Anatomy & Cell Biology

Online ISSN: 2093-3673
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(A-D) Immunohistochemical localization of cytokeratin (CK)5 in frozen sections of mouse thymus from control animals (A1-3, n=4), and at 3 (B1-3, n=7), 7 (C1-3, n=6), and 14 (D1-3, n=4) days after cyclophosphamide treatment. C, cortex; M, medulla; arrows, CK5+ thymic epithelial cells. Scale bars=100 µm (A1, B1, C1, D1), 50 µm (A2, A3, B2, B3, C2, C3, D2, D3). (E) Data on the relative intensity of CK5 expression in the thymic medulla are also plotted in a bar graph and expressed as mean±standard deviation. CY 3d, CY 1w, and CY 2w represent 3, 7, and 14 days after cyclophosphamide treatment, respectively. *P<0.001 compared with the control, as determined by the t-test.
(A-D) Immunohistochemical localization of cytokeratin (CK)14 in frozen sections of mouse thymus from control animals (A1-3, n=4), and at 3 (B1-3, n=7), 7 (C1-3, n=6), and 14 (D1-3, n=4) days after cyclophosphamide treatment. C, cortex; M, medulla; arrows, CK14+ thymic cortical epithelial cells. Scale bars=100 µm (A1, B1, C1, D1), 50 µm (A2, A3, B2, B3, C2, C3, D2, D3). (E) Data on the relative intensity of CK14 expression in the thymic medulla are also plotted in a bar graph and expressed as mean±standard deviation. CY 3d, CY 1w, and CY 2w represent 3, 7, and 14 days after cyclophosphamide treatment, respectively. *P<0.001 compared with the control, as determined by the t-test.
(A-D) Immunohistochemical localization of cytokeratin (CK)8 in frozen sections of mouse thymus from control animals (A1-3, n=4), and at 3 (B1-3, n=7), 7 (C1-3, n=6), and 14 (D1-3, n=4) days after cyclophosphamide treatment. C, cortex; M, medulla. Scale bars=100 µm (A1, B1, C1, D1), 50 µm (A2, A3, B2, B3, C2, C3, D2, D3). (E) Data on the relative intensity of CK8 expression in the thymic cortex are also plotted in a bar graph and expressed as mean±standard deviation. CY 3d, CY 1w, and CY 2w represent 3, 7, and 14 days after cyclophosphamide treatment, respectively. *P<0.001 compared with the control, as determined by the t-test.
High magnification images of the stellate and globular thymic medullary epithelial cells (mTECs) by immunohistochemical localization of cytokeratin (CK)5, CK8, and CK14 in frozen sections of the mouse thymus from control animals. (A, B) CK5 and CK14 immunoreactivity was observed in virtually all of the stellate mTECs, whereas thymic cortical epithelial cells (cTECs) barely expressed CK5 and CK14 in the thymic cortex of normal mice. (C) CK8 was expressed by a minor subset of mTECs, most of which are globular in appearance, in the thymic medulla of control mice, whereas all the cTECs expressed CK8 in the thymic cortex of normal mice. C, cortex; M, medulla; arrows, CK8+ globular mTECs. Scale bars=50 µm.
(A) Two-color double-label immunofluorescent localization of cytokeratin (CK)8 (green) and CK5 (red) in frozen sections of mouse thymus 3 days after cyclophosphamide treatment. C, cortex; M, medulla; arrows, CK5+CK8+ thymic epithelial cells. Scale bar=50 µm. Data on the relative number of CK5+CK8+ (B) and CK14+CK8+ (C) Cotical thymic epithelial cells (cTECs) are also plotted in a bar graph and expressed as mean±standard deviation. *P<0.001 compared with the control, as determined by the t-test.
Article
The thymus is a central lymphoid organ for T cell development. Thymic epithelial cells (TECs) constitute a major component of the thymic stroma, which provides a specialized microenvironment for survival, proliferation, and differentiation of immature T cells. In this study, subsets of TECs were examined immunohistochemically to investigate their cytokeratin (CK) expression patterns during thymus regeneration following thymic involution induced by cyclophosphamide treatment. The results demonstrated that both normal and regenerating mouse thymuses showed a similar CK expression pattern. The major medullary TECs (mTEC) subset, which is stellate in appearance, exhibited CK5 and CK14 staining, and the minor mTEC subset, which is globular in appearance, exhibited CK8 staining, whereas the vast majority of cortical TECs (cTECs) expressed CK8 during thymus regeneration. Remarkably, the levels of CK5 and CK14 expression were enhanced in mTECs, and CK8 expression was upregulated in cTECs during mouse thymus regeneration after cyclophosphamide-induced acute thymic involution. Of special interest, a relatively high number of CK5(+)CK8(+) TEC progenitors occurred in the thymic cortex during thymus regeneration. Taken together, these findings shed more light on the role of CK5, CK8, and CK14 in the physiology of TECs during mouse thymus regeneration, and on the characterization of TEC progenitors for restoration of the epithelial network and for concomitant regeneration of the adult thymus.
 
Effect of KA treatment on neuronal death and neuro degeneration in the hippocampus and amygdala. (A) Location of hippo campal cell death in a coronal brain section 48 h after KA treatment. (CA3, Cornu ammonis 3 of the hippocampus; CeA, Central nucleus of the amygdala ; BLA, Basolateral nucleus of the amygdala). (B) Photomicrograph of TUNEL and FJBstained cells in the CA3 region of the hippocampus, CeA, and BLA 48 h aft er KA treatment. Scale bars = 200 μm.
Effect of KA treatment on neuronal death and neurodegeneration in the hippocampus and amygdala. (A) Location of hippocampal cell death in a coronal brain section 48 h after KA treatment. (CA3, Cornu ammonis 3 of the hippocampus; CeA, Central nucleus of the amygdala; BLA, Basolateral nucleus of the amygdala). (B) Photo-micrograph of TUNEL and FJB-stained cells in the CA3 region of the hippocampus, CeA, and BLA 48 h after KA treatment. Scale bars = 200 µm.
Effect of KA treatment on p-JNK expression in the hippocampus and amygdala. Western blot showing the time course of p-JNK expression in the hippocampus (A) and amygdala (B) after KA treatment. p-JNK was normalized to α-tubulin and represented as arbitrary units. Data are presented as the mean±SEM. *indicates P<0.05.
Effect of KA treatment on p-14-3-3ζ expression in the hippocampus. (A) Representative micrographs showing p-14-3-3ζ immunoreactivity in mouse hippocampus after KA treatment. (B) Western blot showing the time course of p-14-3-3ζ and total 14-3-3ζ expression in the hippocampus after KA treatment. (C) p-14-3-3ζ was normalized to 14-3-3ζ and represented as ratio. Data are presented as the mean±SEM. *indicates P<0.05. Scale bar = 100 µm. Arrows indicate p-14-3-3ζ-immunostained cells from CA3 region of the hippocampus.
Effect of KA treatment on p-14-3-3ζ expression in the amygdala. (A) Representative micrographs showing p-14-3-3ζ immunoreactivity in the CeA after KA treatment. (B) Western blot showing the time course of p-14-3-3ζ and total 14-3-3ζ expression in the hippocampus after KA treatment. (C) p-14-3-3ζ was normalized to 14-3-3ζ and represented as a ratio. Data are presented as the mean±SEM. *indicates P<0.05. Scale bar=100 µm.
Article
Oxidative stress-induced cell death leads to phosphorylation of 14-3-3ζ at serine 58. 14-3-3ζ is detected at significant levels in cerebrospinal fluid after kainic acid (KA)-induced seizures. Here we examined temporal changes in 14-3-3ζ phosphorylation in the hippocampus and amygdala of mice after KA treatment. Mice were killed at 2, 6, 24, or 48 h after KA (30 mg/kg) injection. We observed an increase in TUNEL and Fluoro-Jade B (FJB)-stained neurons in the hippocampus and amygdala of KA-treated mice. Phospho (p)-14-3-3ζ and p-JNK expression was increased in the hippocampus 2 and 6 h after KA treatment, respectively. In immunohistochemical analysis, p-14-3-3ζ-positive cells were present in the CA3 region of the hippocampus and the central nucleus of amygdala (CeA) of KA-treated mice. Thus, phosphorylation of 14-3-3ζ at serine 58 may play an important role in KA-induced hippocampal and amygdaloid neuronal damage.
 
Immunohistochemical analysis of cytokeratin and connexin in the human fetal lung and abdominal organs: a fetus at 12 weeks. (A, D, G, J) Expression of cytokeratin-19 (CK19); (B, E, H, K) expression of cytokeratin-14 (CK14); (C, F, I, L) expression of connexin-43 (CX43). CK19 immunopositivity is seen in the lung (A), kidney (D), colon (G), and gallbladder (J), whereas these tissues lack CK14 immunoreactivity. The kidney and adrenal cortex (F) show positive results for CX43. Scale bar in (C)=0.5 mm (A-L).
Immunohistochemical results for cytokeratin and connexin in the human fetal kidney: 2 fetuses at 16 weeks. Panels (A) and (B) and panels (C) and (D) display adjacent sections of the same specimen. Connexin-43 (CX43) immunopositivity is seen in the renal tubules of the cortex (A, C), whereas strong immunoreactivity is seen for cytokeratin-19 (CK19) in the collecting ducts and renal pelvis (B, D). Scale bar in (A)=0.5 mm (A-D).
Immunohistochemical analysis for cytokeratin and connexin in the human fetal kidney: a fetus at 11 weeks. Panels (A) and (B) and panels (C) and (D) show adjacent sections. Connexin-43 (CX43) immunopositivity is seen in some of the renal tubules of the cortex (arrows in A and C), whereas cytokeratin-19 (CK19) shows strong immunoreactivity in the collecting ducts and renal pelvis (B, D). Scale bar in (A)=0.2 mm (A-D).
Article
Connexin-43, a major gap junction protein, and cytokeratin-19, one of the intermediate filament keratins, are known to be markers of well-differentiated epithelium. In this study, we investigated the expression of these markers in the head region, lungs, and abdominal organs of 10 human mid-term fetuses. The expression of connexin-43 was found to be restricted to the dura mater, kidney, and adrenal cortex. In the kidney, we found a clear site-dependent difference in the expression pattern of these markers: connexin-43 expression was observed in the tubules of the renal cortex whereas cytokeratin-19 was strongly expressed in the collecting ducts and renal pelvis. This difference remained unchanged throughout the fetal stages examined. Immunoreactivity was not observed for either of the markers in the intrarenal vessels, including the glomeruli, and mesangial cells. Connexin-43 expression seemed to be restricted to the metanephric vesicle-derived structures that differentiate in the urogenital ridge of the splanchnic mesoderm. The adrenal cortex also originates from the same para-aortic mesoderm. In contrast, in the urogenital organs, cytokeratin-19 seemed to be expressed in ducts derived from the urogenital sinus.
 
The dendritic spines at tertiary dendritic tree of a Purkinje cell. Decreased density and length of spines are observed in rolling mouse Nagoya (B) compared to wild type mouse (A). Some spines of rolling dendritic tree projected to irregular direction and intermingled adjacent spines (C) (Scale bar: 5 µm).
Quantitative analyses of the dendritic spines with NIH image. The density (A) and length (B) of spines have been decreased significantly in rolling mice compared to wild type (Student t-test, P<0.05).
Ectopic spines in proximal dendrites of rolling mouse Nagoya. High voltage electron microscope (HVEM) photomicrographs of wild type (A) and rolling (B) Purkinje cells. Numerous spines from proximal dendrtie of rolling mouse Purkinje cells are observed in both HVEM and semithin section (B, C). Spines (black or white stars) from proximal dendrite of rolling Purkinje cell make synapses with climbing fiber (Arrow, D). (Scale bar=10 µm in A, B and C; 1 µm in D).
Parallel fiber varicosities images taken by HVEM (×8,000). Each of the varicosities was the largest varicosity observed in rolling (B) and wild type mice (A) under the same magnification observation. The varicosity of rolling was large and had multiple pale spots, considered synaptic points (Scale bar: 5 µm).
Article
Voltage dependent calcium channels (VDCC) participate in regulation of neuronal Ca(2+). The Rolling mouse Nagoya (Cacna1a(tg-rol)) is a spontaneous P/Q type VDCC mutant, which has been suggested as an animal model for some human neurological diseases such as autosomal dominant cerebellar ataxia (SCA6), familial hemiplegic migraine and episodic ataxia type-2. Morphology of Purkinje cell (PC) dendritic spine is suggested to be regulated by signal molecules such as Ca(2+) and by interactions with afferent inputs. The amplitude of excitatory postsynaptic current was decreased in parallel fiber (PF) to PC synapses, whereas apparently increased in climbing fiber (CF) to PC synapses in rolling mice Nagoya. We have studied synaptic morphology changes in cerebella of this mutant strain. We previously found altered synapses between PF varicosity and PC dendritic spines. To study dendritic spine plasticity of PC in the condition of insufficient P/Q type VDCC function, we used high voltage electron microscopy (HVEM). We measured the density and length of PC dendritic spines at tertiary braches. We observed statistically a significant decrease in spine density as well as shorter spine length in rolling mice compared to wild type mice at tertiary dendritic braches. In proximal PC dendrites, however, there were more numerous dendritic spines in rolling mice Nagoya. The differential regulation of rolling PC spines at tertiary and proximal dendrites in rolling mice Nagoya suggests that two major excitatory afferent systems may be regulated reciprocally in the cerebellum of rolling mouse Nagoya.
 
Article
In rats, ageing results in dysfunctional patterns of micturition and diminished sexual reflexes that may reflect degenerative changes within spinal circuitry. In both sexes the dorsal lateral nucleus and the spinal nucleus of the bulbospongiosus, which lie in the L5-S1 spinal segments, contain motor neurons that innervate perineal muscles, and the external anal and urethral sphincters. Neurons in the sacral parasympathetic nucleus of these segments provide autonomic control of the bladder, cervix and penis and other lower urinary tract structures. Interneurons in the dorsal gray commissure and dorsal horn have also been implicated in lower urinary tract function. This study investigates the cellular localisation of PG-21 androgen receptors, steroid receptor co-activator one (SRC-1) and the phosphorylated form of c-AMP response element binding protein (pCREB) within these spinal nuclei. These are components of signalling pathways that mediate cellular responses to steroid hormones and neurotrophins. Nuclear expression of PG-21 androgen receptors, SRC-1 and pCREB in young and aged rats was quantified using immunohistochemistry. There was a reduction in the number of spinal neurons expressing these molecules in the aged males while in aged females, SRC-1 and pCREB expression was largely unchanged. This suggests that the observed age-related changes may be linked to declining testosterone levels. Acute testosterone therapy restored expression of PG-21 androgen receptor in aged and orchidectomised male rats, however levels of re-expression varied within different nuclei suggesting a more prolonged period of hormone replacement may be required for full restoration.
 
