Sphingomyelin (SM) is a major sphingolipid in mammalian cells that forms specific lipid domains in combination with cholesterol (Chol). Using molecular-dynamics simulation and density functional ...theory calculation, we identified a characteristic Raman band of SM at ∼1643 cm−1 as amide I of the SM cluster. Experimental results indicate that this band is sensitive to the hydration of SM and the presence of Chol. We showed that this amide I Raman band can be utilized to examine the membrane distribution of SM. Similarly to SM, ceramide phosphoethanolamine (CerPE) exhibited an amide I Raman band in almost the same region, although CerPE lacks three methyl groups in the phosphocholine moiety of SM. In contrast to SM, the amide I band of CerPE was not affected by Chol, suggesting the importance of the methyl groups of SM in the SM-Chol interaction.
This experimental study was performed using human ligamentum flavum-derived cells (HFCs).
To investigate the intracellular signaling mechanism of interleukin-6 (IL-6) secretion in transforming growth ...factor-β (TGF- β)-stimulated HFCs.
Lumbar spinal stenosis (LSS) is a prevalent disease among the elderly, characterized by debilitating pain in the lower extremities. Although the number of patients with LSS has increased in recent years, the underlying pathomechanism remains unclear. Clinical examinations typically rely on magnetic resonance imaging to diagnose patients, revealing ligamentum flavum hypertrophy. Some studies have suggested an association between ligamentum flavum hypertrophy and inflammation/fibrosis, and expression of TGF-β and IL-6 has been observed in surgically obtained ligamentum flavum samples. However, direct evidence linking TGF-β and IL-6 expression in HFCs is lacking.
HFCs were obtained from patients with LSS who had undergone decompression surgery. The cells were stimulated with TGF-β and pretreated with either the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580 or the p44/42 MAP kinase inhibitor FR180204. IL-6 secretion in the cell culture medium and IL-6 messenger RNA (mRNA) expression levels were analyzed using an enzyme-linked immunoassay and real-time polymerase chain reaction, respectively.
TGF-β administration resulted in a dose- and time-dependent stimulation of IL-6 release. Treatment with SB203580 and FR180204 markedly suppressed TGF-β-induced IL-6 secretion from HFCs. Moreover, these inhibitors suppressed IL-6 mRNA expression in response to TGF-β stimulation.
Our findings indicate that TGF-β induces IL-6 protein secretion and gene expression in HFCs through the activation of p38 or p44/42 MAP kinases. These results suggest a potential association between IL-6-mediated inflammatory response and tissue hypertrophy in LSS, and we provide insights into molecular targets for therapeutic interventions targeting LSS-related inflammation through our analysis of the MAP kinase pathway using HFCs.
The smiley-face rod method has been reported to be a successful technique for reducing slippage and repairing pars defects in lumbar spondylolisthesis. However, we encountered a patient who developed ...right L5 radiculopathy with muscle weakness after use of the smiley-face rod method. The patient was a 19-year-old female judo player who had undergone direct repair surgery using the smiley-face rod method for terminal-stage lumbar spondylolysis. Postoperatively, she developed paresthesia on the lateral side of the right thigh with weakness of the right tibialis anterior and extensor hallucis longus. Computed tomography showed right foraminal stenosis at L5 with the floating lamina shifted ventrally and apophyseal ring fracture. In this case, the spondylolysis fracture angle differed between the left and right sides, with the fracture line on the right side running more sagittally. As a result, the floating lamina was shifted ventrally on the right side by compression and the right L5 intervertebral foraminal space was narrowed due to the ventral shift in the floating lamina and the apophyseal ring bone fragment. The shape of the fracture line should be examined carefully before surgery to avoid this technical pitfall. J. Med. Invest. 69 : 308-311, August, 2022
Nitric oxide (NO) plays diverse and significant roles in biological processes despite its cytotoxicity, raising the question of how biological systems control the action of NO to minimize its ...cytotoxicity in cells. As a great example of such a system, we found a possibility that NO-generating nitrite reductase (NiR) forms a complex with NO-decomposing membrane-integrated NO reductase (NOR) to efficiently capture NO immediately after its production by NiR in anaerobic nitrate respiration called denitrification. The 3.2-Å resolution structure of the complex of one NiR functional homodimer and two NOR molecules provides an idea of how these enzymes interact in cells, while the structure may not reflect the one in cells due to the membrane topology. Subsequent all-atom molecular dynamics (MD) simulations of the enzyme complexmodel in amembrane and structure-guided mutagenesis suggested that a few interenzyme salt bridges and coulombic interactions of NiR with the membrane could stabilize the complex of one NiR homodimer and one NOR molecule and contribute to rapid NO decomposition in cells. The MD trajectories of the NO diffusion in the NiR:NOR complex with the membrane showed that, as a plausible NO transfer mechanism, NO released from NiR rapidly migrates into the membrane, then binds to NOR. These results help us understand the mechanism of the cellular control of the action of cytotoxic NO.
