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Morphological integration and functional assessment of transplanted neural progenitor cells in healthy and acute ischemic rat eyes.

We have functionally and morphologically characterized the retina and optic nerve after neural progenitor cell transplants to healthy rat eyes and eyes damaged by acute elevation of intraocular pressure (IOP). Green fluorescent protein-expressing adult rat hippocampal progenitor cells (AHPCs) were transplanted by intravitreal injection into healthy eyes and eyes damaged with acute ocular hypertension. Pupil light reflexes (PLR) and electroretinograms (ERGs) were recorded preoperatively and postoperatively. Eyes were subsequently prepared for immunohistochemical analysis and confocal imaging. Transplanted AHPCs were found in 8 of 15 (53%) acute ischemic eyes 62 days after surgery and 5 of 10 (50%) healthy eyes 32 days after grafting. Analysis of PLR and ERG function in acute ischemic eyes revealed no statistically significant difference compared to controls after transplantation for all observed functional parameters. Transplant into healthy rat eyes revealed no PLR or ERG amplitude deficits between transplanted and non-transplanted (control) eyes. Morphological and immunohistochemical analysis revealed that transplanted AHPCs survived and differentiated in both normal and injured retinal environments. Morphological integration occurred primarily within the inner retinal layers of the acute ischemic eyes. AHPCs were found to express neuronal and glial markers following transplantation. Transplanted AHPCs have the ability to integrate and differentiate in ischemia damaged retinas. PLR and ERG analysis revealed no significant difference in functional outcome in transplant recipient eyes.

Acute Disease↗

The development of synaptic function and integration in the central auditory system.

The development of inhibitory synaptic transmission is difficult to assess because the afferents usually arise from intrinsic neurons that are difficult to stimulate independently. The postnatal maturation of excitatory and inhibitory synaptic function was compared in the gerbil lateral superior olive (LSO), where it is possible to stimulate physically discrete afferent projections. Intracellular recordings obtained in a brain slice preparation revealed that transmission was prominent at birth. The EPSPs and IPSPs were up to 2 orders of magnitude longer than in more mature animals. Brief trains of electrical stimulus pulses led to a temporal summation of postsynaptic potentials (PSPs) in 1-14 d animals resulting in prolonged depolarizations or hyperpolarizations. In neonates, the depolarization could exceed 1 sec following a 70 msec stimulus train. The IPSPs in neonates were often of sufficient amplitude to evoke a rebound depolarization or action potential. The number of converging afferents was estimated from the quantized increases in PSP size. There was a significant decrease with age, suggesting that both inhibitory and excitatory afferents were eliminated during the first 3 postnatal weeks. The integration of action potentials with IPSPs was examined with conjoint stimuli to the two afferent pathways, and demonstrated that the effective IPSP duration decreased approximately 20-fold during the first 3 postnatal weeks. The magnitudes and durations of electrical stimulus-evoked PSPs suggest that spontaneous discharge of afferents to the LSO could have a substantial impact on their development, even prior to the response to airborne sound at 12 d. Furthermore, the synaptic responses obtained at 12-14 d postnatal indicated that both amplitude and temporal processing remain compromised. These immature synaptic properties would be expected to compound the inadequacies present in the cochlea and cochlear nucleus.

Animals↗

Embryotoxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD): the embryonic vasculature is a physiological target for TCDD-induced DNA damage and apoptotic cell death in Medaka (Orizias latipes).

Vertebrate embryos are extremely sensitive to environmental contaminants known as planar halogenated hydrocarbons (PHHs). The physiological targets that mediate PHH-induced embryotoxicity are not known. We have characterized embryotoxicity in medaka (Orizias latipes) caused by 2,3,7,8-tetrachlorodibezo-p-dioxin (TCDD), the prototypic PHH. DNA degradation in cells of the embryonic vasculature and loss of functional integrity of the medial yolk vein were demonstrated in TCDD-exposed embryos. Pharmacological intervention with piperonyl butoxide inhibited TCDD-induced DNA degradation, restored the functional integrity of the medial yolk vein, and protected against the embryotoxicity of TCDD. Treatment of TCDD-exposed embryos with the antioxidant N-acetylcysteine also provided significant protection against the embryotoxicity of TCDD. These results demonstrate that DNA damage and consequent cell death in the embryonic vasculature are key physiological mediators of TCDD-induced embryotoxicity.

