Trabecular structural difference between the superior and inferior regions of the vertebral body: a cadaveric and clinical study.

Background: Osteoporotic vertebral compression fractures commonly involve the superior vertebral body; however, their associated causes have not yet been clearly established. This study aimed to determine the trabecular structural differences between the superior and inferior regions of the vertebral body using cadaveric and clinical studies. Materials and methods: First, five vertebrae were collected from three human cadavers. The trabecular structures of the superior and inferior regions of each vertebral body were analyzed using micro-computed tomography (micro-CT), finite element analysis (FEA), and biomechanical test. Based on the results of the ex vivo study, we conducted a clinical study. Second, spine CT images were retrospectively collected. Bone volume and Hounsfield unit were analyzed for 192 vertebral bodies. Finally, after sample size calculation based on the pilot study, prospectively, 200 participants underwent dual-energy X-ray absorptiometry (DXA) of the lateral spine. The bone mineral densities (BMDs) of the superior and inferior regions of each lumbar vertebral body were measured. The paired t-test and Wilcoxon signed-rank test were used for the statistical analyses, and p-value < 0.05 was considered significant. Results: Cadaver studies revealed differences between the superior and inferior trabecular bone structures. The bone volume ratio, BMD, and various other trabecular parameters advocated for decreased strength of the superior region. Throughout the biomechanical study, the limitations of the compression force were 3.44 and 4.63 N/m2 for the superior and inferior regions, respectively. In the FEA study, the inferior region had a lower average displacement and higher von Mises stress than the superior region. In the clinical spine CT-based bone volume and BMD study, the bone volume was significantly higher in the inferior region than in the superior region. In the lateral spine DXA, the mean BMD of the superior region of vertebral bodies was significantly lower compared with that of the inferior region. Conclusion: The superior trabecular structure of the lumbar vertebral bodies possesses more biomechanical susceptibility compared with the inferior trabecular structure, confirming its dominant role in causing osteoporotic vertebral fractures. Physicians should also focus on the BMD values of the superior region of the vertebral body using lateral spine DXA to evaluate osteoporosis.

Association of Thymidylate Synthase (TS) Gene Polymorphisms with Incidence and Prognosis of Coronary Artery Disease.

Coronary artery disease (CAD) is a prevalent cardiovascular condition characterized by the accumulation of plaque within coronary arteries. While distinct features of CAD have been reported, the association between genetic factors and CAD in terms of biomarkers was insufficient. This study aimed to investigate the connection between genetic factors and CAD, focusing on the thymidylate synthase (TS) gene, a gene involved in DNA synthesis and one-carbon metabolism. TS plays a critical role in maintaining the deoxythymidine monophosphate (dTMP) pool, which is essential for DNA replication and repair. Therefore, our research targeted single nucleotide polymorphisms that could potentially impact TS gene expression and lead to dysfunction. Our findings strongly associate the TS 1100T>C and 1170A>G genotypes with CAD susceptibility. We observed that TS 1100T>C polymorphisms increased disease susceptibility in several groups, while the TS 1170A>G polymorphism displayed a decreasing trend for disease risk when interacting with clinical factors. Furthermore, our results demonstrate the potential contribution of the TS 1100/1170 haplotypes to disease susceptibility, indicating a synergistic interaction with clinical factors in disease occurrence. Based on these findings, we propose that polymorphisms in the TS gene had the possibility of clinically useful biomarkers for the prevention, prognosis, and management of CAD in the Korean population.

Effects of a small molecule R-spondin-1 substitute RS-246204 on a mouse intestinal organoid culture.

Organoids, a multi-cellular and organ-like structure cultured in vitro, can be used in a variety of fields such as disease modeling, drug discovery, or cell therapy development. When organoids derived from Lgr5 stem cells are cultured ex vivo, recombinant R-spondin-1 protein should be added at a high concentration for the initiation and maintenance of the organoids. Because the addition of large amounts of R-spondin-1 greatly increases the cost of organoids, the organoids grown with R-spondin-1 are not practical for large-scale drug screening and for the development of therapeutic agents. In this study, we tried to find a R-spondin-1 substitute compound that is able initiate small intestinal organoids without the use of the R-spondin-1 protein; thus, using organoid media that each included one compound from among an 8,364 compound library instead of R-spondin-1, we observed whether organoids were established from the crypts of the small intestine. As a result, we found one compound that could promote the initial formation and growth of enteroids in the medium without R-spondin-1 and named it RS-246204. The enteroids grown with RS-246204 had a similar differentiation capacity as well as self-renewal capacity as the enteroids grown with R-spondin-1. Furthermore, the RS-246204-derived enteroids could successfully produce the forskolin induced swelling and the organoid based epithelial to mesenchymal transition model. This compound could be used for developing a cost-efficient culturing method for intestinal organoids as well as for exploring Lgr5 signaling, intestinal stem cell physiology and therapeutics for GI tract diseases.

