OBJECTIVE: To investigate serum 25-hydroxyl vitamin D (25(OH)D) and vitamin D-binding protein (VDBP) concentrations in women with endometriosis according to the severity of disease. METHODS: Women with mild endometriosis (n = 9) and advanced endometriosis (n = 7), as well as healthy controls (n = 16), were enrolled in this observational study. Serum total 25(OH)D concentrations were analyzed using the Elecsys vitamin D total kit with the Cobas e602 module. Concentrations of bioavailable and free 25(OH)D were calculated. Concentrations of VDBP were measured using the Human Vitamin D BP Quantikine ELISA kit. Variables were tested for normality and homoscedasticity using the Shapiro-Wilk test and Leven F test, respectively. Correlation analysis was used to identify the variables related to total 25(OH)D and VDBP levels. To assess the effects of total 25(OH)D and VDBP levels in the three groups, multivariate generalized additive modeling (GAM) was performed. RESULTS: Gravidity and parity were significantly different across the three groups. Erythrocyte sedimentation rate (ESR) and CA-125 levels increased as a function of endometriosis severity, respectively (p= 0.051, p= 0.004). The correlation analysis showed that total 25(OH)D levels were positively correlated with gravidity (r = 0.59, p< 0.001) and parity (r = 0.51, p< 0.003). Multivariate GAM showed no significant relationship of total 25(OH)D levels with EMT severity after adjusting for gravidity and ESR. However, the coefficient of total 25(OH)D levels with gravidity was significant (1.87; 95% confidence interval, 0.12–3.63; p= 0.040). CONCLUSION: These results indicate that vitamin D and VDBP levels were not associated with the severity of endometriosis.
Blood Sedimentation ; Endometriosis ; Enzyme-Linked Immunosorbent Assay ; Female ; Gravidity ; Humans ; Observational Study ; Parity ; Vitamin D ; Vitamin D-Binding Protein ; Vitamins

PURPOSE: Vitamin D deficiency is reported to be more common in type 1 diabetes patients and might be associated with the increased urinary loss of vitamin D binding protein (VDBP) consequent to impaired 25-hydroxyvitamin D (25(OH)D) circulation. We aimed to evaluate the possible increased urinary loss of VDBP, a correlation between VDBP and circulating 25(OH)D level, and risk factors influencing low vitamin D level in pediatric type 1 diabetes patients without microalbuminuria. METHODS: This is a cross-sectional study of subjects who visited Seoul National University Children’s Hospital between January and March 2013. Forty-two type 1 diabetes patients and 29 healthy controls were included. Biochemical parameters including serum and urine VDBP concentrations were analyzed. RESULTS: There was no significant difference in the frequency of vitamin D deficiency or serum 25(OH)D level between the 2 groups. The serum and urine VDBP concentrations did not show any difference between the 2 groups. Serum 25(OH) D level did not correlate with serum or urine VDBP. Multivariate regression analysis revealed that daylight outdoor hours (β=2.948, P=0.003) and vitamin D intake (β=2.865, P=0.003) affected the 25(OH)D level; the presence of type 1 diabetes or urinary VDBP excretion was not significant. CONCLUSIONS: In pediatric type 1 diabetes patients, urinary VDBP excretion did not contribute to low serum 25(OH)D level in the setting of normoalbuminuria. The factors associated with 25(OH)D level during winter periods were daylight outdoor hours and vitamin D intake. Further studies including both micro- and macroalbuminuria patients with type 1 diabetes are warranted.
Albuminuria ; Child ; Cross-Sectional Studies ; Diabetes Mellitus, Type 1 ; Ergocalciferols ; Humans ; Risk Factors ; Seoul ; Vitamin D ; Vitamin D Deficiency ; Vitamin D-Binding Protein* ; Vitamins*

OBJECTIVES: We attempted to compartmentalize the periaqueductal gray (PAG) of the rabbit in terms of the different distribution patterns between NADPH-diaphorase (NADPHd)- and calbindin D28K (CB)-positive neurons. METHODS: Immunohistochemistry and immunofluorescent labelling for CB and histochemistry for NADPHd were carried out on coronally-sectioned midbrain slices of the rabbit. RESULTS: NADPHd-positive neurons were selectively localized in the dorsolateral (DL), the middle one-third of the lateral (L), the dorsal half of the ventrolateral (VLd) PAG, and the supraoculomotor cap nucleus (Su3C). Clusters of CB-immunoreactive perikarya marked the dorsal half of DL (DLd), Su3C, the ventral one-third of L, and the ventral half of the ventrolateral (VLv) PAG. Double labelling for NADPHd and CB revealed that two markers labelled different neuronal groups in DLd and Su3C subdivisions. CONCLUSION: The present data suggest that NADPHd and CB can be regarded as reliable neurochemical markers to reveal the longitudinally-columnar organization within the PAG and to subdivide each columnar area.
Calcium-Binding Protein, Vitamin D-Dependent ; Immunohistochemistry ; Mesencephalon ; Neurons ; Periaqueductal Gray

