Apparent diffusion coefficients and T2 relaxation time measurements to evaluate disc degeneration. A quantitative MR study of young patients with previous vertebral fracture.

PURPOSE: To assess the suitability of apparent diffusion coefficient (ADC) measurements to evaluate degeneration processes of the vertebral disc and to compare the results with T2 relaxation time measurements in both degenerated and normal intervertebral discs. MATERIAL AND METHODS: Fourteen young patients (8.8-20.8 years old) who had had a vertebral compression fracture at least 1 year earlier, underwent MR studies with diffusion imaging in three orthogonal directions and T2 relaxation time measurements. ADC values and T2 relaxation times of both degenerated and normal intervertebral discs were compared to the values of 20 healthy young asymptomatic control subjects. RESULTS: In the degenerated discs of patients, the ADCx and ADCy values were decreased compared to earlier determined values of healthy controls. ADC values in the z-direction in degenerated discs did not differ significantly from the values of controls. T2 relaxation times were shorter in the degenerated discs of patients compared to the values of controls. The greatest changes in both these values were observed in degenerated discs followed by discs with normal signal intensity adjacent to primary trauma area and secondary trauma area. CONCLUSION: We suggest that decreased ADC values reflect the lost integrity of the intervertebral disc. ADC measurements at MR may prove sensitive depicting of early degenerative changes in vertebral discs.

Slow vasomotor fluctuation in fMRI of anesthetized child brain.

Signal intensity changes in fMRI during rest caused by vasomotor fluctuations were investigated in this work. Resting-state baseline fluctuations were evaluated in 12 children anesthetized with thiopental. Five subjects had fluctuations related to subvoxel motion. In seven subjects without significant motion, slow signal fluctuation at 0.025-0.041 Hz near one or more primary sensory cortices was observed. In each subject the amplitude and frequency of the fluctuations were stable. It is hypothesized that thiopental, which reduces blood pressure and flow in the cortex, alters the feedback in neurovascular coupling leading to an increase in the magnitude and a reduction in the frequency of these fluctuations. The use of anesthesia in fMRI may provide new insight into neural connectivity and the coupling of blood flow and neural metabolism.

Apparent diffusion coefficient in thoracolumbar intervertebral discs of healthy young volunteers.

Apparent diffusion coefficient values (ADC) of healthy intervertebral discs of young volunteers in the thoracolumbar spine were determined using a single-shot EPI sequence. ADC(z) was in the lumbar spine slightly higher than ADC(x) or ADC(y). In vivo diffusion measurements of intervertebral discs may offer a novel diagnostic tool to evaluate disc diseases in early phases.

Interactions of Hsp90 with histones and related peptides.

The 90 kDa heat shock protein (Hsp90) induces the condensation of the chromatin structure [Csermely, P., Kajtár, J., Hollósi, M., Oikarinen, J., and Somogyi, J. (1994) Biochem. Biophys. Res. Commun. 202, 1657-1663]. In our present studies we used surface plasmon resonance measurements to demonstrate that Hsp90 binds histones H1, H2A, H2B, H3 and H4 with high affinity having dissociation constants in the submicromolar range. Strong binding of the C-terminal peptide of histone H1 containing the SPKK-motif and a pentaeicosa-peptide including the Hsp90 bipartite nuclear localization signal sequence was also observed. However, a lysine/arginine-rich peptide of casein, and the lysine-rich platelet factor 4 did not display a significant interaction with Hsp90. Histones and positively charged peptides modulated the Hsp90-associated kinase activity. Interactions between Hsp90, histones, and high mobility group (HMG) protein-derived peptides raise the possibility of the involvement of Hsp90 in chromatin reorganization during steroid action, mitosis, or after cellular stress.

Volume rendering using seed-filling acceleration: supporting cut planes by fast reseeding.