Animals↗

Why insight fails.

Psychoanalysts rely on insight to promote therapeutic change. However, even when a great deal of insight has been attained, significant change may not be forthcoming in some patients, more often in borderline-level patients. The author examines such a case with particular reference to E. Kris' major points on insight, including the role of the ego's integrative functions in the attainment and utilization of insight. The author contends that failure to utilize insight is overdetermined and not necessarily owing to impairments in the integrative functions or to resistance in all its expressions. The author postulates that failure to make progress is due to a combination of the strength of the unmodulated drives, general ego weakness, poor early object relations, particularly preverbal experience, and constitution, in interaction. The whole of the personality is implicated in the inability to utilize insight.

Aged↗

Physicochemical properties and amino acid sequence of sheep brain galectin-1.

A beta-galactoside-specific soluble 14-kD lectin from sheep brain was isolated, sequenced, and compared with similar galectins from other species. Percent identities of amino acid sequence and the carbohydrate recognition domain (CRD) revealed that the isolated galectin shares all the absolutely preserved and critical residues of the mammalian galectin-1 subfamily. The isolated sheep brain galectin (SBG) showed more than 90% amino acid sequence (92%) and carbohydrate recognition domain identity (96%) with human brain galectin-1. Conformational changes were found induced by interaction of the protein with its specific disaccharide and oxidizing agent (hydrogen peroxide). Upon oxidation a drastic change in the environment of aromatic residues and conformation of the galectin was observed with the loss of hemagglutination activity, while no significant change was observed upon addition of D-lactose (Gal(beta1-4)Glc) in the far-UV and near-UV spectra, suggesting no significant modification in the secondary as well as tertiary structures of sheep brain galectin. But the functional integrity of the CRD is found to be affected in the presence of oxidizing agent, indicating intramolecular disulfide bonds and requirement of complete polypeptide chain for functional integrity of the carbohydrate recognition domain.

Amino Acid Sequence↗

"Pastoral crisis intervention": toward a definition.

The pastoral community represents a large and often untapped resource in times of crisis. It possesses a unique aggregation of characteristics that makes it uniquely valuable amidst the turmoil of a psychological crisis. In critical incidents such as terrorism, mass disasters, violence, the loss of loved ones, and any events wherein human actions result in injury, destruction, and/or death, the pastoral community may possess especially powerful restorative attributes. Unfortunately, heretofore, there has existed no generally recognized and accepted manner in which the healing factors inherent in pastoral care have been functionally integrated with the well-formulated principles of crisis intervention. This paper represents an initial effort to elucidate how the principles of pastoral care may be functionally integrated with those of crisis intervention. The amalgam shall heretofore be referred to as "pastoral crisis intervention" and is defined herein.

Aggression↗

Verbal memory outcome in patients with normal preoperative verbal memory and left mesial temporal sclerosis.

PURPOSE: Previous studies have shown that structural integrity (i.e., presence/absence of mesial temporal sclerosis (MTS)) of the left mesial temporal lobe is associated with verbal memory outcome following left anterior temporal lobectomy (ATL). However, the functional integrity of the left temporal lobe, as exemplified by preoperative verbal memory performance, has also been associated with verbal memory outcome following surgery. We investigated the risk of verbal memory loss in patients with known structural abnormality (i.e., left mesial temporal sclerosis by MRI) and normal preoperative verbal memory performance who undergo left ATL. METHODS: Seventeen patients with left temporal lobe epilepsy, MRI-based exclusive left MTS, and normal preoperative verbal memory were identified. Normal verbal memory was defined as performance on both Acquisition (learning across trials 1-5) and Retrieval (long delayed free recall) portions of the California Verbal Learning Test (CVLT) above a T score of 40 (>16%ile). Postoperative verbal memory outcome was established by incorporating standardized regression-based (SRB) change scores. RESULTS: Postoperative declines across both CVLT Retrieval T scores and Acquisition T scores (average 20% and average 15% declines from baseline scores, respectively) were measured for the group. The average CVLT Retrieval SRB change score was -2.5, and the average CVLT Acquisition SRB change score was -1.0. A larger proportion of patients demonstrated postoperative declines on Retrieval scores than Acquisition scores (64.7% vs 17.6%, respectively). CONCLUSIONS: Even in the presence of left MTS, patients exhibiting normal presurgical verbal memory are at risk for verbal memory declines following ATL. These results suggest that the functional integrity of the left mesial temporal lobe may play an important role in the verbal memory outcome in this patient group.

