Substance P-like immunoreactivity in human cerebrospinal fluid

In a study to elucidate molecular mechanisms in pain, substance P-like immunoreactivity (SP-LI) was measured in lumbar CSF from 75 patients with lower back pain. Two samples--one before and one after lidocaine treatment--were obtained from each patient, and total SP-LI was measured in unfractionated (no HPLC) samples. SP-LI data were separated into three categories: placebo responders, pharmacological responders, and pharmacological non-responders. A significant difference was observed between the total SP-LI measurement of first and second samples of pharmacological non-responders. Distribution of SP-LI immunoreactive molecular species in two CSF patient samples (no ODS) was analyzed with a combination of reversed phase (RP) HPLC and RIA. Immunoreactivity in collected HPLC fractions was measured at calibrated retention times of synthetic SP-sulfoxide (SP-O), SP, and SP. Qualitative and quantitative differences in those HPLC-RIA metabolic profiles were observed within and between those two patients' samples. These data indicate that the type and amount of SP metabolism and SP precursor-processing differs in CSF between these two representative patients and within the short amount of time elapsed between acquiring these two samples.     

Focus: Rare Disease: A Case of Spinal Infectious Osteomyelitis Versus Gout: Advanced Imaging with Dual Energy CT

A 67-year-old male presented to the hospital for lower back pain and left lower extremity radiculopathy. Although the patient was afebrile and white blood cell count was normal, MRI was concerning for discitis/osteomyelitis at L4-L5. Subsequently, the patient developed a right knee joint effusion and underwent an arthrocentesis that was notable for the presence of urate crystals. A systemic urate crystal arthropathy was proposed as a potential etiology for the patient’s back pain and radiculopathy. Dual energy CT of the lumbar spine was performed, a technique which determines material composition by comparing the photon attenuation of the substance from two different x-ray energy levels. Results revealed the presence of monosodium urate crystals in the intervertebral discs. This technique is proposed as a noninvasive way to evaluate for gout in atypical locations or those difficult to sample and may replace an invasive intervertebral disc/endplate aspiration and/or biopsy. Dual energy CT should be considered in patients with elevated serum uric acid and concern for spinal involvement of gout.     

PLIF与TLIF手术治疗老年单节段腰椎退变性疾病的临床疗效对比

目的:探讨改良后路腰椎体间融合术(PLIF)与改良经椎间孔入路腰椎体间融合术(TLIF)手术治疗老年单节段腰椎退变性疾病的临床疗效。方法:收集2009年1月-2015年1月期间我院收治的经确诊为老年单节段腰椎退变性疾病的80例患者,按照随机数字表法分为观察组和对照组,各40例;观察组患者采用TLIF手术,对照组患者采用PLIF手术;比较两组患者手术前后腰背部疼痛程度(VAS评分)与活动功能(ODI评分),临床指标及并发症发生率。结果:观察组患者手术时间、术中出血量及术后引流量均明显短于或少于对照组患者(P0.05);两组患者手术前腰背部VAS评分与ODI评分比较差异无统计学意义(P0.05),手术后6个月,两组患者腰背部VAS评分与ODI评分均明显低于手术前(P0.05),而组间比较差异无统计学差异(P0.05);观察组术后并发症总发生率为5.00%(2/40),显著低于对照组的22.50%(9/40),差异有统计学差异(P0.05)。结论:PLIF与TLIF手术治疗老年单节段腰椎退变性疾病患者在改善腰背部疼痛程度与腰椎活动功能中的疗效相当,但TLIF手术有助于显著缩短手术时间,降低术中出血量与术后引流量,降低术后并发症发生风险,值得临床推广应用。     

Identification of neuropeptide FF-related peptides in human cerebrospinal fluid by mass spectrometry

Several neuropeptide FF (NPFF)-related peptides, known as modulators of the opioid system, have been previously characterized in bovine and rodent brain. Reverse-phase high pressure liquid chromatography (HPLC) fractions of a human with normal pressure hydrocephalus cerebrospinal fluid (CSF), co-migrating with NPFF-related synthetic peptides, were characterized by capillary HPLC coupled on-line to nanospray ion trap tandem mass spectrometry. Two peptides present in the pro-NPFF(A) precursor, NPAF (AGEGLNSQFWSLAAPQRF-NH2) and NPSF (SLAAPQRF-NH2), were identified. The monitoring of NPFF-related peptides in human CSF can be helpful to understand their roles in pain sensitivity.     

