Layer-specific dendritic regression of pyramidal cells with ageing in the human prefrontal cortex

Huntingtin is the protein product of the gene for Huntington's disease (HD) and carries a polyglutamine repeat that is expanded in HD (>36 units). Huntingtin-associated protein (HAP1) is a neuronal protein and binds to huntingtin in association with the polyglutamine repeat. Like huntingtin, HAP1 has been found to be a cytoplasmic protein associated with membranous organelles, suggesting the existence of a protein complex including HAP1, huntingtin, and other proteins. Using the yeast two-hybrid system, we found that HAP1 also binds to dynactin P150(Glued) (P150), an accessory protein for cytoplasmic dynein that participates in microtubule-dependent retrograde transport of membranous organelles. An in vitro binding assay showed that both huntingtin and P150 selectively bound to a glutathione transferase (GST)-HAP1 fusion protein. An immunoprecipitation assay demonstrated that P150 and huntingtin coprecipitated with HAP1 from rat brain cytosol. Western blot analysis revealed that HAP1 was enriched in rat brain microtubules and comigrated with P150 and huntingtin in sucrose gradients. Immunofluorescence showed that transfected HAP1 colocalized with P150 and huntingtin in human embryonic kidney (HEK) 293 cells. We propose that HAP1, P150, and huntingtin are present in a protein complex that may participate in dynein-dynactin-associated intracellular transport.     

Analyses of mutation and loss of heterozygosity of coding sequences of the entire transforming growth factor beta type II receptor gene in sporadic human gastric cancer

Mutations in the transforming growth factor beta type II receptor (TGFbetaRII) gene have been detected in several human cancer types exhibiting microsatellite instability. Using intron primers previously reported for examination of the entire coding region of the TGFbetaRII gene, 29 sporadic gastric cancers were screened with non-radioactive single strand conformation polymorphism and subsequent DNA sequencing analysis. Mutations of the TGFbetaRII gene were detected in three out of 29 tumors (10%). Two cases showed deletions in a polyadenine tract in both alleles and was positively associated with replication error. One case had an insertion of GA dinucleotide sequence in one allele. Mutations of the TGFbetaRII gene were restricted to exon 3 and other coding regions were not affected. Loss of heterozygosity was detected by analyzing a polymorphic site in intron 2. Three out of nine (33%) informative cases, which were all of intestinal type and advanced cases, showed loss of heterozygosity but neither TGFbetaRII mutation nor replication error was found in these cases. Immunoreactivity of TGFbetaRII in tumor tissues was reduced to a different extent in the gastric cancer with genetically abnormal transforming growth factor. Although the numbers studied are small, homozygous (A)10 deletion or loss of heterozygosity of TGFbetaRII is involved in tumorigenesis and progression of at least some part of sporadic gastric cancer.     

Isolation of a second recombinant human respiratory syncytial virus monoclonal antibody fragment (Fab RSVF2-5) that exhibits therapeutic efficacy in vivo

A second human respiratory syncytial virus (RSV)-neutralizing monoclonal antibody was isolated and its binding site was identified. Fab F2-5 is a broadly reactive fusion (F) protein-specific recombinant Fab generated by antigen selection from a random combinatorial library displayed on the surface of filamentous phage. In an in vitro plaque-reduction test, the Fab RSVF2-5 neutralized the infectivity of a variety of field isolates representing viruses of both RSV subgroups A and B. The Fab recognized an antigenic determinant that differed from the only other human anti-F monoclonal antibody (RSV Fab 19) described thus far. A single dose of 4.0 mg of Fab RSVF2-5/kg of body weight administered by inhalation was sufficient to achieve a 2000-fold reduction in pulmonary virus titer in RSV-infected mice. The antigen-binding domain of Fab RSVF2-5 offers promise as part of a prophylactic regimen for RSV infection in humans.     

Identification of rCop-1, a new member of the CCN protein family, as a negative regulator for cell transformation

By using a model system for cell transformation mediated by the cooperation of the activated H-ras oncogene and the inactivated p53 tumor suppressor gene, rCop-1 was identified by mRNA differential display as a gene whose expression became lost after cell transformation. Homology analysis indicates that rCop-1 belongs to an emerging cysteine-rich growth regulator family called CCN, which includes connective-tissue growth factor, CYR61, CEF10 (v-src inducible), and the product of the nov proto-oncogene. Unlike the other members of the CCN gene family, rCop-1 is not an immediate-early gene, it lacks the conserved C-terminal domain which was shown to confer both growth-stimulating and heparin-binding activities, and its expression is lost in cells transformed by a variety of mechanisms. Ectopic expression of rCop-1 by retroviral gene transfers led to cell death in a transformation-specific manner. These results suggest that rCop-1 represents a new class of CCN family proteins that have functions opposing those of the previously identified members.     

Practical management of treatment-resistant depression

Patients receiving antidepressant monotherapy may be partially or totally resistant to treatment in 10 to 30 percent of cases. In patients who have experienced only partial treatment results, the clinician should first consider optimizing antidepressant dosage or lengthening therapy. Antidepressant drug substitution should generally be reserved for use in patients who haven't responded at all (nonresponders). Combining two or more antidepressants is generally not recommended, as this approach may obscure adequate monotherapy evaluation and lead to significant adverse effects or drug-drug interactions. Use of electroconvulsive therapy is recommended in patients with psychotic and severe refractory depression. Augmentation therapy is often efficacious in patients who exhibit a partial antidepressant response. Lithium and thyroid hormone have been the most extensively studied augmentative agents but, more recently, pindolol and buspirone have also been used for this purpose.     

