The role of familiarity in three-dimensional view-transferability of face identity adaptation

Recent studies show that face adaptation effects partially transfer across three-dimensional viewpoint change. Here we investigated whether the degree of adaptation transfer is mediated by experience with a face. We manipulated face familiarity and measured identity aftereffects both within- and across-viewpoint. Familiarity enhanced the overall strength of identity adaptation as well as the degree to which adaptation transferred across-viewpoint change. These findings support the idea that transfer effects in adaptation vary as a function of experience with particular faces, and suggest the use of adaptation as a tool for tracking face representations as they develop.     

Coated microneedles for drug delivery to the eye

PURPOSE: To test the hypothesis that coated microneedles can deliver drugs into the eye via intrascleral and intracorneal routes in a minimally invasive manner. METHODS: Solid metal microneedles measuring 500 to 750 microm in length were coated with model drugs, protein, and DNA; inserted into nonpreserved human cadaveric sclera; and imaged. Microneedles coated with sodium fluorescein were then inserted into rabbit cornea in vivo. After needle removal, fluorescein concentration in the anterior segment of the rabbit eye was measured for 24 hours. Similar experiments were performed using pilocarpine-coated microneedles, and the rabbit pupil size was monitored afterward. RESULTS: In vitro insertion tests showed that microneedles were mechanically strong enough to penetrate into human cadaveric sclera and that the drug coating rapidly dissolved off the needles within the scleral tissue within 30 seconds after insertion. In vivo delivery from fluorescein-coated microneedles showed that fluorescein concentrations in the anterior chamber were 60 times greater than those achieved by topical application without microneedles. Similarly, microneedle delivery of pilocarpine caused rapid and extensive rabbit pupil constriction. There were no measurable inflammatory responses caused by microneedle insertion. CONCLUSIONS: This study demonstrated for the first time that coated microneedles can deliver drugs into the eye via intrascleral and intracorneal routes. This minimally invasive approach may avoid the complications associated with intraocular injection and systemic administration.     

Screening of genes associated with termination of the critical period of visual cortex plasticity in rats

PURPOSE: To investigate the molecular mechanism involved in the termination of the critical period of visual cortex plasticity in the rat. METHODS: The rats were divided into two groups, one group was dark-reared for 67 days after birth, while the other group was dark-reared for 60 days and then put under a normal light/dark cycle for 7 days. A subtracted cDNA library was constructed from the visual cortex, and differentially expressed genes were screened by nested PCR, reverse Northern hybridization, sequencing, and homology analysis. RESULTS: Fourteen genes were found to be up-regulated in the visual cortex. These included 13 known genes and a novel fragment (Genbank submission EB174193). Of the known genes, three genes encoding beta -tubulin, myelin basic protein, and cyclophilin were previously reported to be associated with visual cortex plasticity. CONCLUSION: A set of candidate genes related to the termination of the critical period was identified using the subtracted cDNA library. This work provides an important basis for understanding the molecular mechanisms involved in the termination of plasticity in the visual cortex.     

Nuclear factor-kappaB p65 and upregulation of interleukin-6 in retinal ischemia/reperfusion injury in rats

We previously demonstrated that endogenous interleukin-6 (IL-6) is upregulated and may be neuroprotective after retinal ischemia. The purpose of this study is to investigate the role of nuclear factor kappa-B (NF-kappaB) in regulating IL-6 expression after ischemia. NF-kappaB p65 mRNA levels were significantly elevated between 2 and 12 h after the insult. A high number of NF-kappaB p65 positive cells were detected in the inner retina at 12 h after ischemia. Activated nuclear NF-kappaB p65 and IL-6 were colocalized in cells, which were also marked by a microglial/phagocytic cell marker (ED1) in the inner retina. Carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG-132, a proteasome inhibitor, which inhibits IkappaB degradation and hence prevents the activation and translocation of NF-kappaB into the nucleus) abolished the increase in NF-kappaB p65 mRNA levels after the insult, while there was no effect by helenalin (an inhibitor which inhibits NF-kappaB activity by alkylation of the p65 subunit, thereby blocking its binding to the target DNA). However, MG-132 and/or helenalin significantly diminished the increase in IL-6 mRNA levels after the insult. Small interfering RNAs (siRNAs, inhibit target gene expression through the sequence-specific destruction of the target messenger RNA) against NF-kappaB p65 significantly reduced the increase in NF-kappaB p65 mRNA levels as well as IL-6 mRNA levels after ischemia. The number of retinal ganglion cells (RGCs) was also significantly decreased using the inhibitors of NF-kappaB compared with those of the controls after ischemia. These findings support the hypothesis that upregulation of endogenous retinal IL-6 in retinal I/R injury in microglial/phagocytic cells is controlled predominantly by NF-kappaB p65.     

Characterization of cerebral tissue by MRI map ISODATA in embolic stroke in rat

ISODATA using MRI parameter-weighted images has been previously employed to characterize ischemic cell damage after stroke in rats. In an effort to increase the objectivity and to further automate the ISODATA, MRI parameter maps were now employed. Male Wistar rats were subjected to embolic stroke and received treatment via a femoral vein at 4 h post-stroke. The control rats received saline and were sacrificed at 6, 24 and 48 h after stroke, respectively. Treated rats received rtPA alone or were treated with a combination of rtPA and an antibody, 7E3 F(ab′)₂, against the glycoprotein receptor that binds the platelet to fibrin. These rats were sacrificed at 24, or 48, h post-stroke. T₁, T₂ and diffusion maps were employed for map ISODATA analysis. H&E histological analysis of coronal sections of tissue was performed and compared with map ISODATA from the corresponding sections. ISODATA signatures were highly correlated (R ∼ 0.80, P < 0.0001) with the ischemic cell damage analyzed at 6, 24 and 48 h post-stroke. At 24 and 48 h after stroke, ISODATA lesion sizes were highly correlated (R > 0.97, P < 0.001) with lesion sizes measured histologically. The combination treatment of rtPA and 7E3 F(ab′)₂ reduced both infarction size (P < 0.002) and average signature (P < 0.03) at 48 h after stroke, compared to saline-treated animals. No significant difference was found between saline and rtPA-alone-treated rats. The map ISODATA successfully provides objective and automated quantitation of the ischemic damage in both size and severity in an embolic stroke model of rat with and without a therapeutic intervention.     

