Intervertebral Disc Calcification in Childhood – A Case Report and Review of the Literature

Summary  The authors report the case of a 10-year-old girl with intervertebral disc calcifications from the levels C6/C7 to Th1/Th2, presenting with a herniated calcified intervertebral disc at the C7/Th1 level, causing spinal cord compression with subsequent progressive paresis and sensory loss of her left leg. After anterior cervical discectomy and fusion the neurological deficits completely resolved within 2 weeks. It can be concluded that calcification of an intervertebral disc is a rare syndrome in childhood, causing progressive neurological deficit only in a few reported cases. Although the treatment of choice is conservative, surgery is required in patients who develop progressive neurological deficit.     

Molecular Basis of Intervertebral Disc Degeneration and Herniations: What Are the Important Translational Questions?

Background

Intervertebral disc degeneration is a common condition with few inexpensive and effective modes of treatment, but current investigations seek to clarify the underlying process and offer new treatment options. It will be important for physicians to understand the molecular basis for the pathology and how it translates to developing clinical treatments for disc degeneration. In this review, we sought to summarize for clinicians what is known about the molecular processes that causes disc degeneration.

Results

A healthy disc requires maintenance of a homeostatic environment, and when disrupted, a catabolic cascade of events occurs on a molecular level resulting in upregulation of proinflammatory cytokines, increased degradative enzymes, and a loss of matrix proteins. This promotes degenerative changes and occasional neurovascular ingrowth potentially contributing to the development of pain. Research demonstrates the molecular changes underlying the harmful effects of aging, smoking, and obesity seen clinically while demonstrating the variable influence of exercise. Finally, oral medications, supplements, biologic treatments, gene therapy, and stem cells hold great promise but require cautious application until their safety profiles are better outlined.

Conclusions

Intervertebral disc degeneration occurs where there is a loss of homeostatic balance with a predominantly catabolic metabolic profile. A basic understanding of the molecular changes occurring in the degenerating disc is important for practicing clinicians because it may help them to inform patients to alter lifestyle choices, identify beneficial or harmful supplements, or offer new biologic, genetic, or stem cell therapies.     

The Effect of Interleukin-1α Polymorphisms on Bone Mineral Density and the Risk of Vertebral Fractures

Interleukin-1α (IL-1α) stimulates bone resorption via osteoclasts. Mononuclear cells from patients with osteoporosis show increased IL-1α production, and IL-1α mRNA is more often detected in bone biopsies from osteoporotic compared to normal postmenopausal women. Polymorphisms have been identified in the IL-1α gene; however, none of these has been examined for an effect on bone phenotypes in Caucasians. We investigated if the polymorphisms in the IL-1α gene affect the risk of osteoporotic fractures, bone mineral density (BMD), and bone turnover in 462 osteoporotic patients and 336 normal controls. Based on previous studies of polymorphisms in the gene and data from the International Hap-Map Project, four polymorphisms needed examination in order to investigate the effect of known polymorphisms in the IL-1α gene. We examined C⁻¹²⁰²-T(rs1800794), C^–889-T(rs1800587), T^{155 + 209}-C(rs2071373), C^{155 + 320}-T(rs2856838), and G³⁹⁸-T(rs 17561) by Taqman and restriction fragment-length polymorphism assays. BMD was examined by dual-energy X-ray absorptiometry. Bone turnover was evaluated by serum osteocalcin, serum carboxy-terminal propeptide of human type I procollagen, serum bone-specific alkaline phosphatase, serum carboxy-terminal telopeptide of type I collagen, and urinary hydroxyproline/creatinine. Genotype distributions were in Hardy-Weinberg equilibrium. All polymorphisms were in strong linkage disequilibrium. The C allele of the C^{155 + 320}-T polymorphism tended to be more common among patients with vertebral fractures (P = 0.06) and patients with BMD T score <–2.5 (P = 0.05). Furthermore, haplotype 1 was associated with reduced risk of having BMD T score <–2.5 (P = 0.02). None of the other polymorphisms or haplotypes was associated with fracture risk or BMD T score <–2.5. BMD and bone turnover were not associated with any of the genetic variants. In conclusion, all the polymorphisms within the IL-1α gene are in strong linkage disequilibrium and not convincingly associated with fracture risk, BMD, or bone turnover.     

