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Wavelet variations on the Shannon sampling theorem.

The Shannon sampling theorem asserts that a continuous square-integrable function on the real line which has a compactly supported Fourier transform is uniquely determined by its restriction to a uniform lattice of points whose density is determined by the support of the Fourier transform. This result can be extended to the wavelet representation of functions in two ways. First, under the same type of conditions as for the Shannon theorem, the scaling coefficients of a wavelet expansion will determine uniquely the given square-integrable function. Secondly, for a more general function, there is a unique extension from a given set of scaling coefficients to a full wavelet expansion which minimizes the local obstructions to translation invariance in a variational sense.

Fourier Analysis↗

Simple epithelium keratins are required for maintenance of hepatocyte integrity.

Keratin 8 (K8)-deficient adult mice develop a severe disease of the gastrointestinal tract characterized mainly by colorectal hyperplasia and inflammation. Given that hepatocytes contain K8/K18 heteropolymers only, this animal model was used to assess the contribution of these simple epithelium keratins to hepatocyte structural and functional integrity. Homozygous mutant (HMZ), heterozygous, and wild-type (WT) mice were examined for hepatocyte structural and metabolic features and their survival to partial hepatectomy. Except for the presence of few necrotic foci, no other tissular or cellular alterations were observed in nonhepatectomized HMZ mouse livers; glycogen and lipid peroxidation levels were essentially normal, but a small reduction in bile flow was observed. In response to a single pentobarbital injection, HMZ mice had longer sleeping times than heterozygous and WT mice. After a two-thirds partial hepatectomy under pentobarbital anesthesia, all HMZ mice died within a few hours, whereas those anesthetized with ether survived for 1 to 2 days. One hour after hepatectomy after pentobarbital anesthesia, many hepatocytes contained erythrocytes and large vacuoles in the cytoplasm, which suggests damage at the plasma membrane level in response to a sudden increase in portal blood flow. In line with these findings, an uptake of trypan blue by HMZ but not WT mouse hepatocytes was observed during a 10 ml/minute perfusion via the portal vein with a dye-supplemented buffer. Subsequent cellular dispersion led to viable WT mouse hepatocytes but largely nonviable HMZ mouse hepatocytes. Better viability was obtained at lower perfusion rates. Partially hepatectomized heterozygous mice developed liver steatosis, a condition that was not associated with a change in K8 content but perhaps linked to the presence of the neo gene. Transgenic HMZ mouse rescue experiments with a full-length K8 gene confirmed that the phenotypic alterations observed in partially hepatectomized HMZ mice were caused by the disruption of the K8 gene. Taken together, these findings demonstrate that simple epithelium keratins are essential for the maintenance of hepatocyte structural and functional integrity.

Animals↗

The cleaved N-terminal signal sequence of the cardiac Na(+)-Ca2+ exchanger is not required for functional membrane integration.

The cardiac Na(+)-Ca2+ exchanger differs from most other polytopic membrane proteins in that the amino terminus is cleaved during integration into the endoplasmic reticulum membrane. In this study, the cleaved N-terminal signal sequence of the exchanger was deleted (DelSS) or rendered uncleavable by mutation of the cleavage site (MutSS). Functional analysis of the mutants expressed in Xenopus laevis oocytes and sf9 insect cells demonstrates that DelSS exchanger catalyzes Na(+)-dependent Ca2+ transport at wild-type levels, while activity of MutSS exchanger is reduced to approximately 60% of wild-type in oocytes and 20% in sf9 cells. These results indicate that neither the presence nor the cleavage of the signal peptide is required for functional assembly of the exchanger protein in the membrane. Furthermore, these observations support the concept that internal topogenic signals play the major role in membrane insertion of the Na(+)-Ca2+ exchanger.

Animals↗

The tyrosine kinase receptor met and its ligand HGF are co-expressed and functionally active in HHV-8 positive primary effusion lymphoma.

