Nervous system exposure of different classes of nanoparticles: A review on potential toxicity and mechanistic studies.

Nanoparticles (NPs) are generally defined as very small particles in the size range of 1-100 nm. Due to the rapid development of modern society, many new materials have been developed. The widespread use of NPs in medical applications, the food industry and the textile industry has led to an increase in NPs in the environment and the possibility of human contact, which poses a serious threat to human health. The nervous system plays a leading role in maintaining the integrity and unity of the body and maintaining a harmonious balance with the external environment. Therefore, based on two categories of organic and inorganic NPs, this paper systematically summarizes the toxic effects and mechanisms of NPs released into the nervous system. The results showed that exposure to NPs may damage the nervous system, decrease learning and cognitive ability, and affect embryonic development. Finally, a remediation scheme for NPs entering the body via the environment is also introduced. This scheme aims to reduce the neurotoxicity caused by NPs by supplementing NPs with a combination of antioxidant and anti-inflammatory compounds. The results provide a valuable reference for future research in this field.

Causal relationship between bone mineral density and intervertebral disc degeneration: a univariate and multivariable mendelian randomization study.

BACKGROUND: Although previous studies have suggested a possible association between bone mineral density (BMD) and intervertebral disc degeneration (IDD), the causal relationship between them remains unclear. Evidence from accumulating studies indicates that they might mutually influence one another. However, observational studies may be affected by potential confounders. Meanwhile, Mendelian randomization (MR) study can overcome these confounders to assess causality. OBJECTIVES: This Mendelian randomization (MR) study aimed to explore the causal effect of bone mineral density (BMD) on intervertebral disc degeneration (IDD). METHODS: Summary data from genome-wide association studies of bone mineral density (BMD) and IDD (the FinnGen biobank) have been acquired. The inverse variance weighted (IVW) method was utilized as the primary MR analysis approach. Weighted median, MR-Egger regression, weighted mode, and simple mode were used as supplements. The Mendelian randomization pleiotropy residual sum and outlier (MR-PRESSO) and MR-Egger regression were performed to assess horizontal pleiotropy. Cochran's Q test evaluated heterogeneity. Leave-one-out sensitivity analysis was further conducted to determine the reliability of the causal relationship. Multivariate MR (MVMR) analyses used multivariable inverse variance-weighted methods to individually and jointly adjust for four potential confounders, body mass index (BMI), Type2 diabetes, hyperthyroidism and smoking. A reverse MR analysis was conducted to assess potential reverse causation. RESULTS: In the univariate MR analysis, femoral neck bone mineral density (FNBMD), heel bone mineral density (eBMD), lumbar spine bone mineral density (LSBMD), and total body bone mineral density (TB BMD) had a direct causal effect on intervertebral disc degeneration (IDD) [FNBMD-related analysis: OR(95%CI) = 1.17 (1.04 to 1.31), p = 0.008, eBMD-related analysis: OR(95%CI) = 1.06 (1.01 to 1.12), p = 0.028, LSBMD-related analysis: OR(95%CI) = 1.20 (1.10 to 1.31), p = 3.38E-7,TB BMD-related analysis: OR(95%CI) = 1.20 (1.12 to 1.29), p = 1.0E-8]. In the MVMR analysis, it was revealed that, even after controlling for confounding factors, heel bone mineral density (eBMD), lumbar spine bone mineral density (LSBMD), and total body bone mineral density (TB BMD) still maintained an independent and significant causal association with IDD(Adjusting for heel bone mineral density: beta = 0.073, OR95% CI = 1.08(1.02 to 1.14), P = 0.013; Adjusting for lumbar spine bone mineral density: beta = 0.11, OR(95%CI) = 1.12(1.02 to 1.23), P = 0.03; Adjusting for total body bone mineral density: beta = 0.139, OR95% CI = 1.15(1.06 to 1.24), P = 5.53E - 5). In the reverse analysis, no evidence was found to suggest that IDD has an impact on BMD. CONCLUSIONS: The findings from our univariate and multivariable Mendelian randomization analysis establish a substantial positive causal association between BMD and IDD, indicating that higher bone mineral density may be a significant risk factor for intervertebral disc degeneration. Notably, no causal effect of IDD on these four measures of bone mineral density was observed. Further research is required to elucidate the underlying mechanisms governing this causal relationship.

