Effect of Osteoporosis on Clinical and Radiological Outcomes Following One-Level Anterior Cervical Discectomy and Fusion.

STUDY DESIGN: Retrospective study. PURPOSE: This study aimed to evaluate how osteoporosis affected the clinical and radiological outcomes of patients who underwent anterior cervical discectomy and fusion (ACDF) with plating. OVERVIEW OF LITERATURE: The incidence of complications associated with implants is high when ACDF is performed in patients with poor bone quality. METHODS: In total, 101 patients without (T-score ≥1.0, group A) and 25 with (T-score ≤-2.5, group B) osteoporosis who underwent single-level ACDF with plating were followed up for >2 years. The clinical and radiological outcomes were compared between the two groups. The fusion rate and implant-related complications were evaluated. RESULTS: Although clinical outcomes such as visual analog scale scores for the arm (2.0±2.3 vs. 2.4±2.9, p=0.490) and neck pain (1.4±1.9 vs. 1.8±2.2, p=0.343) and neck disability index (7.7±7.1 vs. 9.9±7.5, p=0.225) were slightly higher in group B, no statistically significant difference was noted. Cage subsidence (13.9% vs. 16.0%, p=0.755) and plate migration (7.9% vs. 8.0%, p=1.000) rates did not differ between the two groups. The fusion rate at 1 year postoperatively was higher in group A than in group B (80.3% vs. 68.2%, p=0.139) and slightly increased in both groups (94.6% vs. 86.4%, p=0.178) at the final follow-up. CONCLUSIONS: Osteoporosis did not significantly affect the rate of cage subsidence or plate migration after cervical fusion. After ACDF, increased cage subsidence and implant migration rates had no significant effect on clinical outcomes.

Reliability and Diagnostic Accuracy of Standard Dermatomes and Myotomes for Determining the Pathologic Level in Surgically Verified Patients With Cervical Radiculopathy.

OBJECTIVE: This study aimed to investigate the reliability and diagnostic accuracy of typical dermatomes and myotomes for determining the pathologic level in surgically verified patients with cervical radiculopathy. METHODS: Patients who underwent single-level surgery due to cervical radiculopathy with at least a 60% reduction in preoperative symptoms or recovery of muscle power after surgery were included. The observed clinical symptoms (pain, paresthesia, motor weakness) were compared to those of typical cervical dermatomes and myotomes. RESULTS: Among the 227 patients reviewed, 142 (62.6%) had a standard dermatomal pattern, and 74 of 110 (67.3%) had a standard myotomal pattern. The myotome of C5/6 radiculopathy showed much more variance than those of other cervical segments. Among the patients with severe motor weakness (muscle strength ≤ grade 3 or obvious muscle atrophy), all those with involvement of root C5, C7, and C8 showed a typical pattern (C4/5: 13 of 13 patients, C6/7: 5 of 5 patients, C7/T1: 3 of 3 patients), while only 2 of the 6 patients (33.3%) with severe motor weakness caused by C5/6 radiculopathy fit the typical pattern. CONCLUSION: Among various symptoms, cervical myotome is of great value in determining the pathological level. However, it should be noted that there is high variability in human dermatomes and myotomes, especially for motor weakness due to C6 root compression, which is more variable than others.

Fate of Ossification of Posterior Longitudinal Ligament Following Anterior Cervical Fusion: Progression of Cervical Ossification of Posterior Longitudinal Ligament After Vertebral Body Sliding Osteotomy or Laminoplasty.

OBJECTIVE: In the treatment of ossification of posterior longitudinal ligament (OPLL)-induced cervical myelopathy, laminoplasty (LMP) is the most widely used surgical procedure. However, the progression of ossification masses is a well-known complication of LMP. This study aimed to investigate whether the novel anterior cervical decompression technique (vertebral body sliding osteotomy; VBSO) based on anterior column fusion suppresses the progression of OPLL compared with motion-preserving posterior decompression surgery (LMP). METHODS: All 77 consecutive patients (VBSO group, n = 33; LMP group, n = 44) who underwent VBSO or LMP for cervical OPLL at our institute between January 2012 and November 2017 were included. A total of 62 and 86 cervical motion segments in the VBSO and LMP groups were investigated, respectively. The OPLL thickness was measured twice (immediate postoperative and final follow-up), and the change of OPLL thickness was compared between the 2 groups. RESULTS: The increase in OPLL thickness in the VBSO group (-0.18 ± 0.24 mm) was significantly smaller than that in the LMP group (1.0 ± 0.9 mm, P < 0.001). Interestingly, in some patients, suppressed OPLL progression and decreased OPLL thickness were observed. CONCLUSIONS: The solid fusion of the anterior column by VBSO is associated with the lower incidence of OPLL growth and potential for growth arrest.