Effect of resveratrol (RES) treatment on AMP-activated protein kinase (AMPK) activation in RAW 264.7 cells. (A) Concentration-dependent effects of RES on RAW 264.7 cell viability. Cells were incubated with different concentrations (0.1 or 10 µM) of RES for 3-h. (B) Western blot analysis for AMPK activation in different concentrations of RES. p-AMPK was normalized to β-actin and represented as arbitrary units. Data are expressed as mean±SEM of the ratio of p-AMPK protein expression for three independent experiments. P<0.05 by ANOVA. *P<0.005,†P<0.001 vs. control.
Effect of resveratrol (RES) treatment on AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) phosphorylation in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Cells were incubated with 0.5 µM RES for 3-h, 1 mM 5-aminoimidazole-4-carboxamide ribose (AICAR) for 1-h, and 10 µM compound C (CC) for 1-h. Cells were then treated with 50 ng/ml LPS for 3-h. Cell lysates were subjected to Western blot analysis with p-AMPK and total AMPK (A) and p-ACC and total ACC (B) antibodies. p-AMPK or p-ACC was normalized to total AMPK or ACC and represented as arbitrary units, respectively. Data are expressed as mean±SEM for three independent experiments. P<0.05 by ANOVA. *P<0.005, †P<0.001 vs. control (CTL) or LPS-treated cells, respectively.
Effect of resveratrol (RES) treatment on tumor necrosis factor (TNF)-α and TNF receptor (TNFR) expression in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Cells were incubated with 0.5 µM RES for 3-h, 1 mM 5-aminoimidazole-4-carboxamide ribose (AICAR) for 1-h, and 10 µM compound C (CC) for 1-h. Cells were then treated with 50 ng/ml LPS for 3-h. (A) Cell supernatant TNF-α concentrations measured by enzyme-linked immunosorbent assay. Cell lysates were subjected to Western blot analyses with TNF-α (B) and TNFR (C) antibodies, respectively. TNF-α and TNFR1 were normalized to β-actin and represented as arbitrary units. Data are expressed as mean±SEM for three independent experiments. P<0.05 by ANOVA. *P<0.005, †P<0.001 vs. the respective control (CTL) or LPS-treated cells, respectively.
Effect of resveratrol (RES) treatment on nuclear factor κB (NF-κB) translocation in lipopolysaccharide (LPS)-treated RAW 264.7 cells. (A) Representative photomicrographs of immunolabeling for NF-κB (red) and nuclear counterstaining with DAPI (blue) in control (CTL), LPS, LPS+RES, LPS+5-aminoimidazole-4-carboxamide ribose (AICAR), and LPS+compound C (CC)-treated RAW 264.7 cells. LPS treatment induced increased colocalization of NF-κB and DAPI (purple) in LPS-treated cells. However, RES treatment reduced NF-κB translocation, as shown by decreased colocalization with DAPI. Arrows indicate translocation from the cytoplasm to the nucleus. Scale bar=100 µm. (B) Nuclear translocation of cytoplasmic NF-κB from each group was assessed by Western blotting. (C) Quantitative analysis of NF-κB levels calculated from Western blots. Cytoplasmic (cyto) or nuclear (nu) NF-κB was normalized to β-actin or lamin A and represented as arbitrary units, respectively. Data are expressed as mean±SEM for three independent experiments. P<0.05 by ANOVA. *P<0.05, †P<0.001 vs. the respective control (CTL) or LPS-treated cells, respectively.
Effect of resveratrol (RES) treatment on cyclooxygenase (COX)-2 expression in lipopolysaccharide (LPS)-treated RAW 264.7 cells. (A) Cells were incubated with 0.5 µM RES for 3-h, 1 mM 5-aminoimidazole-4-carboxamide ribose (AICAR) for 1-h, and 10 µM compound C (CC) for 1-h. Cells were then treated with 50 ng/ml LPS for 3-h. Cell lysates were subjected to Western blot analysis with a COX-2 antibody. (B) Quantitative analysis of COX-2 levels calculated from Western blots. COX-2 was normalized to β-actin and represented as arbitrary units. Data are expressed as mean±SEM for three independent experiments. P<0.05 by ANOVA. *P<0.001 vs. the respective control (CTL) or LPS-treated cells, respectively.
Article
AMP-activated protein kinase (AMPK), an enzyme involved in energy homeostasis, regulates inflammatory responses, but its precise mechanisms are not fully understood. Recent evidence has shown that resveratrol (RES), an AMPK activator, reduces prostaglandin E(2) production in lipopolysaccharide (LPS)-treated microglia. Here, we examined the effect of RES on nuclear factor kappa B (NF-κB) dependent cyclooxygenase (COX)-2 activation in LPS-treated RWA 264.7 macrophages. We found that treatment with RES increased AMPK activation. AMPK and acetyl CoA carboxylase phosphorylation were attenuated in cells treated with LPS+RES, compared to cells treated with LPS alone. RES inhibited tumor necrosis factor (TNF)-α and TNF receptor 1 in LPS-treated cells. Finally, RES inhibited LPS-induced NF-κB translocation into the nucleus and COX-2 expression. Moreover, the effects of 5-aminoimidazole-4-carboxamide ribose and compound C were consistent with the effects of RES in LPS-treated cells. Taken together, these results suggest that the anti-inflammatory action of RES in RAW 264.7 macrophages is dependent on AMPK activation and is associated with inhibition of the LPS-stimulated NF-κB-dependent COX-2 signaling pathway.
 
Article
The high resistance to infections in lizard wounds suggests that these reptiles possess effective antimicrobial peptides in their tissues. The present immunocytochemical study shows the cellular localization of beta-defensin 27 in tail tissues and in the blood, a defensin previously identified in the lizard Anolis carolinensis through biomolecular methods. Beta-defensin-27 immunoreactivity is only observed in some large granules mainly contained in heterophilic granulocytes that are sparse within the dermis of the skin or in the isolated blood. This peptide is absent in other cell types of the skin, in keratinocytes and in subdermal muscle tissue of the tail in normal conditions. Pre-corneous keratinocytes of the regenerating tail epidermis are unlabeled or show a weak labeling for the peptide only in sparse cytoplasmic areas or in the extracellular spaces among corneocytes of the wound and regenerating epidermis. The study suggests that beta-defensin 27 is normally stored in granulocytes present in the blood or in connective tissues while in the epidermis keratinocytes do not show the presence of this peptide unless these cells are stimulated from injury to produce and likely release beta-defensins.
 
Article
Mast cells participate in allergies and inflammation by secreting a variety of pro-inflammatory mediators. Curcumin, the active component of turmeric, is a polyphenolic phytochemical with anti-tumor, anti-inflammatory, anti-oxidative, and anti-allergic properties. The effects of curcumin on compound 48/80-induced mast cell activation and passive cutaneous anaphylactoid reactions are unknown. In this report, we investigated the influences of curcumin on the passive cutaneous anaphylactoid response in vivo and compound 48/80-induced mast cell activation in vitro. The mechanism of action was examined by calcium uptake measurements and cAMP assays in mast cells. Curcumin significantly attenuated the mast cell-mediated passive cutaneous anaphylactoid reaction in an animal model. In agreement with this in vivo activity, curcumin suppressed compound 48/80-induced rat peritoneal mast cell (RPMC) degranulation and histamine release from RPMCs. Moreover, compound 48/80-elicited calcium uptake into RPMCs was reduced in a dose-dependent manner by curcumin. Furthermore, curcumin increased the level of intracellular cAMP and significantly inhibited the compound 48/80-induced reduction of cAMP in RPMCs. These results corroborate the finding that curcumin may have anti-allergic activity.
 
Article
Alpha-lipoic acid (LA), a naturally occurring dithiol compound, is an essential cofactor in metabolic reactions involved in energy utilization. LA improves glycemic control, reduces diabetic polyneuropathies, atherosclerosis, and allergic inflammation. The effects of LA on mast cell-mediated anaphylactic reactions, however, are unknown. LA dose-dependently inhibited systemic and passive cutaneous anaphylaxis-like reactions in mice induced by compound 48/80, a condensation product of N-methyl-p-methoxyphenethylamine and formaldehyde. Pretreatment with LA, prior to induction of the systemic anaphylaxis-like reaction with compound 48/80, reduced plasma histamine levels in a dose-dependent manner. In our in vitro study, LA decreased histamine release from rat peritoneal mast cells (RPMCs) triggered by compound 48/80. Moreover, an increase in calcium uptake activated by compound 48/80 was inhibited by LA. LA also significantly elevated intracellular cyclic adenosine-3',5' monophosphate (cAMP) levels in RPMCs. This inhibition of mediator release from RPMCs may be due to inhibition of calcium uptake and augmentation of intracellular cAMP levels. Based on these results, we suggest that LA may be a potential remedy for allergy-related diseases.
 
Reverse transcription-polymerase chain reaction with IL-6Rα (251 base pairs), Gp130 (326 base pairs), and GAPDH (98 base pairs) primers and electrophoresis on 1.5% agarose gel. IL-6Rα, interleukin 6 receptor α; Gp130, glycoprotein 130; GAPDH, glyceraldehyde-3-phosphatedehydrogenase.
Interleukin 6 receptor α (IL-6Rα) expression in the fallopian tube. IL-6Rα mRNA expression was higher in the fallopian tubes of women with ectopic pregnancies than in the fallopian tubes of the nonpregnant women in the luteal phase of their menstrual cycle (control group). Expression of IL-6Rα mRNA was lower in the fallopian tubes of women with ectopic pregnancies than in the fallopian tubes of normal pregnant women, but this difference was not significant (P>0.05). *indicates a significant difference from the control group (P<0.05).
Glycoprotein 130 (Gp130) expression in the fallopian tube. Expression of Gp130 mRNA was lower in the fallopian tubes of women with ectopic pregnancies than in the fallopian tubes of nonpregnant woman in the luteal phase of their menstrual cycle (control group). Expression of Gp130 mRNA was higher in the women with ectopic pregnancies than in normal pregnant women, although this difference was not statistically significant (P>0.05). *indicates a significant difference from the control group (P<0.05).
Oligonucleotide sequences designated for this study
Article
Women with tubal ectopic pregnancies have high levels of circulating interleukin 6 (IL-6). IL-6 treatment in vitro significantly reduces the ciliary activity of tubal epithelium. The effects of IL-6 on target cells occur via the formation of a high-affinity complex with its receptors IL-6Rα and glycoprotein 130 (Gp130). IL-6Rα is specifically expressed in the cilia of the epithelial cells. In this study, we performed a quantitative reverse transcriptase polymerase chain reaction to determine the mRNA expression of IL-6Rα and Gp130 in the fallopian tubes obtained from 12 women with ectopic pregnancies, 12 women with normal pregnancies, and 12 healthy nonpregnant women in the luteal phase of their menstrual cycle. Fallopian tubes were evaluated from specimens taken during tubal ligation in normal pregnancies and nonpregnant fertile women or during tubal surgery in ectopic pregnancies. We observed that IL-6Rα mRNA expression in fallopian tubes was increased in ectopic pregnancy compared with that in the midluteal phase. We also found that the Gp130 mRNA expression was significantly lower in fallopian tubes from ectopic pregnancies than in those from nonpregnant women during the midluteal phase of their menstrual cycle, although its expression was noticeably high in fallopian tubes in the midluteal phase, which suggests that high Gp130 levels may possibly contribute to embryo transport into the uterus.
 
Astrocytes in the CA1 area of the hippocampus in all groups. (A) Control. (B) Sham (scopolamine). (C) Treatment 40. (D) Treatment 80. (E) Pretreatment 40. (F) Pretreatment 80 (PTAH staining). Scale bars=20 µm.
Mean and SD of astrocytes number in CA1 area of hippocampus
Mean and SD of astrocytes number in CA3 area of hippocampus
Mean and SD of astrocytes number in DG area of hippocampus
Article
The regular extract of Ginkgo biloba has been shown to possess neuroprotective properties in disorders like hypoxia, ischemia, seizure activity and peripheral nerve damage. Also, G. biloba has received attention as a potential cognitive enhancer for the treatment of Alzheimer's disease, but there is not any documentation about the effect of an extract of G. biloba on astrocytes. Therefore, the aim of this study was examined the effects of G. biloba extract on the rat's hippocampal astrocytes after scopolamine based amnesia. In this study, 36 adult male Wistar rats were used. Rats were randomly distributed into control, sham, protective and treatment groups. The rats in the sham group only received scopolamine hydrobromide (3 mg/kg) intraperitoneally. The rats in the protective and treatment groups received G. biloba extract (40, 80 mg/kg) for 7 days intraperitoneally before and after scopolamine injection. Forty eight hours after the last injection, the brains of the rats were withdrawn and fixed with paraformaldehide, and then after histological processing, the slices were stained with phosphotungstic acid-haematoxylin for astrocytes. Data were analyzed by the analysis of variance (ANOVA) post hoc Tukey test; P<0.05 was considered significant. Results showed that scopolamine can reduce the number of astrocytes in all areas of hippocampal formation compared with the control. However, G. biloba extract can compensate for the reduction in the number of astrocytes in the hippocampus before or after the encounter with scopolamine. We concluded that a pretreatment and treatment injection of G. biloba extract can have a protective effect for astrocytes in all areas of hippocampal formation.
 
Classification of variations in hepatic arterial supply by Michels [7] and Hiatt et al. [8]
Photograph showing multiple anomalies of celiac trunk (CT) and superior mesenteric artery (SMA) and their branches with stomach and part of common hepatic artery (CHA)/left hepatic artery (LHA) removed (represented by a dotted line). CA, cystic artery; GB, gallbladder; GDA, gastroduodenal artery; LGA, left gastric artery; RHA, replaced right hepatic artery; SA, splenic artery.
Schematic representation of Fig. 1 showing multiple variations in arterial anatomy as observed in present case. CA, cystic artery; CHA, common hepatic artery; CT, celiac trunk; GDA, gastroduodenal artery; LGA, left gastric artery; LHA, left hepatic artery; RGA, right gastric artery; RHA, replaced right hepatic artery; SA, splenic artery; SMA, superior mesenteric artery.
Classification of variations in hepatic arterial supply by Michels [7] and Hiatt et al. [8]
Article
Vascular anomalies are frequently encountered in abdomen. But they are usually asymptomatic and diagnosed accidently during angiography or surgery leading into severe complications. Thus knowledge of angioarchitecture in abdomen, whether normal or variant, is considered prerequisite for successful, uncomplicated surgeries and interventional radiology. This case report describes one of such varying branching pattern of celiac trunk and superior mesenteric artery. During routine abdominal dissection, gastroduodenal artery was seen arising from celiac trunk along with its usual three branches. Common hepatic artery continued as left hepatic artery after giving rise the right gastric artery and a tortuous replaced right hepatic artery arose from superior mesenteric artery. An unusually long cystic artery arose from left hepatic artery and gave rise to 2-3 small anastomotic branches towards hepatic flexor of colon, in addition to its normal gallbladder supply. Awareness of such variations would certainly be helpful in upper abdominal surgeries.
 
Left adrenal (LA) gland and left kidney (LK). Adrenal gland was absent in right side.  
Persistent ductus arteriosus. Pulmonary trunk (PT) connects to aorta by persistent ductus arteriosus (arrow). BC, brachiocephalic trunk; LCc, left common carotid; LSc, left subsabclavian.  
Article
We report on an extremely rare case of multiple absences of the branches of abdominal aorta with congenital absence of the portal vein, unilateral adrenal agenesis and persistent ductus arteriosus in an adult female cadaver. Specifically, instead of celiac trunk, superior and inferior mesenteric arteries, solely a single arterial trunk aroused from the anterior aspect of abdominal aorta, inferior phrenic and ovarian arteries were absent in both sides. Left kidneys drained by two veins. There were not superior, splenic and mesenteric veins, while left renal vein received an additional vein, which run downward and drained primarily all parts of digestive tract and its associated glands (portal vein did not exist). Right adrenal gland was absent. To the best of our knowledge, it is the only reported case with such widespread anomalies. We think the importance of this case is beyond the surgical consideration and needs more profound developmental studies.
 