Adolescent↗

Effects of hypothermia on the function, membrane integrity, and cytoskeletal structure of hepatocytes.

To increase the storage time of livers for transplantation, a better understanding of hypothermia-induced hepatocyte damage is necessary. To this end, we have characterized the effects of hypothermia on long-term function and cytoskeletal organization of hepatocytes cultured in the collagen sandwich configuration, which maintains the expression of liver-specific functions for several weeks. In these studies, cultured hepatocytes (maintained at 37 degrees C for 7 days) were exposed to 4 degrees C in Leibovitz-15 (L15), University of Wisconsin (UW) solution, or L15 supplemented with 2.5 g% polyethylene glycol (PEG) for various time periods followed by a return to normothermia. When L15 medium was used, the long-term albumin secretion rate of cultured hepatocytes was decreased by 50% after 4 h, and by 95% after 24 h of exposure to 4 degrees C. Amorphous precipitates of F-actin and fragmented short microtubules were also observed after 4 and 12 h of hypothermia, respectively. Similar results were obtained when hepatocytes were stored in UW solution. However, in L15 supplemented with PEG, no significant reduction in long-term albumin secretion rates and intact actin and microtubule morphology was observed even after 24 h of exposure to 4 degrees C. The membrane integrity and long-term albumin secretion of hepatocytes stored in the presence of PEG were decreased to approximately 50% only after 48 h of exposure to 4 degrees C. Thus, PEG may be a useful additive in preservation solutions for hepatocytes in hepatocyte-based liver support systems and for intact tissue as well.

Actins↗

Rheb interacts with Raf-1 kinase and may function to integrate growth factor- and protein kinase A-dependent signals.

Rheb is a recently described member of the Ras family that was originally identified as an immediate-early gene in brain but is also widely expressed in other tissues. Here we demonstrate that Rheb interacts with and appears to regulate Raf-1 kinase, an essential component of the H-Ras signaling pathway. In direct contrast to H-Ras, however, the interaction of Rheb with Raf-1 is potentiated by growth factors in combination with agents that increase cyclic AMP (cAMP) levels. Protein kinase A-dependent phosphorylation of serine 43 within the regulatory domain of Raf-1 reciprocally potentiates its interaction with Rheb and decreases its interaction with H-Ras. A single amino acid in the G2 effector domain is critical for the differential properties of Rheb. Since Rheb is an immediate-early gene, our studies suggest that Rheb functions in concert with H-Ras to dynamically integrate cAMP and growth factor signaling.

3T3 Cells↗

Acetylcholine release from intrahippocampal septal grafts is under control of the host brain.

The activity of intrahippocampal transplants of cholinergic neurons was monitored by microdialysis in awake, freely moving rats. Fetal septal-diagonal band tissue was implanted into rats with a complete transection of the fimbria-fornix cholinergic pathway either as a cell suspension injected into the hippocampus or as a solid graft implanted in the lesion cavity. The grafts restored baseline acetylcholine release in the graft-reinnervated hippocampus to normal or supranormal levels. The graft-derived acetylcholine release was dependent on intact axonal impulse flow, and it was markedly increased during behavioral activation by sensory stimulation or by electrical stimulation of the lateral habenula. The results demonstrate that the septal grafts, despite their ectopic location, can become functionally integrated with the host brain and that the activity of the transplanted cholinergic neurons can be modulated from the host brain during ongoing behavior. Anatomical observations, using immunohistochemistry and retrograde tracing, indicate that direct or indirect brainstem afferents to the graft could mediate this functional integration. Host afferent control of the graft may thus play a role in the recovery of lesion-induced functional deficits seen with these types of transplants.