Ipsilateral Pedicle Stress Fracture After Endoscopic Foraminal Decompression for Patient with Isthmic Spondylolisthesis.

BACKGROUND: Minimally invasive endoscopic spine surgery is useful for the treatment of various spinal conditions. Although surgery-related complications such as dural injury, exiting nerve root injury, incomplete decompression, and hematoma have been reported, there are few reports of late complications after endoscopic surgery. CASE DESCRIPTION: A 51-year-old man complained of radiating pain to the right leg. The patient underwent endoscopic foraminal decompression under the diagnosis of foraminal stenosis with isthmic type spondylolisthesis (L5-S1). The lower extremity radiating pain was improved after surgery. Six weeks after surgery, the patient's symptoms recurred. The patient experienced a sudden onset of severe low back pain, which was aggravated by any motion of the lumbar spine. Computed tomography scan and magnetic resonance imaging revealed a fracture line with a sclerotic margin at the base of the right pedicle at the L5 level. Because the symptoms significantly interfered with his normal activities of daily living, the patient was treated with a total laminectomy, followed by posterior instrumented fusion. As the indication for endoscopic spinal surgery is widening, endoscopic decompression surgery is being performed for patients with low-grade lumbar spondylolisthesis, regardless of the presence of advanced spinal instability. However, endoscopic decompression surgery may cause damage to the posterior facet joint, which may have worsened the instability and lead to late complications such as progression of spondylolisthesis and pedicle stress fracture. CONCLUSIONS: The surgeon should carefully review risk factors such as isthmic type spondylolisthesis before planning spine surgery and minimize facet joint damage during endoscopic decompression.

Safety and Efficacy of a Novel Anterior Decompression Technique for Ossification of Posterior Longitudinal Ligament of the Cervical Spine.

INTRODUCTION: Anterior cervical corpectomy and fusion (ACCF) for cervical ossification of the posterior longitudinal ligament (OPLL) is associated with a high incidence of surgery-related complications. A novel anterior decompression technique (vertebral body sliding osteotomy [VBSO]) has been developed to prevent such complications. This study attests the efficacy and safety of VBSO versus those of standard ACCF. METHODS: Patients requiring surgery for cervical OPLL underwent VBSO (24 patients) or ACCF (38 patients). Operating time, estimated blood loss, neurologic outcomes, complications, and various radiographic parameters were investigated. RESULTS: The VBSO group showed a shorter mean operating time and less estimated blood loss versus the ACCF group. Sixteen patients in the ACCF group experienced various complications, namely neurologic deficit (two patients), cerebrospinal fluid leakage (four patients), graft migration (three patients), and pseudarthrosis (seven patients). In the VBSO group, only pseudarthrosis was reported (two patients). CONCLUSIONS: VBSO provides similar neurologic outcomes with a shorter operating time and less bleeding compared with ACCF. Surgeons do not need to directly manipulate the OPLL mass or dissect the interspace between the OPLL and dura mater. Therefore, this technique may decrease the incidence of surgery-related complications. STUDY DESIGN: Retrospective comparative study.

Anterior tibial displacement on preoperative stress radiography of ACL-injured knee depending on knee flexion angle.