A comparative study on the clinical efficacy of percutaneous endoscopic lumbar discectomy and conventional open surgery in the treatment of lumbar disc herniation.

Objective: To analyze the efficacy of single-channel percutaneous endoscopic lumbar discectomy (PELD) and conventional open surgery in the treatment of lumbar disc herniation (LDH). Methods: This is a retrospective study. A total of 66 patients with LDH admitted to Tianjin Medical University from June 2017 to June 2018 were divided into two groups: the observation group (single-channel PELD) and the control group (posterior lumbar interbody fusion), with 33 cases in each group. The two groups were compared in terms of visual analogue scale(VAS), oswestry disability index (ODI), Japanese Orthopaedic Association Score(JOA), perioperative indicators, clinical efficacy, postoperative complications, changes in inflammatory factors and serum T lymphocyte subsets. Results: The operation time, incision length, intraoperative blood loss, time in bed, hospital stay in the observation group were all lower than those in the control group. At 7d after treatment, the improvement of ODI, VAS and JOA in the observation group were better than that in the control group. At the last follow-up, there was no significant difference in Cobb angle and lumbar lordosis angle between the two groups. The levels of serum IL-1, IL-6 and TNF-α in the observation group were lower than those in the control group. The degree of reduction of serum CD3+ and CD4+ in the observation group were higher than those in the control group. And the level of elevation of CD8+ in the observation group was lower than that in the control group. Moreover, there was no significant difference in CD4+/CD8+ level between the two groups. The excellent rate of surgical results in the observation group was higher than that in the control group. Complications occurred in both groups, with no significant difference between the two groups. Conclusions: Single-channel PELD can achieve superior clinical efficacy over conventional open surgery in the treatment of LDH.

Application of self-anchored lateral lumbar interbody fusion in lumbar degenerative diseases.

STUDY DESIGN: This is a retrospective study. OBJECTIVE: The aim of the study was to evaluate the efficacy of self-anchored lateral lumbar interbody fusion (SA-LLIF) in lumbar degenerative diseases. METHODS: Forty-eight patients with lumbar degenerative disease between January 2019 and June 2020 were enrolled in this study. All patients complained of low back and leg pain, which were aggravated during standing activities and alleviated or disappeared during lying. After general anesthesia, the patient was placed in the right decubitus position. The anterior edge of the psoas major muscle was exposed through an oblique incision of approximately 6 cm, using an extraperitoneal approach. The psoas major muscle was then properly retracted dorsally to expose the disc. After discectomy, a suitable cage filled with autogenous bone graft from the ilium was implanted. Two anchoring plates were inserted separately into the caudal and cranial vertebral bodies to lock the cage. Clinical efficacy was evaluated using the visual analog scale (VAS) and Oswestry Disability Index (ODI). Lumbar lordosis, intervertebral disc height, spondylolisthesis rate, cage subsidence and fusion rate were also recorded. RESULTS: A total of 48 patients were enrolled in this study, including 20 males and 28 females, aged 61.4 ± 7.3 (range 49-78) years old. Surgery was successfully performed in all patients. Lumbar stenosis and instability were observed in 22 cases, disc degenerative disease in eight cases, degenerative spondylolisthesis in nine cases, degenerative scoliosis in six cases, and postoperative revision in three cases. In addition, five patients were diagnosed with osteoporosis. The index levels included L2-3 in three patients, L3-4 in 13 patients, L4-5 in 23 patients, L2-4 in three patients, and L3-5 in six patients. The operation time was 81.1 ± 6.4 (range 65-102) min. Intraoperative blood loss was 39.9 ± 8.5 (range 15-72) mL. No severe complications occurred, such as nerve or blood vessel injuries. The patients were followed up for 11.7 ± 2.3 (range 4-18) months. At the last follow-up, the VAS decreased from 6.2 ± 2.3 to 1.7 ± 1.1, and the ODI decreased from 48.4% ± 11.2% to 10.9% ± 5.5%. Radiography showed satisfactory postoperative spine alignment. No cage displacement was found, but cage subsidence 2-3 mm was found in five patients without obvious symptoms, except transient low back pain in an obese patient. The lumbar lordosis recovered from 36.8° ± 7.9° to 47.7° ± 6.8°, and intervertebral disc height recovered from 8.2 ± 2.0 mm to 11.4 ± 2.5 mm. The spondylolisthesis rate decreased from 19.9% ± 4.9% to 9.4% ± 3.2%. The difference between preoperative and last follow-up was statistically significant (P<0.05). CONCLUSION: SA-LLIF can provide immediate stability and good results for lumbar degenerative diseases with a standalone anchored cage without posterior internal fixation.