Primary effusion lymphoma (PEL) harbors consistent infection by human herpesvirus-8, preferentially develops in immunodeficient patients and selectively localizes to the serous body cavities. Histogenetic analysis has suggested that PEL originates from post-germinal center, pre-terminally differentiated B cells sharing phenotypic features with plasma cells. Here we have investigated the expression status and functional integrity of the Met tyrosine kinase receptor and of its ligand hepatocyte growth factor (HGF). Thirteen PEL (nine cell lines and four primary specimens) were analyzed for Met and HGF expression and function by multiple assays. For comparison, a panel of 34 high grade B cell non-Hodgkin lymphomas (NHL) other than PEL was also investigated. Co-expression of Met and HGF was found in all PEL analyzed, whereas it was restricted to 1/34 B cell NHL other than PEL (P < 0.001; chi2 test). The Met protein expressed by PEL displays biochemical characteristics typical of Met expressed by other cell types and is capable of tyrosine autophosphorylation. By using a combination of immunological and biological assays, production and secretion of a functional HGF species was identified in all PEL cell lines analyzed. HGF stimulation of PEL cells rapidly induces Met tyrosine phosphorylation, demonstrating the functional integrity of the Met/HGF loop. Because of the well known mitogenic and motogenic properties of Met/HGF interactions, these data may bear implications for PEL growth and dissemination. Among B cell neoplasms, Met/HGF co-expression selectively clusters with PEL and, as demonstrated by previous studies, with multiple myeloma plasma cells, thus reinforcing the notion that PEL displays biologic similarities with tumors derived from late stages of B cell differentiation.

Gene Expression Regulation, Neoplastic↗

The binding of natural variants of human factor IX to endothelial cells.

The Gla-domain of human factor IX contains a specific element required for the binding of factor IX to an endothelial cell surface protein. We have investigated the dependence of this interaction on the structural integrity of the adjacent hydrophobic stack and epidermal growth factor-like domains. The ability of purified natural variants of human factor IX to compete with wild-type factor IX binding to the endothelial cell surface was used to obtain apparent Ki values of the variants. Our data suggest that the functional integrity of the Gla domain, enabling factor IX to specifically interact with an endothelial cell surface protein, depends on the structural and functional integrity of both the hydrophobic stack domain and the first epidermal growth factor-like domain.

Blotting, Western↗

Characterization of autoantigenic sites on isolated dog heart mitochondria.

1. Anti-heart mitochondria autoantibodies were developed in serum from dogs following experimental myocardial infarction. 2. Heart mitochondria frozen and thawed repeatedly in a sucrose/Tris-chloride buffer retained both their functional integrity as measured by the respiratory control ratio and their ability to serve as an antigen in a complement fixation test. Mitochondria frozen and thawed in a potassium chloride/Tris-chloride buffer lost both their functional integrity and their autoantigenic activity after one freeze-thaw cycle. 3. Extraction of the heart mitochondria with acetone/water mixtures to remove phospholipids from the membrane led to a complete loss of the ability of the mitochondria to react in the complement fixation test but did not affect the ability of the membranes to bind autoantibody in absorption experiments. 4. Treatment of the mitochondrial membranes with increasing concentrations of trypsin caused a loss of up to approximately 50% of the membrane protein with a gradual decrease in the autoantigenic activity of the membrane without impairment of the ability of the membrane to bind autoantibody. 5. Removal of up to 90% of the sialic acid of the mitochondrial membrane with neuraminidase resulted in a considerable increase in the complement-fixing autoantigenic activity of the membrane without changing the apparent ability of the membrane to bind autoantibody in absorption experiments. 6. Exposure of mitochondrial membranes to autoantibody and complement caused an inhibition of both an inner mitochondrial membrane enzyme, i.e. cytochrome oxidase (48%) and an outer mitochondrial membrane enzyme, i.e. NADH cytochrome c reductase (rotenone insensitive) (37%).

Animals↗

Integration of functional brain mapping in image-guided neurosurgery.

Magnetoencephalographic (MEG) brain mapping was performed in 90 patients with lesions associated with eloquent sensorimotor cortex. The MEG-derived sensorimotor mapping information was utilised for risk analysis and planning. Subsequently, these patients underwent either stereotactic volumetric resection, stereotactic biopsy or non-surgical management of their lesions. In seventeen patients, the MEG sensorimotor localization was integrated into an operative stereotactic database (consisting of CT, MRI and digital angiography) to be used in an interactive fashion during computer-assisted stereotactic volumetric resection procedures. The spatial relationship between the MEG derived functional anatomy, the structural/radiological anatomy and the pathology could then be viewed simultaneously, thereby affording a safer trajectory and approach. In addition, the real-time availability of functional mapping information in an interactive fashion helped reduce surgical risk and minimise functional morbidity. All of these patients had resection of their lesions with no change in their neurological status. In conclusion, MEG is a non-invasive, accurate, and reproducible method for pre-operative assessment of patients with lesions associated with eloquent sensory and motor cortex. The interactive use of MEG functional mapping in the operating room can allow for a safer approach and resection of these eloquent cortex lesions.

Adolescent↗

Development of an organ culture experimental system to investigate collagen synthesis in corpora cavernosa.