PURPOSE: To compare side-to-side difference (SSD) of anterior tibial translation in instrumented stress radiography for each series of anterior cruciate ligament (ACL)-injured subjects according to knee flexion angle. METHODS: Forty subjects who were suspected of having significant ACL injury by manual Lachman test and MRI were recruited for this prospective study. These subjects took stress radiographs for both knees with corresponding knee flexion of 10° (series M1) and 30° (series M2) using Telos stress device. Mean SSDs of M1 and M2 were compared. Sensitivities of M1 and M2 were assessed using the SSD ≥ 3 mm or ≥ 5 mm as a cutoff value. RESULTS: Mean SSDs in series M1 and M2 were 4.22 ± 3.72 mm and 3.25 ± 3.30 mm, respectively (p < 0.001). When 3 mm of SSD was used as a cutoff value, sensitivities of series M1 and M2 were 47.5% (19/40) and 32.5% (13/40), respectively (p = 0.171). When 5 mm of SSD was used as a cutoff value, sensitivities of series M1 and M2 were 45.0% (18/40) and 22.5% (9/40), respectively (p = 0.033). CONCLUSIONS: Anterior tibial translation on stress radiographs using a Telos device is more prominent when knee flexion angle is 10° compared to that when knee flexion angle is 30°. However, stress radiography using Telos device, either at 10° or 30° of knee flexion, might not be suitable to make decision on surgical treatment due to relatively low sensitivities.

Efficacy of Escherichia coli-derived recombinant human bone morphogenetic protein-2 in posterolateral lumbar fusion: an open, active-controlled, randomized, multicenter trial.

BACKGROUND CONTEXT: The efficacy and safety of recombinant human bone morphogenetic protein-2 (rhBMP-2) as a bone graft substitute in spinal fusion has been widely researched. However, no study of the efficacy and safety of Escherichia coli-derived rhBMP-2 (E.BMP-2) with a hydroxyapatite (HA) carrier has been proposed. PURPOSE: This study aimed to compare the efficacy and safety of fusion materials between E.BMP-2 and autogenous iliac bone graft in posterolateral fusion (PLF). STUDY DESIGN/SETTING: An open, active-controlled, randomized, multicenter trial was carried out. PATIENT SAMPLE: This study included 93 patients who underwent single-level lumbar or lumbosacral PLF. OUTCOME MEASURES: The primary outcome measure was computed tomography (CT)-based fusion rate at 12 and 24 weeks. Secondary outcome measures were fusion grade by radiographs and CT at 12 and 24 weeks and changes in Oswestry Disability Index (ODI), Short Form-36 (SF-36) Health Survey, and visual analogue scale (VAS). METHODS: Patients who underwent 1-level PLF (between L1 and S1) for severe spinal stenosis or grade 1 spondylolisthesis were randomized to receive E.BMP-2 with an HA carrier (E.BMP-2 group) or autogenous iliac bone graft (AIBG group). Thin-section CT (<2 mm), VAS, ODI, and SF-36 were obtained pre- and postoperatively at 12 and 24 weeks. Outcome measures were compared between the groups. RESULTS: A total of 100 patients were enrolled in this trial. Among them, 93 patients underwent planned surgery. Preoperative demographic and clinical data showed no difference between groups. CT-based fusion rates were 100.0% (41/41) for the E.BMP-2 group and 90.2% (46/51) for the AIBG group (p=.062) at 12 weeks and 100.0% (41/41) and 94.1% (48/51) (p=.251) at 24 weeks, respectively. Fusion grade based on radiographs and CT showed non-inferiority of the E.BMP-2 group compared with the AIBG group. All clinical parameters improved postoperatively. However, there was no difference in changes in VAS, ODI, or SF-36 between the groups. No serious adverse event related to E.BMP-2 was found. CONCLUSIONS: The fusion rate of E.BMP-2 was comparable with that of AIBG following PLF. Good clinical efficacy and safety of E.BMP-2 in spinal fusion were also revealed. It was also suggested that HA shows suitability as a carrier for E.BMP-2. Thus, E.BMP-2 with an HA carrier can be an alternative bone graft material in spinal fusion.

Bone Union Rate Following Instrumented Posterolateral Lumbar Fusion: Comparison between Demineralized Bone Matrix versus Hydroxyapatite.