Interplay between polymorphisms in the endothelial nitric oxide synthase (eNOS) gene and metabolic syndrome in determining the risk of ischemic stroke in Koreans.

BACKGROUND: Endothelial nitric oxide synthase (eNOS) gene variants are known to play a role in atherosclerotic development. However, whether interplay between eNOS polymorphisms and metabolic syndrome (MetS) affects ischemic stroke (IS) risk has yet to be discovered. We investigated whether the combined effects of eNOS polymorphisms and MetS influence ischemic stroke risk in Koreans. METHODS: We genotyped the eNOS -922A>G, -786T>C, 4a4b, and 894G>T polymorphisms in 531 IS cases and 502 controls using polymerase chain reaction-amplified DNA. We then investigated whether the presence of MetS and the number of MetS risk factors worked with eNOS polymorphisms to influence IS risk. RESULTS: IS patients had a significantly higher prevalence of MetS than controls [adjusted odds ratio (AOR)=2.943, 95% confidence interval (CI), 2.256-3.840, P<0.0001], and MetS prevalence did not differ between stroke subtypes. The 894GT+TT genotypes were positively associated with IS (AOR=1.670, 95% CI, 1.208-2.308, P=0.002), and the -786TC+CC genotypes showed co-morbidity with MetS (AOR=1.448, 95% CI, 1.401-2.015, P=0.028). Among subjects with three or more MetS risk factors, the highest AOR value (28.490, 95% CI, 3.162-256.688) was observed for the eNOS -786TC+CC genotypes. CONCLUSIONS: The eNOS 894T allele and interplay between the eNOS -786C allele and MetS may predispose Koreans to IS.

Therapeutic effect of BDNF-overexpressing human neural stem cells (HB1.F3.BDNF) in a rodent model of middle cerebral artery occlusion.

Ischemic stroke mainly caused by middle cerebral artery occlusion (MCAo) represents the major type of stroke; however, there are still very limited therapeutic options for the stroke-damaged patients. In this study, we evaluated the neurogenic and therapeutic potentials of human neural stem cells (NSCs) overexpressing brain-derived neurotrophic factor (HB1.F3.BDNF) following transplantation into a rodent model of MCAo. F3.BDNF human NSCs (F3.BDNF) were transplanted into the contralateral side of striatum at 7 days after MCAo, and the transplanted animals were monitored up to 8 weeks using animal MRI and various behavioral tests before they were sacrificed for immunohistochemical analysis. Interestingly, animal MRI results indicate that the majority of contralaterally transplanted neural stem cells were migrated to the peri-infarct area, showing a pathotropism. Transplanted animals exhibited significant behavioral improvements in stepping, rotarod, and modified neurological severity score (mNSS) tests. We also found that the transplanted human cells were colocalized with nestin, DCX, MAP2, DARPP-32, TH, GAD65/67-positive cells, of which results can be correlated with neural regeneration and behavioral recovery in the transplanted animals. More importantly, we were able to detect high levels of human BDNF protein expression, presumably derived from the transplanted F3.BDNF. Taken together, these results provide strong evidence that human neural stem cells (F3.BDNF) are effective in treating stroke animal models.

Surface mapping of motor points in biceps brachii muscle.