Horizontal sections of a 13mm crown-r ump leng th embr yo (approximately 5 weeks). Panels (A) and (G) are the most superior and inferior sides of the figure, respectively. Intervals between panels are 0.1 mm (A, B), 0.4 mm (B, C), 0.2 mm (C, D) and 0.1 mm (D-G). Panels (A) and (B) show levels immediately below the tracheal bifurcation: the esophagus (ES) is located on the anterior side of the aorta (AO). The asterisk in panel (A) indicates the apex of the pleural cavity. The umbilical vein (UV) runs upward along the inferior part of the pericardium (C). The diaphragm has not yet developed and the pleural and peritoneal cavities communicate via the pleuroperitoneal canal. Thus, in panel (D), the left lung (LL) faces the adrenal (AD) and liver (L) without interrupting any membranous structure. (C-E) The esophagus is connected with the aorta by a mesentery-like structure ( i.e., the mesoesophag us; arrow). The pneumatoenteric recesses (PER; recesses of the pleuroperitoneal canal) are present at the inferior end of the mesoesophagus (F, G). The stomach (ST) is separated from the liver (L) by peritoneal recesses (black stars in panels D-G): and by the future omental bursa (OB, open stars). A terminal portion of the UV does not simply correspond to the inferior vena cava due to a lack of communication with the adrenal vein [5]. BR, bronchus; H, heart; M, mesonephros; PC, pericardial cavity; RL, right lung ; ST, stomach; VC, vertebral column; VN, vagus nerve. All panels were prepared at the same magnification. Scale bar=1 mm. 
Sag ittal sections of a 22mm crown-r ump leng th embr yo (approximately 7 we eks). Panels (A) and (F) are the left-and rightmost sides of the figure, respectively. Intervals between panels are 0.1 mm (A, B), 0.3 mm (B, C) and 0.1 mm (C-F). Panels (A-E) were prepared at the same magnification, while panel (F) is at a higher magnification to show details of the fascia and space. Scale bar=1 mm. Panels (A) and (B) contain the right end of the left lung (LL), which is connected to the esophagus (ES) by a mesentery-like structure (open stars). In panel (C), the loose mesenchymal tissue (black stars) between the esophagus and aorta (AO) corresponds to the mesoesophagus. A recess of the peritoneal cavity (arrows) separates the stomach (S T ) and abdominal esophagus from the adrenal (AD) and celiac ganglion (GL). The diaphragm (DI) has not yet extended to the space behind the AD (asterisks in panels A and B). At the superior aspect of the diaphragm, the esophagus is surrounded by a recess of the pleural cavity (arrowheads in panels C-F). The liver caudate lobe (LC) protrudes into the omental bursa (OB) in panels (D-F). DP, dorsal pancreas; G, gonad; H, heart; IVC, inferior vena cava; K, kidney; L, liver; LCCV, left common cardinal vein; M, mesonephros; PC, pericardial cavity; SC, spinal cord; SPN, splanchnic nerve; TR, trachea; UV, umbilical vein; VC, vertebral column; VN, vagus nerve. 
Tilted horizontal sections of a 155-mm crown-rump length fetus (approximately 18 weeks). Panels (A) and (E) are the most superior and inferior sides of the figure, respectively. Intervals between panels are 16.5 mm (A, B), 2.8 mm (B, C), 2.4 mm (C, D), and 1.6 mm (D, E). Panel (A) shows a level immediately below the tracheal bifurcation, with the esophagus (ES) located on the right side of the aorta (AO). In panels (B) and (C), the mesoesophagus (arrows) connects the aorta and esophagus. Arrowheads in panel (B) indicate bilateral recesses of the pleural cavity in the pulmonary ligament. The abdominal esophagus is attached to the diaphragm (asterisk in panel D). A recess of the peritoneal cavity (open stars in panel C), present between the stomach and left adrenal, provides a sac-like enlargement (open star in panel D). Another slit-like recess is also seen (arrows in panels D and E). The liver caudate lobe (LC) protrudes into the omental bursa (OB). AD, adrenal gland (fetal adrenal cortex); BR, bronchus; DI, diaphragm; IVC, inferior vena cava; L, liver; LC, liver caudate lobe; LL, left lung; RL, right lung; ST, stomach; TH, thymus. All panels were prepared at the same magnification. Scale bar=1 mm. 
A schematic representation showing recesses of the celomic, pleural and peritoneal cavities protruding into the mesoesophagus. Panel (A) (at and before 5 weeks) exhibits a primitive form in which the esophagus is embedded in a posterior mesenchymal tissue. Panel (B) (5 weeks) displays an initial mesoesophagus with the pneumatoenteric recess (a curved arrow with a): the mesentery-like shape is sculptured by the developing lungs. Panel (C) (7-18 weeks) shows a pleural recess around the lower thoracic esophagus (a curved arrow with b) as well as a peritoneal recess behind the abdominal esophagus (a curved arrow with c) between the left adrenal (AD) and stomach (ST). Asterisk indicates disappearance of the posterior mesenchymal tissue, including the mesoesophagus, due to cell death. Those recesses are similar to a Korean traditional symbol of the nature in shape, i.e., Taegeuk, as shown in the lower part of the figure. 
Article
A term "mesoesophagus" has been often used by surgeons, but the morphology was not described well. To better understand the structures attaching the human abdominal and lower thoracic esophagus to the body wall, we examined serial or semiserial sections from 10 embryos and 9 fetuses. The esophagus was initially embedded in a large posterior mesenchymal tissue, which included the vertebral column and aorta. Below the tracheal bifurcation at the fifth week, the esophagus formed a mesentery-like structure, which we call the "mesoesophagus," that was sculpted by the enlarging lungs and pleural cavity. The pneumatoenteric recess of the pleuroperitoneal canal was observed in the lowest part of the mesoesophagus. At the seventh week, the mesoesophagus was divided into the upper long and lower short parts by the diaphragm. Near the esophageal hiatus, the pleural cavity provided 1 or 2 recesses in the upper side, while the fetal adrenal gland in the left side was attached to the lower side of the mesoesophagus. At the 10th and 18th week, the mesoesophagus remained along the lower thoracic esophagus, but the abdominal esophagus attached to the diaphragm instead of to the left adrenal. The mesoesophagus did not contain any blood vessels from the aorta and to the azygos vein. The posterior attachment of the abdominal esophagus seemed to develop to the major part of the phrenoesophageal membrane with modification from the increased mass of the left fetal adrenal. After postnatal degeneration of the fetal adrenal, the abdominal esophagus might again obtain a mesentery. Consequently, the mesoesophagus seemed to correspond to a small area containing the pulmonary ligament and aorta in adults.
 
Photographs of the abdominal aortic aneurysm (AAA) (A) and intracranial artery aneurysms (B) in the A3 segment of anterior cerebral artery and bifurcation of the middle cerebral artery (asterisks). ICA, internal carotid artery; IMA, inferior mesenteric artery.
Article
We found multiple aneurysms in the intracranial arteries and abdominal aorta of an 87-year-old Korean female cadaver, whose cause of death was reported as "cholangiocarcinoma." An abdominal aortic aneurysm was observed in the infrarenal aorta, where the inferior mesenteric artery arose. The intracranial aneurysms were found in the A3 segment of the anterior cerebral artery and at the bifurcation of the middle cerebral artery. This case provides an example of the very rare association of peripheral intracranial aneurysms with an abdominal aortic aneurysm. Clinicians as well as anatomists should recognize the potential association between these two aneurysm types.
 
Targeted activation of β-catenin in the dentoalveolar complex. (A) In immunohistochemistry, β-catenin is localized in the Am, Od, and Ob of developing mouse dentoalveolar complex at P8. (B) β-galactosidase activities are also found in the Od and Ob of Col1a1-cre:R26R double transgenic mouse dentoalveolar complex at P8. (C) Col1a1-cre:Catnb+/lox(ex3) mutant mice exhibit severe growth retardation with short height and small body weights. (D) Differences in body weights between WT and MT mice are appeared in 3 weeks after birth and are clearer in the mice at 4 weeks-old. (E) Genotype analysis of mutant mice after intercross with Col1a1-cre and Catnb+/lox(ex3) mice. (F-I) Gross appearance of incisors in WT and MT mice at 4 weeks-old. Both of upper and lower incisors are normally erupted in WT mice, whereas upper incisors are erupted but lower incisors (white arrow) are not erupted in MT mice. (J, K) Molars of lower jaw (white arrowheads) are not erupted into oral cavity in MT mice in contrast to normally erupted in WT mice. Am, ameloblasts; Od, odontoblasts; Ob, osteoblasts; WT, wild type; MT, mutant. *P<0.05. Scale bar=200 µm (A, B).
Disturbances in tooth formation and eruption failure of molars in Col1a1-cre:Catnb+/lox(ex3) mice. (A, B) In microradiography, general dimensions of craniofacial skeleton in MT mice are smaller than those in WT littermates. Particularly, mandible of MT is severely retarded and resulted in malocclusion. (C, D) In the mid-sagittal view, mandibular incisor is shorter and smaller than that of WT mice. (E, F) Molars of MT mice are impacted within mandible and root formation is impaired while those of WT mice are normally formed and erupted into the oral cavity. (G, H) In H&E-stained sagittal sections of the mandibles, bone deposition is remarkably increased and molars are not erupted in the MT mice. (I, J) In MT mice, β-catenin expression is upregulated in the odontoblasts and osteoblasts. MT, mutant; WT, wild type; H&E, hematoxylin and eosin. Scale bar=100 µm (G-J).
Mineralization defects in dentin of Col1a1-cre:Catnb+/lox(ex3) mice. (A, B) Immunohistochemistry reveal that β-catenin is localized in the Od and pulp cells of mandibular first molar of WT mice, which is restrictedly increased in the odontoblasts of MT mice. (C, D) Phex is localized in the odontoblasts of WT mice, but which is clearly reduced in the odontoblasts of MT mice. (E, F) In WT mice, Dspp is localized in the dentin as well as odontoblasts, but it is obviously decreased in the predentin and odontoblasts of MT mice. Dspp remains only in thin mineralized dentin of MT mice. (G, H) Localization of Fgf23 is not found in the dentin and odontoblasts of WT mice but which is clearly observed in the predentin of MT mice. (I-L) Bgn and Dmp1 are mainly localized in the predentin and a part of dentin in WT mice, which also decreased in the predentin of MT mice. Od, odontoblasts; WT, wild type; MT, mutant; Phex, phosphate regulating endopeptidase homologue on the X chromosome; Dspp, dentin sialophosphoprotein; Fgf23, fibroblast growth factor-23; Bgn, biglycan; Dmp1, dentin matrix protein-1. Scale bar=100 µm (A-L).
Deformities in the roots and periodontium of Col1a1-cre:Catnb+/lox(ex3) mice. (A, B) In contrast to WT mice, RD is thin and hypomineralized in MT mice. In addition, PS is narrower due to excessive formation of alveolar bone in MT mice. (C, D) In the RD of MT mice, Dspp is decreased in the dentin. (E-J) In WT mice, Bsp, Dmp1 and Fgf23 are specifically localized in the matrix of acellular cementum (arrows) and alveolar bone, which is decreased in the MT mice. (K, L) Osteopontin is localized in the cementum and periodontal ligaments of WT mice. It is also decreased in the cementum and periodontal ligaments of MT mice. WT, wild type; RD, root dentin; H&E, hematoxylin and eosin; MT, mutant; PS, periodontal space; Dspp, dentin sialophosphoprotein; Bsp, bone sialoprotein; Dmp1, dentin matrix protein-1; Fgf23, fibroblast growth factor-23; Opn, osteopontin. Scale bar=50 µm (A-L).
Molecular changes related with increased bone mass in the mandibles of Col1a1-cre:Catnb+/lox(ex3) mice. (A, B) In the mandible of MT mice, bone mass is increased but number of osteoblasts are decreased to compare with those of WT mice. (C-F) Bsp expression is slightly increased with increase of bone mass, but Tnap is clearly decreased in the MT. (G-J) Bgn is significantly increased in the mandible of MT mice, while Phex is almost disappeared in MT mice. (K, L) Increased expression of Dmp1 is also observed in MT mice. WT, wild type; H&E, hamatoxylin and eosin; MT, mutant; Bsp, bone sialoprotein; Tnap, tissue-nonspecific alkaline phosphatase; Bgn, biglycan; Phex, phosphate regulating endopeptidase homologue on the X chromosome; Dmp1, dentin matrix protein-1. Scale bar=50 µm (A-L).
Article
Wnt/β-catenin signaling plays a critical role in bone formation and regeneration. Dentin and cementum share many similarities with bone in their biochemical compositions and biomechanical properties. Whether Wnt/β-catenin signaling is involved in the dento-alveolar complex formation is unknown. To understand the roles of Wnt/β-catenin signaling in the dento-alveolar complex formation, we generated conditional β-catenin activation mice through intercross of Catnb(+/lox(ex3)) mice with Col1a1-cre mice. In mutant mice, tooth formation and eruption was disturbed. Lower incisors and molars did not erupt. Bone formation was increased in the mandible but tooth formation was severely disturbed. Hypomineralized dentin was deposited in the crown but roots of molars were extremely short and distorted. In the odontoblasts of mutant molars, expression of dentin matrix proteins was obviously downregulated following the activation of β-catenin whereas that of mineralization inhibitor was increased. Cementum and periodontal ligament were hypoplastic but periodontal space was narrow due to increased alveolar bone formation. While cementum matrix proteins were decreased, bone matrix proteins were increased in the cementum and alveolar bone of mutant mice. These results indicate that local activation of β-catenin in the osteoblasts and odontoblasts leads to aberrant dento-alveolar complex formation. Therefore, appropriate inhibition of Wnt/β-catenin signaling is important for the dento-alveolar complex formation.
 
Left retroperitoneal space dissection shows the abnormal drainage of a retro-aortic bifid left renal vein into the inferior vena cava. The variant termination of the 1st and 3rd lumbar veins into the left renal vein is also seen. (A) The abdominal aorta is dissected to show the bifid termination of the retro-aortic left renal vein. (B) The left kidney and structures that pass through its hilum are shown in their reflected position. AA, abdominal aorta; D, diaphragm; FLLV, first left lumbar vein; IVC, inferior vena cava; LK, left kidney; LRA, left renal artery; LRV, left renal vein; LSRG, left suprarenal gland; LSRV, left suprarenal vein; LTV, left testicular vein; PM, psoas major; Pm, psoas minor; QL, quadrates lumborum; RDC, right diaphragmatic crus; RK, right kidney; RRA, right renal artery; RTV, right testicular vein; SC, sympathetic chain; TLLV, third left lumbar vein; U, ureter; VC, vertebral column.
Right retroperitoneal space dissection shows; (A) the abnormal drainage of additional testicular vein into the right renal vein. (B) Light micrograph illustrating a histological section of testicular vein before drainage into the right renal vein. (C) Right testicular artery with accompanying veins in a common sheath (right, schematic illustration; left, histological section) (B, ×200; C, ×100). ATV, additional testicular vein; D, diaphragm; HV, hepatic vein; IVC, inferior vena cava; PM, psoas major; RK, right kidney; RRV, right renal vein; RSRG, right suprarenal gland; RTA, right testicular artery; U, ureter.
Retroperitoneal space dissection. (A) The greatly enlarged spleen forced the left kidney below the level of the right kidney. (B) The isolated enlarged spleen. LK, left kidney; RK, right kidney; S, spleen; SA, splenic artery; SV, splenic vein.
Article
Knowledge of the renal vascular anatomy may greatly contribute to the success of surgical, invasive and radiological procedures of the retroperitoneal region. Here, morphometric and histological studies of a human cadaveric specimen presented a complex, anomalous pattern of renal veins. The left renal vein had an oblique retro-aortic course and received two lumbar veins. It bifurcated near its drainage point into the inferior vena cava. The right renal vein received the right testicular vein. In addition, the left kidney was located at a low position. The spleen was enlarged. The present case is unique and provides information that may help surgeons or angiologists to apply safer interventions.
 