Effect of tumor necrosis factor-α and interferon-γ on the growth of human prostate cancer cell lines

Human recombinant tumor necrosis factor- (rTNF-, 10^-12–10^-8 M) inhibited the proliferation of androgen-dependent LNCaP cells by 32–56%. In contrast, proliferation of androgen-independent PC-3 and JCA-1 cells was only slightly inhibited, or not inhibited at all, respectively. Human recombinant interferon- (rIFN-, 500 U/ml) decreased proliferation of PC-3 and JCA-1 cells by 35% and 53%, respectively, but had no effect on LNCaP cells. Interestingly, the combination of rIFN- and TNF- had greater antiproliferative effects on JCA-1 cells than treatment with either cytokine alone. However, the antiproliferative effects of this combination were similar to those observed for PC-3 or LNCaP cells treated with rIFN- or TNF- alone, respectively. These data suggest that some forms of androgen-independent prostate cancer may benefit from a combination therapy of IFN- and TNF-, while the use of IFN- alone may be more efficacious in others.     

Hypoxic Regulation of Mitochondrial Metabolism and Mitophagy in Nucleus Pulposus Cells Is Dependent on HIF-1α–BNIP3 Axis

Nucleus pulposus (NP) cells reside in an avascular and hypoxic microenvironment of the intervertebral disc and are predominantly glycolytic due to robust HIF-1 activity. It is generally thought that NP cells contain few functional mitochondria compared with cells that rely on oxidative metabolism. Consequently, the contribution of mitochondria to NP cell metabolism and the role of hypoxia and HIF-1 in mitochondrial homeostasis is poorly understood. Using mitoQC reporter mice, we show for the first time to our knowledge that NP cell mitochondria undergo age-dependent mitophagy in vivo. Mechanistically, in vitro studies suggest that, under hypoxic conditions, mitochondria in primary NP cells undergo HIF-1α-dependent fragmentation, controlled by modulating the levels of key proteins DRP1 and OPA1 that are involved in mitochondrial fission and fusion, respectively. Seahorse assays and steady state metabolic profiling coupled with [1-2-¹³C]-glucose flux analysis revealed that in hypoxia, HIF-1α regulated metabolic flux through coordinating glycolysis and the mitochondrial TCA cycle interactions, thereby controlling the overall biosynthetic capacity of NP cells. We further show that hypoxia and HIF-1α trigger mitophagy in NP cells through the mitochondrial translocation of BNIP3, an inducer of receptor-mediated mitophagy. Surprisingly, however, loss of HIF-1α in vitro and analysis of NP-specific HIF-1α null mice do not show a decrease in mitophagic flux in NP cells but a compensatory increase in NIX and PINK1-Parkin pathways with higher mitochondrial number. Taken together, our studies provide novel mechanistic insights into the complex interplay between hypoxia and HIF-1α signaling on the mitochondrial metabolism and quality control in NP cells. © 2020 American Society for Bone and Mineral Research.     

Effect of the angle between the optical axis of the endoscope and the instruments’ plane on monitor image and surgical performance

Background: The aim of the study was to examine the effect of the angle between optical axis of the endoscope and the instruments plane (OAIP) on the monitor display angle (approach angle between the two instruments on a two-dimensional monitor) and endoscopic task performance. Methods: Two experiments were carried out in the Advanced Dundee Endoscopic Psychomotor Tester (ADEPT) using a standard two-dimensional video endoscopic system. In the first experiment, the monitor display angle was measured during use, with OAIP angles ranging between 0 and 80° (at 10° increments) and manipulation angles varying between 45 and 120° (at 15° increments). In the second experiment, 10 surgeons tied 500 intracorporeal knots with OAIP angles of 0° (optical axis in the same plane as the instruments), +15° and +30° (viewing above the instruments plane), and –15 and –30° (optical axis looking from below instruments plane). The end points were the execution time and knot quality score (KQS). Results: In the first experiment, instruments entered the visual field from lateral sides of the monitor with an apparent 180° monitor display angle with a 0° OAIP angle, whereas the monitor display angle approached the actual manipulation angle between the two needle drivers when an 80° OAIP angle was used. In the second experiment, the instruments appeared to enter the image field from the side of the surgeon when the endoscope viewed the instruments from above, whereas instruments entered the field from the opposite side to the surgeon when the endoscope viewed instruments plane from below. As a result, use of 0 and +15° OATP angles yielded significantly shorter execution times: 70 s compared to 83, 93, and 77 s for +30, –30, and –15° OAIP angles (p < 0.001), with KQS of 39° and 40 vs 38, 36, and 34%, respectively (p = 0.257). Conclusions: The angle between the optical axis of the endoscope and instruments plane determines how the instruments appear to enter the operative field. The monitor display angle between the instruments is different from the real manipulation angle unless the OAIP angle is near 80°. The apparent entry of instruments into the operative field becomes intuitive for the surgeon only if the endoscope is viewing from above or in the same plane as the instruments. Hence, the best performance for endoscopic knot tying is obtained with this configuration, and execution time increases significantly with viewing from below the instruments plane.     