OBJECTIVE: To localize the site of motor points within human biceps brachii muscles through surface mapping using electrophysiological method. METHOD: We recorded the compound muscle action potentials of each lattice of the biceps brachii in 40 healthy subjects. Standardized reference lines were made as the following: 1) a horizontal reference line (elbow crease) and 2) a vertical reference line connecting coracoid process and mid-point of the horizontal reference line. The Compound muscle action potentials were mapped in reference to the standardized reference lines. The locations of motor points were mapped to the skin surface, in the ratio to the length of the vertical and the half of the horizontal reference lines. RESULTS: The motor point of the short head of biceps was located at 69.0±4.9% distal and 19.1±9.5% medial to the mid-point of horizontal reference line. The location of the motor point of the long head of the biceps was 67.3±4.3% distal and 21.4±8.7% lateral. The motor point of the short head of the biceps was located more medially and distally in the male subjects compared to that in the female (p<0.05). CONCLUSION: This study showed electrophysiological motor points of the biceps brachii muscles through surface mapping. This data might improve the clinical efficacy and the feasibility of motor point targeting, when injecting botulinum neurotoxin in biceps brachii.

In vivo Tracking of Human Neural Stem Cells Following Transplantation into a Rodent Model of Ischemic Stroke.

BACKGROUND AND OBJECTIVES: Ischemic stroke caused by middle cerebral artery occlusion (MCAo) is the major type of stroke, but there are currently very limited options for cure. It has been shown that neural stem cells (NSCs) or neural precursor cells (NPCs) can survive and improve neurological deficits when they are engrafted in animal models of various neurological diseases. However, how the transplanted NSCs or NPCs are act in vivo in the injured or diseased brain is largely unknown. In this study, we utilized magnetic resonance imaging (MRI) techniques in order to understand the fates of human NSCs (HB1.F3) following transplantation into a rodent model of MCAo. METHODS AND RESULTS: HB1.F3 human NSCs were pre-labeled with ferumoxides (Feridex(®))-protamine sulfate complexes, which were visualized and examined by MRI up to 9 weeks after transplantation. Migration of the transplanted cells to the infarct area was further confirmed by histological methods. CONCLUSIONS: Based on these observations, we speculate that the transplanted NSCs have the extensive migratory ability to the injured site, which will in turn contribute to functional recovery in stroke.

In vivo tracking of human mesenchymal stem cells in experimental stroke.

To understand the fates of human mesenchymal stem cells (hMSCs) following transplantation into a rodent model of middle cerebral artery occlusion (MCAo), magnetic resonance imaging (MRI) techniques were employed, hMSCs were labeled with ferumoxides (Feridex)--protamine sulfate complexes, which were visualized and examined by MRI up to 10 weeks following transplantation. Migration of the transplanted cells to the infarcted area was further confirmed by histological methods. We found that the hMSCs transplanted in MCAo models possess the capacity to migrate to the infarcted area extensively in both ipsilateral and contralateral injections, exhibiting a pathotropism. We also analyzed the detailed migration patterns of transplanted hMSCs. We speculate that the extensive migratory ability of hMSCs may represent a therapeutic potential for developing efficient cell transplantation strategies in stroke.

Influence of combined methionine synthase (MTR 2756A > G) and methylenetetrahydrofolate reductase (MTHFR 677C > T) polymorphisms to plasma homocysteine levels in Korean patients with ischemic stroke.

PURPOSE: Methionine synthase (MTR) and 5,10-methylenetetrahydrofolate reductase (MTHFR) are the main regulatory enzymes for homocysteine metabolism. The present case- control study was conducted to determine whether there is an association between the MTR 2756A > G or MTHFR 677C > T polymorphism and plasma homocysteine concentration in Korean subjects with ischemic stroke. MATERIALS AND METHODS: DNA samples of 237 patients who had an ischemic stroke and 223 age and sex-matched controls were studied. MTR 2756A > G and MTHFR 677C > T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Frequencies of mutant alleles for MTR and MTHFR polymorphisms were not significantly different between the controls and cases. The patient group, however, had significantly higher homocysteine concentrations of the MTR 2756AA and MTHFR 677TT genotypes than the control group (p=0.04 for MTR, p=0.01 for MTHFR). The combined MTR 2756AA and MTHFR 677TT genotype (p= 0.04) and the homocysteine concentrations of the patient group were also higher than those of the controls. In addition, the genotype distribution was significant in the MTHFR 677TT genotype (p=0.008) and combined MTR 2756AA and MTHFR 677TT genotype (p=0.03), which divided the groups into the top 20% and bottom 20% based on their homocysteine levels. CONCLUSION: The results of the present study demonstrate that the MTR 2756A > G and MTHFR 677C > T polymorphisms interact with elevated total homocysteine (tHcy) levels, leading to an increased risk of ischemic stroke.