Hierarchical clustering display of data from a 24-h time course of abortion following treatment with lipopolysaccharide. Hierarchical clustering of the complete set of 7,426 clones. Individual genes or clones are represented in rows, and time points are in columns. Individual cells are colored based on the log of the fluorescent ratio, with black representing a ratio of 1, or no change in expression relative to a control reference. Increasing ratios indicative of increased expression are represented by increasing red intensity, whereas decreasing ratios are represented by increasing green intensity and reveal decreased gene expression.
Intensity ratio histograms showing the numbers (y-axis) of genes up- or down-regulated in lipopolysaccharide 2 h- (A), 6 h- (B), 12 h- (C), and 24 h- (D) treated mice uteri. The x-axis represents the log values for the intensity ratios (e.g., log22 indicates a two-fold change).
Validation of the microarray data. Gene expression patterns representing different characteristic patterns were assessed by real-time polymerase chain reaction (PCR), and compared with the expression values obtained by microarray analysis. The genes assessed were: (A) Ccl4 (Mm244263), (B) Mx2 (Mm14157), (C) Gbp2 (Mm24038), and (D) Gbp3 (Mm1909). The oligonucleotides used for real-time PCR analysis are listed in Table 1. LPS, lipopolysaccharide.
Primer sequences used for real-time reverse transcription polymerase chain reaction
List of selected genes with significant changes (>8 times) in expression levels between control and LPS-treated uteri
Article
To identify genes that participate in the abortion process, normal pregnant uteri were compared to lipopolysaccharide (LPS)-induced abortion uteri. At day 6 of pregnancy, mice were treated with LPS at various time points to induce an abortion. Total RNAs were applied to a cDNA microarray to analyze genes with altered expression. At the early stage (2 hours) of LPS-induced abortion, upregulated genes were mainly composed of immune responsive genes, including Ccl4, Ccl2, Cxcl13, Gbp3, Gbp2, Mx2, H2-Eb1, Irf1 and Ifi203. Genes related to toll-like receptor signaling were also overexpressed. At late stages of abortion (12-24 hours), many genes were suppressed rather than activated, and these were mainly related to the extracellular matrix, cytoskeleton, and anti-apoptosis. Altered expression of several selected genes was confirmed by real time reverse transcription-polymerase chain reaction. The results demonstrated that many known genes were altered in the LPS-treated pregnant uterus, implying that the molecular mechanisms of the genes involved in LPS-induced abortion are complicated. Further analysis of this expression profile will help our understanding of the pathophysiological basis for abortion.
 
Th e palmaris longus types was classifi ed according to the degree of the prominence of the two tendons. P, palmaris longus; F, fl exor carpi radialis; Line, visible present; Dashed-line, palpable present.
Th e frequency of the absence of the palmaris longus
The palmaris longus types was classified according to the degree of the prominence of the two tendons. P, palmaris longus; F, flexor carpi radialis; Line, visible present; Dashed-line, palpable present.
The frequency of the absence of the palmaris longus
The frequency of the pattern of the Palmaris longus according to 3 methods
Article
The palmaris longus (PL) is a slender, spindle-shaped weak flexor of the wrist. Congenital absence of the PL is estimated to occur in 15% among individuals worldwide. However, the frequency of its absence varies considerably among different population groups and with different detection techniques. In the present study, the presence of the PL tendon was examined in a Korean population (n=269) using three clinical tests, namely the Traditional Test, Mishra's Test II, and the Gangata Test. We classified subjects into six types based on whether inspection or palpation was required to determine the presence of the PL and flexor carpi radialis. The most reliable test was determined using Kendall's coefficient of concordance. Our results showed that the PL tendon was absent in 4.1% of the subjects in our study, and bilateral and unilateral absences were 2.2% and 1.8%, respectively. Statistical analysis revealed that these tests had similar reliability for assessing the PL tendon, and the Traditional Test showed the highest effectiveness, at 93%. Therefore the Traditional Test was found to be the most effective for revealing the PL in this Korean population.
 
Undisturbed position of abdominal viscera after reflection of anterior abdominal wall. Greater omentum has been reflected upwards. Note the inverted U shaped loop of ascending and descending colons. Jejunum and ileum are occupying the upper left part of the abdomen and the sigmoid colon is crossing from right to left.  
Descending colon (DC) has been pulled to the left to show the descending mesocolon (DMC). Note the continuity of the descending mesocolon with the ascending mesocolon (AMC), and sigmoid mesocolon (SMC). Ascending colon (AC) and sigmoid colon (SC) are also seen.  
The descending colon (DC) has been pulled to the right to show the descending mesocolon (DMC). Note the ileum passing behind the prominent margin of the descending mesocolon. Sigmoid colon (SC) and sigmoid mesocolon (SMC) are also seen.  
A simplified illustration of the anomalies observed. The caecum (C), ascending colon (AC), ascending mesocolon (AMC), descending colon (DC), descending mesocolon (DMC), sigmoid colon (SC), sigmoid mesocolon (SMC), jejunum ( JJ), and ileum (IL) have been labelled.  
Article
Congenital anomalies such as positional anomalies of the right half of the colon are more common when compared to its left half. We report a rare case of congenital anomaly where the transverse colon was totally absent. Ascending colon continued as descending colon at the right colic flexure. Ascending and descending colons formed an inverted U shaped loop which was situated in the right half of the abdomen. The sigmoid colon began from the descending colon, on the right side of the midline and coursed to the left iliac fossa. The terminal part of ascending colon and entire descending colon had a persistent mesocolon. The jejunum and ileum were situated in the upper left part of the abdominal cavity. This anomaly can cause volvulus of the colon at any stage of life. Furthermore, the knowledge of this anomaly is very useful for radiologists, gastroenterologists and surgeons.
 
Photograph of the right axilla showing the absence of musculocutaneous nerve. BA, brachial artery; BBr, biceps brachii muscle; CB, coracobrachialis muscle; CTMCAF, common trunk for medial cutaneous nerve of arm and forearm; LPN, lateral pectoral nerve; MN, median nerve; MPN, medial pectoral nerve; NCB, nerve to coracobrachialis; UN, ulnar nerve; arrow, communication between lateral and medial pectoral nerves; 1, nerve to biceps brachii from MN.
Photograph showing the variant branches of median nerve on the right side. BA, brachial artery; BBr, biceps brachii; Br, brachialis; CTMCAF, common trunk for medial cutaneous nerve of arm and forearm from medial cord; MCNA, medial cutaneous nerve of arm; MCNF, medial cutaneous nerve of forearm; MN, median nerve; 1, nerve to biceps brachii from MN; 2, branch to brachialis from common trunk of MN; 3, lateral cutaneous nerve of forearm from common trunk of MN.
Photograph of the left axilla showing the absence of musculocutaneous nerve. BA, brachial artery; BBr, biceps brachii muscle; CB, coracobrachialis; CTMCAF, common trunk for medial cutaneous nerve of arm and forearm; MN, median nerve; NCB, nerve to coracobrachialis; UN, ulnar nerve; arrow, single long trunk from MN; arrowhead, additional lateral root of MN from lateral cord; 1, nerve to biceps brachii from the single long trunk of MN.
Photograph showing the variant branches of median nerve on the left side. BBr, biceps brachii; CTMCAF, common trunk for medial cutaneous nerve of arm and forearm from medial cord; MCNA, medial cutaneous nerve of arm; MCNF, medial cutaneous nerve of forearm; MN, median nerve; UN, ulnar nerve; arrow, single long trunk from MN; 1, nerve to biceps brachii; 2, branch to brachialis; 3, lateral cutaneous nerve of forearm.
Article
A 43-year-old female cadaver showed a complete bilateral absence of the musculocutaneous nerve. The anterior compartment muscles of both arms were supplied by median nerve excepting the coracobrachialis which was innervated by a direct branch from the lateral cord of brachial plexus. The median nerve, after supplying the biceps and brachialis muscles, gave onto the lateral cutaneous nerve of the forearm. The median nerve also showed variation on the left side where it was formed by two lateral roots and one medial root. Variations of the brachial plexus are of great interest to anatomists, clinicians and surgeons, in that they may be incorporated in their day to day practice. Our present case may be noted for its clinical and surgical significance in the variations of brachial plexus which can be useful for diagnostic purposes.
 
The Cavalieri method. The volume of the medial prefrontal cortex (MPFC) and its subdivisions; anterior cingulated (Cg1), prelimbic (PrL), and infralimbic areas (IL) were estimated using Cavalieri's method. (A) The MPFC was sectioned executively. Nine to twelve sections were sampled (here 9). "t" is the distance between the sampled sections. (B) A grid of points was superimposed on the sampled sections. "a(p)" is the area per point and is calculated by multiplying "ΔX" by "ΔY". The arrow indicates the point that is the right upper corner of the cross. Cresyl-violet staining.
Estimation of numerical density. (A) A schematic drawing of a microscopic slide with a section of the medial prefrontal cortex mounted on it. "h" is the height of the optical disector and "t" is the real thickness of the section. In each field, the first 5 µm of the section thickness was ignored (guard zone: gz), and cell counting was done in the next 10 µm of the section thickness "h". (B, C) Sampling of cells using optical disector. The unbiased counting frame was superimposed on the images. Just the cells whose nucleoli did not appear in the beginning of the disector height (B) and appeared at the following optical scan (C) were counted. The cells whose nucleoli were completely or partly inside the counting frame or touching the upper and right lines were counted as "ΣQ-". Cresyl-violet staining.
Mean±SD of the total number of the neurons, oligodendrocytes, and the volume (mm3) of medial prefrontal cortex (MPFC) in 30 days morphine treated rats (MOR), without or with 30 days abstinence (ABS), and saline-treated controls (SAL) (n=6)
Article
Quantitative studies to date on the effects of opioid consumption and abstinence on the nervous system using modern stereological methods have not received enough attention. In addition, they have yielded controversial results. The present study was conducted to investigate the effects of morphine, with or without abstinence, on the neurons and oligodendrocytes of the medial prefrontal cortex (MPFC) in rats using quantitative stereological methods. The male rats were divided into four groups: the first (saline [SAL]) and second (morphine [MOR]) groups were treated with saline and an escalating dose of morphine (5-20 mg/kg) for 30 days, respectively; the third (SAL+abstinence [ABS]) and fourth (MOR+ABS) groups were treated in the same manner as the previous groups plus they had a 30-day abstinence period. The results showed that the volume of the MPFC and its subdivisions decreased by approximately 15% in the MOR group compared with that in the SAL group (P<0.05). In addition, the volume decreased by approximately 24% in the MOR+ABS group compared with that in the SAL+ABS group (P<0.05). The number of neurons in the MOR and MOR+ABS groups decreased by approximately 44% and 35%, respectively, compared with that in their corresponding control groups. Moreover, the number of the oligodendrocytes in the MOR and MOR+ABS groups decreased by approximately 41% and 37%, respectively. No significant difference was noted in the number of cells in the MOR and MOR+ABS groups. In conclusion, morphine consumption leads to a permanent reduction in the number of neurons and oligodendrocytes, and no additional neuron and oligodendrocyte loss occurs after abstinence.
 
A dissection of the forearm and carpus in one left upper extremity. The other muscles have been reflected to show the accessory muscle. PT, pronator teres muscle (cutting); ME, medial epicondyle; FCR, flexor carpi radialis muscle; PL, palmaris longus musle; FDS, flexor digitorum superficialis musle; FCU, flexor carpi ulnaris muscle; AFCU, accessory flexor carpi ulnaris musle; PA, palmar aponeurosis; FR, flexor retinaculum; P, pisiforme bone.
Schematic diagram of the Fig. 1. PT, pronator teres muscle (cutting); ME, medial epicondyle; FCR, flexor carpi radialis muscle; PL, palmaris longus musle; FDS, flexor digitorum superficialis musle; FCU, flexor carpi ulnaris muscle; AFCU, accessory flexor carpi ulnaris musle; PA, palmar aponeurosis; FR, flexor retinaculum; P, pisiforme bone.
Article
Muscular variations of the flexor compartment of forearm are usual and can result in multiple clinical conditions limiting the functions of forearm and hand. The variations of the muscles, especially accessory muscles may simulate soft tissue tumors and can result in nerve compressions. During a routine dissection of the anterior region of the forearm and hand, an unusual muscle was observed on the left side of a 65-year-old male cadaver. The anomalous muscle belly arose from the medial epicondyle approxiamately 1 cm posterolateral to origin of normal flexor carpi ulnaris muscle (FCU), and from proximal part of the flexor digitorum superficialis muscle. It inserted to the triquetral, hamate bones and flexor retinaculum. Passive traction on the tendon of accessory muscle resulted in flexion of radiocarpal junction. The FCU which had one head, inserted to the pisiform bone hook of hamate and palmar aponeurosis. Its contiguous muscles displayed normal morphology. Knowledge of the existence of muscle anomalies as well as the location of compression is useful in determining the pathology and appropriate treatment for compressive neuropathies. In this study, a rare accessory muscle has been described.
 
Article
Pancreatic islets are responsible for blood glucose homeostasis. Reduced numbers of functional (insulin-secreting) beta-cells in pancreatic islets underlies diabetes. Restoration of the secretion of the proper amount of insulin is a goal. Beta-cell mass is increased by neogenesis, proliferation and cell hypertrophy, and is decreased by beta-cell death primarily through apoptosis. Many hormones and nutrients affect beta-cell mass, and glucose and free fatty acid are thought to be the most important determinants of beta-cell equilibrium. A number of molecular pathways have been implicated in beta-cell mass regulation and have been studied. This review will focus on the role of the principle metabolites, glucose and free fatty acid, and the downstream signaling pathways regulating beta-cell mass by these metabolites.
 
Neuron-specific enolase-positive cell number in the mucus or serous lobule in the sublingual gland 
Submandibular gland obtained from an 83-year-old female cadaver. (A) The distribution of the mucous and serous glands (periodic acid Schiff staining). The mucous gland acini are res tricted to a small area in each lobule. Panels (B) and (C) show higher-magnification views of the squares marked B and C in panel (A), respectively (immunohistochemistry for neuron-specific enolase or neuron-specific enolase [NSE]). There is no clear difference in the density of NSEpositive neural elements between the area rich in mucous acini (B) and that rich in serous acini (C). Scale bars=1 mm (A), 0.1 mm (B, C). 
Innervation of the sublingual and submandibular glands obtained from a 78-year-old male cadaver. Using near sections, panels (A), (C), (E), (G), and (I) (or B, D, F, H, and J) display the sublingual (or submandibular) gland. Panels (C), (E), (G), and (I) (or D, F, H, and J) are higher-magnification views of the square in panel (A) (or panel B). Panels (A) and (B) show periodic acid Schiff staining : the sublingual gland contains an area of serous acini or lobules (serous in panel A), whereas in the submandibular gland the area of mucous acini is very limited (mucous in panel B). Immunohistochemistry for S100A protein (Z0311) (C, D), neuron-specific enolase (NSE) (E, F), neuronal nitric oxide synthase (NOS) (G, H), and tyrosine hydroxylase (TH) (I, J). Some of the myoepithelial cells and ductal cells are positive for S100 (arrowheads in panel C). Interlobar nerves express both NOS (arrows in panels G and H) and TH (arrows in panels I and J), but some ducts are also positive for both markers (open stars in panels G, H, and J). Scale bars=1 mm (A, B), 0.1 mm (C-J). 
Immunohistochemistry of myoepithelial cells in the lingual and submandibular glands from an 83-yearold female cadaver. The same specimen as that shown in Figs. 1 and 2. Using near sections, panels (A), (C), (E), and (G) (or B, D, F, and H) display the sublingual (or submandibular) gland. Open stars indicate the same acinus in corresponding panels in the sublingual gland, while closed stars indicate the corresponding sites in the submandibular gland. In both glands, most of the myoepithelial cells are positive for smooth muscle actin (SMA) (arrows in panels G and H), but negative for either S100 protein (mainly S100B, N1573) (A, B), glial fibrillary acidic protein (GFAP) (C, D) or peripheral myelin protein 22 (PMP) (E, F). Thin nerve fibers in the arterial wall express GFAP (arrowheads in panels C and D). Blood cells (arrows in panel F) as well as some of the submandibular ductal cells express PMP (circle marked I in panel F). Panel (I) shows a highermagnification view of the PMP-positive duct in panel (F), and panel ( J) shows positive ducts in another section of the same specimen. In interlobular tissues of both glands, thick myelinated nerve fibers express PMP strongly (K). Scale bars=0.1 mm (A-K). 
Immunohistochemistry of myoepithelial cells in the lingual and submandibular glands from a 78-yearold male cadaver. The same specimen as that shown in Fig. 3. Using near sec tions, panels (A), (C), (E), and (G) (or B, D, F and H) display the sublingual (or submandibular) gland. Open stars indicate the same acinus in corresponding panels in the subling ual g land, while closed stars indicate the corresponding sites in the submandibular gland. In both glands, most of the myoepithelial cells are positive for smooth muscle actin (SMA) (arrows in panels G and H), but ne gative for either neuron specific enolase (NSE) (A, B), S100 protein (mainly S100B, N1573) (C, D) or glial fibrillary acidic protein (GFAP) (E, F). Thin nerve elements around the acini tend to express NSE strong ly, while thick nerves are stained dark for S100. Thin nerve fibers in the arte rial wall express GFAP (arrows in panels E and F). Scale bars=0.1 mm (A-H). 
Article
We examined morphological differences between the sublingual and submandibular glands with special reference to their innervation. The sublingual gland contained abundant periodic acid Schiff-positive mucous acini: some lobules were composed of purely mucous acini, while others were purely serous or mixed. However, in the submandibular gland, the area of mucous acini was very limited. Notably, in the sublingual gland, immunohistochemistry for neuron-specific enolase demonstrated that the serous acini carried a higher density of nerve elements than the mucous acini. However, no such difference was evident in the submandibular gland, possibly due to the small areas of the mucous acini. In both types of gland, neuronal nitric oxide synthase-positive parasympathetic nerves as well as tyrosine hydroxylase-positive sympathetic nerves were observed in the interlobular tissue, but we were unable to trace these thin fibers to the acini. Myoepithelial cells expressed smooth muscle actin, but were negative for S100B protein, glial fibrillary acidic protein and neuron-specific enolase. However, antibody against S100A stained some of the myoepithelial cells and ductal cells in the sublingual gland. Cells positive for peripheral myelin protein 22 were seen in some of the ductal cells in the submandibular gland, but not in the sublingual gland. Therefore, with regard to the neurogenic features of the gland cells, S100B reactivity might disappear first in postnatal life, whereas S100A reactivity is likely to remain as aging progresses. The sublingual gland in elderly individuals seems to provide a good model for comparison of the nerve supply between mucous and serous acini.
 