An organ culture model of rabbit corpus cavernosum has been developed to investigate fibrillar collagen synthesis in intact organ. Rabbit peni were removed en bloc, the corpora cavernosa were dissected, sliced into 5 mm pieces and incubated in culture media up to 24 h. Tissue viability and metabolic and functional integrity were assessed by (a) lactate dehydrogenase (LDH) release, (b) protein synthesis, and (c) ability to respond to exogenously added cytokines. Explants, maintained for 24 h in organ culture, exhibited minimal LDH release and maintained functional integrity determined from protein synthesis and ability to synthesize collagen in response to TGF-beta(1). These data suggest that explants of rabbit corpus cavernosum in organ culture represent a viable model to investigate connective tissue protein biosynthesis in vitro.

Animals↗

Actions of cyclic adenosine monophosphate on the cytodifferentiation of ovarian cells: studies in cultured swine granulosa cells using a novel exogenous adenylate cyclase from Bordetella pertussis.

The regulation by cAMP of cholesterol side-chain cleavage activity and the synthesis of immunoisolated cytochrome P-450scc and adrenodoxin proteins was investigated in primary cultures of swine ovarian (granulosa) cells. Administration of a novel adenylate cyclase toxin isolated from Bordetella pertussis increased granulosa-cell cAMP accumulation up to 200-fold over basal. These effects were additive with those of FSH, forskolin, and cholera toxin. In contrast, bacterial extracts BP 347 and BP 348 from mutant strains of B. pertussis that lack either all virulent factors or the adenylate cyclase toxin and hemolysin were devoid of effect. Granulosa-cell cAMP accumulation supported by active bacterial adenylate cyclase was accompanied by 2- to 11-fold, time-dependent increases in [35S]methionine incorporation into immunospecific cytochrome P-450scc and adrenodoxin. These increases in the synthesis of cholesterol side-chain cleavage proteins were associated with enhanced pregnenolone production in response to exogenous sterol substrate, 25-hydroxycholesterol, and augmented progesterone secretion both in the absence and presence of exogenous lipoprotein. Moreover, the effects of Bordetella adenylate cyclase toxin on granulosa cell steroidogenesis were functionally integrated with other regulatory responses, since the non-cAMP dependent effector, estradiol 17-beta, interacted synergistically with bacterial adenylate cyclase in stimulating progesterone production. We conclude that exogenous adenylate cyclase isolated from B. pertussis can be functionally integrated into the cAMP-dependent effector pathway of granulosa cells with a resulting increase in intracellular cAMP concentrations, augmented biosynthesis of progesterone and pregnenolone, enhanced synthesis of immunospecific cytochrome P-450scc and adrenodoxin, and synergistic interactions with a non-cAMP-dependent ovarian effector hormone (estradiol).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylate Cyclase Toxin↗

[Myocardial function in the late period after an operation for aortocoronary shunting].

Thirty patients with coronary heart disease were examined prior to and 18.3 +/- 3 months after surgery, with 66% of the shunts proving to be patent. All patients were divided into 3 groups: group I--14 (47%) patients with patent shunts, group II--10 (33%) patients with partially patent shunts and group III--6 (20%) patients with occluded shunts. The group I and II patients displayed an improvement in the segmentary contractility and the integral function of the left ventricle as compared to the preoperative values. The group III patients showed a negative time-course of the segmentary and integral function of the left ventricle. In all cases of the impaired patency of the aortocoronary shunts there was progression of the atherosclerotic process in the basin of the shunted artery.

Adult↗

MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor.

The lipopolysaccharide (LPS) receptor is a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2. Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor.

Antigens, Surface↗

An integrated reverse functional genomic and metabolic approach to understanding orotic acid-induced fatty liver.

In functional genomics, DNA microarrays for gene expression profiling are increasingly being used to provide insights into biological function or pathology. To better understand the significance of the multiple transcriptional changes across a time period, the temporal changes in phenotype must be described. Orotic acid-induced fatty liver disease was investigated at the transcriptional and metabolic levels using microarrays and metabolic profiling in two strains of rats. High-resolution 1H-NMR spectroscopic analysis of liver tissue indicated that Kyoto rats compared with Wistar rats are predisposed to the insult. Metabolite analysis and gene expression profiling following orotic acid treatment identified perturbed metabolic pathways, including those involved in fatty acid, triglyceride, and phospholipid synthesis, beta-oxidation, altered nucleotide, methyl donor, and carbohydrate metabolism, and stress responses. Multivariate analysis and statistical bootstrapping were used to investigate co-responses with transcripts involved in metabolism and stress responses. This reverse functional genomic strategy highlighted the relationship between changes in the transcription of stearoyl-CoA desaturase 1 and those of other lipid-related transcripts with changes in NMR-derived lipid profiles. The results suggest that the integration of 1H-NMR and gene expression data sets represents a robust method for identifying a focused line of research in a complex system.

