Association of Tarlov cyst with cauda equina syndrome and spinal cord infarction following caudal epidural block: A case report.

RATIONALE: Caudal epidural block (CEB), which injects drugs into the epidural space through a sacral hiatus, is considered a safer alternative to other approaches. Serious complications, such as cauda equina syndrome or spinal cord infarction, have been reported very rarely, but their coexistence after CEB, which may be related to the ruptured perineural cyst, also known as a Tarlov cyst, was not reported. PATIENT CONCERNS: A 40-year-old male patient presented with bilateral lower extremity radicular pain. CEB was performed without image guidance. The patient exhibited sensory deficits below L2, no motor function (0-grade), hypotonic deep tendon reflexes, and no pathological reflexes. DIAGNOSES: Spinal cord infarction, cauda equina syndrome, and sacral level perineural cyst with hemorrhage. INTERVENTION: High doses of steroids and rehabilitation were performed. OUTCOMES: The patient was discharged after 28 days with persistent bilateral leg paralysis and sensory deficits below the L2 level. The patient demonstrated no neurological improvement. LESSONS: Magnetic resonance imaging, including the sacral area, should be performed before performing CEB, to confirm the presence of a perineural cyst.

Feasibility and Effectiveness of a Motion Tracking-Based Online Fitness Program for Office Workers.

The development of technology-based home fitness has emerged from the booming digital healthcare market and recent demands for at-home fitness and health equipment due to the COVID-19 pandemic. Digital healthcare company Alyce Healthcare recently developed Weelo, which is a web-based online fitness program. Weelo recommends an exercise protocol through machine-learning-enabled recognition of the user's motion and provides visual and auditory feedback. We evaluated whether Weelo improves physical and mental well-being to assess its capabilities and effectiveness. Thirty-two participants performed a total of 20 exercise sessions following the Weelo guide on a laptop. The participants were evaluated using a before and after exercise program, body composition, handgrip strength, six-minute walk test, modified star excursion balance test, short form 36, fatigue severity scale, Beck depression index, and a satisfaction survey. Overall, there was a significant improvement in muscle strength, endurance, and balance ability, as well as an improved quality of life and significant reduction in fatigue and depression. Participants showed high motivation to continue following the Weelo exercise program. In conclusion, utilizing Weelo improved physical and mental well-being and is considered to be an individual-use indoor exercise program that serves as an alternative to traditional face-to-face exercise.

Concurrent Index-to-Little Finger Dorsal Dislocations of the Carpometacarpal Joints with Carpal Bone Fractures.

A 32-year-old man presented with simultaneous dorsal dislocations of the index-to-little finger carpometacarpal (CMC) joint with carpal bone fractures. Closed reduction was unsuccessful even after general anesthesia. During open dorsal approach, we found interposed joint capsule in the CMC joints and after removal of the joint capsule open reduction was easily achieved. We placed four Kirschner wires through the CMC joint. Furthermore, the fractured dorsal fragments of the trapezoid and hamate were fixed with mini screw in each. During 1-year followup, the patient showed good recovery and no evidence of posttraumatic arthritic changes in plain X-ray. We recommend to fix the dorsal fragment of the carpal bone with screws as well as the transarticular fixation of the CMC joint in case of concurrent CMC joint fracture-dislocation of all four fingers.

In vivo transplantation of 3D encapsulated ovarian constructs in rats corrects abnormalities of ovarian failure.

Safe clinical hormone replacement (HR) will likely become increasingly important in the growing populations of aged women and cancer patients undergoing treatments that ablate the ovaries. Cell-based HRT (cHRT) is an alternative approach that may allow certain physiological outcomes to be achieved with lower circulating hormone levels than pharmacological means due to participation of cells in the hypothalamus-pituitary-ovary feedback control loop. Here we describe the in vivo performance of 3D bioengineered ovarian constructs that recapitulate native cell-cell interactions between ovarian granulosa and theca cells as an approach to cHRT. The constructs are fabricated using either Ca++ or Sr++ to crosslink alginate. Following implantation in ovariectomized (ovx) rats, the Sr++-cross-linked constructs achieve stable secretion of hormones during 90 days of study. Further, we show these constructs with isogeneic cells to be effective in ameliorating adverse effects of hormone deficiency, including bone health, uterine health, and body composition in this rat model.

