Diagnosis of arylsulfatase A deficiency in intact cultured cells using a fluorescent derivative of cerebroside sulfate

A fluorescent derivative of cerebroside sulfate (12-(1-pyrene)dodecanoyl-sphingosylgalactosyl-0-3-sulfate (P12-sulfatide) has been synthesized as a potential substrate for the determination of cerebroside sulfatidase (or arylsulfatase A) activity. It was administered into cultured human skin fibroblasts and thereby utilized for the diagnosis of arylsulfatase A deficiency. Cultured skin fibroblasts from normal individuals and healthy persons suffering from a pseudoarylsulfatase A deficiency (PD) degraded the P12-sulfatide, while in cells derived from a metachromatic leukodystrophy (MLD) patient it remained essentially intact. This contrasts with in vitro determinations of enzymatic activity, where the MLD or PD-derived arylsulfatase A exhibit similar deficiency, in spite of a profoundly different clinical course. Administration of the fluorescent sulfatide into the intact cells permitted a sensitive and rapid diagnosis of MLD and its distinction from the PD-phenomenon. This might be of particular importance for cases in which a rapid diagnosis is required and for prenatal diagnosis of fetuses from families afflicted with both MLD and pseudo-deficiency mutant genes.     

Mechanical Characterization of an In Vitro Device Designed to Quantitatively Injure Living Brain Tissue

Due to the nonlinear, viscoelastic material properties of brain, its mechanical response is dependent upon its total strain history. Therefore, a low strain rate, large strain will likely produce a tissue injury unique from that due to a high strain rate, moderate strain. Due to a lack of current understanding of specific in vivo physiological injury mechanisms, a priori assumptions cannot be made that a low strain rate injury induced by currently employed in vitro injury devices is representative of clinical, nonimpact, inertial head injuries. In the present study, an in vitro system capable of mechanically injuring cultured tissue at high strain rates was designed and characterized. The design of the device was based upon existing systems in which a clamped membrane, on which cells have been cultured, is deformed. However, the present system incorporates three substantial improvements: (1) noncontact measurement of the membrane deflection during injury; (2) precise and independent control over several characteristics of the deflection; and (3) generation of mechanical insults over a wide range of strains (up to 0.65) and strain rates (up to 15s^–1). Such a system will be valuable in the elucidation of the mechanisms of mechanical trauma and determination of injury tolerance criteria on a cellular level utilizing appropriate mechanical injury parameters.     

A Strain Device Imposing Dynamic and Uniform Equi-Biaxial Strain to Cultured Cells

The objective of this study is to design a new apparatus to allow the control of the magnitude and frequency of dynamic stretch applied uniformly to cells cultured on a silicon elastic membrane. The apparatus is designed to produce equi-biaxial dynamic stretches with area changes ranging from 0% to 55% and frequencies ranging from 0 to 2 Hz. Homogeneous finite strain analysis using triangles of markers was performed to compute the symmetric two-dimensional Lagrangian strain tensor on the membrane. Measurements of strain in both static and dynamic conditions showed that the shear component of the strain tensor (E_rc) was near zero, and that there was no significant difference between radial (E_rr) and circumferential (E_cc) components, indicating the attainment of equi-biaxial strain. Bovine aortic endothelial cells were transiently transfected with a chimeric construct in which the luciferase reporter is driven by TPA-responsive elements (TRE). The transfected cells cultured on the membrane were stretched. The luciferase activity increased significantly only when the cells were stretched by 15% or more in area. Cells in different locations of the membrane showed similar induction of luciferase activities, confirming that strain is uniform and equi-biaxial across the membrane. © 1998 Biomedical Engineering Society. PAC98: 8780+s, 8745-k, 8722-q     

Nonhomogeneous Deformation in the Anterior Leaflet of the Mitral Valve

In the mitral valve, regional variations in structure and material properties combine to affect the biomechanics of the entire valve. Previous biaxial testing has shown that mitral valve leaflet tissue is highly extensible, and exhibits nonlinear, anisotropic material properties. In this study, experimental measurements of mitral valve leaflet deformation under quasi-static pressure loading were performed on isolated porcine hearts. Biplane video images of markers placed on the anterior leaflet surface were used to reconstruct the 3D position of the markers at several pressure levels over the physiological range. A least-squares finite-element method was used to fit parametric models to the markers and to calculate the deformation over the surface. The results showed that the leaflet deformations were anisotropic, exhibiting a large nonhomogeneous radial stretch and a small circumferential stretch. This information can be used to better understand how the valve deforms under physiological loading, and to help design treatments for valve problems, such as mitral regurgitation.     