TTC (2,3,5-triphenyltetrazolium chloride) staining of brain slice from bregma -2.30 mm 3 h after permanent middle cerebral artery occlusion (pMCAO). Tissues of the penumbra (marked with white arrows) represented the red zone near the infarction zone in the ipsilateral hemisphere over a series of brain sections (A). Cresyl violet staining demonstrated that viable cells in the penumbra (marked with short black arrows) were significantly decreased 3 to 6 h after pMCAO (B, C). Moreover, viable cells were not detected 24 h after pMCAO which resembled the ischemic core (D). Scale bar=50 µm.
Expression of Na+/K+ adenosine triphosphatase (Na+/K+-ATPase) in rats with pMCAO and in sham operated control rats. (A) Immunoblot was reacted with affinity purified anti-Na+/K+-ATPase antibody, revealing a 110 KDa product. (B) Densitometric analysis revealed that focal cerebral ischemia produced a significant decrease of Na+/K+-ATPase expression in the penumbra 24 h after pMCAO as compared to sham-operated rats (78±6%, n=4, *P<0.05).
Expression of Na+/Ca2+ exchanger 1 (NCX1) in rats with pMCAO and sham operated control rats. (A) Immunoblot with affinity purified anti-NCX1 antibody, revealing a 120 KDa product. (B) Densitometric analysis revealed that focal cerebral ischemia produced a significant increase of NCX1 expression in the ischemic penumbra 24 h after pMCAO compared with sham-operated rats (144±2%, n=4, *P<0.05).
Expression of N-methyl-D-aspartate (NMDA) receptor 2A and 2B subunits (NR2A and NR2B) in rats with pMCAO and in sham operated control rats. (A) Immunoblot was reacted with affinity purified anti-NR2A and NR2B antibodies, revealing 175 and 180 KDa products. (B) Densitometric analysis revealed that focal cerebral ischemia produced a decrease of NR2A (3 h: 153±9%; 6 h: 74±13%; 24 h: 40±7%, n=4, *P<0.05) while the expression of NR2B was increased (3 h: 37±2%; 6 h: 72±12%; 24 h: 120±16%, n=4, *P<0.05) depending on the duration of ischemia as compared with those of sham-operated controls.
Expression of neuronal nuclei (NeuN), glial fibrillary
acidic protein (GFAP), and 2',3'-cyclic nucleotide 3'-phosphodiesterase monoclonal antibody (CNPase) in rats with pMCAO and in sham operated control rats. (A) Immunoblot was reacted with affinity purified anti-NeuN, anti-GFAP, and anti-CNPase antibodies, revealing 46-48, 50, and 46 KDa products. (B) Densitometric analysis revealed that focal cerebral ischemia produced a time-dependent decrease of NeuN (3 h: 52±6%; 6 h: 38±12%; 24 h: 29±8%, n=5,*P<0.05) and GFAP (3 h: 69±5%; 6 h: 56±12%; 24 h: 53±14% n=5 *P<0.05) in the penumbra as compared with sham-operated controls. However, the expression of CNPase in the penumbra was not changed after pMCAO.
Article
Instrumental role of Na(+) and Ca(2+) influx via Na(+)/K(+) adenosine triphosphatase (Na(+)/K(+)-ATPase) and Na(+)/Ca(2+) exchanger 1 (NCX1) is examined in the N-Methyl-D-aspartate (NMDA) receptor-mediated pathogenesis of penumbra after focal cerebral ischemia. An experimental model of 3, 6, and 24 h focal cerebral ischemia by permanent occlusion of middle cerebral artery was developed in rats. The changes in protein expression of Na(+)/K(+)-ATPase and NCX1 as well as functional subunits of NMDA receptor 2A and 2B (NR2A and NR2B) in the penumbra were assessed using by quantitative immunoblottings. The most prominent changes of Na(+)/K(+)-ATPase (78±6%, n=4, (*)P<0.05) and NCX1 (144±2%, n=4, (*)P<0.05) in the penumbra were developed 24 h after focal cerebral ischemia. The expression of NR2A in the penumbra was significantly increased (153±9%, n=4, (*)P<0.05) whereas the expression of NR2B was significantly decreased (37±2%, n=4, (*)P<0.05) as compared with sham-operated controls 3 h after focal cerebral ischemia. However, the expression of NR2A and NR2B in the penumbra was reversed 24 h after focal cerebral ischemia (NR2A: 40±7%; NR2B: 120±16%, n=4, (*)P<0.05). Moreover, the decreased expression of neuronal nuclei (NeuN) in the penumbra was most prominent than that of glial fibrillary acidic protein (GFAP) 24 h after focal cerebral ischemia. These findings imply that intracellular Na(+) accumulation via decreased Na(+)/K(+)-ATPase exacerbate the Ca(2+) overload cooperated by the increased NCX1 and NR2B-containing NMDA receptor which may play an important role in the pathogenesis of the penumbra.
 
Article
Among the several rotenoids, amorphigenin is isolated from the leaves of Amopha Fruticosa and it is known that has anti-proliferative effects and anti-cnacer effects in many cell types. The main aim of this study was to investigate the effects of amorphigenin on osteoclast differentiation in vitro and on LPS treated inflammatory bone loss model in vivo. We show here that amorphigenin inhibited RANKL-induced osteoclast differentiation from bone marrow macrophages in a dose dependent manner without cellular toxicity. Anti-osteoclastogenic properties of amorphigenin were based on a down-regulation of c-fos and NFATc1. Amorphigenin markedly inhibited RANKL-induced p38 and NF-κB pathways, but other pathways were not affected. Micro-CT analysis of the femurs showed that amorphigenin protected the LPS-induced bone loss. We concluded that amorphigenin can prevent inflammation-induced bone loss. Thus we expect that amorphigenin could be a treatment option for bone erosion caused by inflammation.
 
Distribution and synthesis of advanced glycation end products (AGE)-albumin in human microglial cells and rat brain. Triple-labeled confocal microscopy was used to study the distribution and relative levels of albumin (ALB, green), AGE (red) and a specific marker of microglial cells (Iba1, blue) in the human microglial cell line, human primary microglial cells, and entorhinal cortex of rat brain after amyloid β (Aβ) treatment. Scale bar=50 µm.
Relationship between advanced glycation end products (AGE)-albumin (ALB) and amyloid β (Aβ) contributing to Aβ aggregation. (A) Co-localization of albumin (blue), AGE (red), and ThT fluorescence (green) in amyloid plaques in Aβ-treated rat brains and human Alzheimer disease (AD) brains were evaluated by triple-labeled fluorescent microscopic images. (B) In teraction between AGE-ALB and Aβ, contributing to increased synthesis and aggregation of Aβ. Proximity ligation O-Link analysis was performed by fluorescent microscopic images to determine the relative extents of interaction (red spots) between ALB and Aβ in human microglial cells and rat brains before and after Aβ treatment, as well as human brains from normal and AD individuals, respectively. The average numbers of blob per cell from each sample are compared in bar graphs. Scale bars=50 µm.
Relative changes in advanced glycation end products (AGE) and receptor protein for AGEs (RAGE) interactions. (A) Proximity ligation assay was performed using fluorescent microscopy images to determine the relative extents of interaction (red spots) between AGE and RAGE in the rat brains before or after amyloid β (Aβ) treatment. Scale bar=50 µm. (B) The average number of blobs per cell from each sample is summarized in bar graphs.
Induction of the neuronal mitogen-activated protein kinases (MAPK) pathway by advanced glycation end products (AGE)-albumin (ALB). Double confocal microscopy images simultaneously show relative levels of extracellular signal-regulated kinases 1 and 2 (ERK1/2), pERK1/2, p38K, pp38K, stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK), pSAPK/JNK (green), or DAPI (blue) in human primary neuronal cells before and after AGE-ALB treatment for 6 h. Scale bar=50 µm.
Article
A holy grail of curing neurodegenerative diseases is to identify the main causes and mechanisms underlying neuronal death. Many studies have sought to identify these targets in a wide variety of ways, but a more important task is to identify critical molecular targets and their origins. Potential molecular targets include advanced glycation end products (AGEs) that can promote neuronal cell death, thereby contributing to neurodegenerative disorders such as Alzheimer disease or Parkinson disease. In this study, we showed that AGE-albumin (glycated albumin) is synthesized in microglial cells and secreted in the human brain. Our results provide new insight into which microglial cells can promote the receptor for AGE-mediated neuronal cell death, eventually leading to neurodegenerative diseases.
 
(A-F) Facial motoneurons stained with cresyl violet on the contralateral (A) and ipsilateral (B-F) sides. Significant loss of facial motoneurons was clearly identified from 16 weeks (w) post-lesion (E, F). Cont, contralateral. Scale bar=100 µm (A-F). (G) Survival rates of ipsilateral facial motoneurons compared to the corresponding contralateral side. *P<0.05, †P<0.01.
(A-L) Localization of activating transcription factor 3 (ATF3) (A-F) and phosphorylated c-Jun (pc-Jun) (G-L) in axotomized facial motoneurons. (A, G) Contralateral, (B-F, H-L) ipsilateral side. Both ATF3 and pc-Jun were localized in the nuclei of numerous, normal-sized neurons by 2 weeks (w) post-lesion (B, C, H, I), then the numbers of ATF3- and pc-Jun-positive nuclei decreased progressively (D-F, J-L). At later time points, long-lasting activation of ATF3 and c-Jun was found predominantly in degenerating neurons with severely shrunken soma and condensed nuclei (arrowheads in insets of E, F, K, L) but not in neurons with normal morphology and soma size (arrows in insets of E, F, K, L). Cont, contralateral. Scale bars in (F, L)=100 µm (A-L); insets of (F, L)=20 µm (B, E, F, H, K, L). (M) Percentages of ATF3- and pc-Jun-positive neurons compared to the corresponding ipsilateral facial motoneurons.
Co-localization of activating transcription factor 3 (ATF3) and phosphorylated c-Jun (pc-Jun) in the axotomized facial motoneurons at 2 weeks post-lesion. ATF3 and pc-Jun were predominantly co-localized in the same neurons (arrows), with the exception of a few cells that were either pc-Jun-positive (double arrowheads) or ATF3-positive (arrowheads). Scale bar=20 µm.
(A-F) OX6 immunohistochemistry on the contralateral (A) and ipsilateral (B-F) sides. A few OX6-positive microglia were identified in the ipsilateral facial motor nucleus as early as 1 week (w) post-lesion (wpl) (B). The number of OX6-positive microglia increased significantly by 8 wpl (C, D) and subsequently decreased, but a few persisted up to 24 wpl (E, F). All OX6-positive microglia were ramified and closely adhered to or surrounded facial motoneurons (arrows in insets of C, D, F). (G-I) Double labeling of FluoroGold (FG) and OX6 at 2 wpl. OX6-positive microglia intimately adhered to the axon (arrow in G), dendrites (arrows in H), and cell bodies (arrows in I) of FG-positive, axotomized facial motoneurons. Cont, contralateral. Scale bars in (F)=100 µm (A-F), (I)=50 µm (G-I); inset of (F)=20 µm (C, D, F).
(A-F) Double labeling of OX6 (brown) and ED1 (black) followed by counterstaining with cresyl violet in the facial motor nucleus at various time points. All OX6-positive microglia contained ED1-positive particles or spherical bodies (arrows). These microglia closely adhered to the degenerating facial motoneurons throughout all degenerative stages (A-F). Some microglia were labeled by ED1 but not by OX6 (arrowheads). (G-I) Triple labeling of neuronal nuclei (purple), phosphorylated c-Jun (black), and OX6 (brown) in the ipsilateral facial motor nucleus at 8 weeks post-lesion. Numerous pc-Jun-positive facial motoneurons were closely surrounded and phagocytosed by OX6-positive microglia (arrows in H, I). d, days; w, weeks. Scale bars in (F)=30 µm (A-F); (G)=100 µm; (I)=50 µm (H, I).
Article
Activating transcription factor 3 (ATF3) and c-Jun play key roles in either cell death or cell survival, depending on the cellular background. To evaluate the functional significance of ATF3/c-Jun in the peripheral nervous system, we examined neuronal cell death, activation of ATF3/c-Jun, and microglial responses in facial motor nuclei up to 24 weeks after an extracranial facial nerve axotomy in adult rats. Following the axotomy, neuronal survival rate was progressively but significantly reduced to 79.1% at 16 weeks post-lesion (wpl) and to 65.2% at 24 wpl. ATF3 and phosphorylated c-Jun (pc-Jun) were detected in the majority of ipsilateral facial motoneurons with normal size and morphology during the early stage of degeneration (1-2 wpl). Thereafter, the number of facial motoneurons decreased gradually, and both ATF3 and pc-Jun were identified in degenerating neurons only. ATF3 and pc-Jun were co-localized in most cases. Additionally, a large number of activated microglia, recognized by OX6 (rat MHC II marker) and ED1 (phagocytic marker), gathered in the ipsilateral facial motor nuclei. Importantly, numerous OX6- and ED1-positive, phagocytic microglia closely surrounded and ingested pc-Jun-positive, degenerating neurons. Taken together, our results indicate that long-lasting co-localization of ATF3 and pc-Jun in axotomized facial motoneurons may be related to degenerative cascades provoked by an extracranial facial nerve axotomy.
 
Article
Spatially and temporally programmed expression of the Hox genes along the antero-posterior (A-P) axis is essential for correct pattern formation during embryonic development. An accumulating body of evidence indicates the pivotal role of spatial chromatin organization for the coordination of gene regulation. Recently, chromosome conformation capture (3C) technique has been developed and opened a new way to study chromosomal interactions in the nucleus. In this study, we describe 3C method we applied in F9 embryonic teratocarcinoma cells and demonstrate that the chromosomal interactions at Hox loci are successfully detected. Interestingly, at Hoxc loci, the abundance of intrachromosomal interactions with neighboring fragments was drastically decreased when the genes are expressed. These results indicate the possibility of the dynamic pattern of chromosomal interaction in association with the transcriptional regulation of Hox genes.
 
