73alpha decreases the p300-mediated acetylation of NF-YC, p300 competes with p73alpha for binding NF-YB, and P/CAF competes with p73alpha for binding NF-YB and NF-YC. Furthermore, p73alpha, but not the oncogenic DeltaNp73alpha, binds directly to HDAC1. We performed chromatin immunoprecipitation with antibodies against p73, DeltaNp73, NFYB, p300 and HDAC1 at different periods after serum stimulation in serum-starved NIH3T3 cells. A marked decrease of DeltaNp73, NF-YB and p300 was detected 6 hours after serum stimulation when the expression of PDGFRB decreased. Conversely, HDAC1 was found bound at its maximum and the anti-p73 detecting both TAp73 and DeltaNp73 was found at all time points, indicating that p73, but not DeltaNp73, remains bound at this time. Double immunofluorescence staining of TAp73 and HDAC1 revealed that both of these molecules exist in the nucleus at this time point, supporting the presence of endogenous interaction. These results suggest that p73 and DeltaNp73 behave as physiological regulators for the transcription of the PDGFRB promoter.

The suprachiasmatic nucleus (SCN) has long been recognized as the central mammalian circadian pacemaker that controls behavioral and physiological processes. The role of the SCN in circadian rhythms has been the subject of a wide range of physiological and behavioral studies, although the influence of homeostasis rhythms (such as fluctuating hormone levels) on the SCN of the hypothalamus is not entirely clear. The present study was undertaken to examine the morphological interactions between astroglial and neuronal elements in the SCN of mice after either a short-term excessive testosterone treatment (ETT) or castration, using glial fibrillary acidic protein (GFAP), and immediate early gene c-fos as well as calbindin-D28k (CB) immunohistochemistry. Both ETT and castration resulted in a significant increase in the accumulation of reactive astrocytes and Fos-imunoreactivity (IR), especially in the dorsomedial (DM) sub-region of the SCN. However, CB-IR neurons in the examined brain regions showed little change. These findings indicate that the DM sub-region of the SCN may be a possible center of hormonal regulation via a hypothalamic neuroendocrine circuit, and that a non-photic stimuli mechanism might play a role in circadian rhythm regulation.

We have examined alterations in the hypothalamo-pituitary GH-somatic growth axis and the hypothalamo-pituitary LH-ovarian axis in a line of transgenic ICR mice expressing human GH (hGH) under the influence of the whey acid protein promoter. Transgenic female mice weighed twice as much as control females and were infertile. The size of the anterior pituitary (AP) was 1/3 that of the controls. In transgenic mice, acinar cells in the mammary and mandibular glands displayed hGH-immunoreactivity, and plasma hGH was detected by radioimmunoassay. In the medial basal hypothalamus (MBH) of transgenic females, the immunoreactive-GHRH level was decreased (P

Pituitary glands of chicken, from stages 20 (70 ∼ 72 h of incubation) to 46 (20 days) of Hamburger and Hamilton (1951), were studied by immunocytochemical and histological stainings and India ink injection into blood vessels. Using the distribution pattern of 6 types of immunoreactive adenohypophyseal cells and the location of pituitary stalk as guideposts, we found how specific areas in the epithelium of Rathke’s pouch differentiate into specific regions of the adenohypophysis at 20 days. In the sagittal plane, the walls of Rathke’s pouch were tentatively divided into the upper part (A1 + A2) and lower part (A3) of the anterior wall, and the posterior wall (P1 + P2 + P3). The cephalic lobe was mainly assembled by the proliferation of parenchymal cells in the areas A2 + A3 + P2 of Rathke’s pouch epithelia at 3 days of incubation. The caudal lobe was derived from A1 + P1 + P3. The pars tuberalis was derived from A1 + A2. Thus, the avian adenohypophysis is established at 13 days, though the blood supply to the pars distalis is established at 20 days. Therefore, the cephalic lobe and caudal lobe of the pars distalis and the pars tuberalis of the chicken adenohypophysis are derived from specific areas of the cell cords of Rathke’s pouch at 3 days of incubation.