Serotonin in human lumbar cerebrospinal fluid: a reassessment

An inter-laboratory comparison study was carried out in order to ascertain mean levels of serotonin (5-HT) in human lumbar cerebrospinal fluid (CSF). Analyses were performed using high performance liquid chromatography (HPLC) coupled with either electrochemical (LC-EC) or fluorometric (LC-F) detection. With the detection limits obtained (7-8 pg/ml for LC-EC, 7-15 pg/ml for LC-F) 5-HT was not usually detected in human lumbar CSF. The findings indicate that the true mean concentration of CSF 5-HT is less than 10 pg/ml. This upper limit is substantially lower than all previous reports of 5-HT concentrations in normal human lumbar CSF. The extremely low concentrations of 5-HT present in CSF make it unlikely that CSF 5-HT will be of clinical utility in assessing central serotonergic function.     

Partial characterization of a novel endogenous opioid in human cerebrospinal fluid

Human cerebrospinal fluid (CSF) contains many uncharacterized endogenous opioids, in addition to the known enkephalins, endorphins, and dynorphins. These opioids may be separated by gel filtration chromatography and identified by radioreceptor assay for opioid activity. One region of the chromatographic elution profile, designated "Peak B" has previously been shown to be related to the pain status of chronic pain patients. We now report that human Peak B isolated from the CSF of pain-free elective surgery patients is present at a typical concentration equivalent in activity to 1.4 pmol of morphine sulfate per ml of CSF measured by radioreceptor assay. At a dose of 0.06 and 0.12 pmol morphine sulfate equivalents of CSF (MSE), injected into the cerebroventricular system of the mouse, Peak B produced an antinociceptive effect, the intensity and duration of which was dose-dependent and which was antagonized by naloxone. The mouse vas deferens (MVD) preparation was inhibited by Peak B in a manner that was sensitive to antagonism by naloxone only at low (less than 1.0 microM) but not at higher (greater than 6.0 microM) concentrations of the antagonist. Peak B activity in the MVD assay was unaffected by treatment with trypsin or alpha-chymotrypsin.     

Effect of naloxone administration upon the diurnal concentrations of oxytocin in the cerebrospinal fluid of rhesus and cynomolgus monkeys.

A diurnal pattern in oxytocin concentrations is present in cerebrospinal fluid (CSF) removed from the spinal subarachnoid space of monkeys, with elevated levels occurring in the early light hours. In order to investigate the possible role of endogenous opioid peptides in the generation of this oxytocin rhythm, we administered naloxone (0.4 mg/kg/h x 48 h) to rhesus and cynomolgus monkeys and examined the effects on the diurnal pattern of oxytocin in CSF collected from the lumbar subarachnoid spinal space. Monkeys maintained on jacket/tether/swivel systems and in a 12 h light: 12 h dark cycle (lights on 07.00-19.00 h) were implanted with temporary spinal subarachnoid catheters. CSF was continuously collected from the lumbar subarachnoid space and assayed for oxytocin. Oxytocin concentrations in CSF showed a diurnal variation with peak and nadir concentrations during light and dark hours, respectively. The lumbar CSF concentrations of oxytocin were not significantly different during naloxone vs. saline infusion. Plasma oxytocin concentrations, measured in the same animals, displayed no diurnal variation and were not significantly different during naloxone vs. saline infusion. We conclude that naloxone administration for 48 h does not perturb the diurnal variation in oxytocin concentrations in the CSF of monkeys. Mu opioid receptors are unlikely to be involved in modulating the diurnal rhythm of oxytocin in the CSF of monkeys.     

CSF distribution of morphine, methadone and sucrose after intrathecal injection

The lumbar to cisternal CSF distribution of morphine and methadone were compared to C-14 sucrose, a standard marker of CSF bulk flow, after lumbar subarachnoid injections in a sheep preparation. Morphine appeared and peaked simultaneously with C-14 sucrose in cisternal CSF at 90 to 190 minutes. The mean peak cisternal CSF morphine concentrations were sustained for 30-40 minutes, and averaged 148 ng/ml, representing 0.3% of the administered dose. Methadone was not detectable in cisternal CSF up to 240-300 minutes after lumbar subarachnoid administration. The C-14 sucrose/morphine ratio was increased an average of 6.7 times in cisternal CSF as compared to the ratio of the two compounds injected into the lumbar subarachnoid space. These studies demonstrate that morphine, a hydrophilic opioid, given intrathecally moves rostrally and appears in cisternal CSF by bulk flow. Furthermore the rostral redistribution of morphine is associated with the clearance of morphine from CSF. Methadone, a lipophilic opioid, appears to be completely cleared from CSF before it reaches the cisterna magna. These pharmacokinetic studies support a contribution of supraspinal sites to the analgesic and adverse effects produced by morphine given by spinal routes of administration. In contrast methadone appears to exert its effects predominantly at spinal sites.     