Acetylcholine↗

Usability of the hypoosmotic swelling "Water-test"--a simple method to assess sperm membrane integrity in mouse spermatozoa.

The functional integrity of the membrane of sperm from 3 strains of mice (BALB/c, CBA/KW, and C57BL/KW) was assessed by studying the swelling reaction of the sperm when suspended in a medium of distilled water. The results were correlated with analysis of sperm motility and those of Eosin-Y staining. The results obtained from the Eosin-Y test and "Water-test" differed (P < 0.01) between strains C57BL/KW and both BALB/C, and CBA/KW. A good correlation was observed in all strains of mice between the values of the "Water-test" and those for Eosin-Y staining (P < 0.01).

Animals↗

Transplantation of fetal dopamine neurons in Parkinson's disease: PET [18F]6-L-fluorodopa studies in two patients with putaminal implants.

Two patients with Parkinson's disease who underwent implantation of fetal mesencephalic tissue into the putamen were serially studied using positron emission tomography and [18F]6-L-fluorodopa ([18F]dopa). The uptake of [18F]dopa is related to the functional integrity of the presynaptic dopaminergic system. Preoperative studies revealed a marked decrease in putamen [18F]dopa uptake, with lesser involvement of the caudate. Two and 4 months, respectively, after operation, both patients demonstrated functional improvement, as described elsewhere. One patient was scanned 5, 8, and 13 months after the operation and the other was scanned 7 and 12 months after the operation. In both patients, [18F]dopa uptake increased within the operated putamen despite a progressive decrease in tracer uptake in the unoperated striatal structures. We believe that this increased uptake of [18F]dopa at the implantation site represents functional integrity within a surviving neural graft. While there has been little further clinical improvement beyond the fifth postoperative month, the uptake of [18F]dopa at the operation site in both patients has progressively increased. The kinetic data provide evidence of disease progression in the unoperated striatum, which, balanced against increasing graft function, may explain why clinical improvement reached a plateau within months after surgery.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Exploring a hospital-wide database: integrating statistical functions with ClinQuery.

Routinely collected clinical data are a valuable source of information for clinical epidemiology, health services research, and outcomes research. To explore, graph, and analyze such a large amount of material easily, one needs three kind of tools: a database management system, graphic displays, and statistical software. Although commercial software packages with these three functions do exist, it may be difficult to find one that fulfills the multiple requirements of a large hospital, allowing researchers to use the hospital information system to study various hypotheses. To address this issue we have integrated new exploratory modules with ClinQuery, a user-friendly computer program for online searching of clinical data. In this paper we describe some of the newly implemented graphics and statistical functions, along with practical examples of the way they may be used.

Boston↗

Phenotypic aberrations of CD3+ and CD4+ cells and functional impairments of lymphocytes at low-level occupational exposure to lead.

To study the immunological effects of low-level occupational exposure to lead, we have examined the phenotypic parameters and functional integrity of peripheral blood lymphocytes in a group of firearms instructors and compared the data to those obtained from healthy unexposed controls. Our results indicate that, among individuals with mildly elevated blood lead levels (> 25 micrograms/dl), detrimental effects on the host's immune functions occur. Such dysfunctions are multicellular as to mechanism and dose dependent in nature. The absolute percentage and number of CD3+ and CD4+ cells were significantly reduced, while the values for CD8+ cells were unchanged. Functional integrity of T cells as determined by responses to mitogens was impaired while that of T-cell-dependent B-lymphocyte function appeared to be within normal range at all stages of maturation. These data suggest the presence of a defective feedback loop which regulates T-cell-dependent functions and cell-to-cell cooperation. Furthermore, the marked deficiency in MLC responses that was also observed suggests that the adverse effect of lead may be due to its high affinity for the TCR and HLA-DR surface receptors thereby interfering with antigen processing from monocytes to T lymphocytes.