Effects of the cage height and positioning on clinical and radiographic outcome of lateral lumbar interbody fusion: a retrospective study.

BACKGROUND: The proper cage positioning and height in lateral lumbar interbody fusion (LLIF). This study evaluated their effects on clinical and radiographic outcome measures in patients undergoing LLIF. METHODS: This single-center retrospective study analyzed the characteristics and perioperative data of patients who underwent LLIF between January 2019 and December 2020. Radiographic (lumbar lordosis [LL], foraminal height, disc height [DH], segmental angle [SA], cross-sectional area [CSA] of thecal sac) and clinical (Oswestry Disability Index and Visual Analog Scale) outcomes were assessed preoperatively, postoperatively, and at the last follow-up. The effects of cage height and positioning on these parameters were also investigated. RESULTS: With a mean follow-up of 12.8 months, 47 patients with 70 operated level were analyzed. Data demonstrated that postsurgical clinical and radiographic outcome measures were significantly better than before surgery(P < 0.05). Cage height and positioning showed no significant difference with regarding to clinical outcome(P > 0.05). Subgroup analysis of the cage positioning showed that DH and SA were better restored by the final follow-up in patients with anteriorly placed cages than those with posteriorly placed cages (P < 0.05). Cages of posterior position showed significantly upgrading cage subsidence (P = 0.047). Cage height subgroup analysis showed that the preoperative forminal height, DH, and SA in the 11-mm cage group were significantly lower than in the 13-mm cage group; however, these parameters were comparable in the two groups postoperatively and at the final follow-up (P > 0.05). Furthermore, the postoperative and final follow-up degrees of DH, SA, and LL have improved in the 11-mm cage group more than the 13-mm cage group. The preoperative, postoperative, and final follow-up LL values in the 11-mm cage group were lower than in the 13-mm cage group(P < 0.01). CONCLUSIONS: Cage height and positioning did not affect the clinical outcomes in the present study. Cages in anterior position showed better restoration in DH, SA and decreased the incidence of cage subsidence. A comparable radiographic outcome can be achieved by inserting an appropriate cage height based on preoperative radiography.

Targeted fully endoscopic visualized laminar trepanning approach under local anaesthesia for resection of highly migrated lumbar disc herniation.

PURPOSE: To introduce a new fully endoscopic visualized laminar trepanning approach with a periendoscopic trephine under local anesthesia for resection of highly migrated lumbar disc herniation (LDH) and report the clinical outcomes of one year follow-up. METHODS: Twenty-one patients with highly migrated LDH who underwent percutaneous endoscopic lumbar discectomy via the laminar trepanning approach from June 2019 to August 2020 were retrospectively reviewed. Patient-Reported Outcomes Measurement Information System (PROMIS) Short Forms-Pain Interference (PI) and Physical Function (PF) were selected as outcome measures. The operating duration and complication were documented. RESULTS: The average age of the 21 patients (15 males, 6 females) was 37.8 ± 6.0 years (29-52 years). Disc migration originated from L4/5 in 19 patients, L5/S1 in two patients. The mean operative duration was 54.1 ± 9.0 minutes (42-79 min). All patients were followed up to 12 months after the operation. PROMIS PI T-scores decreased significantly from pre-operatively mean 68.6 ± 2.4 to 54.4 ± 1.9 (P < 0.001) and 47.1 ± 4.3 (P < 0.001) at six weeks and 12 months, respectively. PROMIS PF T-scores improved significantly from pre-operatively mean 26.7 ± 4.7 to 44.3 ± 4.2(P < 0.001) and 58.4 ± 4.0 (P < 0.001) at six weeks and 12 months, respectively. No complications and disc herniation recurrences occurred. CONCLUSION: The targeted full endoscopic laminar trepanning under local anesthesia with a visualized periendoscopic trephine offers a safe, efficient and cost-effective option for the resection of highly migrated LDH.