The effect of collagen hydrogel on 3D culture of ovarian follicles.

The in vivo function and phenotype of ovarian follicle cells are determined by many factors. When these cells are removed from the in vivo microenvironment and grown in a 2D in vitro environment, the function of the follicular cells is difficult to preserve. A collagen hydrogel was used to examine the hormone and oocyte maturation of ovary follicles in a 3D culture system. Ovarian follicles from rats were isolated and cultured in various concentration of type I collagen hydrogels ranging from 1% to 7% (weight/volume). Differences in cell survival, follicle growth and development, sex hormone production, and oocyte maturation were seen with the modifications in the collagen hydrogel density and elasticity. The results show the significance of the collagen hydrogel properties on phenotype and function maintenance of the ovarian follicles in a 3D culture system.

Syngeneic Myoblast Transplantation Improves Muscle Function in a Murine Model of X-Linked Myotubular Myopathy.

X-linked myotubular myopathy (XLMTM) is an isogenic muscle disease characterized by progressive wasting of skeletal muscle, weakness, and premature death of affected male offspring. Recently, the XLMTM gene knock-in mouse, Mtm1 p.R69C, was found to have a similar phenotype as the Mtm1 gene mutation in humans (e.g., central nucleation of small myofibers, attenuated muscle strength, and motor unit potentials). Using this rodent model, we investigated whether syngeneic cell therapy could mitigate muscle weakness. Donor skeletal muscle-derived myoblasts were isolated from C57BL6 wild-type (WT) and Mtm1 p.R69C (KI) mice for transplantation into the gastrocnemius muscle of recipient KI mice. Initial experiments demonstrated that donor skeletal muscle-derived myoblasts from WT and KI mice remained in the gastrocnemius muscle of the recipient KI mouse for up to 4 weeks posttransplantation. KI mice receiving syngeneic skeletal muscle-derived myoblasts displayed an increase in skeletal muscle mass, augmented force generation, and increased nerve-evoked skeletal muscle action potential amplitude. Taken together, these results support our hypothesis that syngeneic cell therapy may potentially be used to ameliorate muscle weakness and delay the progression of XLMTM, as application expands to other muscles.

Myogenic-induced mesenchymal stem cells are capable of modulating the immune response by regulatory T cells.

Cell therapy for patients who have intractable muscle disorders may require highly regenerative cells from young, healthy allogeneic donors. Mesenchymal stem cells are currently under clinical investigation because they are known to induce muscle regeneration and believed to be immune privileged, thus making them suitable for allogeneic applications. However, it is unclear whether allogeneic and myogenic-induced mesenchymal stem cells retain their immunomodulatory characteristics. Therefore, our aim was to evaluate the effects of mesenchymal stem cell differentiation on the immune characteristics of cells in vitro. We investigated the immunologic properties of mesenchymal stem cells after myogenic induction. Mesenchymal stem cells were obtained from C57BL/6 mice and the C3H/10T1/2 murine mesenchymal stem cell line. Two different 5-aza-2'-deoxycytidine doses (0.5 and 3 µM) were evaluated for their effects on mesenchymal stem cell skeletal myogenic differentiation potential, immune antigen expression, and mixed lymphocytic reactions. Using a mixed lymphocytic reaction, we determined the optimal splenocyte proliferation inhibition dose. The induction of regulatory T cells was markedly increased by the addition of 3 µM 5-aza-2'-deoxycytidine-treated mesenchymal stem cells. Myogenic-induced mesenchymal stem cells do not elicit alloreactive lymphocyte proliferative responses and are able to modulate immune responses. These findings support the hypothesis that myogenic-induced mesenchymal stem cells may be transplantable across allogeneic barriers.