Poly(ADP-ribose) polymerase 1 (PARP1) inhibition reduces necrosis induced by cisplatin in kidney proximal tubular cells after cisplatin injury. After 18 hours of starvation, HK-2, LLC-PK1, and MCT cells were treated with 400 mM cisplatin for 8 hours. The cells were also treated with high or low dose of PJ34 at 2 hours after treatment with cisplatin. (A) PARP activity was measured by a universal PARP assay kit. (B) Necrosis in HK-2 cells was detected by propidium iodid (PI) staining. Nuclei were counterstained with DAPI. Scale bars=50 μm. (C) The percentage of PI-positive cells was assessed in 10 fields (×400) per well. Error bars represent SD (n=3 experiments). **P<0.01 versus control. † P<0.05, † † P<0.01, † † † P<0.001 versus vehicle. 
Poly(ADP-ribose) polymerase 1 (PARP1) inhibition reduces plasma membrane disruption and HMGB1 release induced by cisplatin in kidney proximal tubular cells. After 18 hours of starvation, HK-2 cells were treated with 400 μM cisplatin for 8 hours. The cells were also treated with high or low dose of PJ34 at 2 hours after treatment with cisplatin. After 8 hours of cisplatin injury, the cells and media were harvested. (A) Lactate dehydrogenase (LDH) release was measured enzymatically using a CytoTox 96 Non-Radioactive Cytotoxicity Assay kit. (B) The levels of released HMGB1 in cell culture media were measured using Western blot analysis. Protein bands were quantified using Lab Works analysis software (Ultra-Violet Products). Error bars represent SD (n=3 experiments). **P<0.01 versus control. † P<0.05, † † P<0.01, † † † P<0.001 versus vehicle. 
Poly(ADP-ribose) polymerase 1 (PARP1) inhibition ameliorates ATP depletion induced by cisplatin in kidney proximal tubular cells. After 18 hours of starvation, HK-2 cells were treated with 400 μM cisplatin for 8 hours. The cells were also treated with high or low dose of PJ34 at 2 hours after treatment with cisplatin. The level of ATP was measured by an ATP assay kit (BioVision). Error bars represent SD (n=3 experiments). **P<0.01, ***P<0.001 versus control. † † P<0.01, † † † P<0.001 versus vehicle.
Poly(ADP-ribose) polymerase 1 (PARP1) inhibition reduces oxidative stress induced by cisplatin in kidney proximal tubular cells. After 18 hours of starvation, HK-2 cells were treated with 400 μM cisplatin for 8 hours. The cells were also treated with high or low dose of PJ34 at 2 hours after treatment with cisplatin. The levels of 8-OHdG (A) and lipid hydroperoxide (B) were measured by assay kits (Cayman). Error bars represent SD (n=3 experiments). **P<0.01 versus control. † P<0.05, † † P<0.01, † † † P<0.001 versus vehicle. 
Article
Treatment with cisplatin for cancer therapy has a major side effect such as nephrotoxicity; however, the role of poly (ADP-ribose) polymerase 1 (PARP1) in necrosis in response to cisplatin nephrotoxicity remains to be defined. Here we report that cisplatin induces primary necrosis through PARP1 activation in kidney proximal tubular cells derived from human, pig and mouse. Treatment with high dose of cisplatin for 4 and 8 hours induced primary necrosis, as represented by the percentage of propidium iodide-positive cells and lactate dehydrogenase release. The primary necrosis was correlated with PARP1 activation during cisplatin injury. Treatment with PJ34, a potent PARP1 inhibitor, at 2 hours after injury attenuated primary necrosis after 8 hours of cisplatin injury as well as PARP1 activation. PARP1 inhibition also reduced the release of lactate dehydrogenase and high mobility group box protein 1 from kidney proximal tubular cells at 8 hours after cisplatin injury. Oxidative stress was increased by treatment with cisplatin for 8 hours as shown by 8-hydroxy-2'-deoxyguanosine and lipid hydroperoxide assays, but PARP1 inhibition at 2 hours after injury reduced the oxidative damage. These data demonstrate that cisplatin-induced PARP1 activation contributes to primary necrosis through oxidative stress in kidney proximal tubular cells, resulting in the induction of cisplatin nephrotoxicity and inflammation.
 
Lysosomal acidification is re qu ire d f or d emy e l inati on. (A) Differential interference microscopy of teased nerve fibers after 3 days of culture in vitro (3DIV). Scale bar=200 μm. (B) Quantitative result showing myelin ovoid index. (C, D) Western blot analysis showing the levels of myelin ba sic protein (M B P) (C) and lysosomal associated membrane protein 1 (LAMP1) (D) in cultured nerve explants. "C" indicates uninjured sciatic nerves. CQ, chloroquine; BFA, bafilomycin A.
Transient induction of lysosomal associated membrane protein 1 (LAMP1) in demyelinating Schwann cells. (A) LAMP1 was localized in perinuclear areas (arrows) in control sciatic nerves. (B) Sciatic nerve crush-induced LAMP1 in demyelinating Schwann cells, particularly around myelin ovoids 3 days (3d) after injury. The upper panels are double staining against LAMP1 and S100 (arrow), and the lower panels are double staining against LAMP1 and myelin basic protein (MBP). Asterisk in the middle panels are myelin ovoids demonstrated by differential interference microscopy. Asterisks in the lower panels are MBP-positive clumps surrounded by LAMP1 immunoreactivity. Arrows indicate a LAMP1-negative intact myelin sheath. (C) Downregulation of LAMP1 in demyelinating Schwann cells 7 days (7d) after injury (asterisks). Arrows indicate LAMP1 and CD68-positive macrophages. Macrophages were found inside or outside the endoneurial tube labeled with antibody against laminin (LAM). (D) Three weeks (3W) after injury, remyelinating Schwann cells (labeled with MBP or LAM) were devoid of LAMP1 staining, whereas many LAMP1-positive macrophages were still present around remyelinating Schwann cells. DIC, differential interference contrast. Scale bars=100 µm.
Lysosomal acidification is required
for demyelination. (A) Differential interference microscopy of teased nerve fibers after 3 days of culture in vitro (3DIV). Scale bar=200 µm. (B) Quantitative result showing myelin ovoid index. (C, D) Western blot analysis showing the levels of myelin basic protein (MBP) (C) and lysosomal associated membrane protein 1 (LAMP1) (D) in cultured nerve explants. "C" indicates uninjured sciatic nerves. CQ, chloroquine; BFA, bafilomycin A.
Lysosomal acidification inhibitors prevent p75 nerve growth factor receptor (p75NGFR) induction. (A) Immunofluorescence microscopy showing p75NGFR expression in teased nerve fibers. Neurofilament (NF) staining showed extensive fragmentation at 3 days culture in vitro (3DIV), regardless of the inhibition of lysosomal acidification. (B) Western blot analysis showing the levels of p75NGFR in cultured nerve explants at 3DIV. (C) Reverse transcription-polymerase chain reaction experiment showing the induction of p75NGFR mRNA in cultured nerve explants at 3 days. (D) Western blot analysis showing the levels of p75NGFR in the sciatic nerves of tumor necrosis factor-α null mice (TNF-KO). (E) Immunofluorescence microscopy showing p75NGFR and lysosomal associated membrane protein 1 (LAMP1) expression in teased nerve fibers 3 days (3d) after injury. "C" indicates uninjured sciatic nerves. BFA, bafilomycin A; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; 3d, three days after crush; WT, wild-type; Con, control. Scale bars=100 µm.
Article
Myelinated Schwann cells in the peripheral nervous system express the p75 nerve growth factor receptor (p75NGFR) as a consequence of Schwann cell dedifferentiation during Wallerian degeneration. p75NGFR has been implicated in the remyelination of regenerating nerves. Although many studies have shown various mechanisms underlying Schwann cell dedifferentiation, the molecular mechanism contributing to the re-expression of p75NGFR in differentiated Schwann cells is largely unknown. In the present study, we found that lysosomes were transiently activated in Schwann cells after nerve injury and that the inhibition of lysosomal activation by chloroquine or lysosomal acidification inhibitors prevented p75NGFR expression at the mRNA transcriptional level in an ex vivo Wallerian degeneration model. Lysosomal acidification inhibitors suppressed demyelination, but not axonal degeneration, thereby suggesting that demyelination mediated by lysosomes may be an important signal for inducing p75NGFR expression. Tumor necrosis factor-α (TNF-α) has been suggested to be involved in regulating p75NGFR expression in Schwann cells. In this study, we found that removing TNF-α in vivo did not significantly suppress the induction of both lysosomes and p75NGFR. Thus, these findings suggest that lysosomal activation is tightly correlated with the induction of p75NGFR in demyelinating Schwann cells during Wallerian degeneration.
 
Laminar fl ow inhibits 15d-PGJ 2-induced endothelial apoptosis. Confl uent HUVECs were cultured under L-fl ow (12 dyne/cm 2 ) or a static condition for 24 h. Cells were then exposed to 10 μM 15d-PGJ 2 for 8 h. (A) Cell lysates were applied for immunoblotting with anti-PARP, anticleaved caspase 3, and anti-tubulin antibodies. Data are representative of results from three separate experiments. (B) Cell viability was determined by using MTT assay. Data are expressed as the mean±SD from three independent experiments. *P<0.01. Morphological changes of HUVECs were examined by microscopy under control (C), 15d-PGJ 2 (D), or 15d-PGJ 2 under L-fl ow (E). Scale bars=100 μm (C-E). 15d-PGJ 2 , 15-deoxyΔ(12,14)-prostaglandin J 2 ; HUVECs, human umbilical vein endothelial cells; L-fl ow, steady laminar fl ow; PARP, poly (ADP-ribose) polymerase; OD, optical density.
Laminar flow inhibits 15d-PGJ2-induced endothelial apoptosis. Confluent HUVECs were cultured under L-flow (12 dyne/cm2) or a static condition for 24 h. Cells were then exposed to 10 µM 15d-PGJ2 for 8 h. (A) Cell lysates were applied for immunoblotting with anti-PARP, anti-cleaved caspase 3, and anti-tubulin antibodies. Data are representative of results from three separate experiments. (B) Cell viability was determined by using MTT assay. Data are expressed as the mean±SD from three independent experiments. *P<0.01. Morphological changes of HUVECs were examined by microscopy under control (C), 15d-PGJ2 (D), or 15d-PGJ2 under L-flow (E). Scale bars=100 µm (C-E). 15d-PGJ2, 15-deoxy-Δ(12,14)-prostaglandin J2; HUVECs, human umbilical vein endothelial cells; L-flow, steady laminar flow; PARP, poly (ADP-ribose) polymerase; OD, optical density.
L-flow inhibits 15d-PGJ2-induced endothelial apoptosis via p53 down regulation. (A) HUVECs were exposed to L-flow followed by treatment with 15d-PGJ2 for 8 h. Protein expression was determined by Western blotting using anti-p53 and anti-tubulin antibodies. (B) HUVECs were transfected with either control or p53 siRNA (si-p53) for two days. Cells were then exposed to 10 µM 15d-PGJ2 for 8 h. The expression of cleaved caspase 3, p53, and tubulin was detected by Western blotting with the respective antibodies. (C) Cell viability was determined by MTT assay. Data are expressed as the mean±SD from three independent experiments. *P<0.01. L-flow, steady laminar flow; 15d-PGJ2, 15-deoxy-Δ(12,14)-prostaglandin J2; HUVECs, human umbilical vein endothelial cells.
The ERK5-CHIP signal module protects endothelial apoptosis in response to 15d-PGJ2. (A) HUVECs were pretreated with DMSO or 10 µM BIX02189, a specific inhibitor of MEK5, before applying L-flow. Cells were then exposed to 10 µM 15d-PGJ2 for 8 h. The protein expression of p53, phosphorylated ERK5, and tubulin were determined by immunoblotting with specific antibodies. (B) HUVECs were transfected with siRNA against human CHIP (si-CHIP) or control RNA and then followed by transducing the adenovirus encoding con stitutively active form of MEK5 alpha (Ad-CA-MEK5α). Cells were then exposed to 10 µM 15d-PGJ2 for 8 h. Cell lysates were subjected to Western blot analysis with anti-cleaved caspase 3, anti-phospho ERK5, anti-CHIP, and anti-tubulin antibodies. (C) Cell viability was determined by MTT assay. Data are expressed as the mean±SD from three independent experiments. *P<0.01. ERK5, extracellular signal-regulated kinase 5; CHIP, C terminus of Hsc70-interacting protein; 15d-PGJ2, 15-deoxy-Δ(12,14)-prostaglandin J2; HUVEC, human umbilical vein endothelial cells; OD, optical density; NS, non-significant.
ERK5 activation induces p53 ubiquitination via increasing CHIP ubiquitin ligase activity. (A) After transduction with Ad-CA-MEK5α or the Ad-LacZ control, HUVECs were exposed to 15d-PGJ2 for 4 h. The level of p53 mRNA was determined by using real time quantitative RT-PCR as described in the Materials and Methods section. (B) The endogenous level of p53 ubiquitination was determined by immunoprecipitation analysis with anti-p53 antibody, followed by immunoblotting with an anti-ubiquitin antibody. (C) CHIP ubiquitin ligase activity was determined by an in vitro ubiquitination assay kit (BostonBiochem) with a GST-fused p53 recombinant protein as a substrate. HUVECs were transduced with Ad-CA-MEK5α and then followed by treatment with 10 µM BIX02189. Cell lysates were applied for immunoprecipitation analysis with an anti-CHIP antibody. Immunoprecipitated CHIP was incubated with recombinant proteins including ubiquitin and E1/E2 enzyme mixture, for 60 min at 37℃. CHIP ubiquitin ligase activity was determined by immunoblotting with an anti-ubiquitin antibody. The protein amount of GST-p53 and immunoprecipitated CHIP was detected by Western blot analysis with anti-p53 and anti-CHIP antibodies. Data are representative of three independent experiments. ERK5, extracellular signal-regulated kinase 5; CHIP, C terminus of Hsc70-interacting protein; Ad-CA-MEK5α, adenovirus encoding constitutively active form of MEK5 alpha; 15d-PGJ2, 15-deoxy-Δ(12,14)-prostaglandin J2; RT-PCR, reverse transcriptase-polymeratse chain reaction; HUVEC, human umbilical vein endothelial cells; IP, immunoprecipitation; IB, immunoblot; NS, non-significant.
Article
Atherosclerosis is readily observed in areas where disturbed flow is formed, while the atheroprotective region is found in areas with steady laminar flow (L-flow). It has been established that L-flow protects endothelial cells against endothelial dysfunction, including apoptosis and inflammation. It has also been reported that extracellular signal-regulated kinase 5 (ERK5) regulated endothelial integrity and protected endothelial cells from vascular dysfunction and disease under L-flow. However, the molecular mechanism by which L-flow-induced ERK5 activation inhibits endothelial apoptosis has not yet been determined. Transcription factor p53 is a major pro-apoptotic factor which contributes to apoptosis in various cell types. In this study, we found that 15-deoxy-Δ(12,14)-prostaglandin J(2) induced p53 expression and that endothelial apoptosis was reduced under the L-flow condition. This anti-apoptotic response was reversed by the biochemical inhibition of ERK5 activation. It was also found that activation of ERK5 protected endothelial apoptosis in a C terminus of Hsc70-interacting protein (CHIP) ubiquitin ligase-dependent manner. Moreover, molecular interaction between ERK5-CHIP and p53 ubiquitination were addressed with a CHIP ubiquitin ligase activity assay. Taken together, our data suggest that the ERK5-CHIP signal module elicited by L-flow plays an important role in the anti-apoptotic mechanism in endothelial cells.
 