The hippocampus makes new memories and is involved in mental cognition, and the hippocampal dentate gyrus (DG) is critical because neurogenesis, which occurs throughout life, occurs in the DG. We observed the differentiation of neuroblasts into mature neurons (granule cells) in the DG of C57BL/6 mice at various early postnatal (P) ages: P1, P7, P14, and P21 using doublecortin (DCX) immunohistochemistry (IHC) for neuroblasts and calbindin D-28k (CB) IHC for granule cells. DCX-positive cells decreased in the DG with age; however, CB+ cells increased over time. At P1, DCX and CB double-labeled (DCX+CB+) cells were scattered throughout the DG. At P7, DCX+CB+ cells (about 92% of CB+ cells) were seen only in the granule cell layer (GCL) of the dorsal blade. At P14, DCX+CB+ cells (about 66% of CB+ cells) were found in the lower half of the GCL of both blades. In contrast, at P21, about 18% of CB+ cells were DCX+CB+ cells, and they were mainly located only in the subgranular zone of the DG. These results suggest that the developmental pattern of DCX+CB+ cells changes with time in the early postnatal stages.
Animals ; Calcium-Binding Protein, Vitamin D-Dependent ; Cognition ; Dentate Gyrus ; Hippocampus ; Immunohistochemistry ; Mice ; Neurogenesis ; Neurons

The distributions of calretinin (CR)- and parvalbumin (PV)-immunoreactive neurons in the main olfactory bulb (MOB) of the goat were examined in this study. As in other animals, the goat MOB has a characteristic laminar structure with laminar types and distribution patterns in each layer. CR-immunoreaction was observed in all layers of the MOB, except for the olfactory nerve layer. Most of CR-immunoreactive neurons were observed in the glomerular and granule cell layers. Relatively small number of CR-immunoreactive neurons was detected in other layers. These CR-immunoreactive neurons were interneurons. PV-immunoreaction was detected in all layers. In contrast to CR, olfactory nerve bundles were immunostained with PV. Most of PV-immunoreactive neurons were distributed in the glomerular and granule cell layers. PV-immunoreactive neurons were interneurons. This result suggests that CR and PV may play important roles in the olfactory signal modulation through interneurons in the goat MOB.
Animals ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Goats ; Immunohistochemistry ; Interneurons ; Neurons ; Olfactory Bulb ; Olfactory Nerve ; Smell

Calbindin D-28K (CALB) is one of the calcium-binding proteins which is assumed to be buffering, transport of Ca2+, and regulation of various enzyme systems. In the spinal cord, a subpopulation of calbindin-immunoreactive neurons located in the ventral portion of lamina VII, medial to the motoneuron column, has recently been proposed to be Renshaw cells (RCs), that mediate recurrent inhibition of spinal alpha-motoneurons, based on the anatomical location. In this study, we have performed to investigate the correlation between RCs containing high levels of CALB and motoneurons in the ventral horn of lumbar spinal cord of the ataxic pogo mice, that characterized by a failures of interlimb coordination, and prolonged excessive tone of hindlimb extensor muscles. We have shown that CALB immunoreactive RCs was significantly decreased in the ventral horn of lumbar spinal cord of the ataxic pogo mice (p.0.05), when compared with the control mice. Whereas, CALB immunoreactivity expression levels were no difference in the dorsal horn. Furthermore, CALB protein was significantly decreased in the lumbar spinal cord of the ataxic pogo mice (p.0.01). However, there were no difference in the cervical and thoracic spinal cord of the between control and pogo mice. These results suggest that motoneurons of ventral horn of the lumbar spinal cord might be more excited state, results in the decreased CALB immunoreactive RCs have not mediated a motoneuron excitability, in the atxic mice, pogo.
Animals ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Hindlimb ; Horns ; Mice ; Muscles ; Neurons ; Spinal Cord

Avian influenza (AI) is an infectious, low pathogenic virus that is endemic all over the world and poses a potential threat to the poultry industry. Vaccination is a widely used effective method to prevent avian influenza virus. Here we employed a comparative proteomics approach [two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight (MALDI-TOF)] to characterize proteome in the sera from the specific pathogen free (SPF) chickens, the vaccinated chickens, and the naturally infected chickens. We identified total 58 proteins that were differentially expressed in the sera of three groups. Among them ovotransferrin and vitamin D-binding protein were more expressed in the sera of naturally infected chickens compare with other groups. Our results suggested that the level of these two proteins in the serum may help to discriminate the naturally infected chicken from the vaccinated chicken.
Animals ; Chickens ; Conalbumin ; Electrophoresis ; Influenza in Birds ; Poultry ; Proteins ; Proteome ; Proteomics ; Specific Pathogen-Free Organisms ; Vaccination ; Viruses ; Vitamin D-Binding Protein