Simple and versatile molecular method of copy-number measurement using cloned competitors.

Variations and alterations of copy numbers (CNVs and CNAs) carry disease susceptibility and drug responsiveness implications. Although there are many molecular methods to measure copy numbers, sensitivity, reproducibility, cost, and time issues remain. In the present study, we were able to solve those problems utilizing our modified real competitive PCR method with cloned competitors (mrcPCR). First, the mrcPCR for ERBB2 copy number was established, and the results were comparable to current standard methods but with a shorter assay time and a lower cost. Second, the mrcPCR assays for 24 drug-target genes were established, and the results in a panel of NCI-60 cells were comparable to those from real-time PCR and microarray. Third, the mrcPCR results for FCGR3A and the FCGR3B CNVs were comparable to those by the paralog ratio test (PRT), but without PRT's limitations. These results suggest that mrcPCR is comparable to the currently available standard or the most sensitive methods. In addition, mrcPCR would be invaluable for measurement of CNVs in genes with variants of similar structures, because combination of the other methods is not necessary, along with its other advantages such as short assay time, small sample amount requirement, and applicability to all sequences and genes.

Engineered multilayer ovarian tissue that secretes sex steroids and peptide hormones in response to gonadotropins.

Although hormone replacement therapy is an option for the loss of ovarian function, hormone delivery through pharmacological means results in various clinical complications. The present study was designed to deliver sex steroids by a functional construct fabricated using encapsulation techniques. Theca and granulosa cells isolated from ovaries of 21-day old rats were encapsulated in multilayer alginate microcapsules to recapitulate the native follicular structure. Cells encapsulated in two other schemes were used as controls to assess the importance of the multilayer structure. The endocrine functions of the encapsulated cells were assessed in vitro for a period of 30 days. Encapsulated cells showed sustained viability during long-term in vitro culture with those encapsulated in multilayer capsules secreting significantly higher and sustained concentrations of 17 ß-estradiol (E(2)) than the two other encapsulation schemes (p < 0.05, n = 6) in response to follicle-stimulating hormone (FSH) and luteinizing hormone (LH). In addition, cells in the multilayer microcapsules also secreted activin and inhibin in vitro. In contrast, when granulosa and theca cells were cultured in 2D culture, progesterone (P(4)) secretion increased while E(2) secretion decreased over a 30-day period. In summary, we have designed a multilayer engineered ovarian tissue that secretes sex steroids and peptide hormones and responds to gonadotropins, thus demonstrating the ability to recapitulate native ovarian structure ex vivo.

Abnormalities in the uninvolved foot in children with spastic hemiplegia.

BACKGROUND: Although the uninvolved foot in patients with hemiplegia has been thought to be normal, we frequently observed valgus deformity of the uninvolved foot among those patients. The purpose of this study was to evaluate by dynamic pedobarograph the prevalence and pattern of foot deformity in the uninvolved limb among children with hemiplegia. METHODS: In this study, we included 119 patients with hemiplegia (67 males and 52 females) who underwent gait analysis from 2001 to 2008. The mean age at evaluation was 9.2 ± 3.4 years (range, 5.1 to 19.8 y). Patient demographics, passive range of motion, kinematics, kinetics, and dynamic pedobarographic data were obtained from the medical records. Coronal index [(CI): the impulse percentage under the medial column minus the impulse percentage under the lateral column] was calculated from the pedobarographic data. RESULTS: Of 119 feet, 60 feet (50.4%) had a normal CI, whereas 52 feet (43.7%) showed a valgus CI. Only 7 feet (5.9%) had a varus CI. Compared with the patients with a normal CI, patients who had a valgus CI had increased ankle dorsiflexion at initial contact (-0.9 ± 4.1 vs. 0.8 ± 4.7, P=0.048) and knee extension moment (0.6 ± 0.31 vs. 0.73 ± 0.28, P=0.036) of the uninvolved foot. Actual limb-length discrepancy did not differ between the 2 groups (P=0.556). CONCLUSIONS: Valgus foot deformity of the uninvolved foot is common among children with hemiplegia. It is associated with increased ankle dorsiflexion and knee extension moments of the uninvolved side. Longer follow-up will be needed to evaluate the effect of the valgus foot deformity of the uninvolved limb on the gait in patients with hemiplegia. LEVEL OF EVIDENCE: Level III. Diagnostic.