Background:Transforaminal full-endoscopic spine surgery (FESS) is the least invasive spinal surgery and can be performed under local anesthesia. In Japan, the population is rapidly aging and the ...number of spinal surgeries performed in the elderly is also increasing. Object:In this report, we describe 3 patients aged 90 years or older in whom we performed FESS under local anesthesia. Case:The first case was a 90-year-old man who presented with severe leg pain. He had multiple medical comorbidities and was unsuitable for general anesthesia. We performed FESS. After surgery, the leg pain resolved with full recovery of muscle strength. He was discharged with no perioperative complications. The second case was a 90-year-old man who presented with severe leg pain. MRI showed a herniated nucleus pulposus and foraminal stenosis at L4/5. We performed FESS. The leg pain improved immediately after surgery. The third case was a 91-year-old woman in whom we diagnosed left L5 radiculopathy due to foraminal stenosis at L5/S1. After surgery, her leg pain was relieved. Conclusion:FESS is a good surgical procedure for elderly patients who are in a poor general condition because it is minimally invasive and can be performed under local anesthesia with early mobilization. J. Med. Invest. 71 : 169-173, February, 2024
A cadaveric study.
To investigate the anatomical features of segmental arteries and veins in the anterior part of the spinal column to prevent segmental vessel injury.
The lateral transpsoas approach ...to the lumbar intervertebral discs (IVD) is associated with the risk of segmental vessel injury. Previous studies have described the vascular anatomy on the lateral part of the vertebral body. However, there are no studies that describe the segmental vessels on its anterior aspect. Here, we report the important anatomical features of the segmental arteries and veins that can intersect the anterior part of the IVD. These vessels are considered at risk of vascular injury when placing the anterior retractors during lateral lumbar interbody fusion or cutting the anterior longitudinal ligament during anterior column realignment.
Five formalin-embalmed human cadavers were used. We assessed the proportion of segmental arteries and veins that intersected the IVD in the L2-L5 range and their course on the anterior part of the spinal column.
The segmental arteries and veins commonly intersect the anterior part of the IVD (artery, 28.1%; vein, 42.1%). Seven of 10 (70%) segmental arteries at L2 intersected the IVD, but only one artery intersected the IVD at L3 and L4. The proportions of segmental veins that intersected the IVD were 60%, 50%, and 16.7% at L2, L3, and L4, respectively.
The segmental arteries and veins frequently intersect the IVD in the anterior part of the spinal column. Therefore, it is necessary to consider these individual anatomical features to prevent vascular damage during lateral lumbar interbody fusion surgery.
Human ligamentum flavum-derived cells (HFCs) were obtained from surgical samples for a basic experimental study.
We sought to evaluate the inflammatory response of human ligamentum flavum cells to ...investigate hypertrophic changes occurring in the ligamentum flavum.
Lumbar spinal stenosis (LSS) is a disease commonly observed in the elderly. The number of patients with LSS has increased over time, yet the pathomechanisms of LSS still have not been fully elucidated. One of the clinical features of LSS is hypertrophy of the ligamentum flavum, which results in narrowing of the lumbar spinal canal. Some reports have suggested that ligamentum flavum hypertrophy is associated with inflammation and fibrosis; meanwhile, the p38 mitogen-activated protein (MAP) kinase is involved in the hypertrophy of human ligamentum flavum cells.
HFCs were obtained from patients with LSS who underwent surgery. HFCs were stimulated by tumor necrosis factor-α (TNF-α) and a p38 MAP kinase inhibitor, SB203580. Phosphorylation of the p38 MAP kinase was analyzed by western blotting. The concentration of interleukin-6 (IL-6) in the conditioned medium was measured by enzyme-linked immunoassay and IL-6 messenger RNA expression levels were determined by real-time polymerase chain reaction.
TNF-α induced the phosphorylation of p38 MAP kinase in a time-dependent manner, which was suppressed by the p38 MAP kinase inhibitor, SB203580. TNF-α also stimulated IL-6 release in both a time- and dose-dependent manner. On its own, SB203580 did not stimulate IL-6 secretion from HFCs; however, it dramatically suppressed the degree of IL-6 release stimulated by TNF-α from HFCs.
This is the first report suggesting that TNF-α stimulates the gene expression and protein secretion of IL-6 via p38 MAP kinase in HFCs. A noted association between tissue hypertrophy and inflammation suggests that the p38 MAP kinase inflammatory pathway may be a therapeutic molecular target for LSS.
A new perturbation theory based measure of mode coupling strengths in potential energy surfaces is introduced.
A measure of the importance of two- and three-mode couplings in molecular potential ...energy surfaces is introduced based on perturbation theory arguments. The measure requires a low-order polynomial representation of the potential energy function. The method is appropriate for reducing the computational cost of potential energy surface generation as well as vibrational wave function calculations. Example applications on grid-based potentials for formaldehyde and ethylene oxide are reported.
Infrared (IR) and Raman spectra of a sphingomyelin (SM) bilayer have been calculated for the amide I, II and A modes and the double-bonded CC stretching mode by a weight averaged approach, based on ...an all-atom molecular dynamics (MD) simulation and a vibrational structure calculation. Representative structures and statistical weights of SM clusters connected by hydrogen bonds (HBs) are observed in MD trajectories. After constructing smaller fragments from the SM clusters, the vibrational spectra of the target modes were calculated by normal mode analysis with a correction for anharmonicity, using density functional theory. The final IR and Raman spectra of a SM bilayer were obtained as the weight averages over all SM clusters. The calculated Raman spectrum is in excellent agreement with a recent measurement, providing a clear assignment of the peak in question observed at 1643 cm(-1) to the amide I modes of a SM bilayer. The analysis of the IR spectrum has also revealed that the amide bands are sensitive to the water content inside the membrane, since their band positions are strongly modulated by the HB between SM and water molecules. The present study suggests that the amide I band serves as a marker to identify the formation of SM clusters, and opens a new way to detect lipid rafts in the biological membrane.
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