OBJECTIVE: Seed filling in view lattice is a method for accelerating volume rendering. Our previously published results on using seed filling had the limitation that they did not support cut planes. Cut planes are one of the main advantages of using volume rendering over surface rendering. In this article we describe the seed-filling acceleration technique and propose two fast reseeding processes that may be performed for each frame. MATERIALS AND METHODS: The reseeding process performs a connected component search on the cut plane within the volume data set. The amount of processing done by a connected component search algorithm is proportional to N(2), assuming the data set size is N(3). We implemented two connected component search algorithms: one for orthogonal cut planes and another for oblique cut planes. We measured the time taken by the cutting and estimated the effect on the rendering performance when the cut plane was moved interactively. RESULTS: Our measurements show that rendering rates of several frames per second can be achieved with a 266-MHz Pentium II PC, even when the object is interactively modified with cut planes during the rendering. CONCLUSIONS: We have shown that the seed-filling algorithm is also applicable in situations where the displayed data is modified interactively using cut planes or objects. Applications in this regard include, for example, computer-aided surgery systems, where the instrument may be used to interactively perform the simulated cut in the data set.

Effect of smoking on the prevalence of insulin resistance-associated cardiovascular risk factors among Finnish men in military service.

BACKGROUND: Obesity, central obesity, hypertension, dyslipidemia and hyperinsulinemia tend to cluster in the same individuals as a metabolic syndrome; smoking has an adverse effect on insulin resistance. The aim of our study was to examine the occurrence of clusters of insulin-resistance-associated abnormalities and the effect of smoking on this clustering in young men. METHODS: In 1995 we invited all the 1268 servicemen attending military service in the Ostrobothnian Brigade, Finland, for screening of the metabolic syndrome. The first phase consisted of an interview concerning smoking and measurements of blood pressure, weight, height, waist and hip circumferences. If diastolic pressure was > 85 mmHg, body mass index > 27 or waist-to-hip ratio > 0.98, these participants were invited to blood samples for the measurements of fasting serum lipids, plasma glucose and insulin. These results were obtained from 144 screening-positive men (120%) and from their 79(7%) randomly selected controls. RESULTS: The metabolic syndrome, defined as plasma insulin > or = 13.0 mU/l and serum triglycerides > or = 1.70 mmol/l and/or total cholesterol/high-density lipoprotein cholesterol > 5.0, was present in 10% (n = 1 4) of the screening-positive participants. None of the randomly selected controls had the metabolic syndrome. The metabolic syndrome was present in 12% (n = 11) of 93 smokers and in 2% (n = 3) of 1 28 non-smokers (P= 0.004). CONCLUSIONS: We conclude that clusters of metabolic abnormalities associated with insulin resistance are already present in young men, and that the prevalence of these clusters in smokers is sixfold compared with non-smokers.

Steroid-involved transcriptional regulation of human genes encoding prostatic acid phosphatase, prostate-specific antigen, and prostate-specific glandular kallikrein.

We have compared the steroid regulation of human genes encoding prostatic acid phosphatase (hPAP), prostate-specific antigen (hPSA), and prostate-specific glandular kallikrein (hK2) at the level of transcription. Reporter constructs of hPAP promoter covering the region -734/+467 were functional in both prostatic (LNCaP and PC-3) and nonprostatic (CV-1) cell lines in transient transfections. hPAP -231/+50 with eight identified transcription factor-binding sites showed the highest, and hPAP -734/+467 showed the lowest transcriptional activity in CV-1 cells. The hPAP promoter could not be induced with androgen, glucocorticoid, or progesterone, contrary to the hPSA (-620/+40) and hK2 (-493/+27) promoters in PC-3 cells cotransfected with the respective steroid receptor expression vector. Therefore, steroids cannot directly regulate hPAP gene expression via receptor binding to steroid response elements at -178 and +336, which have been shown to have androgen receptor-binding ability in vitro. Glucocorticoid was the most powerful activator of the hPSA construct at 10-nM steroid concentrations. On the contrary, glucocorticoid stimulation of the transcriptional activity of the hK2 construct was the weakest among the tested steroids. The results indicate that the steroid response elements in the proximal promoters of hPSA and hK2 genes are not androgen specific, offering the molecular basis for the expression of these genes outside the prostate in tissues containing steroid receptors.