Administration, Oral↗

IL-8/CXC ligand 8 survives neonatal gastric digestion as a result of intrinsic aspartyl proteinase resistance.

The human fetus and neonate swallow biologically significant quantities of IL-8/CXC ligand 8 (CXCL8) in amniotic fluid and breast milk, and this remains measurable through simulated neonatal gastric and proximal intestinal digestions. We sought to confirm the structural and functional integrity of IL-8/CXCL8 in digestates and determine the mechanisms underlying this protease resistance. We observed that in comparison with BSA, IL-8/CXCL8 is highly resistant to pepsin and can be detected intact in assays for structural, immunologic, and functional integrity. In a computational molecular docking simulation, IL-8/CXCL8 was observed to fit poorly in the pepsin active site. On the basis of simulated mutation analyses, we hypothesized that this protease resistance is due to disulfide bond-related tertiary folding in IL-8/CXCL8. This was confirmed on chemical reduction of these groups.

Animals↗

Comodulation masking release: effects of varying the level, duration, and time delay of the cue band.

The phenomenon of comodulation masking release (CMR) was studied in a series of experiments. When the relative level of the correlated cue band was more than about 10 dB less than that of the masker band, the CMR was abolished. When the duration of the tonal signal was varied with continuous maskers and cues, the course of the standard temporal-integration function (about -10 dB/decade) was followed by both the correlated-cue and the uncorrelated-cue conditions. In a burst masker paradigm employing several burst durations, the data for the correlated-cue condition closely followed the previously determined temporal-integration function. Finally, when the cue band was time delayed more than about 1.6 ms, the CMR began to decline, and it was abolished somewhere between 3 and 15 ms of delay, depending upon the subject. This latter outcome was essentially the same for masker and cue bands of both 75 and 100 Hz in width; in neither instance was there evidence of a cyclic, autocorrelation-like pattern following the period of the envelope. Supplementary experiments revealed two facts: The detectability of a masked narrow-band signal is not improved by the simultaneous presence of a correlated (or uncorrelated) noise band, and a small CMR can be obtained under conditions of forward masking.

Acoustic Stimulation↗

Passive motion therapy in temporomandibular joint disorders: the use of a new hydraulic device and case reports.

To understand the importance of motion on the structural and functional integrity of the temporomandibular joint (TMJ), the effects of immobilization and mobilization on synovial joints are reviewed. Impaired motion has profound deleterious effects on the synovial membrane, articular cartilage, muscles, ligaments, bone, and periarticular connective tissue. Conversely, motion has been shown to be essential to the maintenance of the structural and functional integrity of synovial joints. As in other synovial joints, the relative lack of the stimulus of mobility of the TMJ contributes to pathogenic mechanisms. Restoration of joint mobility is an important principle in the management of TMJ disorders. Illustrative cases are presented in which passive motion therapy, using a new device with a hydraulic mechanism, was successfully used for rehabilitation of TMJ function.

Adolescent↗

The function and integrity of the neurovascular unit rests upon the integration of the vascular and inflammatory cell systems.

The neurovascular unit is composed of a microvascular endothelium, neuron, and glial cell elements that are in physical proximity to the endothelium. The vascular system provides oxygen, glucose, and hormones for brain cells and guides the cells to appropriately respond to the local environment. Conversely, the brain cells, especially glial cells, can regulate the function of blood vessels in response to local requirements. The disruption of the neurovascular coordination was observed in a variety of inflammation-related diseases in brain, such as infectious diseases, stroke, vascular dementia, and multiple sclerosis. Inflammatory responses resulting from infections or injury of the brain activate the endothelium and glial cells to various degrees depending on the type, titer, or strength and duration of exposure to the agents or insults. The activation of endothelial and microglial cells may be modulated by the action of cytokines or other substances secreted from these cells. In an effort to understand the pathogenesis and find rational treatments against inflammatory disorders in brain, studies have been separately carried out using either endothelial cells or microglia. Increasing evidence, however, indicates that a crosstalk between these two cell types is important for the brain inflammation. Here, we review recent advances that provide insights into the coordinated interaction between the vascular and microglial systems, including the role of the specialized endothelium in regulating the immune response that occurs within CNS, the influence of microglial cells on the properties of endothelial cells, and the effects of endothelium on the state of microglial activation.