Article
Vitamin C, one of essential micronutrients, has been reported to modulate the humoral immune responses in some mammals. We investigated whether vitamin C might modulate this response in mice by directly affecting B cells. Splenic B cells were isolated and activated by CD40- and B cell receptor-ligation in vitro. The cells were cultured with a pretreatment of vitamin C from 0 to 1 mM of concentrations. Vitamin C slightly increased apoptosis of B cells dose-dependently and behaved as an antioxidant. We found that in vivo administration of vitamin C by intraperitoneal injection affected isotype switching as previously reported: the titer of antigen-specific IgG1 antibody was decreased, while that of IgG2a was unaffected. Somewhat different from those observed in vivo, in vitro exposure to vitamin C slightly decreased isotype switching to IgG1 and increased isotype switching to IgG2a. Pretreatment with vitamin C in the safe range did not affect either proliferation of cultured B cells or the expression of CD80 and CD86 in those cells. Taken together, in vivo results suggest that vitamin C acts to modulate isotype switching in the mouse. However, because of our in vitro results, we suggest that the modulation exerted by vitamin C in vivo is by indirectly affecting B cells, perhaps by directly influencing other immune cells such as dendritic cells.
 
Structure and recombinant adenovirus transfection in cultured cells and cerebellar injections in Sprague-Dawley rats. (A) Cell type specific promoter and tracer transgene of recombinant adenovirus. (B) HEK293 cells were cultured in 10-cm culture dishes. (C) Five days after transfection with the pAdeasy-TH-RFP-CMV-GFP construct, strong GFP expression was observed in HEK293 cells. (D) Three days after injection of the TH-RFP-CMV-GFP recombinant adenovirus into the cerebellum of male Sprague-Dawley rats, GFP was highly expressed in the cerebellar cortex. g, granular layer; P, Purkinje cell layer; m, molecular layer; (E) RFP was not expressed in the cerebellar cortex. Scale bars=100 µm.
Transgene expression 3 days after injection of the TH-RFP-CMV-GFP recombinant adenovirus into the locus coeruleus. (A) Distribution of Hoechst 33342-stained nuclei in the midbrain. (B) GFP was highly expressed in the injected side of midbrain. (C) RFP was expressed in several neurons of the LC. (D) RFP expression colocalized with GFP-positive neurons. Scale bar=100 µm.
Transgene expression 3 days after TH-WGA-CMV-GFP recombinant adenovirus injection into the VTA. (A) GFP was highly expressed in the injected side. (B) Magnified image of the indicated region of (A) (square). (C) WGA immunohistochemistry showed the presence of WGA-immunoreactive neurons in regions adjacent to the injected site. Scale bar=100 µm.
Transgene expression 3 days after TH-WGA-CMV-GFP recombinant adenovirus injection into the LC. (A) Distribution of Hoechst 33342-positive nuclei in the midbrain. (B) GFP was highly expressed in the injected side of the LC and in adjacent regions. Some GFP-positive fibers were distributed in the lower lateral side of midbrain. (C) WGA immunohistochemistry showed that several neurons were positive for WGA staining. (D) Merged image of (A, B) and (C). (E~H) Magnified images of the indicated region of (D) (square). (E) Distribution of Hoechst 33342 stained nuclei. (F) GFP-expressing cells. (G) WGA-immunoreactive neurons. (H) Merged image. (I) WGA immunohistochemistry using DAB development showed WGA-immunoreactive neurons in the injected side of the locus coeruleus. (J) Contralateral side of the locus coeruleus. (K) Magnified image of the indicated region of (I) (square). (L) Magnified image of the indicated region of (J) (square). Scale bar=100 µm.
Transgene expression 3 days after TH-WGA-CMV-GFP recombinant adenovirus injection in the VTA. (A) Distribution of Hoechst 33342-positive nuclei in the injected side of the VTA. (B) GFP was highly expressed in the injected side of the VTA. Many GFP-positive fibers were distributed among the GFP-positive cell bodies. (C) WGA immunohistochemistry showed that several neurons were WGA positive. WGA-positive fibers and cell bodies were distributed in adjacent regions. (D) Merged image of (A, B) and (C). Z-stack image of (E) GFP and WGA colocalized in the same cells. Scale bar=100 µm.
Article
Selective labeling of small populations of neurons of a given phenotype for conventional neuronal tracing is difficult because tracers can be taken up by all neurons at the injection site, resulting in nonspecific labeling of unrelated pathways. To overcome these problems, genetic approaches have been developed that introduce tracer proteins as transgenes under the control of cell-type-specific promoter elements for visualization of specific neuronal pathways. The aim of this study was to explore the use of tracer gene expression for neuroanatomical tracing to chart the complex interconnections of the central nervous system. Genetic tracing methods allow for expression of tracer molecules using cell-type-specific promoters to facilitate neuronal tracing. In this study, the rat tyrosine hydroxylase (TH) promoter and an adenoviral delivery system were used to express tracers specifically in dopaminergic and noradrenergic neurons. Region-specific expression of the transgenes was then analyzed. Initially, we characterized cell-type-specific expression of GFP or RFP in cultured cell lines. We then injected an adenovirus carrying the tracer transgene into several brain regions using a stereotaxic apparatus. Three days after injection, strong GFP expression was observed in the injected site of the brain. RFP and WGA were expressed in a cell-type-specific manner in the cerebellum, locus coeruleus, and ventral tegmental regions. Our results demonstrate that selective tracing of catecholaminergic neuronal circuits is possible in the rat brain using the TH promoter and adenoviral expression.
 
Article
Stress has long been known to be a causative factor of various disease states. In this study, we investigated the effects of repeated restraint stress on platelet endothelial cell adhesion molecule-1 (PECAM-1), a very important mediator in inflammation, immunoreactivity and protein levels as well as neuronal damage, in the gerbil hippocampus after 5 minutes of transient cerebral ischemia. Transient ischemia-induced neuronal death was shown in CA1 pyramidal cells 4 days after ischemia/reperfusion. However, repeated restraint stress protected neuronal death induced by ischemic damage. In the ischemia-group, PECAM-1 immunoreactivity and its protein levels were significantly increased in all the hippocampal subregions 4 days after ischemia/reperfusion. However, PECAM-1 immunoreactivity and its protein levels did not change significantly in the hippocampus of the stress-ischemia-group compared to the sham-groups. These results indicate that repeated restraint stress protects neuronal damage induced by transient cerebral ischemia, and this may be associated with maintenance of PECAM-1levels.
 
The phospholylation of insulin receptor substrate (IRS)-1 in streptozotocin (STZ) induced dementia rat model. To confirm the dysfunction of brain insulin system in STZ-induced dementia rat model known as common animal model to study Alzheimer's disease in vivo, we conducted immunohistochemistry using phospho IRS-1 antibody. (A) In the hippocampus, phospholylation of IRS-1 decreased in the STZ induced dementia group compared with sham (control group). (B) In the cortex, phospholylation of IRS-1 decreased in the STZ induced dementia group compared with sham (control group). Green color, phospho IRS-1; red color, propidium iodide (PI).
Adiponectin related diseases and pathologies
Article
Adiponectin is an adipocytokine released by the adipose tissue and has multiple roles in the immune system and in the metabolic syndromes such as cardiovascular disease, Type 2 diabetes, obesity and also in the neurodegenerative disorders including Alzheimer's disease. Adiponectin regulates the sensitivity of insulin, fatty acid catabolism, glucose homeostasis and anti-inflammatory system through various mechanisms. Previous studies demonstrated that adiponectin modulates memory and cognitive impairment and contributes to the deregulated glucose metabolism and mitochondrial dysfunction observed in Alzheimer's disease. Here, we aim to summarize recent studies that suggest the potential correlation between adiponectin and Alzheimer's disease.
 
Establishment of human mesenchymal stem cells (MSCs) from adipose tissue. MSCs were isolated from human fat tissue. (A) On day 3 of culture, cells adhered to the bottom of the culture dish and appeared spindle-shaped. On day 3, cells cultured in EMG-2 media were nearly 90% confluent (left panel), while cells in mixed media were about 50% confluent (right panel). (B) The obtained cells were stained with antibodies against various surface markers, as indicated, and analyzed by flow cytometric analysis. Cell populations were negative for hematopoietic markers (CD34, CD45, and CD117), and positive for stem cell markers (CD29, CD44, CD90, and CD105). (C) Cells were induced to differentiate to adipocytes (left panel), chondrocytes (middle panel), or osteoblasts (right panel) using corresponding differentiation medium, fixed with 4% paraformaldehyde at an appropriate time point, and stained with Oil Red-O, Alcian blue, or Alizarin red S, respectively.
Suppressive effects of human adipose tissue-derived mesenchymal stem cells (hAd-MSCs) on the proliferation of T cells. Human peripheral blood mononuclear cells (PBMCs) (A), mouse splenocytes (B), and mouse splenic T cells (C) were stimulated with antibodies against human or mouse CD3 and CD28, respectively, and cultured in the presence or absence of hAd-MSCs (1:10 ratio of MSCs:target cells) for 2 days. 3[H]-thymidine was added to the culture, and radioactivity was determined after 18 hours. The presence of hAd-MSCs markedly suppressed T cell proliferation of both human and mouse cells. Results are representative of three independent experiments. CPM, count per minute. *P<0.001.
Expression of immunomodulatory molecules in activated human adipose tissue-derived mesenchymal stem cells (hAd-MSCs). (A) Peripheral blood mononuclear cells (PBMCs) or mouse T cells were activated with concanavalin A (ConA), with antibodies against human or mouse CD3 and CD28, or with phorbol myristate acetate/ionomycin (PMA+I) for 3 days, and the supernatants were thus obtained. hAd-MSCs were stimulated with each supernatant for 24 hours and harvested. Total RNA was purified from these cells, and reverse transcription polymerase chain reaction analysis was performed for the molecules as indicated. (B) The average values of the intensities of the polymerase chain reaction bands from three replicate experiments measured by densitometry are shown. Lines at the top of each bar represent the magnitude of standard deviation. COX-2, cyclooxygenase-2; iNOS, inducible NO synthase; IDO, indoleamine 2,3-dioxygenase.
Restoration of the suppressive effect of human adipose tissue-derived mesenchymal stem cells (hAd-MSCs) by the cyclooxygenase-2 (COX-2) inhibitor NS-398. (A) Mouse splenic T cells were co-cultured with hAd-MSCs in the presence or absence of 0.5 mM NS-398 (left panel), or in the presence or absence of 0.5 mM 1-MT (indoleamine 2,3-dioxygenase [IDO] inhibitor), or 1 mM L-NAME (right panel) for 2 days. 3[H]-thymidine was added to the co-culture, and radioactivity was determined after 18 hours. The presence of hAd-MSCs markedly suppressed the T cell proliferation of mouse T cells, which was almost completely recovered by the addition of the COX-2 inhibitor (left panel), while the addition of the IDO inhibitor or inducible NO synthase inhibitor did not affect the suppressive function of hAd-MSCs (right panel). (B) hAd-MSCs were stimulated with the culture supernatant of mouse T cells or with each recombinant mouse cytokine including tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interlukin (IL)-4, or IL-5 at a concentration of 1 ng/ml. Cells were harvested after 24 hours, total RNA was extracted, and reverse transcription polymerase chain reaction was performed for COX-2 expression. Only the hAd-MSCs stimulated with culture supernatant or TNF-α expressed the molecule. Each experiment was repeated more than three times with representative results depicted here. CPM, count per minute; DMSO, dimethyl sulfoxide, vehicle control; Sup, culture supernatant of mouse T cells; *P<0.01.
Primer sets used in this study
Article
Mesenchymal stem cells (MSCs) of human origin have been frequently applied to experimental animal models to evaluate their immunomodulatory functions. MSCs are known to be activated by cytokines from T cells, predominantly by interferon-γ (IFN-γ), in conjunction with other cytokines such as tumor necrosis factor-α (TNF-α) and interlukin-1β. Because IFN-γ is not cross-reactive between human and mouse species, the manner in which human MSCs administered in experimental animals are activated and stimulated to function has been questioned. In the present study, we established MSCs from human adipose tissue. They successfully suppressed the proliferation of not only human peripheral blood mononuclear cells but also mouse splenic T cells. When these human MSCs were stimulated with a culture supernatant of mouse T cells or recombinant murine TNF-α, they expressed cyclooxygenase-2 (COX-2), but not indoleamine 2,3-dioxygenase. The dominant role of COX-2 in suppressing mouse T cell proliferation was validated by the addition of COX-2 inhibitor in the co-culture, wherein the suppressed proliferation was almost completely recovered. In conclusion, human MSCs in a murine environment were activated, at least in part, by TNF-α and mainly used COX-2 as a tool for the suppression of in vitro T cell proliferation. These results should be considered when interpreting results for human MSCs in experimental animals.
 
Article
Using immunohistochemical staining for alpha-smooth muscle actin (α-SMA), glial fibrillary acidic protein (GFAP), S100 protein (S100), p63, cytokeratin 14 (CK14), and cytokeratin 19 (CK19), we studied acinar and myoepithelial cells of major and minor salivary glands obtained from 14 donated cadavers (78-92 years old) and 5 donated fetuses (aborted at 15-16 weeks of gestation). CK and p63 expression was investigated only in the adult specimens. SMA was detected in all adult glands as well as in fetal sublingual and pharyngeal glands. GFAP expression was seen in a limited number of cells in adult glands, but was highly expressed in fetal pharyngeal glands. S100-positive myoepithelial-like cells were present in adult minor glands as well as in fetal sublingual and pharyngeal glands. Expression of p63 was evident in the ducts of adult glands. CK14 immunoreactivity was observed in a limited number of glandular cells in adults, in contrast to consistent expression of CK19. In both adults and fetuses, a mosaic expression pattern was usually evident for each of the examined proteins. A difference in immunoreactivity for the nerve markers GFAP and S100 was observed between the major and minor glands. Thus, in the present histologic study, we distinguished between the specific gland types on the basis of their immunohistochemical staining. A mosaic expression pattern suggested that the immunoreactivity against nerve protein markers in myoepithelial cells could not be due to the persistence of neural crest remnants or the physiological status of the gland, such as age-related degeneration.
 
Article
The occurrence of a brachial plexus united into a single cord is very rare. During routine dissection of an elderly female cadaver, the brachial plexus united into a single cord was observed bilaterally. On the left side, C4, C5, and C6 roots combined to form the upper trunk, the C7 root continued as the middle trunk, and C8 and T1 united to form the lower trunk. All three trunks almost immediately fused to form a single cord. On the right side, C5 and C6 roots joined to form the upper trunk, which divided into anterior and posterior divisions. C7, C8, and T1 roots combined to form the lower trunk. The anterior and posterior divisions united with the lower trunk to form a single cord. On both sides, the subclavian artery was superior to the single cord. Supraclavicular brachial plexus injuries in such individuals may have serious clinical manifestations.
 