Effect of isoflurane on expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA In the hippocamapus of immature rats

Obiective To investigate the effect ofisoflurane on expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA in the hippocampus of immature rats.Methods sixty-four 7-clay-old SD rats were randomly assigned into 2 groups(n=32 each):control group(group C)and isoflurane group(group S).group S was exposed to 1.5% isoflurane for 6 h while group C to air.Fore animals were killed before anesthesia(T0,baseline),at 2,4,6 h(T1-3)of isoflurane anesthesia and 4,6,12 and 24 h after anesthesia(T4-7).The hippocampi were immediately removed for determimation of the expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA by RT-PCR.Results Compared with group C,the expression of IL-1β mRNA at T1-5,IL-6 mRNA at T2.3 and TNF-α mRNA at T1-6 in the hippocampus was upregulated in group S.Conclusion The expression of IL-1β mRNA,IL-6 mRNA and TNF-β mRNA was elevated in the hippocampus of immature rats after being exposed to isoflurane.     

Effect of isoflurane on expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA In the hippocamapus of immature rats

Obiective To investigate the effect ofisoflurane on expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA in the hippocampus of immature rats.Methods sixty-four 7-clay-old SD rats were randomly assigned into 2 groups(n=32 each):control group(group C)and isoflurane group(group S).group S was exposed to 1.5% isoflurane for 6 h while group C to air.Fore animals were killed before anesthesia(T0,baseline),at 2,4,6 h(T1-3)of isoflurane anesthesia and 4,6,12 and 24 h after anesthesia(T4-7).The hippocampi were immediately removed for determimation of the expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA by RT-PCR.Results Compared with group C,the expression of IL-1β mRNA at T1-5,IL-6 mRNA at T2.3 and TNF-α mRNA at T1-6 in the hippocampus was upregulated in group S.Conclusion The expression of IL-1β mRNA,IL-6 mRNA and TNF-β mRNA was elevated in the hippocampus of immature rats after being exposed to isoflurane.     

Regulative effect of P38MAPK on release of TNFα and NO from alveolar macrophages under endotoxin stimulation

OBJECTIVE: To evaluate activation of P38 mitogen-activated protein kinase (P38MAPK) in alveolar macrophage (AM), release of TNFalpha and NO from cells, and their relationship following lipopolysaccharide (LPS) stimulation. METHODS: AM was isolated from branch alveolar lavage fluid (BALF). The activation of P38MAPK was assayed by Western blot. SB203580, a specific inhibitor of P38MAPK, was used with gradient concentration to evaluate the regulative effect of P38MAPK on the release of TNFalpha and NO from AM. RESULTS: P38MAPK was activated by LPS (100 ng/ml) with peak activation at 30 minutes. The activation of P38MAPK was inhibited by SB203580. The secretion of TNFalpha and NO stimulated with LPS increased (P<0.01) and was inhibited by SB203580 significantly. CONCLUSIONS: The results indicate that P38MAPK is involved in the secreting process of TNFalpha and NO following LPS stimulation. P38MAPK may be an important site for controlling the secretion of both inflammatory mediators during lung inflammatory disorders.     

The Protective Effect of Aprotinin and α-Tocopherol on Ischemia-Reperfusion Injury of the Rat Liver

Background

Liver injury caused by ischemia-reperfusion (I/R) processes is a complication of hepatic resection surgery and transplantation, particularly using grafts from marginal donors. Despite improvements in organ preservation and advances in surgical techniques, I/R injury remains a significant clinical problem. In this study, we investigated whether aprotinin provided protection against the adverse effects of I/R injury in liver tissue.

Methods

Forty rats were randomized into four groups (n = 10): group I: (control group) I/R + no medication; group II: sham-operated group + no medication or I/R; group III: I/R + aprotinin; group IV: I/R + α-tocopherol. Malondialdehyde (MDA) was measured in the liver tissue and superoxide dismutase (SOD), catalase (CAT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), as well as lactate dehydrogenase (LDH) in rat serum.

Results

Administration of aprotinin and α-tocopherol before I/R resulted in significant reductions of MDA levels compared to the I/R alone group (group I; P = .01 and P < .01, respectively). Administration of aprotinin or α-tocopherol prior to I/R resulted in significant increases in SOD and CAT levels compared with the I/R group (P < .05 each). Compared to the I/R group, significant decreases in plasma AST, ALT, and LDH levels were observed both in the aprotinin and in the α-tocopherol group (P < .05). Histological evaluation revealed the injury grade to be relatively lower among groups III and IV compared to group I.

Discussion

In conclusion, rat hepatic structures in aprotinin and α-tocopherol administered groups were well protected. Therefore, aprotinin may provide protection against the adverse effects of I/R injury in liver transplantation.     