Defects of cohesin loader lead to bone dysplasia associated with transcriptional disturbance.

Cohesin loader nipped-B-like protein (Nipbl) is increasingly recognized for its important role in development and cancer. Cornelia de Lange Syndrome (CdLS), mostly caused by heterozygous mutations of Nipbl, is an autosomal dominant disease characterized by multiorgan malformations. However, the regulatory role and underlying mechanism of Nipbl in skeletal development remain largely elusive. In this study, we constructed a Nipbl-a Cas9-knockout (KO) zebrafish, which displayed severe retardation of global growth and skeletal development. Deficiency of Nipbl remarkably compromised cell growth and survival, and osteogenic differentiation of mammalian osteoblast precursors. Furthermore, Nipbl depletion impaired the cell cycle process, and caused DNA damage accumulation and cellular senescence. In addition, nucleolar fibrillarin expression, global rRNA biogenesis, and protein translation were defective in the Nipbl-depleted osteoblast precursors. Interestingly, an integrated stress response inhibitor (ISRIB), partially rescued Nipbl depletion-induced cellular defects in proliferation and apoptosis, osteogenesis, and nucleolar function. Simultaneously, we performed transcriptome analysis of Nipbl deficiency on human neural crest cells and mouse embryonic fibroblasts in combination with Nipbl ChIP-Seq. We found that Nipbl deficiency caused thousands of differentially expressed genes including some important genes in bone and cartilage development. In conclusion, Nipbl deficiency compromised skeleton development through impairing osteoblast precursor cell proliferation and survival, and osteogenic differentiation, and also disturbing the expression of some osteogenesis-regulatory genes. Our study elucidated that Nipbl played a pivotal role in skeleton development, and supported the fact that treatment of ISRIB may provide an early intervention strategy to alleviate the bone dysplasia of CdLS.

Comparative radiological outcomes and complications of sacral-2-alar iliac screw versus iliac screw for sacropelvic fixation.

PURPOSE: To compare the outcomes of sacropelvic fixation (SPF) using sacral-2-alar iliac (S2AI) screw with SPF using iliac screw (IS). METHODS: A comprehensive search of PubMed, EMBASE, Cochrane Central Register of Controlled Trials and Scopus was performed for comparative studies between S2AI and IS for SPF. Two independent investigators selected qualified studies and extracted data indispensably. With 95% confidence intervals (CI), the odds ratio (OR) was applied to dichotomous outcomes and standardized mean difference (SMD) was applied to continuous outcomes for each item. RESULTS: We included data from thirteen studies involving 722 patients (S2AI, 357 patients; IS, 365 patients). In the pediatric population, the S2AI group had a smaller pelvic obliquity (PO) than the IS group at final follow-up (SMD, - 0.38; 95% CI, - 0.72 to - 0.04). Patients who underwent S2AI screws showed reduced rates of re-operation (S2AI, 13%; IS, 28%), implant failure (S2AI, 12%; IS, 26%) [screw loosening (S2AI, 8%; IS, 20%); screw breakage (S2AI, 2%; IS, 12%)], implant prominence (S2AI, 2%; IS, 14%), pseudarthrosis (S2AI, 3%; IS, 15%), wound infection (S2AI, 8%; IS, 22%) and less blood loss (S2AI, 2035.4 ml; IS, 2708.4 ml). CONCLUSION: Radiological outcomes indicate an effective maintenance of the correction and arrest of progression of deformity by S2AI, which is equal or better than IS. SPF with S2AI screw has obviously lower incidence of postoperative complications and less blood loss. Given these advantages, the S2AI screw seems to be a beneficial alternative to IS.

Treatment of simple bone cysts of the humerus by intramedullary nailing and steroid injection.