STUDY DESIGN: Retrospective study. PURPOSE: To compare the union rate of posterolateral lumbar fusion (PLF) using demineralized bone matrix (DBM) versus hydroxyapatite (HA) as bone graft extender. OVERVIEW OF LITERATURE: To our knowledge, there has been no clinical trial to compare the outcomes of DBM versus HA as a graft material for PLF. METHODS: We analyzed prospectively collected data from consecutive 79 patients who underwent instrumented PLF. Patients who received DBM were assigned to group B (n=38), and patients who received HA were assigned into group C (n=41). The primary study outcome was fusion rate assessed with radiographs. The secondary outcomes included pain intensity using a visual analogue scale, functional outcome using Oswestry disability index score, laboratory tests of inflammatory profiles and infection rate. RESULTS: One year postoperatively, bone fusion was achieved in 73% in group B and 58% in group C without significant difference between the groups (p=0.15). There were no differences between the groups with respect to secondary outcomes. CONCLUSIONS: DBM would provide noninferior outcomes compared to the HA as a fusion material for PLF, and could be a notable alternative.

The importance of proximal fusion level selection for outcomes of multi-level lumbar posterolateral fusion.

BACKGROUND: There are few studies about risk factors for poor outcomes from multi-level lumbar posterolateral fusion limited to three or four level lumbar posterolateral fusions. The purpose of this study was to analyze the outcomes of multi-level lumbar posterolateral fusion and to search for possible risk factors for poor surgical outcomes. METHODS: We retrospectively analyzed 37 consecutive patients who underwent multi-level lumbar or lumbosacral posterolateral fusion with posterior instrumentation. The outcomes were deemed either 'good' or 'bad' based on clinical and radiological results. Many demographic and radiological factors were analyzed to examine potential risk factors for poor outcomes. Student t-test, Fisher exact test, and the chi-square test were used based on the nature of the variables. Multiple logistic regression analysis was used to exclude confounding factors. RESULTS: Twenty cases showed a good outcome (group A, 54.1%) and 17 cases showed a bad outcome (group B, 45.9%). The overall fusion rate was 70.3%. The revision procedures (group A: 1/20, 5.0%; group B: 4/17, 23.5%), proximal fusion to L2 (group A: 5/20, 25.0%; group B: 10/17, 58.8%), and severity of stenosis (group A: 12/19, 63.3%; group B: 3/11, 27.3%) were adopted as possible related factors to the outcome in univariate analysis. Multiple logistic regression analysis revealed that only the proximal fusion level (superior instrumented vertebra, SIV) was a significant risk factor. The cases in which SIV was L2 showed inferior outcomes than those in which SIV was L3. The odds ratio was 6.562 (95% confidence interval, 1.259 to 34.203). CONCLUSIONS: The overall outcome of multi-level lumbar or lumbosacral posterolateral fusion was not as high as we had hoped it would be. Whether the SIV was L2 or L3 was the only significant risk factor identified for poor outcomes in multi-level lumbar or lumbosacral posterolateral fusion in the current study. Thus, the authors recommend that proximal fusion levels be carefully determined when multi-level lumbar fusions are considered.

A Prospective Randomized Clinical Trial Comparing Bone Union Rate Following Anterior Cervical Discectomy and Fusion Using a Polyetheretherketone Cage: Hydroxyapatite/B-Tricalcium Phosphate Mixture versus Hydroxyapatite/Demineralized Bone Matrix Mixture.

STUDY DESIGN: Prospective randomized noninferiority trial. PURPOSE: To evaluate whether the union rate of anterior cervical discectomy and fusion (ACDF) using a polyetheretherketone (PEEK) cage filled with a mixture of hydroxyapatite (HA) and demineralized bone matrix (DBM) is inferior to that of a mixture of ß-tricalcium phosphate (ß-TCP) and HA. OVERVIEW OF LITERATURE: There have been no clinical trials investigating the outcomes of a mixture of HA and DBM in a PEEK cage in ACDF. METHODS: Eighty-five eligible patients were randomly assigned to group B (n=43), in which a PEEK cage with a mixture of HA and DBM was used, or group C (n=42), in which a PEEK cage with a mixture of HA and ß-TCP was used. The primary study endpoint was the fusion rate, which was assessed with dynamic radiographs and computed tomography (CT) scans. Secondary endpoints included pain intensity using a visual analogue scale, functional outcome using a neck disability index score, laboratory tests of inflammatory profiles, and the infection rate. RESULTS: Seventy-seven patients (38 in group B and 39 in group C) were included in the final analysis. One year postoperatively, bone fusion was achieved in 87% of group B patients and 87% of group C patients on dynamic radiographs, and 87% of group B patients and 72% of group C patients on CT scans (p=1.00 and 0.16, respectively). There were also no between-groups differences with respect to the secondary endpoints. CONCLUSIONS: A HA/DBM mixture inside a PEEK cage can provide noninferior outcomes compared to a HA/TCP mixture in ACDF.