Pretreatment effect of adenosine on activation of NF—κB and level of TNF—α during myocardial ischemia and reperfusion in rats

Ithasbeenshownthatischemiaandreperfusion(I/R)increasecytokinelevelsincludingTNF α ,IL 1,IL 6 ,IL 8,IF γ ,andintercellularadhesionmolecule 1(ICAM 1)inthemyocardium .1TNF αandIL 6arethoughttobeimportantintheprogressionofmyocardialdysfunction .2 AdenosineinhibitsmyocardialTNF αproductionin postischenmicrat ,reduceshumanmyocardialinjuryafterI/R ,3anddecreasesLipopolysaccharide (LPS )inducedcardiacandmacrophageTNF a production .4 However ,themolecularmechanismsofadenosineinthedown …     

TNFα pathway blockade ameliorates toxic effects of FSGS plasma on podocyte cytoskeleton and β3 integrin activation

Background

In the absence of mutant genes encoding components of the podocyte slit diaphragm, about 30–50 % of children with primary glucocorticoid-resistant focal segmental glomerulosclerosis (FSGS) develop recurrent proteinuria and slowly progressive FSGS lesions following renal transplantation. Recurrence of FSGS in the allograft strongly suggests a circulating factor that disturbs normal podocyte biology. To date, the nature of the circulating factor is unclear, and there is no cure for the recurrent form of FSGS (R-FSGS).

Methods

Cultured differentiated human podocytes were exposed to the plasmapheresis effluent or blood plasma samples from pediatric patients with recurrent or primary FSGS; in some cases, podocytes were pre-incubated with specific antibodies to block the tumor necrosis factor-alpha (TNFα) signaling pathway. Integrity of focal adhesion complexes and actin cytoskeleton were investigated by immunofluorescent microscopy.

Results

Plasmapheresis effluent from an R-FSGS child or fresh plasma from two children with primary FSGS rapidly disturbed the cytoskeleton of normal human podocytes in vitro. Plasma from a child with R-FSGS also activated β3 integrin and dispersed focal adhesion complexes. The effects were reversed by pre-incubation with antibodies against TNFα or either of the two TNFα receptors. When our patient with R-FSGS became resistant to plasmapheresis, we initiated treatment with twice weekly etanercept injections and then infliximab. Within 3?weeks of regular anti-TNFα therapy, the patient achieved sustained partial remission of proteinuria, allowing us to wean her off plasmapheresis completely.

Conclusions

We suggest that in some FSGS patients, disruption of the podocyte cytoskeleton and β3 integrin-mediated podocyte attachment are driven by the TNFα pathway.     

Differential effects of sevoflurane on the metastatic potential and chemosensitivity of non-small-cell lung adenocarcinoma and renal cell carcinoma in vitro

Background

Increasing evidence suggests that perioperative factors including anaesthetics influence cancer recurrence and metastasis after surgery. This study investigated the influence of sevoflurane on the response of lung and renal cancer cells to cisplatin, with focus on transforming growth factor-beta (TGF-β) and osteopontin (OPN) that are both closely associated with cancer tumorigenesis and metastasis.

Effect of α-tocopherol,taurine and selenium on the attenuation of ischemia/reperfusion injury of splanchnic organs

Background: Splanchnic artery occlusion shock is caused by increased capillary permeability and cellular injury precipitated by oxygen derived free radicals following ischemia and reperfusion of splanchnic organs. The purpose of this study was to assess the role of several wellknown oxygen-derived free radical scavengers in ameliorating or preventing this syndrome. Study design: Anesthetized rats were subjected to periods of occlusion of the visceral arteries and reperfusion. Tocopherol, taurine, selenium or a ‘cocktail’ of these three agents was injected subcutaneously for 4 consecutive days prior to operation. Mean arterial blood pressure was measured throughout the experimental period. Fluorometry and technetium-99m pyrophosphate counting of the visceral organs were performed as well as a histologic grading system for intestinal viability. Results: Final mean arterial blood pressure associated with the ‘cocktail’ and selenium groups was 79.1 ± 27.4 mmHg and 83.6 ± 17.8 mmHg, respectively. These values were significantly higher than the control group, 40.8 ± 11.4 mmHg (P < 0.05). Similar patterns of the benefit of selenium in contrast with the other groups were obtained with fluorescein perfusion, radioisotopic activity and histologie analysis. Conclusion: Pretreatment with selenium of splanchnic ischemia and reperfusion in the rat improves mean arterial blood pressure and microcirculatory visceral perfusion. Further analysis of the precise protective mechanism of selenium for reperfusion injury will enable visceral organs to withstand the consequences of increased capillary leakage and oxidant injury.