BACKGROUND: Simple bone cysts (SBCs) are common benign lytic bone lesions in children. This study focused on exploring a clinical treatment method, minimally invasive intramedullary decompression and drainage with elastic stable intramedullary nailing (ESIN) combined with intralesional injections of steroids, and evaluated its effectiveness, complications and morbidity through functional and radiographic outcomes. METHODS: The postoperative recovery of 18 children who suffered from SBCs of humerus was evaluated (mean follow-up, 40 months) from January 2009 to December 2016. These patients (11 males, 7 females; 8 in the left, 10 in the right; mean age, 10.9 years old) were treated with minimally invasive intramedullary decompression and drainage with ESIN combined with intralesional injections of steroids. The diagnosis was based on not only pre-operative typical medical images (X-rays/CT/MRI) but also surgical findings and pathological diagnosis. Radiological and functional outcomes were evaluated according to Capanna and Musculoskeletal Tumor Society (MSTS) score. The interclass differences were analyzed by t-test. RESULTS: According to Capanna and MSTS criteria, after treatment, 14 patients made full recoveries which was presented by all the cysts filled with bone tissue, and 4 patients made partially recoveries, which were presented by cystic spaces partially filled with low density bone. All the cysts responded to treatment method, and there was no cyst recurrence. All except 2 patients had good functional results. One of the two patients had irritation of the end of the nail and one patient had a valgus deformity. CONCLUSIONS: Treatment for SBCs of humerus by minimally invasive intramedullary decompression and drainage with ESIN combined with intralesional injections of steroids is safe, effective and convenient. The clinical effect is satisfactory and worth popularizing.

Ribosomal DNA copy number loss and sequence variation in cancer.

Ribosomal DNA is one of the most variable regions in the human genome with respect to copy number. Despite the importance of rDNA for cellular function, we know virtually nothing about what governs its copy number, stability, and sequence in the mammalian genome due to challenges associated with mapping and analysis. We applied computational and droplet digital PCR approaches to measure rDNA copy number in normal and cancer states in human and mouse genomes. We find that copy number and sequence can change in cancer genomes. Counterintuitively, human cancer genomes show a loss of copies, accompanied by global copy number co-variation. The sequence can also be more variable in the cancer genome. Cancer genomes with lower copies have mutational evidence of mTOR hyperactivity. The PTEN phosphatase is a tumor suppressor that is critical for genome stability and a negative regulator of the mTOR kinase pathway. Surprisingly, but consistent with the human cancer genomes, hematopoietic cancer stem cells from a Pten-/- mouse model for leukemia have lower rDNA copy number than normal tissue, despite increased proliferation, rRNA production, and protein synthesis. Loss of copies occurs early and is associated with hypersensitivity to DNA damage. Therefore, copy loss is a recurrent feature in cancers associated with mTOR activation. Ribosomal DNA copy number may be a simple and useful indicator of whether a cancer will be sensitive to DNA damaging treatments.

Biomimetic angle-ply multi-lamellar scaffold for annulus fibrosus tissue engineering.

Constructing a biomimetic scaffold that replicates the complex architecture of intervertebral disc annulus fibrosus (AF) remains a major goal in AF tissue engineering. In this study, a biomimetic angle-ply multi-lamellar polycaprolactone/silk fibroin (PCL/SF) AF scaffold was fabricated. Wet-spinning was used to obtain aligned PCL/SF microfiber sheets, and these were excised into strips with microfibers aligned at +30° or -30° relative to the strip long axis. This was followed by stacking two strips with opposing fiber alignment and wrapping them concentrically around a mandrel. Our results demonstrated that the scaffold possessed spatial structure and mechanical properties comparable to natural AF. The scaffold supported rabbit AF cells adhesion, proliferation, infiltration and guided oriented growth and extracellular matrix deposition. In conclusion, our angle-ply multi-lamellar scaffold offers a potential solution for AF replacement therapy and warrants further attention in future investigations.

L-leucine partially rescues translational and developmental defects associated with zebrafish models of Cornelia de Lange syndrome.

Cohesinopathies are human genetic disorders that include Cornelia de Lange syndrome (CdLS) and Roberts syndrome (RBS) and are characterized by defects in limb and craniofacial development as well as mental retardation. The developmental phenotypes of CdLS and other cohesinopathies suggest that mutations in the structure and regulation of the cohesin complex during embryogenesis interfere with gene regulation. In a previous project, we showed that RBS was associated with highly fragmented nucleoli and defects in both ribosome biogenesis and protein translation. l-leucine stimulation of the mTOR pathway partially rescued translation in human RBS cells and development in zebrafish models of RBS. In this study, we investigate protein translation in zebrafish models of CdLS. Our results show that phosphorylation of RPS6 as well as 4E-binding protein 1 (4EBP1) was reduced in nipbla/b, rad21 and smc3-morphant embryos, a pattern indicating reduced translation. Moreover, protein biosynthesis and rRNA production were decreased in the cohesin morphant embryo cells. l-leucine partly rescued protein synthesis and rRNA production in the cohesin morphants and partially restored phosphorylation of RPS6 and 4EBP1. Concomitantly, l-leucine treatment partially improved cohesinopathy embryo development including the formation of craniofacial cartilage. Interestingly, we observed that alpha-ketoisocaproate (α-KIC), which is a keto derivative of leucine, also partially rescued the development of rad21 and nipbla/b morphants by boosting mTOR-dependent translation. In summary, our results suggest that cohesinopathies are caused in part by defective protein synthesis, and stimulation of the mTOR pathway through l-leucine or its metabolite α-KIC can partially rescue development in zebrafish models for CdLS.