Nucleotide and calcium-induced conformational changes in histone H1.

The principal constituents of chromatin, histone H1 (H1) and the nucleosome have essential roles in regulation of eukaryotic gene expression. However, mechanisms for the H1-dependent inactivation and for the ATP-dependent chromatin remodeling upon activation are largely unelucidated. Using circular dichroism (CD) analysis we show that ATP and other nucleotides and Ca2+ induce structural changes in H1. ATP and Ca2+ also induce changes when H1 is interacting with DNA, and the changes in H1 are accompanied by alterations in its DNA interaction. These results suggest that nucleotide and Ca2+ binding may be important for H1-mediated chromatin changes.

Computed tomography data based rapid prototyping model of the temporal bone before cochlear implant surgery.

Rapid prototyping (RP) technique allows automatic fabrication of 3D model parts. This method was applied to make a temporal bone model before cochlear implant surgery. A helical CT scan is used to acquire high resolution data from the middle and the inner ear of the patient. From the scanning data bone structures and soft tissues can be separated because their different grayscale pixel values. By using a guided image processing tool the desired parts of the anatomy can be extracted and 3D data created. The segmented data are processed to the form suitable for creating a high accuracy RP model. The RP model is made in the stereolithography (SLA) process by means of a computer guided HeCd laser beam inducing polymerisation of acrylic solution as it passes layer by layer over the surface of the polymer solution. In this prototype model the anatomy of the temporal bone can be clearly visualised, including, e.g., mastoid cells, tympanic cavity, bony canal of facial nerve, and round and oval windows. The inner ear spaces including vestibule, semicircular canals and cochlear turn are also shaped. The transparent acrylic material allows bonelike mechanical handling. The RP model can be dissected and used in individual surgical planning and simulation prior to cochlear implantation.

Coordination of transcription of the human 17 beta-hydroxysteroid dehydrogenase type 1 gene (EDH17B2) by a cell-specific enhancer and a silencer: identification of a retinoic acid response element.

Human 17 beta-hydroxysteroid dehydrogenase type 1 (17HSD type 1) catalyzes primarily the reductive reaction of estrone to the biologically more active form, estradiol. The enzyme is highly expressed in the human placenta and the ovary and, in addition, in certain estrogen target cells, such as breast epithelial cells. To elucidate the transcriptional control of the EDH17B2 gene, the gene encoding 17HSD type 1, we fused a series of 5'-deletion mutants of the EDH17B2 gene into chloramphenicol acetyl transferase reporter gene vectors. An enhancer region was identified within the bases -661 to -392 and it increased, in both orientations, thymidine kinase promoter activity more than 200-fold in JEG-3 choriocarcinoma cells. This enhancer region was also functional in another choriocarcinoma cell line, JAR, although to a lesser extent. In BT-20 and T-47D breast cancer cells the enhancer region increased thymidine kinase promoter activity to some degree but not as efficiently as expected on the basis of endogenous enzyme expression. No such enhancer activity was observed in 17HSD type 1 nonexpressing cell lines. The retinoic acid responsive element, which was located between bases -503 and -487 in the EDH17B2 enhancer, bound retinoid acid receptor alpha retinoid X receptor alpha complex and transmitted retinoic acid induction on transcription in JEG-3 and T-47D cells. Finally, a silencer, functional in all the cell lines tested, was localized in the region from -392 to -78. Deletion of the region lad to a 4-fold increase in reporter gene expression. Altogether, our findings suggest that transcriptional control of the EDH17B2 gene is coordinated by the cell-specific enhancer and the silencer.

The ATP/ADP binding domains of actin are conserved in nucleosome complexed with histone H1 and high mobility group-17 protein.