Amniotic fluid-derived stem cells in regenerative medicine research.

The stem cells isolated from amniotic fluid present an exciting possible contribution to the field of regenerative medicine and amniotic fluid-derived stem (AFS) cells have significant potential for research and therapeutic applications. AFS cells are multipotent, showing the ability to differentiate into cell types from all three embryonic germ layers. They express both embryonic and adult stem cell markers, expand extensively without feeder cells, double in 36 h, and are not tumorigenic. The AFS cells can be maintained for over 250 population doublings and preserve their telomere length and a normal karyotype. They differentiate easily into specific cell lineages and do not require human embryo tissue for their isolation, thus avoiding the current controversies associated with the use of human embryonic stem (ES) cells. The discovery of the AFS cells has been recent, and a great deal of work remains to be performed on the characterization and use of these cells. This review describes the various differentiated lineages that AFS cells can form and the future of these promising new stem cells in regenerative medicine research.

Prevalence and patterns of scoliosis in children with multiple pterygium syndrome.

BACKGROUND: Although scoliosis is a predominant feature of multiple pterygium syndrome (MPS), the pattern of deformity and the progression of the curvature have not been well described. The purpose of this study was to assess the prevalence of scoliosis among patients with MPS, and to characterize the abnormalities of the vertebrae and to assess the progression of the scoliosis. METHODS: From 1969 to 2008, we identified 19 patients with MPS but only 16 patients (8 boys and 8 girls) had complete data. Medical records and radiographs of these 16 patients were reviewed. Magnetic resonance imaging was performed in 8 patients to evaluate intraspinal anomalies. Functional mobility score was used to assess the ambulatory ability. RESULTS: Of 16 patients, 13 patients (81.3%) had scoliosis. The mean age when the scoliosis was first noticed was 3.3±2.6 years (range, 1 mo to 8.2 y). The mean Cobb angle at first visit was 37.4±18.1 degrees (range, 14.0 to 75.0 degrees). With a mean follow-up of 4.0±4.9 years, the Cobb angle at the last visit was 43.3±19.1 degrees (range, 20.0 to 72.0 degrees). Congenital scoliosis was observed in 7 patients (3 unilateral unsegmented bar, 3 fusion of the cervical spine, 1 block vertebrae), whereas neuromuscular scoliosis was observed in 1 patient. A common radiographic finding was narrowing of the intervertebral disc space with decreased height of vertebrae in the thoracic area. Intraspinal anomalies were seen in 4 patients (3 tethered spinal cords, 1 syrinx). At the last follow-up, 5 of 13 patients who had scoliosis were able to walk at school without assistance (Functional mobility scale-500 ≥5). CONCLUSIONS: Scoliosis is common among children with MPS. It is frequently accompanied by fusion of the cervical area. Intraspinal anomalies such as tethered cord syndrome and syringomyelia are common associated anomalies. Therefore, it is important to look for intraspinal anomalies. Closed monitoring of the patient's ambulatory ability and bowel and bladder continence is also needed. LEVEL OF EVIDENCE: Level IV, Diagnostic Study.

Transplantation of bone marrow cells reduces CCl4 -induced liver fibrosis in mice.