Alzheimer's disease (AD) is a neurodegenerative disease that proceeds with the age-dependent neuronal loss, an irreversible event which causes severe cognitive and psychiatric devastations. In the present study, we investigated whether the compound, AAD-2004 [2-hydroxy-5-[2-(4-trifluoromethylphenyl)-ethylaminobenzoic acid] which has anti-oxidant and anti-inflammatory properties, is beneficial for the brain of Tg-betaCTF99/B6 mice, a murine AD model that was recently developed to display age-dependent neuronal loss and neuritic atrophy in the brain. Administration of AAD-2004 in Tg-betaCTF99/B6 mice from 10 months to 18 months of age completely repressed the accumulation of lipid peroxidation in the brain. AAD-2004 markedly suppressed neuronal loss and neuritic atrophy, and partially reversed depleted expression of calbindin in the brain of Tg-beta-CTF99/B6. These results suggest that AAD-2004 affords neurodegeneration in the brain of AD mouse model.
Alzheimer Disease ; Animals ; Aspirin ; Atrophy ; Brain ; Calcium-Binding Protein, Vitamin D-Dependent ; Lipid Peroxidation ; Mice ; Neurodegenerative Diseases ; Neurons

BACKGROUND: It is important to differentiate between schwannomas and neurofibromas for the cases in which the histopathologic features overlap. Depending on the tumor type, surgeons can decide on a treatment method and whether to preserve or sacrifice the nerve; the possibility of malignant transformation in the case of neurofibromas also needs to be considered. METHODS: We studied 101 cases of schwannoma and 103 cases of neurofibroma. All the hematoxylin and eosin slides for these cases were reviewed, and tissue microarrays were prepared from the representative areas. Immunohistochemical analysis was performed using antibodies for S-100 protein, calretinin, CD56 and CD34. RESULTS: All the tumors except 3 neurofibromas were positive for the S-100 protein. Calretinin was found in 26.7% of the schwannomas (27/101), but it was not found in any of the neurofibromas. CD56 was positive in 77.2% of the schwannomas (78/101) and in 9.8% of the neurofibromas (10/102). CD34 was positive in 42.5% of the schwannomas (43/101) and in 80.2% of the neurofibromas (81/101). Statistically, calretinin was significantly specific for schwannomas (p<0.001) and CD56 was also sensitive for these tumors (p<0.001). On the other hand, a CD34 expression seemed highly sensitive (p<0.001) for neurofibromas. CONCLUSIONS: We concluded that combined immunohistochemical analysis for calretinin, CD56, and CD34 may be very useful for differentiating schwannomas from neurofibromas.
Antibodies ; Calcium-Binding Protein, Vitamin D-Dependent ; Diagnosis, Differential ; Eosine Yellowish-(YS) ; Hand ; Hematoxylin ; Immunohistochemistry ; Neurilemmoma ; Neurofibroma ; S100 Proteins

Cerebellar Purkinje cells (PCs) play a crucial role in motor functions and their progressive degeneration is closely associated with spinocerebellar ataxias. Although immunohistochemical (IHC) analysis can provide a valuable tool for understanding the pathophysiology of PC disorders, the method validation of IHC analysis with cerebellar tissue specimens is unclear. Here we present an optimized and validated IHC method using antibodies to calbindin D28k, a specific PC marker in the cerebellum. To achieve the desired sensitivity, specificity, and reproducibility, we modified IHC analysis procedures for cerebellar tissues. We found that the sensitivity of staining varies depending on the commercial source of primary antibody. In addition, we showed that a biotin-free signal amplification method using a horseradish peroxidase polymer-conjugated secondary antibody increases both the sensitivity and specificity of ICH analysis. Furthermore, we demonstrated that dye filtration using a 0.22 micrometer filter eliminates or minimizes nonspecific staining while preserving the analytical sensitivity. These results suggest that our protocol can be adapted for future investigations aiming to understand the pathophysiology of cerebellar PC disorders and to evaluate the efficacy of therapeutic strategies for treating these diseases.
Antibodies ; Calcium-Binding Protein, Vitamin D-Dependent ; Cerebellum ; Filtration ; Horseradish Peroxidase ; Purkinje Cells ; Sensitivity and Specificity ; Spinocerebellar Ataxias