Chromatin has been proposed to share a common evolutionary origin and analogous mechanisms of action with various structures of cytoskeleton such as actin filaments. ATP has been shown to facilitate chromatin remodelling, but the exact site of its interaction with chromatin has not been elucidated. The facts, that ATP binds specifically to actin and promotes its polymerization, led us to characterize the possible domains involved in ATP binding in nucleosome. Comparison of the sequences of actin and the protein components of nucleosome suggests that H2A may contain an adenosine binding site similar to the adenosine motif of actin, H1 and/or H2B phosphate/Ca2+ binding sites corresponding to the phosphate 1 motif of actin, HMG17 a phosphate/Ca2+ binding site corresponding to the phosphate 2 motif of actin. The ability to bind ATP may be involved in the organization of chromatin into inactive solenoid-like structures in vivo.

The 90 kDa heat shock protein (hsp90) induces the condensation of the chromatin structure.

The 90 kDa heat shock protein (hsp90) is a member of the "chaperone-complex" of steroid receptors believed to be partially or transiently localized in the cell nucleus. Demonstrating that hsp90 has an ATP binding site and autophosphorylating activity we have observed that histones, especially histone H1, are able to modulate the autophosphorylation of hsp90 [Csermely, P. and Kahn, C.R. (1991) J. Biol. Chem. 266, 4943-4950]. Our present data suggest a direct interaction of hsp90 with histones, showing that hsp90 is able to bind histone-agarose and enhances the binding of histones to DNA. Circular dichroism spectra of rat liver chromatin indicate that hsp90 induces a tighter, condensed state of the chromatin structure which is resistant against disruption by high salt treatment. Interactions of hsp90 with the chromatin may be important in regulating the transcriptional activity of steroid receptors and other transcription factors.

Nucleotide recognition by histone H1 involves specific protein structures.

We have reported previously that histone H1 is capable of binding nucleotides such as ATP, GTP, ADP, and GDP in a specific manner. It is demonstrated here using labeling with the uv-crosslinkable ATP analog 8-azido-[alpha-32P]ATP that this ability is a unique characteristic of H1 among the histone proteins. Phosphate analogs such as AlF-4 efficiently counteract the labeling of H1, while they do not compete for labeling of histones H2A, H2B, H3, and H4. Consistent with the assumption that this labeling is due to specific binding, nucleotides competed for the labeling of H1 in a manner similar to labeling of the catalytic subunit of cAMP-dependent protein kinase, casein kinase-II, and heat shock protein-90, all of which are ATP/GTP-binding proteins. The site of nucleotide interaction was subsequently located in a Gly-rich region of H1 which displays homology with the protein kinases, using either radioactive labeling with nucleotide analogs and endoproteinase Glu-C digestion or synthetic peptides corresponding to the putative binding site. The results imply that specific protein structures are involved in nucleotide binding to H1 and that the ability of H1 to bind nucleotides may provide a mechanism for the regulation of eukaryotic gene expression.

Ultrasound-controlled neuronavigator-guided brain surgery.

The development of a unique neurosurgical navigator is described and a preliminary series of seven cases of intracerebral lesions approached with the assistance of this neuronavigation system under ultrasound control is presented. The clinical series included five low-grade astrocytomas, one chronic intracerebral hematoma, and one porencephalic cyst. Management procedures included biopsy in all cases, drainage of the hematoma, and endoscopy and fenestration for the cyst. The features of the neuronavigation system are interactive reconstructions of preoperative computerized tomography and magnetic resonance imaging data, corresponding intraoperative ultrasound images, versatility of the interchangeable end-effector instruments, graphic presentation of instruments on the reconstructed images, and voice control of the system. The principle of a common axis in the reconstructed images served to align the navigational pointer, biopsy guide, endoscope guide, ultrasound transducer, and surgical microscope to the brain anatomy. Intraoperative ultrasound imaging helped to verify the accuracy of the neuronavigator and check the results of the procedures. The arm of the neuronavigation system served as a holder for instruments, such as the biopsy guide, endoscope guide, and ultrasound transducer, in addition to functioning as a navigational pointer. Also, the surgical microscope was aligned with the neuronavigator for inspection and biopsy of the hematoma capsule to rule out tumor etiology. Voice control freed the neurosurgeon from manual exercises during start-up and calibration of the system.