Spinal load bearing during sitting in an office chair with a tilting back]

Long periods of quiet sitting is considered a cause of low back pain. It is often assumed that spinal loads are high, especially when sitting erect. Modern office chairs with a tiltable back permit changes in the seated posture. In the most reclined position, some new chairs even match a kyphotic form of the lumbar spine. It is assumed that sitting on such a chair reduces low back pain. With the aim of determining spinal loading in different sitting positions, the loads acting on implanted fixation devices were measured telemetrically in two patients. Loads were measured in patients sitting on six different chairs with tiltable backs. In modern chairs, implant loading was always lower than while walking. In the end-tilt position of the chairback, loads were always lower than when the chairback was upright. Even when the lordotic curvature of the lumbar spine was "corrected", loads on the fixator were lower than when the subject was seated in the upright position. In a modern chair, spinal loading is no higher than with non-adjustable office chairs.     

Quantification of 1H NMR spectra of human cerebrospinal fluid: a protocol based on constrained total-line-shape analysis

A protocol for quantitative ¹H NMR analysis of human cerebrospinal fluid (hCSF) was built up and assessed as based on Constrained Total-Line-Shape (CTLS) fitting. In this method, linear constraints were applied to spectral structures. The ¹H NMR spectra of 45 human CSF samples were measured and quantified using the CTLS method. The quantification strategies based on total-line-shape fitting are discussed. The metabolic model for CTLS includes 31 metabolites covering 85% of the total spectral intensity, excluding the protein contribution. Prior to data analysis, the data was divided into patients with no Alzheimer’s disease (AD), but with a normal AD marker profile (the peptide β-amyloid₄₂ and tau protein) present in CSF, and into controls that do not have an AD marker profile in CSF. Unexpectedly large variations in metabolite concentrations within the two patient groups were detected, but an analysis of variance revealed a significant (P = 0.027) difference only in the concentration of creatinine which was higher in patients that had a normal AD marker profile. Multivariate classification tools such as self-organizing maps (SOM) failed in separation of the two classes.     

Episodic secretion of opioid activity in human plasma and monkey CSF: Evidence for a diurnal rhythm

Using a radioreceptor assay, opioiod activity has been determined in human plasma and monkey CSF at two-hour intervals across a 24-hour period. In both human plasma and monkey CSF, opioid activity showed an episodic secretion and a significant variation over time, suggesting a diurnal rhythm with increased levels in the morning. This rhythm is similar to those of adrenocorticotropin and beta-lipotropin (LPH) and reciprocal to the diurnal rhythm of pain sensitivity.     

A high-recovery extraction procedure for quantitative analysis of substance P and opioid peptides in human cerebrospinal fluid

This study reports an improved approach for the determination of neuropeptide levels in human cerebrospinal fluid (CSF). The method is based on sample acidification followed by liquid-liquid extraction (LLE) combined with radioimmunoassay. It was applied to study the recovery and level of some opioid peptides (Met-enkephalin-Arg(6)-Phe(7) and Leu-enkephalin-Arg(6)), substance P and the substance P(1-7) fragment, which are all compounds known to be present in human CSF. The results indicated that the use of LLE highly improved the recovery of these peptides compared to current liquid-solid-phase extraction methods by using silica gel cartridges or mini-columns for ion-exchange chromatography. Peptides added to CSF in concentrations down to 10 fmol/ml were recovered in yields exceeding 80%. The mean recovery of synthetic peptides as recorded by radioimmunoassay in the LLE procedure was significantly improved when HCl was added to the sample. In contrast, when the (125)I-labeled analogues of the peptides were added to CSF samples, the mean recovery of the four labeled peptides using the LLE procedure was markedly reduced in acidified samples. We also found that the inclusion of HCl effectively improved the removal of proteins present in the samples. As an application the levels of substance P and Met-enkephalin-Arg(6)-Phe(7) in CSF samples from patients with chronic pain (fibromyalgia syndrome) were measured using the new procedure. It was possible to confirm a significant difference in the CSF levels of both peptides when comparing patients and controls.     