Adult↗

Desmin: a major intermediate filament protein essential for the structural integrity and function of muscle.

Desmin is a muscle-specific protein and a key subunit of the intermediate filament in cardiac, skeletal and smooth muscles. Desmin filaments are mainly located at the periphery of Z-disk of striated muscles and at the dense bodies of smooth muscle cells, and they have been postulated to play a critical role in the maintenance of structural and mechanical integrity of the contractile apparatus in muscle tissues. This review summarizes the findings in the regulation of the desmin gene and function of the desmin protein. The expression of desmin gene is regulated by a combination of different transcription control regions in muscle cells. The results from mice deficient in desmin reveal the fundamental role of desmin filaments in cell architecture, force transmission and mitochondrial function. Mice lacking desmin postnatally develop a dilated cardiomyopathy, a skeletal myopathy and smooth muscle defects. Some of desmin-related myopathies are attributable to a missense mutations and deletions in the desmin gene. Other desmin-related myopathies, in which the desmin gene is not mutated, could be related to mutations of the genes encoding the proteins that interact with desmin.

Animals↗

Impaired frontostriatal cognitive functioning following posteroventral pallidotomy in advanced Parkinson's disease.

We investigated the long-term effects of posteroventral pallidotomy on tests sensitive to the functional integrity of frontostriatal neural systems in a sample of 11 patients with advanced Parkinson's disease (PD). Patients were assessed within 1 month prior to surgery and at 12 months following pallidotomy. Changes in outcome measures were compared to a control sample of equally performing PD patients receiving nonsurgical medical management assessed over a 12-month period. Measures of cognitive abilities sensitive to frontostriatal functional integrity tested psychomotor processing speed, executive components of working memory, and reasoning. Additional tests of general mental status and semantic memory ability were utilized to assess the specificity of the effect of pallidotomy on cognitive function. Significant declines in performance on all measures sensitive to frontostriatal integrity were found for the surgery group but not the PD control group. No significant changes in performance were found on the measures of general mental status or semantic memory for either the surgery or PD control samples. These results suggest that the posteroventral pallidotomy selectively impairs performance on tests of frontostriatal cognitive abilities.

Adult↗

A technique for isolation perfusion of the canine liver with survival.

A technique of isolation perfusion of the canine liver was developed as a model for treating cancer limited to the liver. Preservation of hepatic functional integrity was dependent on maintenance of near physiologic conditions for the perfusion. Flow to the liver, 1 ml/min/g, was provided by gravity to the portal vein at two thirds total flow, and by pump to the hepatic artery at one third total flow. Utilizing the technique described, six dogs died 4-12 hr after a 1-hr perfusion. The dogs received Ringer's lactate solution IV and had a rising blood lactate to greater than 9 mM and a rising lactate/pyruvate ratio. After changing IV fluids to nonlactate containing solutions, 4/6 dogs survived a 1-hr perfusion at 37 degrees. Blood lactate concentrations remained below 6 mM (alpha less than 0.05) during perfusion, and the lactate/pyruvate ratios remained in a physiologic range on comparison to the nonsurvivors (alpha less than 0.05). Mild elevations occurred in the SGOT and SGPT with the peak on the first postoperative day of 93 +/- 54 and 79 +/- 56, respectively, but returned to normal within 4-7 days. Perfusate chemistries lactate, pyruvate, glucose, total alpha-amino acids, ketone bodies, SGOT, and SGPT demonstrated that hepatic functional integrity was maintained during the 1-hr perfusion. Hepatic function as assessed was no different between survivors and nonsurvivors. Hypoglycemia (glucose less than 2 mM) contributed to the two deaths. Tolerance of the dog to the temporary anhepatic state may be the limiting factor to prolonged perfusions.

Alanine Transaminase↗