Interaction of transcription factor Sp1 with the promoter of the gene for the multifunctional protein disulphide isomerase polypeptide.

Protein disulphide isomerase (PDI) is a unique polypeptide which resides in the lumen of the endoplasmic reticulum and also functions as the beta-subunit of prolyl 4-hydroxylase, as a cellular thyroid hormone-binding protein, as the smaller subunit of the microsomal triacylglycerol transfer protein complex, as a dehydroascorbate reductase and as a protein that binds various peptides in a specific manner. We have recently demonstrated that the promoter of the PDI gene contains six CCAAT boxes and other elements which are needed for efficient transcription. We now demonstrate that purified human recombinant transcription factor Sp1 interacts with two perfect GGGCGG sequences and three other GC-rich elements of the PDI promoter. Sp1 also appears to participate in the regulation of PDI gene expression, since overexpression of Sp1 stimulated PDI promoter activity in HeLa cells and mutations introduced into each of these Sp1-binding sites separately reduced the promoter strength, although even the largest decrease was only about 50%. These results support our view that expression of the gene for this polypeptide with multiple functions is secured by several regulatory elements, some of which are functionally redundant.

Specific non-enzymatic glycation of the rat histone H1 nucleotide binding site in vitro in the presence of AlF4-. A putative mechanism for impaired chromatin function.

We show here that an aluminium derivative, AlF4-, stimulates glycation of histone H1 selectively in the proximity of its nucleotide-binding site. This adduct formation interferes with nucleoside triphosphate hydrolysis by H1 and with nucleotide modulation of H1 DNA binding. The present mode of aluminium action may in part be responsible for its effects on the chromatin structure and expression of tissue-specific genes, and may constitute a mechanism in the pathogenesis of aluminium-induced encephalopathy and in that of Alzheimer's disease, for example.

Neuronavigator-guided cerebral biopsy.

Neuronavigators are new dynamic interactive instruments that use on-line computers to orient imaging data to the surgical field and guide the neurosurgeon to his target. We have been working since 1987 on a neuronavigator that serves not only as a precise pointer, but also as a dynamic arm that can be used to hold instruments, such as biopsy guides. The neuronavigator arm consists of six joints with optical encoders and is attached to the Mayfield headholder. The arm is connected to a workstation running customized 3D image graphics software. Special instruments and surgical technique have been developed. Here, we report on early clinical experience with ten biopsy procedures: 4 low-grade and 3 high-grade astrocytomas, one craniopharyngioma and one chronic intracerebral haematoma and intracerebral cyst, both of the latter with surrounding tumour suspect tissue. In all glioma cases serial biopsies were taken from optimal sites under ultrasound imaging control. Eight cases showed representative tumour tissue, while in two cases neoplasia was ruled out. The neuronavigator proved to be versatile, allowing comprehensive imaging data to be adapted to the surgical field.

DNA binding of histone H1 is modulated by nucleotides.

Histone H1 acts as a general repressor of transcription in eukaryotes by organizing nucleosomes into inaccessible condensed forms of chromatin. The capability of H1 to bind to DNA with some sequence specificity is likely to be critical in the control of these processes. We show here that ATP and several other nucleotides, including non-hydrolyzable derivatives, can inhibit DNA binding of H1. The results also show that ATP differentially affects binding of H1 to DNA in a fashion enhancing nucleotide sequence specificity of the binding. The study suggests a novel mechanism of modulation of H1 activity that has important implications for the role of H1 as a transcriptional regulator.