We have investigated elemental and isotopic compositions of noble gases in the Kobe (CK4) chondrite by a stepwise heating method. The previously observed inconsistency in cosmic-ray exposure ages derived from cosmogenic 21Ne and 38Ar was eliminated in this study, and the averaged value of 38.2 Ma was obtained. Based on neutron-produced 80Kr, a pre-atmospheric size of Kobe is estimated to be larger than ∼23 cm radius, whereas the results from cosmogenic radionuclides indicate a pre-atmospheric radius of ∼10 cm. These results indicate a two-stage exposure history for the Kobe meteorite. The release pattern of the primordial noble gas from Kobe indicates that Kobe has lost its primordial gas by thermal metamorphism. However, the observed Q-Xe concentration is lower than that expected from the HL-Xe as a carbonaceous chondrite, indicating preferential loss of Q-Xe. This cannot be explained solely by thermal metamorphism, suggesting that shock events at high temperature have caused the preferential loss of Q-Xe over HL-Xe in the Kobe meteorite.

Excessive intake of sugar-rich foods leads to metabolic syndrome. D-Psicose (Psi) not commonly found in nature, is noncalorie sweetener with a suppressive effect on the blood glucose level. Thus, Psi has the potential to be utilized as a sucrose (Suc) replacer in sugar-rich foods, including meringue-based confectionery (MBC). In this study, we investigated the effect of Psi on the physical and chemical properties of meringue. Meringue was made by whipping egg white and Suc (at a weight ratio of 1:1) and baking at 93 °C for 2 h. Thirty percent of the total weight of Suc was replaced with D-ketohexoses such as Psi, D-fructose, D-tagatose, and D-sorbose. The meringues containing D-ketohexoses had higher specific volume than the meringue not containing D-ketohexoses (Ct-meringue). Baking of meringue caused differences between Psi and the other D-ketohexose meringues. Meringue containing Psi (P30-meringue) had the highest breaking stress (7.00 × 10(5) N/m(2)) and breaking strain (4.40%), resulting in the crunchiest texture. In addition, P30-meringue also had the highest antioxidant activity (491.84 μM TE/mg-meringue determined by ABTS method) and was the brownest due to a Maillard reaction occurring during baking. The replacement of Suc with Psi improved the characteristics of baked meringue. Thus, Psi was found to be useful in modifying the physical and chemical properties of MBC.

A line of transgenic rats (heterozygotes) carrying a chimeric gene comprising a regulatory portion of murine whey acidic protein and a structural portion of human GH (hGH) genes developed severe obesity with age. To characterize physiological mechanisms that lead to fat accumulation, an array of parameters related to obesity were studied. Blood hGH levels were continuously low, endogenous rat GH secretion was suppressed, and the pulsatility in peripheral GH levels was absent. Plasma glucose, insulin, triglyceride, and FFA levels in the male transgenic rats significantly exceeded those in nontransgenic littermates at 12 and 17 weeks, but not at 7 weeks, of age. All symptoms except hyperlipidemia were restored to normal by treatment with an antidiabetic agent, thiazolidinedione (troglitazone), for 1 week from 17 weeks of age. As phenotypic expression of obesity was already evident before aberration of physiological parameters, it was assumed that animals had a condition in which obesity or hyperlipidemia caused hyperinsulinemia. Gene expression and enzymatic activity of lipoprotein lipase in the adipose tissue in the transgenic rats were not different from those in normal rats. In contrast, the gene expression level of glycerol-3-phosphodehydrogenase was markedly elevated, suggesting that glycerol synthesis was much enhanced in the adipocytes of the transgenic rats. In an ip glucose tolerance test, the transgenic rats were not hyperglycemic at 7 weeks of age; however, the animal became hyperglycemic at 15–17 weeks of age. Finally, treatment with recombinant hGH for 1 week to produce pulsatile secretion reduced the size of epididymal and kidney fat pads and restored normal weight gain. These observations suggest that continuously low peripheral GH levels with the lack of pulsatile secretion resulted in obesity and noninsulin-dependent diabetes mellitus.

The effect of continuous human GH (hGH) secretion on female reproduction was studied in adult female transgenic rats expressing the hGH gene with a mouse whey acidic protein (mWAP) promoter. Two lines of transgenic female rats carrying the mWAP/hGH gene were established and used in the study. One was characterized by relatively high levels of serum hGH (high line), and the other had relatively low levels (low line). High-line female rats had recurring, pseudopregnancy-like estrous cycles accompanied by increased serum progesterone levels for 2 wk after ovulation, and they were fertile. In these rats, luteinization occurred spontaneously without cervical stimulation, probably due to high levels of serum hGH, which has prolactin (PRL)-like activity in the rat. Although low-line female rats had recurring, regular 4-day estrous cycles, they were sterile. In these rats, pseudopregnancy could not be induced by mating or by mechanical cervical stimulation. PRL surges following cervical stimulation were not detected, and PRL secretion was not induced by a dopamine antagonist, metoclopramide. The ovarian tissue in this line had a much higher number of corpora lutea and grew much heavier than in normal littermates, suggesting impairment of PRL-induced structural luteolysis. Suppression of PRL secretion in low-line rats was, at least in part, due to a marked decrease in the number of lactotrophs in the pituitary. The present study shows that the serum hGH level plays a crucial role in regulating luteal function in female transgenic rats expressing the hGH gene.