Light (A, B) and electron (C~E) micrographs showing HRP-labeled vibrissa afferent boutons in the laminae III/IV of the cat trigeminal caudal nucleus (Vc). (A, B) Light micrographs showing the HRP-labeled boutons (arrowheads) in laminae III/IV of Vc. B is an enlargement of boxed area in A. (C) An electron micrograph on a labeled bouton (asterisk) showing a simple synaptic connection with a dendrite (d). (D, E) Electron micrographs on adjacent thin sections showing a labeled bouton (asterisk). The bouton shows an example of complex synaptic arrangement with four dendrites (d1~d4) and presynaptic endings (p1) on adjacent thin sections. Arrows indicate the direction of synaptic transmission. Scale bars, 200 µm in (A); 20 µm in (B) 500 nm in (C~E).
Distribution of the volume of vibrissa afferent boutons and their presynaptic endings (p-endings) in the cat trigeminal caudal nucleus (Vc).
Correlations between the volume of vibrissa afferent boutons and the number of presynaptic endings (p-endings), and between the volume of vibrissa afferent boutons and number of postsynaptic dendrites in the cat trigeminal caudal nucleus (Vc). r, correlation coefficient.
Correlations between the volume of vibrissa afferent boutons and their mitochondrial volume, active zone area, apposed surface area and vesicle density in the cat trigeminal caudal nucleus (Vc). r, correlation coefficient.
Ultrastructural analysis of HRP-labeled vibrissa afferent boutons and their presynaptic endings (p-endings) in the cat trigeminal caudal nucleus
Article
Ultrastructural parameters related to synaptic release and their correlation with synaptic connectivity were analyzed in the low-threshold mechanoreceptive vibrissa afferent boutons in laminae III and IV of the trigeminal caudal nucleus (Vc). Rapidly adapting vibrissa afferents were intra-axonally labeled, and quantitative ultrastructural analyses with serial sections were performed on the labeled boutons and their presynaptic endings (p-endings). The volume of the labeled boutons was widely distributed from small to large ones (0.8~12.3 µm(3)), whereas the p-endings were small and uniform in size. The volume of the labeled boutons was positively correlated with the ultrastructural parameters such as mitochondrial volume (correlation coefficient, r=0.96), active zone area (r=0.82) and apposed surface area (r=0.79). Vesicle density (r=-0.18) showed little correlation to the volume of labeled boutons, suggesting that the total vesicle number of a bouton is proportional to its volume. In addition, the bouton volume was positively correlated with the number of p-endings (r=0.52) and with the number of dendrites postsynaptic to the labeled bouton (r=0.83). These findings suggest that low-threshold mechanoreception conveyed through vibrissa afferents is processed in a bouton size-dependent manner in the Vc, which may contribute to the sensory-motor function of laminae III/IV in Vc.
 
Leonardo da Vinci's depiction of a skull. The image is one of da Vinci's anatomical drawings of a human skull. The left half of the skull is sectioned to reveal the frontal sinus and the maxillary sinus. Of note is the close relation of the two sinuses to the orbit and the teeth of the upper jaw, as understood by Leonardo. Reprinted from Leonardo Da Vinci's drawings [11]. 
Vesalius' images of the skull and the sphenoid bone. The images constitute anatomical drawings from Vesalius' work 'De Humani Corporis Fabrica.' Most of the illustrations in this book were created by Jan Stephan van Calcar, an Italian artist, one of Titian's students. The left half of the picture shows a transverse cross-section of the skull, depicting the calvaria whereas the right half shows the sphenoid bone. The right-hand image shows the frontal sinus as well as the two sphenoid sinuses, which are separated by the sphenoid septum. Reprinted from Andreas Vesalius, De Humani Corporis Fabrica [13]. 
Illustrations from Nathaniel Highmore's book 'Corporis Humani Disquisitio Anatomica.' The maxillary sinus and the projection of the teeth of the upper jaw into the floor of the sinus are clearly shown. The cross-section of the skull in the bottom right of the picture reveals the frontal sinus, sphenoid sinus, ethmoid cells and maxillary sinus. Reprinted from Nathaniel Highmore, Corporis humani disquisitio anatomica, p. 227 [19]. 
Article
Previously, anatomists considered paranasal sinuses as a mysterious region of the human skull. Historically, paranasal sinuses were first identified by ancient Egyptians and later, by Greek physicians. After a long period of no remarkable improvement in the understanding of anatomy during the Middle Ages, anatomists of the Renaissance period-Leonardo da Vinci and Vesalius-made their own contribution. Nathaniel Highmore's name is also associated with the anatomy of paranasal sinuses as he was first to describe the maxillary sinus.
 
Neuronal damages in rat hippocampus (a~f) and cortex (g~i) at 6 (A) and 24 h (B) after transient global ischemia. H-E and TUNEL staining were analyzed for histology, especially in the CA1 region of hippocampus and in the parietal cortex of cerebral cortex. NC, normal control; EC, experimental controls without agmatine treatment; Agm, agmatine treatment groups. Arrows point to necrotic and apoptotic neurons compared with the control. Scale bar=100 µm.
Effects of eNOS and iNOS expression by agmatine treatment following transient global ischemia in rat hippocampus. Representative expressional levels of eNOS (A) and iNOS (C) at 6 h after agmatine treatment (100 mg/kg, i.p), and densitometric data (B, D). Data represent means±SD for n=5/NC, n=3/EC and Agm group per each time point. *P<0.05, vs. normal control; †P<0.05, vs. experimental control at same onset of reperfusion. NC, normal control; EC6, at 6 h after reperfusion without agmatine; Agm6, at 6 h after reperfusion with agmatine; EC24, at 24 h after reperfusion without agmatine; Agm24, at 24 h after reperfusion with agmatine.
Effects of eNOS and iNOS expression by agmatine treatment following transient global cerebral ischemia in rat cerebral cortex. Representative expressional levels of eNOS (A) and iNOS (C) at 24 h after agmatine treatment (100 mg/kg, i.p), and densitometric data (B, D). Data represent means±SD for n=5/NC, n=3/EC and Agm group per each time point. *P<0.05, vs. normal control; †P<0.05, vs. experimental control at same onset of reperfusion. NC, normal control; EC6, at 6 h after reperfusion without agmatine; Agm6, at 6 h after reperfusion with agmatine; EC24, at 24 h after reperfusion without agmatine; Agm24, at 24 h after reperfusion with agmatine.
Photomicrographs of eNOS expression by agmatine treatment (A) and quantification of eNOS positive cells (B, C) in rat brain after global ischemia. Representative expressional levels of eNOS in bregman - 3.8 mm of naïve rats at 6 and 24 h after agmatine treatment (A). The number of eNOS positive cells in the CA 1 sector of hippocampus (B) and in the parietal region of the cerebral cortex (C). Data represent means±SD for n=5/NC, n=3/EC and Agm group per each time point. *P<0.05, vs. normal control; †P<0.05, vs. experimental control at same onset of reperfusion. NC, normal control; EC6, at 6 h after reperfusion without agmatine; Agm6, at 6 h after reperfusion with agmatine; EC24, at 24 h after reperfusion without agmatine; Agm24, at 24 h after reperfusion with agmatine; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum; HiF, hippocampal fissure. Higher magnifications of cells indicated by squares show eNOS positive cells in the endothelium of vessels and capillaries. Arrows point to positive cells that showed higher intensity of immunostaining with hematoxylin in rat brain, compared with the control. Scale bar=50 µm (hippocampus), and 100 µm (cortex).
Regulations of MMPs expression by agmatine treatment following transient global ischemia in rat brain. Representative expressional levels of MMP-2 and MMP-9 6 and 24 h after agmatine treatment (100 mg/kg, i.p) in the hippocampus (A), in the cerebral cortex (C), and densitometric data (B, D). Data represent means±SD for n=5/NC, n=3/EC and Agm group per each time point. *P<0.05, vs. normal control. NC, normal control; EC6, at 6 h after reperfusion without agmatine; Agm6, at 6 h after reperfusion with agmatine; EC24, at 24 h after reperfusion without agmatine; Agm24, at 24 h after reperfusion with agmatine.
Article
Nitric oxide (NO) production by endothelial nitric oxide synthase (eNOS) plays a protective role in cerebral ischemia by maintaining vascular permeability, whereas NO derived from neuronal and inducible NOS is neurotoxic and can participate in neuronal damage occurring in ischemia. Matrix metalloproteinases (MMPs) are up-regulated by ischemic injury and degrade the basement membrane if brain vessels to promote cell death and tissue injury. We previously reported that agmatine, synthesized from L-arginine by arginine decarboxylase (ADC) which is expressed in endothelial cells, has shown a direct increased eNOS expression and decreased MMPs expression in bEnd3 cells. But, there are few reports about the regulation of eNOS by agmatine in ischemic animal model. In the present study, we examined the expression of eNOS and MMPs by agmatine treatment after transient global ischemia in vivo. Global ischemia was induced with four vessel occlusion (4-VO) and agmatine (100 mg/kg) was administered intraperitoneally at the onset of reperfusion. The animals were euthanized at 6 and 24 hours after global ischemia and prepared for other analysis. Global ischemia led severe neuronal damage in the rat hippocampus and cerebral cortex, but agmatine treatment protected neurons from ischemic injury. Moreover, the level and expression of eNOS was increased by agmatine treatment, whereas inducible NOS (iNOS) and MMP-9 protein expressions were decreased in the brain. These results suggest that agmatine protects microvessels in the brain by activation eNOS as well as reduces extracellular matrix degradation during the early phase of ischemic insult.
 
Article
Stroke or cerebrovascular injury is the leading cause of disability and the third leading cause of deaths worldwide. After the initial ischemic injury, sympathetic signals are transmitted to the spleen and a compromised blood-brain barrier, coupled with expression of adhesion molecules by the vascular endothelial cells permits an influx of peripheral immune cells. This influx of peripheral immune cells into the brain exacerbates the local brain inflammatory response, leading to enhanced neurodegeneration. Agmatine is a primary amine formed by decarboxylation of L-arginine synthesized in the mammalian brain. In this study, we determined the effect of agmatine on the immune response in the spleen after transient cerebral ischemia. Twenty-three hours after transient cerebral ischemia, the white pulp area was reduced and the number of CD11b(+) macrophages and CD4(+)CD25(+) regulatory T cells (T reg cells) were increased in the spleens in the experimental group as a result of alteration of the immune response in the spleen, as regulated by inflammatory cytokines. In the agmatine treatment group (100 mg/kg IP), the contraction of white pulp was diminished and the number of CD11b(+) macrophages and CD4(+)CD25(+)T reg cells were decreased. Twenty-three hours after transient cerebral ischemia, the brain infarction area was significantly reduced (5.51±1.63% of the whole brain) in the agmatine treatment group compared to 15.02±4.28% of the whole brain in the experimental control group. These results suggest that agmatine treatment can reduce brain infarction through minimizing neuroinflammation and can lessen the danger of post-stroke infection from depression of the immune system after stroke.
 
MTT assay. Administration of proteasomal synthase inhibitor (PSI) induces PC12-cellular death. Co-administration of WIN55.212.2 protects PC12 cells from cell death. Note that there are statically significant decreases (*) in cytotoxicity in cells co-administered PSI and WIN55.212.2 (P<0.05; data represent mean and standard deviation).
WIN55.212.2 inhibits proteasomal synthase inhibitor (PSI)-induced activation of caspase-3. (A) Representative immunofluorescent micrographs of PC12 cells cultured in the presence of PSI and WIN55.212.2 and stained for activated caspase-3. Note the decreased staining in cultures where PSI and WIN55.212.2 were co-administered, while staining was increased in cultures treated with PSI alone. (B) Upper panel: representative Western blots for cleaved caspase-3 in lysates of PC12 cells cultured in the presence or absence of WIN55.212.2 and PSI (normalized for expression of extracellular signal-regulated kinase 2 [ERK2]). Lower panel: quantitation of cleaved caspase-3 expression in PC12 cells as described above. Note that there are statically significant changes(*) in cleaved caspase-3 expression in cultures treated with PSI alone and in those treated with both PSI and WIN55.212.2 (P<0.05; data represent mean and standard deviation).
WIN55.212.2 inhibits the proteasomal synthase inhibitor (PSI)-induced accumulation of α-synuclein and parkin. (A) Representative immunofluorescent micrographs of PC12 cells cultured in the presence of PSI and WIN55.212.2 and stained for α-synuclein and parkin (arrows). Note the decreased staining in cultures treated with both PSI and WIN55.212.2 relative to that in cultures treated with PSI alone. (B) Upper panel: representative Western blots for α-synuclein and parkin in lysates of PC12 cells cultured in the presence or absence of WIN55.212.2 and PSI. Lower panel: quantitation of α-synuclein and parkin expression in PC12 cells as described above (arrows). Note that there are statically significant changes in α-synuclein and parkin expression in cultures treated with PSI alone compared to those treated with both PSI and WIN55.212.2 (P<0.05; data represent mean and standard deviation). ERK2, extracellular signal-regulated kinase 2.
WIN55.212.2 increases intranuclear expression of nuclear factor κB (NF-κB). Upper panel: representative Western blots for NF-κB in nuclear lysates of PC12 cells cultured in the presence or absence of WIN55.212.2 and proteasomal synthase inhibitor (PSI). Lower panel: quantitation of nuclear NF-κB expression in PC12 cells as described above. Note that there are statistically significant changes in intranuclear NF-κB expression in cultures treated with PSI alone compared to those treated with both PSI and WIN55.212.2 (P<0.05; data represent mean and standard deviation). ERK2, extracellular signal-regulated kinase 2.
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Cannabinoids have been proposed to possess neuroprotective properties; though their mechanism of action remains contentious, they are posited to prevent neurodegenerative disorders, including Parkinson's disease, the pathogenesis of which has not been established. Recent studies have demonstrated that induction of proteasomal dysfunction in animal models results in a phenotype similar to Parkinson's disease. Here, we investigated the neuroprotective function of a synthetic cannabinoid-receptor agonist (WIN55.212.2) in dopaminergic neuronal death induced by a proteasomal synthase inhibitor (PSI), additionally testing the hypothesis that WIN55.212.2 modulates cytoplasmic accumulation of parkin and α-synuclein, a key feature of proteasomal dysfunction in Parkinson's. WIN55.212.2 protects PC12 cells from PSI-induced cytotoxicity, concomitantly inhibiting PSI-induced polyADP ribose polymerase expression and activation of caspase-3. While PSI induces cytoplasmic accumulation of α-synuclein and parkin, WIN55.212.2 counters these effects. Interestingly, however, while PSI induces the activation and nuclear translocalization of nuclear factor κB, WIN55.212.2 potentiates this effect. These data are suggestive that WIN55.212.2 might confer a neuroprotective benefit in PSI-induced proteasomal dysfunction, and could further protect against neuronal degeneration stemming from cytoplasmic accumulation of α-synuclein and parkin. These results indicate that WIN55.212.2 may be a candidate for treatment of neurodegenerative diseases, including Parkinson's disease.
 
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Using computerized tomographic data and three dimensional model, we studied the influence of tibial intramedullary canal axis and other morphologic factors of the tibia on the entry point for tibial intramedullary alignment guides. Various anatomical parameters including tibial anteroposterior dimensions (AP), mediolateral dimensions (ML), aspect ratio (ML/AP), bowing and the intramedullary canal axis were studied. In addition, the entry point for the intramedullary alignment guide for primary and revision total knee arthroplasty were studied. The averaged entry point at the level of the tibial plateau was 5.7±2.2 mm anterior and 4.3±2.0 mm lateral to the classical entry point (P<.001). Furthermore, this entry point was more anterolateral in females when compared to males (P<.001). At a depth 10 mm below the tibial plateau, the entry point was on average 8.8±1.9 mm anterior and 2.9±1.9 mm lateral to the center of the cut surface. With increasing tibial varus the entry point tended to shift laterally at both levels (r=0.49) (P<.001). In Korean, the entry point for tibial intramedullary alignment systems is anterolateral to the classically described entry point. Moreover, the increment of tibial varus necessitates more lateral placement of the entry point. Intraoperatively, the entry point can be localized during primary knee arthroplasty to a point 15.9±2.8 mm anterior to and 1.2±2.8 mm lateral to the lateral tibial spine. For revision knee arthroplasty the point is on average 8.8±1.9 mm anterior and 2.9±1.9 mm lateral to the center of the cut surface of the tibia at a depth of 10 mm from the articular surface.
 
Top-cited authors
Sanjib Ghosh
  • All India Institute of Medical Sciences, Patna
Hassan Azari
  • Barry University
Rouhollah Gazor
  • Guilan University of Medical Sciences
Dae Yong Song
  • Eulji University
Sun Yong Baek
  • Pusan National University