In Vivo Tracking of Human Mesenchymal Stem Cells in Experimental Stroke.

To understand the fates of human mesenchymal stem cells (hMSCs) following transplantation into a rodent model of middle cerebral artery occlusion (MCAo), magnetic resonance imaging (MRI) techniques were employed. hMSCs were labeled with ferumoxides (Feridex®)-protamine sulfate complexes, which were visualized and examined by MRI up to 10 weeks following transplantation. Migration of the transplanted cells to the infarcted area was further confirmed by histological methods. We found that the hMSCs transplanted in MCAo models possess the capacity to migrate to the infarcted area extensively in both ipsilateral and contralateral injections, exhibiting a pathotropism. We also analyzed the detailed migration patterns of transplanted hMSCs. We speculate that the extensive migratory ability of hMSCs may represent a therapeutic potential for developing efficient cell transplantation strategies in stroke.

Enhanced brain targeting efficiency of intranasally administered plasmid DNA: an alternative route for brain gene therapy.

Recently, nasal administration has been studied as a noninvasive route for delivery of plasmid DNA encoding therapeutic or antigenic genes. Here, we examined the brain targeting efficiency and transport pathways of intranasally administered plasmid DNA. Quantitative polymerase chain reaction (PCR) measurements of plasmid DNA in blood and brain tissues revealed that intranasally administered pCMVbeta (7.2 kb) and pN2/CMVbeta (14.1 kb) showed systemic absorption and brain distribution. Following intranasal administration, the beta-galactosidase protein encoded by these plasmids was significantly expressed in brain tissues. Kinetic studies showed that intranasally administered plasmid DNA reached the brain with a 2,595-fold higher efficiency than intravenously administered plasmid DNA did, 10 min post-dose. Over 1 h post-dose, the brain targeting efficiencies were consistently higher for intranasally administered plasmid DNA than for intravenously administered DNA. To examine how plasmid DNA enters the brain and moves to the various regions, we examined tissues from nine brain regions, at 5 and 10 min after intranasal or intravenous administration of plasmid DNA. Intravenously administered plasmid DNA displayed similar levels of plasmid DNA in the nine different regions, whereas, intranasally administered plasmid DNA exhibited different levels of distribution among the regions, with the highest plasmid DNA levels in the olfactory bulb. Moreover, plasmid DNA was mainly detected in the endothelial cells, but not in glial cells. Our results suggest that intranasally applied plasmid DNA may reach the brain through a direct route, possibly via the olfactory bulb, and that the nasal route might be an alternative method for efficiently delivering plasmid DNA to the brain.

Morphometry of the nasal bones and piriform apertures in Koreans.

In order to understand the morphological characteristics of Korean noses, the nasal bones and piriform apertures were measured and classified in the dried skulls of Korean adults. The shapes of the nasal bones were classified into five types (Type A-E). Among the types of the nasal bone, Type A, in which the nasomaxillary sutures initially descended vertically and then obliquely, was present in 43.2% and Type B, in which the nasomaxillary sutures were concave in the middle part, was present in 52.3% of the noses examined. The height of the nasal bone was 25.9 +/- 3.8 mm in males, 24.5 +/- 3.7 mm in females; the width was 9.2 +/- 2.4 mm in males, 8.8 +/- 2.6 mm in females. The height of the piriform aperture was 30.1 +/- 2.6 mm in males, 28.0 +/- 2.8 mm in females; the upper width was 16.8+2.6 mm in males, 17.0 +/- 2.0 mm in females; the lower width was 25.7 +/- 1.7 mm in males, 25.4 +/- 2.1 mm in females. Each measurement was compared with those carried out in other populations.

Skin permeation, biodistribution, and expression of topically applied plasmid DNA.