BACKGROUND: We investigated the reversibility of liver fibrosis induced with a CCl(4) injection and the role of stem cells in reversing the hepatic injury. Furthermore, the most effective cell fraction among bone marrow cells (BMCs) in the repair process was analysed. METHODS: C57BL/6 mice were divided into four groups after 5 weeks of injection of CCl(4) : control, sacrificed after 5 weeks, sacrificed at 10 weeks and sacrificed 5 weeks later after GFP-donor BM transplantation. Liver function tests and real-time polymerase chain reaction (PCR) of markers indicating liver fibrosis were compared between the groups. To identify the most effective BMC fraction that repairs liver injury, the mice were divided into three groups after the injection of CCl(4) for 2 days: granulocyte colony stimulating factor (G-CSF) only, mononuclear cell (MNC) transplantation and Lin-Sca-1+c-kit+haematopoietic stem cell (HSC) transplantation. Eight days after transplantation, the mice were harvested and morphometric, immunohistochemical analyses were performed to compare the expression of extracellular matrix and liver fibrosis-related factors. RESULTS: The liver fibrosis induced by CCl(4) was not spontaneously recovered but was persistent until 10 weeks, but the group injected with BMCs had less fibrosis and better liver function. Mobilization with G-CSF increased the recovery of the injured liver and the best results were seen in those mice administered the MNC fraction and Lin-Sca-1+c-kit+HSC fraction, with no difference between the two groups. CONCLUSION: BMC transplantation and stem cell mobilization with G-CSF effectively treats liver injury in mice. These are promising techniques for autologous transplantation in humans with liver fibrosis.

Bioimaging for the monitoring of the in vivo distribution of infused mesenchymal stem cells in a mouse model of the graft-versus-host reaction.

Cell therapy using MSCs (mesenchymal stem cells) might be effective treatment for refractory GVHD (graft-versus-host disease). However, the fate and distribution of MSCs after transplantation remains unclear. In this study, an animal model was developed to monitor the dynamic distribution of MSCs in mice with GVHD. A GVHD mouse model was established by transplanting C57BL/6 donor bone marrow cells and C57BL/6 EGFP (enhanced green fluorescent protein) splenocytes into lethally irradiated BALB/c nude recipient mice. Donor MSCs were obtained from MHC-identical C57BL/6 RFP (red fluorescent protein) mice and infused into the recipient mice on the same transplantation day. In vivo movement of the donor splenocytes (EGFP) and MSCs (RFP) were evaluated by measuring the biofluorescence (IVIS-Xenogen system). Donor splenocytes and MSCs reached the lungs first, and then the gastrointestinal tract, lymph nodes and skin, in that order; the transit time and localization site of these cells were very similar. In the recipient mouse with GVHD, the number of detectable cells declined with time, as assessed by biofluorescence imaging and confirmed by RT (real-time)-PCR. This bioimaging system might be useful for preclinical testing and the design of therapeutic strategies for monitoring the dynamic distribution of MSCs with GVHD.

Osteoclast activation by receptor activator of NF-kappaB ligand enhances the mobilization of hematopoietic progenitor cells from the bone marrow in acute injury.

Osteoclasts (OCLs) are multinucleated cells that are derived from the monocyte/macrophage hematopoietic lineage in response to receptor activator of NF-kappaB ligand (RANKL) activation. They are specialized cells responsible for physiological bone resorption and as well as pathologic bone loss. In addition to their unique ability to resorb bone, OCLs also play a potential role in the mobilization of hematopoietic progenitor cells from the bone marrow (BM), particularly under various stress stimuli (e.g. hypoxia, injury or inflammation). We investigated the effect of activated OCLs on the stem cell niche and whether this leads to mobilization of hematopoietic progenitors. We induced activated OCLs from the RAW264.7 cell line through stimulation with RANKL and we quantified the levels of the stem cell niche component SDF-1 on the osteblasts and CXCR4 on the bone marrow cells (BMCs) by culturing with supernatants from activated OCLs. In addition, we exposed mice to stress by inducing liver injury with CCl4 followed by injecting RANKL to activate OCLs and compared the effect on the mobilization of hematopoietic progenitor cells from the BM. We found that functional OCLs cleaved SDF-1alpha in the osteoblasts and increased CXCR4 expression in the BMCs. Moreover, under stress in vivo, mobilized hematopoietic progenitor cells were significantly increased after RANKL treatment. These results suggest that OCLs might be involved in alteration of the interaction between SDF-1 and CXCR4 leading to mobilization of hematopoietic progenitor cells from the BM.

