Inducible gene expression from defective herpes simplex virus vectors using the tetracycline-responsive promoter system

Herpes simplex virus-based amplicon vectors have been used for gene transfer into cultured neurons and the adult CNS. Since constitutive expression of a foreign gene or overexpression of an endogenous gene may have deleterious effects, the ability to control temporal expression would be advantageous. To achieve inducible gene expression, we have incorporated the tetracycline-responsive promoter system into amplicon vectors and showed, both in vitro and in vivo, that expression can be modulated by tetracycline. Using the firefly luciferase as the reporter gene, maximal repression by tetracycline in hippocampal cultures was about 50-fold. Withdrawal of tetracycline derepressed gene expression, reaching maximal levels within 10-12 h. In contrast, addition of tetracycline to cultures without prior tetracycline exposure inhibited gene expression rapidly; luciferase activity was reduced to less than 8% within 24 h. In adult rat hippocampus, vectors expressing luciferase or the Escherichia coli lacZ were repressed by tetracycline 9- and 60-fold, respectively. Maximum gene expression from the vectors occurred 2-3 days post-infection and declined thereafter. Such decline impeded further induction of expression by withdrawing tetracycline. This study demonstrates the feasibility of incorporating a powerful inducible promoter system into HSV vectors. The development of such an inducible viral vector system for gene transfer into the adult CNS might prove to be of experimental and therapeutic value.     

Superior colliculus and active navigation: role of visual and non-visual cues in controlling cellular representations of space

To begin investigation of the contribution of the superior colliculus to unrestrained navigation, the nature of behavioral representation by individual neurons was identified as rats performed a spatial memory task. Similar to what has been observed for hippocampus, many superior collicular cells showed elevated firing as animals traversed particular locations on the maze, and also during directional movement. However, when compared to hippocampal place fields, superior collicular location fields were found to be more broad and did not exhibit mnemonic properties. Organism-centered spatial coding was illustrated by other neurons that discharged preferentially during right or left turns made by the animal on the maze, or after lateralized sensory presentation of somatosensory, visual, or auditory stimuli. Nonspatial movement-related neurons increased or decreased firing when animals engaged in specific behaviors on the maze regardless of location or direction of movement. Manipulations of the visual environment showed that many, but not all, spatial cells were dependent on visual information. The majority of movement-related cells, however, did not require visual information to establish or maintain the correlates. Several superior collicular cells fired in response to multiple maze behaviors; in some of these cases a dissociation of visual sensitivity to one component of the behavioral correlate, but not the other, could be achieved for a single cell. This suggests that multiple modalities influence the activity of single neurons in superior colliculus of behaving rats. Similarly, several sensory-related cells showed dramatic increases in firing rate during the presentation of multisensory stimuli compared to the unimodal stimuli. These data reveal for the first time how previous findings of sensory/motor representation by the superior colliculus of restrained/anesthetized animals might be manifested in freely behaving rats performing a navigational task. Furthermore, the findings of both visually dependent and visually independent spatial coding suggest that superior colliculus may be involved in sending visual information for establishing spatial representations in efferent structures and for directing spatially-guided movements.     

Lack of dose-response with Pancrease MT for the treatment of exocrine pancreatic insufficiency in adults

BACKGROUND: Choosing the optimum pancreatic enzyme replacement therapy for patients with exocrine insufficiency remains a problem. An enteric coated enzyme microsphere pancreatic enzyme preparation (Pancrease) has been marketed with several levels of lipase activity, implying that there is a dose-response relationship between dose and effectiveness such that the high potency form appears to be the most cost effective. METHODS: In a randomized, single-blind, cross-over study, we evaluated the effectiveness of a commercial enzyme preparation with different amounts of lipase per dosage unit in adults with exocrine pancreatic insufficiency. Patients received a diet comprising 100 g fat each day for 6 days. With each meal (three per day) they received two capsules of either Pancrease MT4 (8000 unit lipase), Pancrease MT10 (20,000 units lipase), Pancrease MT16 (32,000 units lipase) or placebo. A 72-h quantitative faecal collection was carried out for the last 3 days of the 6-day period. RESULTS: There was a reduction in faecal fat excretion with each of the preparations compared to placebo. The difference failed to reach significance with the 8000 units lipase preparation (P > 0.05) but was significant (P = 0.02) with the 20,000 units lipase and the 32,000 units lipase preparations (faecal fat excretion: placebo = 42.1 +/- 29 g, lipase 8000 = 22.1 +/- 7.3 g, lipase 20,000 = 10.2 +/- 4.5 g and lipase 32,000 = 15.8 +/- 12.5 g, P < for 20,000 units and 32,000 units lipase compared to placebo). CONCLUSION: A dose-response relationship between the amount of lipase administered with each meal and a reduction in faecal fat was not evident. The most potent preparation did not provide additional benefits compared to the less expensive lower potency dosage form.     