Animals↗

Intracarotid Amytal memory test and hippocampal magnetic resonance imaging volumetry: validity of the Wada test as an indicator of hippocampal integrity among candidates for epilepsy surgery.

OBJECT: Intracarotid Amytal testing (the Wada test) has been used to lateralize language and identify patients who may be at risk for memory impairment after temporal lobectomy. The goal of this study was to determine the validity of the Wada test in the assessment of pathological conditions of the hippocampus among candidates for epilepsy surgery. The authors examined the correlation between the functional integrity of the hippocampus, measured using the Wada test, and quantitative measures of hippocampal pathology, determined by obtaining volumetric measurements of the hippocampus with the aid of magnetic resonance (MR) imaging. METHODS: The authors reviewed the relationship between memory scores on the Wada test and hippocampal volumes measured on preoperative MR images in 76 patients who underwent anteromedial temporal lobectomy and amygdalohippocampectomy for the treatment of medically refractory temporal lobe epilepsy. The data were analyzed with respect to their usefulness in lateralizing the seizure focus and predicting the long-term postoperative memory outcome. Right and left hippocampal volume measurements did not correlate with one another (p > 0.1). Similarly, following a left carotid artery injection of Amytal the patients' right hemisphere memory was not significantly related to their left hemisphere memory on the Wada test (p'> 0.1). On the other hand, the patients' right hemisphere memory significantly correlated with their right hippocampal volume (r = 0.51; p < 0.001) and their left hemisphere memory significantly correlated with their left hippocampal volume (r = 0.51; p < 0.001). Both right and left hemisphere memory scores correlated with the hippocampal volumetry ratio (r = 0.47 and r = 0.45, respectively; both p < 0.001). Lateralization of a seizure focus based on hippocampal volumetry results was significantly related to lateralization based on the results of the Wada test (r = 0.49; p < 0.01). The disparity between the Wada memory scores on ipsilateral and contralateral sides was significantly and inversely related to the change in verbal memory following temporal lobectomy (r = -0.28; p < 0.02). The preoperative hippocampal volumetry ratio also significantly and inversely correlated with the change in verbal memory after surgery (r = -0.31; p < 0.01). CONCLUSIONS: The Wada memory test may be a valuable method of measuring the functional integrity of the hippocampus. The systematic study of MR imaging-acquired morphological data and Wada-acquired neuropsychological data may increase our understanding of the location of material-specific memory and the selection of eligible candidates for epilepsy surgery.

Adolescent↗

Restoration of ascending noradrenergic projections by residual locus coeruleus neurons: compensatory response to neurotoxin-induced cell death in the adult rat brain.

There is clinical and experimental evidence that monoamine neurons respond to lesions with a wide range of compensatory adaptations aimed at preserving their functional integrity. Neurotoxin-induced lesions are followed by increased synthesis and release of transmitter from residual monoamine fibers and by axonal sprouting. However, the fate of lesioned neurons after long survival periods remains largely unknown. Whether regenerative sprouting may contribute significantly to recovery of function following lesions which induce cell loss has been questioned. We have previously analyzed the response of locus coeruleus (LC) neurons to systemic administration of the noradrenergic (NE) neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) to adult rats. This drug causes ablation of nearly all LC axon terminals within 2 weeks after administration, followed by a profound loss of LC cell bodies 6 months later. The present study was conducted to determine the fate of surviving LC neurons and to characterize their potential for regenerative sprouting during a 16 month period after DSP-4 treatment. The time-course and extent of LC neuron degeneration were analyzed quantitatively in Nissl-stained sections, and the regenerative response of residual neurons was characterized by dopamine-beta-hydroxylase immunohistochemistry. The results document that LC neurons degenerate gradually after DSP-4 treatment, cell loss reaching on average 57% after 1 year. LC neurons which survive the lesion exhibit a vigorous regenerative response, even in those animals in which cell loss exceeds 60-70%. This regenerative process leads progressively to restoration of the NE innervation pattern in the forebrain, with some regions becoming markedly hyperinnervated. In stark contrast to the forebrain, very little reinnervation takes place in the brainstem, cerebellum and spinal cord. These findings suggest that regenerative sprouting of residual neurons is an important compensatory mechanism by which the LC may regain much of its functional integrity in the presence of extensive cell loss. Furthermore, regeneration of LC axons after DSP-4 treatment is region-specific, suggesting that the pattern of reinnervation is controlled by target areas. Elucidation of the factors underlying recovery of LC neurons after DSP-4 treatment may provide insights into the compensatory mechanisms of central neurons after injury and in disease states.

Animals↗