Clinical and radiological predictive factors to be related with the degree of lumbar back muscle degeneration: difference by gender.

BACKGROUND: The prediction of lumbar back muscle degeneration is important because chronic low back pain and spino-pelvic imbalance have been known to be related to it. However, gender difference should be considered because there are different quality and volume of muscles between genders. The purpose of this study was to search for clinical and radiological factors to predict the degree of lumbar back muscle degeneration according to gender difference. METHODS: We reviewed 112 patients (44 men and 68 women) with spinal stenosis who underwent a decompressive surgery between 1 January 2009 and 31 December 2011. Degrees of lumbar back muscle degeneration were classified into three categories by the fatty infiltration at each L3-4 disc level on the axial view of T1 magnetic resonance imaging (MRI). Age, sex, bone marrow density score, and body mass index (BMI) were obtained from chart reviews. Lumbar lordosis, sacral slope, pelvic tilt (PT), and pelvic incidence were calculated with lumbar spine standing lateral radiographs. The degrees of spinal stenosis and facet arthropathy were checked with MRI. Student t-test, chi-square test, or Fisher exact test were used to compare clinical and radiological parameters between genders. Analysis of variance (ANOVA) and linear regression analysis were used to search for a relationship between lumbar back muscle degeneration and possible predictive factors in each gender group. RESULTS: Many clinical and radiological parameters were different according to gender. The age, BMI, and PT in the female group (p = 0.013, 0.001, and 0.019, respectively) and the PT in the men group (p = 0.018) were predictive factors to be correlated with lumbar back muscle degeneration. CONCLUSIONS: The PT was the important predictive factor for lumbar back muscle degeneration in both, the male and the female group. However, age and BMI were predictive factors in the female group only.

A case of postoperative tuberculous spondylitis with a bizarre course.

Postoperative infections following spine surgery are usually attributable to bacterial organisms. Staphylococcus aureus is known to be the most common single pathogen leading to this infection, and the number of infections caused by methicillin-resistant Staphylococcus aureus is increasing. However, there is a paucity of literature addressing postoperative infection with Mycobacterium tuberculosis. We encountered a case of tuberculous spondylitis after spine surgery. A man had fever with low back pain three weeks after posterior interbody fusion with instrumentation for a herniated intervertebral disc at the L4-L5 level. He had been treated with antibiotics for an extended period of time under the impression that he had a bacterial infection, but his symptoms and laboratory data had not improved. Polymerase chain reaction for Mycobacterium tuberculosis turned out to be positive. The patient's symptoms finally improved when he was treated with antituberculosis medication.

CT20126, a novel immunosuppressant, prevents collagen-induced arthritis through the down-regulation of inflammatory gene expression by inhibiting NF-kappaB activation.

The colchicine-derived CT20126 compound has recently been shown to exert an immune regulatory effect and prolong the survival of allograft skins. In this study, we explored the anti-inflammatory and anti-arthritic effects of CT20126 in vivo and in vitro as well as investigated its underlying action mechanism. CT20126 suppressed the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2, tumor necrosis factor-alpha, and interleukin-1beta as well as the production of nitric oxide and prostaglandin E(2) in lipopolysaccharide (LPS)-treated macrophages as well as LPS-administered mice. This drug also inhibited the production of nitric oxide, prostaglandin E(2), and the chemokines, RANTES, GROalpha, and ENA-78, in cytokine-stimulated human synoviocytes. CT20126 suppressed NF-kappaB activation and iNOS promoter activity, which correlated with its inhibitory effect on phosphorylation-dependent IkappaB kinase activation, IkappaB phosphorylation and degradation, and NF-kappaB nuclear translocation, in LPS-stimulated macrophages. This compound also inhibited LPS-induced NF-kappaB-inducing kinase (NIK) and Akt phosphorylation, which are upstream of NF-kappaB activation. Furthermore, CT20126 significantly decreased the incidence and severity of arthritis as well as inhibited the expression of inflammatory cytokines, chemokines, iNOS, and cyclooxygenase-2 in the paws of collagen-induced arthritic mice. These findings indicate that CT20126 exerts an anti-inflammatory effect through NF-kappaB-responsive inflammatory gene expression by inhibiting the NIK- and Akt-dependent canonical NF-kappaB pathway and can be used as a therapeutic agent for rheumatoid arthritis related to chronic inflammation.