Polymorphous exudates and atypical mononuclear cells in the cerebrospinal fluid of patients with Sj?gren's syndrome

Central nervous system (CNS) complications of Sj?gren's syndrome are now well recognized. To determine if any of the pathologic changes in the CNS in patients with Sj?gren's syndrome were reflected in the cellular composition of cerebrospinal fluid (CSF), we examined the CSF of 14 patients with Sj?gren's syndrome and neurologic symptoms and compared the differential cell counts in those cases with those of 14 control patients with similar neurologic symptoms. Patients with Sj?gren's syndrome had polymorphous (mixed) inflammatory exudates in CSF, composed predominantly of lymphocytes, but including variable numbers of plasma cells, neutrophils and erythrocytes. In addition, the CSF of all patients with Sj?gren's syndrome contained large, atypical, morphologically distinct mononuclear cells. The mean percentage of these cells in the CSF of patients with Sj?gren's syndrome (8.3 +/- 1.9) was significantly higher (p less than 0.001) than that observed in the control patients (0.7 +/- 0.2). These results suggest that involvement by Sj?gren's syndrome may be suspected by noting a polymorphous exudate containing characteristic atypical mononuclear cells in CSF obtained by lumbar puncture.     

Associations of Fatty acids in cerebrospinal fluid with peripheral glucose concentrations and energy metabolism

Rodent experiments have emphasized a role of central fatty acid (FA) species, such as oleic acid, in regulating peripheral glucose and energy metabolism. Thus, we hypothesized that central FAs are related to peripheral glucose regulation and energy expenditure in humans. To test this we measured FA species profiles in cerebrospinal fluid (CSF) and plasma of 32 individuals who stayed in our clinical inpatient unit for 6 days. Body composition was measured by dual energy X-ray absorptiometry and glucose regulation by an oral glucose test (OGTT) followed by measurements of 24 hour (24EE) and sleep energy expenditure (SLEEP) as well as respiratory quotient (RQ) in a respiratory chamber. CSF was obtained via lumbar punctures; FA concentrations were measured by liquid chromatography/mass spectrometry. As expected, FA concentrations were higher in plasma compared to CSF. Individuals with high concentrations of CSF very-long-chain saturated FAs had lower rates of SLEEP. In the plasma moderate associations of these FAs with higher 24EE were observed. Moreover, CSF monounsaturated long-chain FA (palmitoleic and oleic acid) concentrations were associated with lower RQs and lower glucose area under the curve during the OGTT. Thus, FAs in the CSF strongly correlated with peripheral metabolic traits. These physiological parameters were most specific to long-chain monounsaturated (C16∶1, C18∶1) and very-long-chain saturated (C24∶0, C26∶0) FAs. CONCLUSIONS: Together with previous animal experiments these initial cross-sectional human data indicate that central FA species are linked to peripheral glucose and energy homeostasis.     

Highly Sensitive Assay for Dopamine-β-Hydroxylase Activity in Human Cerebrospinal Fluid by High Performance Liquid Chromatography-Electrochemical Detection: Properties of the Enzyme

Abstract: This paper describes a new, sensitive assay for dopamine-β-hydroxylase (DBH) activity in human cerebrospinal fluid (CSF), serum and brain tissues by high performance liquid chromatography (HPLC) with electrochemical detection (ED). Dopamine (DA) was used as a substrate and was incubated under optimal conditions. Norepinephrine (NE) formed enzymatically from DA was isolated by a double-column procedure, the first column of Dowex-50-H⁺ and the second column of aluminum oxide. NE was adsorbed on the second aluminum oxide column and then eluted with 0.5 M-hydrochloric acid and assayed by HPLC-ED. Epinephrine (EN) was added to each incubation mixture as an internal standard, and this assay was therefore highly reproducible. The peak height in HPLC was linear from 500 fmol to 100 pmol of NE and EN. The lower limit of detection for NE formed enzymatically was about 30 pmol, which indicated that the sensitivity of this procedure was comparable to that of radioassay procedures. We applied the method to measurement of the activity of and examination of some of the characteristics of DBH in human CSF. DBH activity in CSF of Parkinsonian patients was lower than that of control patients. The properties of DBH in human CSF were similar to those in serum and adrenal medulla.     