BACKGROUND: Topical application is emerging as a new route of gene delivery. However, the extent of skin permeation and the in vivo fate of topically applied plasmid DNA are not fully understood. METHODS: In vitro permeation of plasmid DNA across human skin and keratinocyte layers was tested using Franz diffusion cells. In vivo absorption and biodistribution of topically applied plasmid in mice were determined using quantitative polymerase chain reaction (PCR). The expression levels of plasmid DNA in various tissues were measured by semiquantitative reverse transcription PCR. RESULTS: In vitro, topically applied DNA was capable of penetrating human skin and keratinocyte layers. Following topical application of plasmid DNA onto murine skin, the levels of plasmid DNA in the serum peaked at 4 hr. At 24 hr post-dose, topically applied DNA existed at higher levels than intravenously administered DNA in almost all tissues, and induced 11.4- and 22-fold higher mRNA expression in muscle and skin, respectively. Moreover, the topical route showed sustained expression of plasmid DNA in the regional lymph nodes over 5 days, whereas the intravenous route did not. CONCLUSIONS: Taken together, our results show that topically applied plasmid DNA is capable of permeating the skin and being expressed for prolonged periods in various tissues including lymph nodes. This suggests that skin may provide an appealing, noninvasive route of delivery for DNA vaccines and other therapeutic genes.

Morphometrical changes of the human uterine tubes according to aging and menstrual cycle.

Studies on the morphological changes in the human uterine tube according to aging and menstrual cycle so far have been limited to microscopic aspects such as cellular changes, mainly due to the inaccessibility of specimens. In this study, postmortem analysis using both macroscopic and microscopic methods was performed using 55 human uterine tubes. The numbers and the degrees of convolution, and the length of the uterine tube had a tendency to decrease according to the increase of age in women by fifties. Under the influence of menopause, the total areas of the tube, mucosal layer and lumen in the ampulla, and lumen in the isthmus and infundibulum were shown to decrease on cross section. However, in the isthmus and infundibulum, the cross sectional area of the tube and mucosal layer did not show statistically significant changes. In the women at reproductive stages, the cross sectional areas of the tube, mucosa and lumen showed variations among different individuals, which may be due to the influence of menstrual cycle rather than the increase of age. No venous engorgement of the tubes was observed at the early proliferative, the mid-secretory and the postmenopausal stage. By contrast, full engorgement was observed at the early secretory stage and the menstrual stage.

Morphometry of nasal bases and nostrils in Koreans.

To investigate morphological characteristics of Korean noses, nasal bases and nostrils were measured and classified in Korean adults. The height and the width of the nasal base, the height and the width of the columella, the length of the long and short nostril axes, and nasal alar angle were measured. The shapes of the nostrils were classified into 7 types by the angle between the right and left long axes of the nostrils. Each measurement was compared with other studies.

Altered expression of the DNA repair protein, N-methylpurine-DNA glycosylase (MPG) in human gonads.

BACKGROUND: The multifunctional mammalian MPG is responsible for a damaged DNA base in the nucleus. The DNA repair enzyme is transported from the cytoplasm to nucleus to repair the DNA base when it is damaged. If the enzyme does not work properly, the damaged DNA may lead to carcinogenesis, cell death, aging or infertility. MATERIALS AND METHODS: This study was performed to determine mRNA expression and intracellular localization of the DNA repair protein, N-methylpurine-DNA glycosylase (MPG), in human ovary and testicular tissues, particularly in epithelial ovarian tumor and spermatogenic (maturation) arrest infertile patients, by RT-PCR and immunohistochemical staining using human MPG monoclonal antibody. RESULTS: MPG mRNA expression in epithelial ovarian tumor and spermatogenic arrest testis tissues was slightly higher than in normal ovarian and testicular tissues, respectively. The present study demonstrated new and unexpected patterns of cellular and subcellular localization of this enzyme. In a normal ovary, immunostaining for MPG was observed in the nucleus of oocyte, granulosa and stromal cells. MPG was stained mostly in the nucleus and faintly-stained in the cytoplasm of normal coelomic epithelium as well as in benign epithelial ovarian tumors. However, the MPG expression of the nucleus in malignant epithelial tumors, both serous and mucinous type, disappeared. The spermatocyte and Leydig cells in normal testis were immunostained only in the cytoplasm. The spermatocyte and Leydig cells in spermatogenic arrest testis tissues showed up both in the nucleus and cytoplasm. The subcellular localization of MPG in the tissues tested was heterogeneous, while the altered MPG expression was found in ovarian tumor and spermatogenic arrest testis. CONCLUSION: These results suggest MPG's role in human gonadal tissues and raise the possibility that the altered mRNA level and intracellular localization could be associated with ovarian tumorigenesis and male infertility.