Effects of somatostatin on the responses of rostrally projecting spinal dorsal horn neurons to noxious stimuli in cats.

Somatostatin (SOM) is a widely distributed peptide in the central nervous system and exerts a variety of hormonal and neural actions. Although SOM is assumed to play an important role in spinal nociceptive processing, its exact function remains unclear. In fact, earlier pharmacological studies have provided results that support either a facilitatory or inhibitory role for SOM in nociception. In the current study, the effects of SOM were investigated using anesthetized cats. Specifically, the responses of rostrally projecting spinal dorsal horn neurons (RPSDH neurons) to different kinds of noxious stimuli (i.e., heat, mechanical and cold stimuli) and to the Adelta-and C-fiber activation of the sciatic nerve were studied. Iontophoretically applied SOM suppressed the responses of RPSDH neurons to noxious heat and mechanical stimuli as well as to C-fiber activation. Conversely, it enhanced these responses to noxious cold stimulus and Adelta-fiber activation. In addition, SOM suppressed glutamate-evoked activities of RPSDH neurons. The effects of SOM were blocked by the SOM receptor antagonist cyclo-SOM. These findings suggest that SOM has a dual effect on the activities of RPSDH neurons; that is, facilitation and inhibition, depending on the modality of pain signaled through them and its action site.

Colchicine-derived compound CT20126 promotes skin allograft survival by regulating the balance of Th1 and Th2 cytokine production.

Colchicine has been shown to regulate the expression of inflammatory gene, but this compound possesses much weaker anti-inflammatory activity. In this study, we synthesized a new colchicine derivative CT20126 and examined its immunomodulatory property. CT20126 was found to have immunosuppressive effects by inhibiting lymphocyte proliferation without cytotoxicity and effectively inhibit the transcriptional expression of the inflammatory genes, iNOS, TNF-alpha, and IL-1beta, in macrophages stimulated by LPS. This effect was nearly comparable to that of cyclosporine A. This compound also significantly suppressed the production of nitric oxide and Th1-related pro-inflammatory cytokines, IL-1beta, TNF-alpha, and IL-2, with minimal suppression of Th2-related anti-inflammatory cytokines IL-4 and IL-10 in the sponge matrix allograft model. Moreover, administration of CT20126 prolonged the survival of allograft skins from BALB/c mice (H-2(d)) to the dorsum of C57BL/6 (H-2(b)) mice. The in vivo immune suppressive effects of CT20126 were similar to that of cyclosporine A. These results indicate that this compound may have potential therapeutic value for transplantation rejection and other inflammatory diseases.

Inhibition of farnesyltransferase prevents collagen-induced arthritis by down-regulation of inflammatory gene expression through suppression of p21(ras)-dependent NF-kappaB activation.