普拉提运动对慢性下腰痛患者的疼痛和腰椎功能的影响

为了揭示普拉提运动对慢性下腰痛患者的疼痛和腰椎功能的影响,本研究选择2017年1月至2018年6月在医院确诊并接受治疗的64例慢性下腰痛患者作为研究对象,根据运动疗法分为对照组(悬吊训练法)和观察组(悬吊训练法结合普拉提运动),采用视觉模拟评分(VAS)评价患者的疼痛程度,采用Oswestry功能障碍指数(ODI)评价患者的腰椎功能障碍情况,采用运动情境动机量表(SSIMS)评价患者的运动参与动机。研究显示,治疗后观察组的VAS评分显著低于对照组(p=0.043)。观察组治疗后的ODI总评分显著低于对照组(p=0.026),观察组患者治疗后的疼痛、生活自理、提物、行走和站立5个维度评分显著低于对照组(p<0.05)。SSIMS量表的4个维度中,鉴别原则得分最高,其次为内部动机,然后是外部调节,最后为缺乏动机。本研究结果提示,普拉提运动可显著降低下腰痛患者的疼痛程度,并改善腰椎功能。此外,普拉提运动具有较好的患者认可度和喜爱度,值得在临床和运动康复训练中应用。     

Activation imbalances in lumbar spine muscles in the presence of chronic low back pain.

Paraspinal electromyographic (EMG) activity was recorded bilaterally from three lumbar levels during 30-s isometric trunk extensions [40 and 80% of maximum voluntary contraction (MVC)] in 20 healthy men and 14 chronic low back pain patients in pain. EMG parameters indicating neuromuscular fatigue and contralateral imbalances in EMG root-mean-square amplitude and median frequency were analyzed. Patients in pain showed less fatigue than controls at both contraction levels and produced only 55% of their MVC. Patients in pain likely did not produce a "true" maximum effort. A low MVC estimate would mean lower absolute contraction levels and less neuromuscular fatigue, thus explaining lower scores in the patients. Contralateral root-mean-square amplitude imbalances were present in both categories of subjects although such imbalances, when averaged across lumbar levels, were significantly larger in patients. Median frequency imbalances were significantly larger in the patients, at segmental as well as across lumbar levels. These results suggest that the presence of pain in these patients caused a redistribution of the activation behavior between synergistic muscles of the lumbar back.     

Atypical responses to morphine in mice: a possible relationship to anorexia nervosa?

According to our previously proposed auto-addiction hypothesis of chronic anorexia nervosa, patients become addicted to an initial period of dieting through endogenous opioid mediated mechanisms. Morphine causes hyperactivity and anorexia in the mouse, symptoms of anorexia nervosa but responses opposite to those of most species including rats and normal human subjects. This suggests that the atypical opioid systems in the mouse may resemble those of the chronic anorexia nervosa patient in contrast to those of most species including the normal human. Characterization of this atypical opioid system may be useful in understanding the pathophysiology of anorexia nervosa.     

腰腹肌运动联合超声波对非特异性下腰痛的治疗效果及机制

为了解腰腹肌运动联合超声波治疗的效果,本研究对腰腹肌运动联合超声波治疗非特异性下腰痛进行了探讨。研究显示,治疗4周后,腰腹肌运动+超声波治疗组的疼痛评分(0.73)显著低于超声波治疗组(1.22)(p<0.05)。腰腹肌运动+超声波治疗组的立位体前屈活动幅度(2.35 cm)显著低于超声波治疗组(6.23 cm)(p<0.05)。腰腹肌运动+超声波治疗组的屈伸肌总功和屈伸肌峰值力矩比显著高于超声波治疗组(p<0.05)。腰腹肌运动+超声波治疗组的Oswestry功能障碍指数问卷表(ODI)评分显著低于超声波治疗组(p<0.05)。治疗4周后,腰腹肌运动+超声波治疗组的血清超氧化物歧化酶(SOD)和过氧化氢酶(CAT)水平显著高于超声波治疗组,而丙二醛(MDA)水平显著低于超声波治疗组(p<0.05)。为了进一步考察运动联合超声波疗法对下腰痛的治疗机制,本研究建立了完全弗氏佐剂诱导的腰椎间盘退变大鼠模型。采用阿利新蓝染色检测大鼠椎间盘纤维环区和髓核区蛋白多糖的水平,并采用免疫组化染色检测蛋白聚糖和Ⅱ型胶原的表达,发现运动联合超声波治疗可明显提高大鼠蛋白多糖、Ⅱ型胶原和蛋白聚糖水平。本研究表明,与超声波治疗相比,腰腹肌运动联合超声波治疗可更大程度地降低非特异性下腰痛患者的疼痛,提高肌肉功能和腰椎功能,增强血清抗氧化能力,并减少氧化应激损伤。此外,该联合疗法可明显提高腰椎间盘退变大鼠椎间盘中的蛋白多糖、Ⅱ型胶原和蛋白聚糖水平。