Amplification of target-specific, ligation-dependent circular probe

In 1973, Narita reported that the rate of demented patients above 65 years was about 2% at the sanatorium--Tama Zenshoen, which was much lower than that of the average aged people (4-5%). The reason for that was thought to be due to the acquirement of strong mind under loneliness and difficulties faced by the patients through social enforcement to live in isolation. From that report on the low percentage of dementia among the Japanese Hansen patients reemed to be widely accepted. Fifteen years later in 1988, Narita reported that the rate of demented patients above 65 years was 3.2% in males and 3.6% in females indicating still a low rate at Tama Zenshoen. In 1994, however, Narita found unexpectedly that the percentage of dementia among patients above 80 at Tama Zenshoen was higher than the average aged population. Therefore in this report, we tried to analyze the reason for this difference. In addition to Tama Zenshoen patients, we also investigated those of another Hansen disease sanatorium namely Nagashima Aiseien which has similar structural organization as Tama Zenshoen. A comparison of dementia between these two was carried out, taking patients above 65 years as the study group (Used GBS scale based on Karasawa's method). In Tama Zenshoen, the rate of patients with dementia was 18.6% in males and 31% in females. Above 80 years those were 54.4% in males and 75.9% in females. The reason for the above high rate of dementia may be probably due to the passage of long years (40 to 50 years) after the enforcement to live in isolation causing the weakening of the spirit. The patients might have got some mental support if they had offspring, but they had no way to have any under the forced contraception. On the contrary, in Nagashima Aiseisen, the percentage of patients with dementia was low, namely 3.6% in males and 9.1% in females. In those patients above 80 years, it was 2.7% in males and 6.8% in females, those were almost similar to the average normal population (6%). Such a big difference was quite surprising. The reason behind this difference, may be due to the construction of the houses for the disabled persons. Both Nagashima Aiseien and Tama Zenshoen, have small individual rooms. In Tama Zenshoen, patients are able to visit their close neighbor, only after changing their shoes or with the aid of wheel chairs, but in Nagashima Aiseien, the corridors connecting quarters allow the patients to move easily to his neighbors with bare feet or even with crawl. Therefore, in Nagashima Aiseien, the number of patients who have social contacts with their neighbors is much greater in number which make them mentally active, thus eliminating the major factors affecting dementia. We can conclude that such communication between friends and co-mates make them alert and keep the spirit in the aged.     

Are shame, guilt, and embarrassment distinct emotions?

182 undergraduates described personal embarrassment, shame, and guilt experiences and rated these experiences on structural and phenomenological dimensions. Contrary to popular belief, shame was no more likely than guilt to be experienced in "public" situations; all 3 emotions typically occurred in social contexts, but a significant proportion of shame and guilt events occurred when respondents were alone. Analyses of participants' phenomenological ratings clearly demonstrated that shame, guilt, and embarrassment are not merely different terms for the same affective experience. In particular, embarrassment was a relatively distant neighbor of shame and guilt, and the differences among the 3 could not be explained simply by intensity of affect or by degree of moral transgression. Finally, participants generally were their own harshest critics in each type of event, evaluating themselves more negatively than they believed others did. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Differential immunohistochemical labeling of Gs, G(il) and 2, and G(o) alpha-subunits in rat forebrain

The anatomical and morphological distribution of the G proteins G(o), G(i1) and 2, and Gs alpha-subunits in rat forebrain sections was determined using immunohistochemical techniques. Diffuse G(o) labeling occurred in the neuropil throughout the cortex, superficial layers of the entorhinal cortex, thalamus, several white matter fiber tracts, and hippocampus. G(i1) and 2 immunoreactivity was also located in the neuropil but produced a more fibrous pattern. Fibrous labeling of G(i1) and 2 was observed in the cortex, amygdala, hippocampal subfield CA3, and several white matter fiber tracts. Both G(o) and G(i1) and 2 labeling was present in the pencil fibers within the striatum and lateral geniculate nucleus. Gs labeling, in contrast to G(o) and G(i1) and 2, was generally cytoplasmic. Cytoplasmic Gs labeling was observed in the thalamus, habenula, dentate, geniculate nucleus, hypothalamus, and hippocampus. Intense Gs labeling was observed in the striatum parenchyma, choroid plexus, and infundibular stem. Based on our results, we conclude that the G proteins G(o), G(i1) and 2, and Gs are anatomically distributed differently throughout the brain. The diffuse neuropil labeling of G(o), fibrous neuropil labeling of G(i1) and 2, and cytoplasmic labeling of Gs strongly suggests that the G proteins are also differentially distributed morphologically within a neuron. The differential anatomical and cellular location of G proteins in the CNS may contribute to the coupling specificity between neurotransmitter receptors and G proteins.