Farnesylation of p21(ras) is an important step in the intracellular signaling pathway of growth factors, hormones, and immune stimulants. We synthesized a potent and selective farnesyltransferase inhibitor (LB42708) with IC(50) values of 0.8 nM in vitro and 8 nM in cultured cells against p21(ras) farnesylation and examined the effects of this inhibitor in the settings of inflammation and arthritis. LB42708 suppressed NF-kappaB activation and iNOS promoter activity by suppressing the I-kappaB kinase activity and I-kappaBalpha degradation. The inhibitor suppressed the expression of inducible NO synthase, cyclooxygenase-2, TNF-alpha, and IL-1beta and the production of NO and PGE(2) in immune-activated macrophages and osteoblasts as well as LPS-administrated mice. Furthermore, in vivo administration of LB42708 significantly decreased the incidence and severity of arthritis as well as mRNA expression of inducible NO synthase, cyclooxygenase-2, TNF-alpha, and IL-1beta in the paws of collagen-induced arthritic mice compared with controls. These observations indicate that the anti-inflammatory and antiarthritic effects of the farnesyltransferase inhibitor may be ascribed to the inhibition of I-kappaB kinase activity and subsequent suppression of NF-kappaB-dependent inflammatory gene expression through the suppression of p21(ras) farnesylation. Together, these findings reveal that the inhibitory effect of LB42708 on p21(ras)-dependent NF-kappaB activation may have potential therapeutic value for arthritis and other inflammatory diseases.

Distinct patterns of cleavage and translocation of cell cycle control proteins in CD95-induced and p53-induced apoptosis.

Apoptotic cell death induced by p53 occurs at a late G1 cell cycle checkpoint termed the restriction (R) point, and it has been proposed that p53-induced apoptosis causes upregulation of CD95. However, as cells with defective in CD95 signaling pathway are still sensitive to p53-induced apoptosis, CD95 cannot be the sole factor resulting in apoptosis. In addition, unlike p53-induced apoptosis, the relationship between CD95-mediated apoptosis and the cell cycle is not clearly understood. It would therefore be worth investigating whether CD95-mediated cell death is pertinent with p53-induced apoptosis in view of cell cycle related molecules. In this report, biochemical analysis showed that etoposide-induced apoptosis caused the induction and the nuclear translocation of effector molecules involved in G1 cell cycle checkpoint. However, there was no such translocation in the case of CD95-mediated death. Thus, although both types of apoptosis involved caspase activation, the cell cycle related proteins responded differently. This argues against the idea that p53-induced apoptosis occurs through the induction of CD95/CD95L expression.

Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation.

Nitric oxide (NO) functions not only as an important signaling molecule in the brain by producing cGMP, but also regulates neuronal cell apoptosis. The mechanism by which NO regulates apoptosis is unclear. In this study, we demonstrated that NO, produced either from the NO donor S-nitroso-N-acetyl-d,l-penicillamine (SNAP) or by transfection of neuronal NO synthase, suppressed 6-hydroxydopamine (6-OHDA)-induced apoptosis in PC12 cells by inhibiting mitochondrial cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation. This protection was significantly reversed by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalon-1-one, indicating that cGMP is a key mediator in NO-mediated anti-apoptosis. Moreover, the membrane-permeable cGMP analog 8-Br-cGMP inhibited 6-OHDA-induced apoptosis. These anti-apoptotic effects of SNAP and 8-Br-cGMP were suppressed by cGMP-dependent protein kinase G (PKG) inhibitor KT5823, indicating that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Both SNAP and 8-Br-cGMP induced endogenous Akt activation and Bad phosphorylation, resulting in the inhibition of Bad translocation to mitochondria; these effects were inhibited by KT5823 and the phosphatidylinositol 3-kinase (PI3K) inhibitors LY294002 and Wortmannin. Our data suggest that the NO/cGMP pathway suppresses 6-OHDA-induced PC12 cell apoptosis by suppressing the mitochondrial apoptosis signal via PKG/PI3K/Akt-dependent Bad phosphorylation.

Regulation of apoptosis by nitrosative stress.

Nitrosative stress can prevent or induce apoptosis. It occurs via S-nitrosylation by the interaction of nitric oxide (NO) with the biological thiols of proteins. Cellular redox potential and non-heme iron content determine S-nitrosylation. Apoptotic cell death is inhibited by S-nitrosylation of the redox-sensitive thiol in the catalytic site of caspase family proteases, which play an essential role in the apoptotic signal cascade. Nitrosative stress can also promote apoptosis by the activation of mitochondrial apoptotic pathways, such as the release of cytochrome c, an apoptosis-inducing factor, and endonuclease G from mitochondria, as well as the suppression of NF-kB activity. In this article we reviewed the mechanisms whereby S-nitrosylation and nitrosative stress regulate the apoptotic signal cascade.