We comprehensively evaluated meat quality in chickens fed a diet consisting of wood charcoal and vinegar (WCV) using food scientific and histological approaches. In culled hens, lipid and fatty acid in Musculus semimembranosus, cooking loss and sensory tests of whole thigh meat, and meat texture of breast meat were observed. In male broilers, cross section of M. semimembranosus was used for observations on muscle area, perimysium, non-collagen total protein and total collagen content, and anti-collagen I and III reactions. In frozen male broilers, conventional morphology of M. semimembranosus as well as chicken anti-collagen III reaction to selected muscles of thigh meat and breast meat were compared between the control and WCV-fed birds. Increased lipid and fatty acids, decreased cooking loss, high score in total evaluation for sensory test of thigh meat, and decreased meat texture values were observed for culled hens fed WCV. The higher values of muscle area, total collagen and collagen III were observed for broilers fed WCV. No perimysium collapse for M. semitendinosus or increased collagen III reactions of M. tensor fasciae latae, the flexor muscle group and M. pectoralis superficialis were observed for frozen muscles in the WCV group. These total results suggest that WCV produces palatable and tender meat by increasing collagen III.

Aims : The effects of ethanol exposure on synaptic structure were investigated in the nucleus of solitary tract (NST) in rats, using the horse-radish peroxidase (HRP) method. Methods : Eight-week-old experimental rats were allowed free access to a liquid diet containing ethanol for 3 weeks, while controls were given an isocaloric diet. Some of the control and experimental animals were given an injection of wheat germ agglutinin conjugated with HRP (WGA-HRP) into the vagus nerve toward the end of the treatment period. After the treatment, the neuropil region of the NST was examined under an electron microscope. Results : We observed that a few terminals were characterized by deep indentation of axodendritic membranes into the post-synaptic neurons. This appeared to be similar to that commonly seen in exocrine glands. Interestingly, the indented portion often contained various sizes of vacuoles and flattened cisternae. HRP-reaction product (RP) transported to terminals was recognized easily as an electron-dense lysosomal substance when lead citrate staining was omitted. Terminals containing HRP-RP also revealed quite a similar structure with indentation of axodendritic membranes as described earlier. The results are considered to confirm that terminals forming ‘apocrine-like structures’ observed in the ethanol-fed animals with no injection of WGA-HRP originate from afferent fibers of the vagus nerve. Conclusion : The present study suggests the possibility that the alteration of the synaptic structure induced by ethanol exposure can lead to the neuronal transcytosis of materials including proteins which is different from the normal vesicular exocytosis involved in chemical synaptic transmission.

This study investigates the combined effect of silicic acid and bamboo vinegar compound liquid (SPV) on the growth and intestinal histological alterations in poultry. Forty-eight 7-day-old male Sanuki Cochin chickens were fed a commercial mash diet supplemented with SPV at 0, 0.1, 0.2, and 0.3% level ad libitum for 112 days. Body weight gain tended to improve with increased concentrations of dietary SPV, although these results were not statistically significant (P

Neuronal transcytosis was observed at the stage when no neurotransmitter was released after the injection of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP; WGA = 22 kDa, HRP = 40 kDa) into the vagus nerve. The co-injection of Rab3A-siRNA with WGA-HRP into the vagus nerve was performed to further examine this phenomenon. This co-injection resulted in the transcytosis of WGA-HRP, both of the passing type, by which it crossed the synapses, and of the secretion type followed by endocytosis of postsynaptic membranes. These findings raised the possibility in vivo that WGA plays an important role in the transcytosis of protein. Therefore, WGA may be a valuable tool for therapeutic drug targeting via transcytosis. The ability of WGA-conjugated Amyloid ß (WGA-Aß) to decrease amyloid deposits in Alzheimer’s disease was investigated. The conjugation of WGA to amyloid-ß (1-40) (Aß; 5 kDa) was confirmed. WGA-Aß was then shown to move to terminals by axonal flow in vivo as well as WGA-HRP. WGA-Aß was also observed in the nodose ganglion cells and terminals after injections of fluorescent Aß (FAß) into the vagus nerve and fluorescent WGA (FWGA) into the common carotid artery. These studies suggested that WGA-Aß could be localized to solitary neurons via transcytosis.