Efficacy and safety of tranexamic acid in reducing blood loss in scoliosis surgery: a systematic review and meta-analysis.

OBJECTIVE: The purpose of this systematic review and meta-analysis of randomized controlled trials (RCTs) and non-RCTs was to gather data to evaluate the efficacy and safety of tranexamic acid (TXA) versus placebo after a scoliosis surgery. METHODS: The electronic databases including Embase, PubMed, CENTRAL (Cochrane Controlled Trials Register), Web of Science, and Google database were searched to identify relevant studies published from the time of the establishment of these databases up to May 2016. This systematic review and meta-analysis was performed according to the PRISMA statement criteria. The primary outcomes were total blood loss, intraoperative blood loss, and hemoglobin after surgery. The second outcome is need for transfusion. Stata 12.0 software was used for the meta-analysis. After testing for publication bias and heterogeneity across studies, data were aggregated for random-effects modeling when necessary. RESULTS: A total of 685 patients (347 patients in the TXA group and 338 in the control group) were finally included for this meta-analysis. The pooled results revealed that administration of TXA can decrease the total blood loss after scoliosis surgery [mean difference (MD) = 682.30, 95% confidence interval (CI) -930.60 to -434.00; P = 0.000] and intraoperative blood loss [(MD) = -535.28; 95% CI -683.74 to -368.82; P = 0.000]. For the hemoglobin (Hb) value after scoliosis surgery, TXA can decrease the Hb value for 0.51 dL [(MD) = 0.51; 95% CI 0.25-0.78; P = 0.000]. There is no statistically significant difference between the TXA versus placebo in terms of the need for transfusion (relative risk = 0.55, 95% CI 0.25-1.20, P = 0.132). CONCLUSION: Based on the current meta-analysis, TXA can decrease the total blood loss and intraoperative blood loss during scoliosis surgery. It is recommended that it be routinely used in scoliosis surgery. High-dose TXA (>20 mg/kg) is more effective than low-dose TXA (<20 mg/kg) in controlling blood loss. However, for the need for transfusion, more high-quality RCTs need to be identified.

A model of acute central cervical spinal cord injury syndrome combined with chronic injury in goats.

PURPOSE: To develop a large animal model for acute central cervical spinal cord injury syndrome (ACCSCIS. METHODS: Twenty-four adult male goats were randomized into four groups including group A with acute compression injury, group B with anterior chronic compression, group C as the test group that received anterior chronic compression by screw and acute compression by posterior balloon insertion, and group D as normal controls that received sham surgery. Neurological function (modified Tarlov motor function), CT, MRI, cortical somatosensory evoked potentials (CSEP), and pathological analysis were evaluated. The data were analyzed statistically. RESULTS: The motor function of the goats in group C was significantly lower than other groups. CSEP before spinal cord compression showed a stable pattern. Spinal cord compression resulted in a gradual decrement in the peak latency and significant increment in the peak amplitude. Cervical spinal canal occupying ratio was significantly lower in group C than the other groups. MRI revealed focal low signal in T1 weighted images and focal high signal in T2 weighted images in group C. Pathological analysis showed more severe lesions in the gray matter than that in the white matter in group C. CONCLUSIONS: The model well simulated the pathogenesis and resembled the clinical characteristics of ACCSCIS. This model seems to have the potential to contribute to the development of effective therapies for ACCSCIS.

Radiographic measurement for transforaminal percutaneous endoscopic approach (PELD).