Mesenchymal stromal cells inhibit graft-versus-host disease of mice in a dose-dependent manner.

BACKGROUND AIMS: Graft-versus-host disease (GvHD) remains a major complication after allogeneic hematopoietic cell transplantation (HCT). Recent literature demonstrates a potential benefit of human mesenchymal stromal cells (MSC) for the treatment of refractory GvHD; however, the optimal dose remains uncertain. We set out to develop an animal model that can be used to study the effect of MSC on GvHD. METHODS: A GvHD mouse model was established by transplanting C3H/he donor bone marrow (BM) cells and spleen cells into lethally irradiated BALB/c recipient mice. MSC were obtained from C3H/he mice and the C3H/10T1/2 murine MSC line. RESULTS: The mRNA expression of Foxp3 in regional lymph nodes (LN) localized with T cells was markedly increased by the addition of C3H10T1/2 cells in a real-time polymerase chain reaction (PCR). Using a mixed lymphocyte reaction, we determined the optimal splenocyte proliferation inhibition dose (MSC:splenocyte ratios 1:2 and 1:1). Three different C3H10T1/2 cell doses (low, 0.5 x 10(6), intermediate, 1 x 10(6), and high, 2 x 10(6)) with a consistent splenocyte dose (1 x 10(6)) were evaluated for their therapeutic potential in an in vivo GvHD model. The clinical and histologic GvHD score and Kaplan-Meier survival rate were improved after MSC transplantation, and these results demonstrated a dose-dependent inhibition. CONCLUSIONS: We conclude that MSC inhibit GvHD in a dose-dependent manner in this mouse model and this model can be used to study the effects of MSC on GvHD.

Prediction of prognosis in children with medulloblastoma by using immunohistochemical analysis and tissue microarray.

OBJECTIVES: Medulloblastoma is the most common malignant neuroepithelial tumor found in children. Several reports have described efforts to identify the prognostic significance of various patterns of pathological and immunohistochemical features in medulloblastoma, but the published data appear to be controversial. The authors therefore attempted to demonstrate these prognostic factors convincingly in a retrospective study performed in patients with medulloblastoma. METHODS: The data used were obtained in 58 patients with medulloblastoma who were > 3 years of age and in whom > 1 year of follow-up was available after the maximal resection, craniospinal irradiation, and chemotherapy treatments. These assessments were performed to compare the immunohistochemical features to cellular differentiation, the proliferation index (PI), the apoptotic index (AI), and oncogenesis revealed by TrkC and c-erbB-3. In addition, the authors tried to determine the prognostic utility of these results in this tumor category. RESULTS: There was no statistically significant correlation between the prognosis and the degree of cell differentiation, but a positive correlation was noted between the PI and the AI in a tumor mass. The number of cases with a PI > 10% was significantly greater in the group of tumors in patients with recurrent medulloblastoma. A close association between the PI as a continuous variable and the progression-free and overall survival was also found. Most importantly, the PI is the only significant prognostic factor for the overall survival of patients with medulloblastoma. CONCLUSIONS: Therefore, the authors suggest that the PI is directly linked to the prognostic factor for medulloblastoma and that immunohistochemical staining is a potentially powerful tool for predicting the prognosis of patients with medulloblastoma.