Nerve growth factor (NGF) as an autocrine or paracrine growth factor plays a critical role in the pathogenesis and progression of human pancreatic cancer. NGF is synthesized as a proform (proNGF) that, when cleaved, releases mature ligand (mNGF). proNGF and mNGF bind to high-affinity tyrosine kinase receptor A (TrkA) and low-affinity receptor p75 to different extents. Histamine is a potent stimulator of NGF in the inflammatory lesion as determined by ELISA. This has generally been attributed to the accumulation of mNGF. To determine the effect of histamine on nerve growth factor/receptor expression in human pancreatic cancer, the present study explored intracellular and extracellular NGF production and p75 and TrkA membrane receptor expression in the PANC-1, KMP-6 and PK-1 cell lines. Histamine enhanced NGF secretion and mRNA expression in PANC-1 and KMP-6 cells, but not in PK-1 cells. proNGF was revealed using Western blotting to be the predominant form of NGF, but was significantly reduced by histamine. p75 receptor binding was increased with histamine treatement, but no significant alteration was observed for TrkA. Proliferating cell nuclear antigen (PCNA), an important indicator of cell proliferation, was significantly reduced by histamine stimulation. H1 and H2 receptors were both observed in the pancreatic cancer cells, and the alterations induced by histamine were counteracted by H1 receptor antagonist pyrilamine; however, the H2 receptor subtype was excluded from this process. These results suggest that histamine induces distinct nerve growth factor/receptor responsiveness via H1 receptor-induced signaling, thus affecting pancreatic cancer cell proliferation.

In normal ontogenetic development, the expression of the sex-determining region of the Y chromosome (SRY) gene, involved in the first step of male sex differentiation, is spatiotemporally regulated in an elaborate fashion. SRY is expressed in germ cells and Sertoli cells in adult testes. However, only few reports have focused on the expressions of SRY and the other sex-determining genes in both the classical organ developing through these genes (gonad) and the peripheral tissue (skin) of adult XY females. In this study, we examined the gonadal tissue and fibroblasts of a 17-year-old woman suspected of having disorders of sexual differentiation by cytogenetic, histological, and molecular analyses. The patient was found to have the 46,X,inv(Y)(p11.2q11.2) karyotype and streak gonads with abnormally prolonged SRY expression. The sex-determining gene expressions in the patient-derived fibroblasts were significantly changed relative to those from a normal male. Further, the acetylated histone H3 levels in the SRY region were significantly high relative to those of the normal male. As SRY is epistatic in the sex-determination pathway, the prolonged SRY expression possibly induced a destabilizing effect on the expressions of the downstream sex-determining genes. Collectively, alterations in the sex-determining gene expressions persisted in association with disorders of sexual differentiation not only in the streak gonads but also in the skin of the patient. The findings suggest that correct regulation of SRY expression is crucial for normal male sex differentiation, even if SRY is translated normally.

Aims To investigate the precise mechanisms underlying the action of estrogenic endocrine disruptors, we evaluated the direct effects of synthetic estrogen diethylstilbestrol (DES) on steroidogenesis in Leydig cells, with particular emphasis on the expression of the cholesterol side-chain cleavage enzyme P450scc. Furthermore, the mechanism underlying the action of DES was compared with that of endogenous estrogen 17β-estradiol (E2), which has a potency equivalent to that of DES. Main methods TTE1 Leydig cells were treated with 5 × 10 − 8 μM to 5 μM DES or E2 for 24 h, and P450scc gene expression and the histone modifications underlying their transcriptional activation were examined using reverse transcription-polymerase chain reaction (RT-PCR) and chromatin immunoprecipitation (ChIP), respectively. Key findings P450scc mRNA expression in the DES-treated and E2-treated cells reduced in inverse proportion to the dose of DES and E2, respectively; however, cAMP stimulation induced a recovery in the expression to a level approximately equal to those in the controls. In the DES-treated cells, ChIP assay revealed histone deacetylation in the P450scc promoter region. Interestingly, E2 did not cause histone deacetylation. Significance In the early stages of steroidogenesis, DES and E2 directly induced a reduction in P450scc mRNA expression in inverse proportion to their doses, and treatment with cAMP restored the decreased P450scc mRNA expression. Furthermore, DES can induce alterations in the histone modification of the P450scc gene, and natural estrogen and synthetic estrogenic compounds such as DES may induce reproductive disorders through different molecular mechanisms.