OBJECTIVE: A radiographic study to analyze the working zone and relationship of the nerve root to their corresponding intervertebral disc for transforaminal percutaneous approaches. METHODS: 100 MRIs of transverse and sagittal views of 37 males, 63 females (average age 45 years), 50 MRIs of coronal views of 22 males, 28 females (average age 42 years), and 100 X-rays, 46 males, 54 females (average age of 44 years) were used for image analysis. All radiologic measurements were obtained independently by two experienced radiologists. On sagittal plane, foraminal height, foraminal diameter, nerve root-disc distance and nerve root-pedicle distance were measured. On transverse plane, foraminal width, nerve root-disc distance, nerve root-facet distance and target angle (J°) were analyzed at the superior (s) and inferior (i) margin of the disc. On coronal plane, nerve root-disc distance and nerve root-pedicle distance were measured at the medial, middle and lateral borders of the pedicle. RESULTS: Sagittal plane; foraminal height and diameter decreased caudally. Transverse plane; foraminal width was larger at the superior margin of the disc. Nerve root-disc distance decreased caudally. The nerve root lied dorsal to the disc at L2-L3 and L3-L4, whereas at L4-L5 and L5-S1 it lied ventrally. Nerve root-facet distance was shortest at the superior margin. Target angles (Js°, Ji°) at L2-L3 and L3-L4 were wider at their superior margin than at their inferior margin. Coronal plane; nerve root-disc distance increased from L2-L3 to L5-S1 whereas nerve root-pedicle distances decreased, thus coursing more vertically. CONCLUSIONS: At lower lumbar levels the exiting nerve root is at risks of injury. Hence, it is advised to enlarge the foramen for safe passage of endoscopic instruments and to minimize the possibility of nerve injury.

Improved transcription and translation with L-leucine stimulation of mTORC1 in Roberts syndrome.

BACKGROUND: Roberts syndrome (RBS) is a human developmental disorder caused by mutations in the cohesin acetyltransferase ESCO2. We previously reported that mTORC1 signaling was depressed and overall translation was reduced in RBS cells and zebrafish models for RBS. Treatment of RBS cells and zebrafish RBS models with L-leucine partially rescued mTOR function and protein synthesis, correlating with increased cell division and improved development. RESULTS: In this study, we use RBS cells to model mTORC1 repression and analyze transcription and translation with ribosome profiling to determine gene-level effects of L-leucine. L-leucine treatment partially rescued translational efficiency of ribosomal subunits, translation initiation factors, snoRNA production, and mitochondrial function in RBS cells, consistent with these processes being mTORC1 controlled. In contrast, other genes are differentially expressed independent of L-leucine treatment, including imprinted genes such as H19 and GTL2, miRNAs regulated by GTL2, HOX genes, and genes in nucleolar associated domains. CONCLUSIONS: Our study distinguishes between gene expression changes in RBS cells that are TOR dependent and those that are independent. Some of the TOR independent gene expression changes likely reflect the architectural role of cohesin in chromatin looping and gene expression. This study reveals the dramatic rescue effects of L-leucine stimulation of mTORC1 in RBS cells and supports that normal gene expression and translation requires ESCO2 function.

Stimulation of mTORC1 with L-leucine rescues defects associated with Roberts syndrome.

Roberts syndrome (RBS) is a human disease characterized by defects in limb and craniofacial development and growth and mental retardation. RBS is caused by mutations in ESCO2, a gene which encodes an acetyltransferase for the cohesin complex. While the essential role of the cohesin complex in chromosome segregation has been well characterized, it plays additional roles in DNA damage repair, chromosome condensation, and gene expression. The developmental phenotypes of Roberts syndrome and other cohesinopathies suggest that gene expression is impaired during embryogenesis. It was previously reported that ribosomal RNA production and protein translation were impaired in immortalized RBS cells. It was speculated that cohesin binding at the rDNA was important for nucleolar form and function. We have explored the hypothesis that reduced ribosome function contributes to RBS in zebrafish models and human cells. Two key pathways that sense cellular stress are the p53 and mTOR pathways. We report that mTOR signaling is inhibited in human RBS cells based on the reduced phosphorylation of the downstream effectors S6K1, S6 and 4EBP1, and this correlates with p53 activation. Nucleoli, the sites of ribosome production, are highly fragmented in RBS cells. We tested the effect of inhibiting p53 or stimulating mTOR in RBS cells. The rescue provided by mTOR activation was more significant, with activation rescuing both cell division and cell death. To study this cohesinopathy in a whole animal model we used ESCO2-mutant and morphant zebrafish embryos, which have developmental defects mimicking RBS. Consistent with RBS patient cells, the ESCO2 mutant embryos show p53 activation and inhibition of the TOR pathway. Stimulation of the TOR pathway with L-leucine rescued many developmental defects of ESCO2-mutant embryos. Our data support the idea that RBS can be attributed in part to defects in ribosome biogenesis, and stimulation of the TOR pathway has therapeutic potential.