Over-expression of p73beta results in apoptotic death of post-mitotic hNT neurons

The p53-related p73 protein is an important mediator of apoptosis, development and tumorigenesis. Previously, we showed that over-expression of the p73β isoform induced apoptosis in proliferating neuronal cells; however, the study did not address the effect of p73 in post-mitotic neurons. To address this question, we used post-mitotic hNT neurons, which have been used as a model of human central nervous system neurons. We found that over-expression of p73β in hNT neurons resulted in apoptosis and an increase in the expression of p57^Kip2 and Bax, but no increase in p53 expression. These results suggest that apoptosis of post-mitotic neurons induced by p73β may involve these mediators. Understanding the regulation of p73 expression will be important for understanding the development of the nervous system and may have implications for the treatment of neurological diseases.     

Chromatin immunoprecipitation in postmortem brain

Methylation and other covalent modifications of nucleosome core histones are key regulators of chromatin structure and function, including epigenetic control of gene expression. For the human brain, however, very little is known about the regulation of histone modifications at specific genomic loci. Furthermore, chromatin immunoprecipitation protocols applicable to postmortem tissue are lacking, and the impact of potential confounds such as autolysis time or tissue pH is unknown. We treated cerebral cortex from human postmortem brain and mice by micrococcal nuclease digestion or, alternatively, by formaldehyde-crosslinking and sonication. We show that the bulk of nucleosomal DNA remains attached to histones during the first 30 h after death. Immunoprecipitation with antibodies against methylated histones was at least 10-fold more effective in unfixed, micrococcal nuclease-digested samples, in comparison to extracts prepared by fixation and sonication. Histone methylation differences across various genomic sites were maintained within a wide range of autolysis times and tissue pH. Therefore, immunoprecipitation of micrococcal nuclease-digested tissue extracts is a feasible approach to profile histone methylation at defined genomic loci in postmortem brain.     

Improvement of cationic albumin conjugated pegylated nanoparticles holding NC-1900, a vasopressin fragment analog, in memory deficits induced by scopolamine in mice

NC-1900, an active fragment analog of arginine vasopressin [arginine vasopressin-(4-9)], has proved to be capable of improving the spatial memory deficits and the impairments in passive avoidance test. In this study, a novel drug carrier for brain delivery, cationic bovine serum albumin conjugated pegylated nanoparticles (CBSA-NPs) holding NC-1900, was developed and its improvement on scopolamine-induced memory deficits was investigated in mice using the platform-jumping avoidance test. CBSA-NPs loaded with NC-1900 in spherical shape and uniform size below 100 nm were prepared by the double emulsion/solvent evaporation procedure, and the zeta potential of CBSA-NPs was about -8mV with the loading capacity of NC-1900 around 0.46%. The in vitro study showed that approximately 10% NC-1900 was released from CBSA-NPs in pH 7.4 phosphate buffer saline (PBS) during 56 h incubation with about 15% NC-1900 released in pH 4.0 PBS during 7 days, indicating the sustained release of this carrier. Furthermore, the half-life of NC-1900 loaded in CBSA-NPs in plasma was about 78 h, which was 4-fold longer than that of free NC-1900 (19 h). The active avoidance behavioral results showed that the s.c. administration of NC-1900 tended to improve memory deficits, but the difference did not present any statistical significance, whereas this peptide failed to produce any positive effects by i.v. administration. However, the i.v. injection of CBSA-NPs loaded with NC-1900 greatly improved memory impairments to a normal level, but the efficacy was slight if the loaded nanoparticles (NPs) were exclusive of the conjugation of CBSA, indicating that CBSA-NP was a promising brain delivery carrier for NC-1900 with CBSA as a potent brain targetor. It was concluded that CBSA-NP loaded with NC-1900 was potentially efficacious in the treatment of memory deficits via i.v. administration.     

Correlation between bacteriology and computed tomography staging for chronic sinusitis

In this study we report on the correlation between bacteriology and disease severity staging by computed tomography (CT) for chronic sinusitis. When patients with chronic sinusitis underwent functional endoscopic sinus surgery (FESS), swab specimens were taken from the ipsilateral middle meati and ethmoid sinuses under endoscope guidance. The severity of chronic sinusitis was evaluated by pre-operative CT scans. The CT scans were staged by the Lund and Mackay system. The scores for the frontal, anterior ethmoid and maxillary sinuses and for the ostiomeatal complex were added.The culture rates were correlated with the added scores. Between November 1998 and January 2003, 79 pairs of specimens were collected from 79 patients whose CT scans were done within a day before FESS. The culture rates of middle-meatus specimens were moderately correlated with the scores, but those of ethmoid sinus specimens were negatively correlated with the scores. If Staphylococcus epidermidis and corynebacteria were considered normal flora, the bacteriology of the middle meatus was highly correlated with the CT scores. This study shows that culture rates of middle-meatal specimens tended to increase with the severity of chronic sinusitis.