Evidence for a subgroup of essential hypertensives with non-suppressible excretion of aldosterone during sodium loading

Summary In patients with grade I and II essential hypertension studied during sodium loading (Na⁺ excretion above 175 meq·d^–1) we found a bimodal behaviour of aldosterone excretion and could distinguish two groups of patients: In the major part of essential hypertensives sodium loading led to a suppression of aldosterone excretion below 6 µg·d^–1, which is the highest control value during sodium loading, with an average of 2.7±1.4 (SD) µg·d^–1. Aldosterone excretion in a second group of patients was not suppressible below 6 µg·d^–1 despite forced sodium loading; it resulted in an average value of 10.0±3.0 (SD) µg·d^–1. During sodium deprivation or free sodium intake, aldosterone excretion in the first group of patients followed exactly the behaviour of normotensive controls, while in the second group of essential hypertensives the correlation of aldosterone excretion and log. Na excretion or log. Na⁺/K⁺ ratio in 24 h urine (r=–0.59) was far below the control value ofr=–0.87. Serum potassium concentration during sodium loading was significantly (p<0.001) lower (3.81±0.44 meq·l^–1) in the essential hypertensives with non-suppressible aldosterone excretion compared to those with suppressible aldosterone excretion (4.26±0.37 meq·l^–1). The blood pressure response to treatment with 200 mg spironolactone·d^–1 was better (p<0.05) in patients with non-suppressible aldosterone excretion compared to the essential hypertensives with normal aldosterone regulation. The plasma renin activity of both groups of patients was not significantly different, however, a tendency prevailed towards lower PRA-values in the patient group with non-suppressible aldosterone excretion during sodium loading.With the technical help of Mrs. R. Schendschilorz and Mrs. G. Suckau     

Metachromatic leukodystrophy caused by a partial cerebroside sulfatase defect

A patient with neuropathy and myopathy since infancy but whose neuropathy had been stable for a number of years showed a profound deficiency of arylsulfatase A in leukocytes and urine. Urine contained material that stained metachromatically and cochromatographed with cerebroside sulfate. In contrast, cultured fibroblasts contained about 10-20% of normal arylsulfatase A with properties identical to properties of normal fibroblast enzyme, except that it showed no cerebroside sulfatase activity. Growing fibroblasts in the cerebroside sulfate loading test had an attenuated rate of sulfatide hydrolysis. A re-examination of the cerebroside sulfatase reaction revealed that while only limited hydrolysis occurred with low concentrations of taurodeoxycholate or cholate (type I activation), significant hydrolysis of the natural substrate did take place with high concentrations of cholate (type II activation). This suggests that there is a partial cerebroside sulfatase defect in this atypical form of metachromatic leukodystrophy.     

A combined 2-deoxyglucose and neurophysiological study of primate somatosensory cortex

The metabolic activity pattern produced in the primary somatosensory cortex (SI) of primates by repetitive delivery of a tactile stimulus is distinctly patchy. The functional significance of these patches, however, remains obscure. This investigation sought to determine the correlation between neural and metabolic activity produced by tactile stimuli and to evaluate the relationship, if any, between the neural activity and metabolic patches evoked by similar stimuli. Experiments were undertaken in which extracellular microelectrode recordings were carried out in animals that subsequently underwent a 2-deoxyglucose (2DG) study. Three types of relations were identified. First, the receptive fields (RF) and modality properties of neurons sampled in locations at which patches of metabolic label were found matched the "place" and "modal" properties of the stimulus used to produce 2DG labeling. Second, in cortical locations where the RF and modality properties of the sampled neurons differed from either the place or modal properties of the stimulus used to evoke the 2DG label, no above-background increases in metabolic labeling were found. Finally, in some cortical locations at which the receptive field and modality properties of the neurons matched those of the 2-deoxyglucose mapping stimulus, no above-background increases in metabolic labeling were found. This outcome leads us to suggest that moment-to-moment changes in neural responsivity, which might remain undetected by conventional receptive field mapping methods, contribute to the patchy pattern of metabolic activity visualized by the 2-deoxyglucose method.     

剑川鼠疫自然疫源地蚤类群落季节动态研究

运用最优分割法(Fisher算法),对剑川鼠疫自然疫源地蚤类群落季节动态进行了研究,分为5个时段,根据结构与功能统一的原理.指出该疫源地的最佳灭蚤时间是2月末3月初。同时指出。如果防制时期选择不当.反而会使有害蚤的功能显现出来。     

肥胖者与非肥胖者24小时动态血压对比观察

目的：比较肥胖者和非肥胖者24h动态血压变化与高血压检出率。方法：采用24h动态血压监测技术,对肥胖者和非肥胖者24h平均收缩压和平均舒张压,白昼与夜间平均收缩压和平均舒张压,血压负荷,平均心率,24h血压波动趋势进行对比观察,结果：肥胖者各项血压指标及平均心率均较非肥胖者有明显升高（P＜0．05或P＜0．01）。结论：肥胖者高血压检出率明显高于非肥胖者,动态血压监测技术对肥胖者血压的评估有一定临床意义。     

X线照射后小鼠淋巴滤泡的动态变化

目的　探讨X线照射对小鼠月国窝淋巴结淋巴滤泡的影响。方法　采用组织学、免疫组织化学及三维重塑技术 ,观察小鼠全身X线照射后月国窝淋巴滤泡的变化。结果　小鼠全身X线照射后月国窝淋巴结重量迅速减小 ,淋巴细胞数量下降 ,淋巴滤泡被破坏。结论　X线照射可破坏淋巴滤泡 ,并经过一定时期可以重新聚集 ;再构筑的淋巴滤泡不是新产生的而是受损淋巴滤泡再生形成的