α-Synuclein disrupts stress signaling by inhibiting polo-like kinase Cdc5/Plk2.

Parkinson disease (PD) results from the slow, progressive loss of dopaminergic neurons in the substantia nigra. Alterations in α-synuclein (aSyn), such as mutations or multiplications of the gene, are thought to trigger this degeneration. Here, we show that aSyn disrupts mitogen-activated protein kinase (MAPK)-controlled stress signaling in yeast and human cells, which results in inefficient cell protective responses and cell death. aSyn is a substrate of the yeast (and human) polo-like kinase Cdc5 (Plk2), and elevated levels of aSyn prevent Cdc5 from maintaining a normal level of GTP-bound Rho1, which is an essential GTPase that regulates stress signaling. The nine N-terminal amino acids of aSyn are essential for the interaction with polo-like kinases. The results support a unique mechanism of PD pathology.

Cohesin proteins promote ribosomal RNA production and protein translation in yeast and human cells.

Cohesin is a protein complex known for its essential role in chromosome segregation. However, cohesin and associated factors have additional functions in transcription, DNA damage repair, and chromosome condensation. The human cohesinopathy diseases are thought to stem not from defects in chromosome segregation but from gene expression. The role of cohesin in gene expression is not well understood. We used budding yeast strains bearing mutations analogous to the human cohesinopathy disease alleles under control of their native promoter to study gene expression. These mutations do not significantly affect chromosome segregation. Transcriptional profiling reveals that many targets of the transcriptional activator Gcn4 are induced in the eco1-W216G mutant background. The upregulation of Gcn4 was observed in many cohesin mutants, and this observation suggested protein translation was reduced. We demonstrate that the cohesinopathy mutations eco1-W216G and smc1-Q843Δ are associated with defects in ribosome biogenesis and a reduction in the actively translating fraction of ribosomes, eiF2α-phosphorylation, and (35)S-methionine incorporation, all of which indicate a deficit in protein translation. Metabolic labeling shows that the eco1-W216G and smc1-Q843Δ mutants produce less ribosomal RNA, which is expected to constrain ribosome biogenesis. Further analysis shows that the production of rRNA from an individual repeat is reduced while copy number remains unchanged. Similar defects in rRNA production and protein translation are observed in a human Roberts syndrome cell line. In addition, cohesion is defective specifically at the rDNA locus in the eco1-W216G mutant, as has been previously reported for Roberts syndrome. Collectively, our data suggest that cohesin proteins normally facilitate production of ribosomal RNA and protein translation, and this is one way they can influence gene expression. Reduced translational capacity could contribute to the human cohesinopathies.

Dihydroartemisinin inhibits the mammalian target of rapamycin-mediated signaling pathways in tumor cells.

Dihydroartemisinin (DHA), an antimalarial drug, has previously unrecognized anticancer activity, and is in clinical trials as a new anticancer agent for skin, lung, colon and breast cancer treatment. However, the anticancer mechanism is not well understood. Here, we show that DHA inhibited proliferation and induced apoptosis in rhabdomyosarcoma (Rh30 and RD) cells, and concurrently inhibited the signaling pathways mediated by the mammalian target of rapamycin (mTOR), a central controller for cell proliferation and survival, at concentrations (<3 µM) that are pharmacologically achievable. Of interest, in contrast to the effects of conventional mTOR inhibitors (rapalogs), DHA potently inhibited mTORC1-mediated phosphorylation of p70 S6 kinase 1 and eukaryotic initiation factor 4E binding protein 1 but did not obviously affect mTORC2-mediated phosphorylation of Akt. The results suggest that DHA may represent a novel class of mTORC1 inhibitor and may execute its anticancer activity primarily by blocking mTORC1-mediated signaling pathways in the tumor cells.