An assay to measure antibiotic efficacy against Staphylococcus epidermidis biofilms on implant surfaces

Infection is a major limitation of implantable devices. Optimal antibiotic therapeutic regimes have not yet been defined. Implant-associated infections have a number of differentiating characteristics, which include the predominance of Staphylococcus epidermidis and other skin bacteria of normally low pathogenicity as the causative agents, together with a relative resistance to host defenses and to antibiotic therapy. These properties have been ascribed to the ability of the bacteria to exist on implant surfaces in the biofilm phase, which is protective. An assay of antibiotic activity using a standardized bacterial biofilm preparation of S. epidermidis is described. The assay is used to evaluate the relative efficacy of antibiotics to sterilize the biofilm, when they are used singly, or in double or triple combinations. The modulating effects of changing antibiotic concentrations and modifying the environment with CAPD variables (fresh and spent dialysis fluid, common PD solution additives) are also measured and the data summarized. It is hoped that, by using this and similar assays, individualized optimal therapeutic regimes of implant-associated infections may be logically planned.     

Centred contact density functions for the statistical analysis of random sets A stereological study on benign and malignant glandular tissue using image analysis

Real structures investigated in the material and biological sciences, such as minerals or tissues, can often be reduced to two phases. In a stochastic approach, the components of such binary structures may be considered as the union of grains — random sets implanted with their centres at random points — and their complementary space, which is called the pore space. The simplest stochastic germ-grain model is the Boolean model of random sets, which we use here instrumentally as a null model (reference model) for comparison with our biological material. After a brief review of basic properties of the Boolean model and related statistical methods, we introduce centred contact density functions as a new approach. Empirical contact density functions are estimated from the empirical contact distribution functions with an image analyser by dilation of the grain phase. Theoretical contact density functions are then predicted from a set of image parameters, under the assumption that the Boolean model holds. A centred contact density function is the difference between the estimated and the predicted contact density function. Apart from a random error term, centred contact density functions amount to zero irrespective of the area fraction of the grain phase, when the germ-grain model is Boolean. As a section of a spatial Boolean model is a planar Boolean model, the method is also applicable in stereological studies where digitized images are obtained from sections of a three-dimensional structure. Centred contact density functions were determined for mastopathic tissue as compared to mammary cancer, and for tumour-free prostatic tissue as compared to prostatic cancer. For each category of specimens, twenty cases with 10 images each were analysed. Benign and malignant glandular tissue of the aforementioned types deviates significantly from the Boolean model. Centred contact density functions show that malignant transformation is connected with profound geometric remodelling of the pore space.     

Psychopathological and behavioral correlates of dopaminergic sensitivity in alcohol-dependent patients

OBJECTIVES: To explore 2 facets of dopamine receptor sensitivity in alcoholics: (1) whether reduced sensitivity of central dopamine receptors is correlated with anxiety, depression, or novelty seeking and (2) whether this reduction is associated with poor treatment outcome. METHOD: Sixty-four alcohol-dependent patients were assessed according to their clinical outcome, sensitivity of central dopamine receptors (apomorphine-induced growth hormone secretion), mood states, and personality traits before and after detoxification. RESULTS: Patients with poor treatment outcome displayed a blunted growth hormone response before, but not after, detoxification. Growth hormone response was not significantly correlated with novelty seeking. Relapsing patients tended to be less depressed than patients who remained abstinent during observation. CONCLUSION: This study did not support the hypothesis that reduced sensitivity of dopamine receptors is associated with anxiety, depressed mood, or high novelty seeking in alcoholism.     

An improved shift-invariant artificial neural network for computerized detection of clustered microcalcifications in digital mammograms

A shift-invariant artificial neutral network (SIANN) has been applied to eliminate the false-positive detections reported by a rule-based computer aided-diagnosis (CAD) scheme developed in our laboratory. Regions of interest (ROIs) were selected around the centers of the rule-based CAD detections and analyzed by the SIANN. In our previous study, background-trend correction and pixel-value normalization were used as the preprocessing of the ROIs prior to the SIANN. A ROI is classified as a positive ROI, if the total number of microcalcifications detected in the ROI is greater than a certain number. In this study, modifications were made to improve the performance of the SIANN. First, the preprocessing is removed because the result of the background-trend correction is affected by the size of ROIs. Second, image-feature analysis is employed to the output of the SIANN in an effort to eliminate some of the false detections by the SIANN. In order to train the SIANN to detect microcalcifications and also to extract image features of microcalcifications, the zero-mean-weight constraint and training-free-zone techniques have been developed. A cross-validation training method was also applied to avoid the overtraining problem. The performance of the SIANN was evaluated by means of ROC analysis using a database of 39 mammograms for training and 50 different mammograms for testing. The analysis yielded an average area under the ROC curve (A(z)) of 0.90 for the testing set. Approximately 62% of false-positive clusters detected by the rule-based scheme were eliminated without any loss of the true-positive clusters by using the improved SIANN with image feature analysis techniques.     

Narrow complex tachycardia with VA block: diagnostic and therapeutic implications

To review our experience with cases of narrow complex tachycardia with VA block, highlighting the difficulties in the differential diagnosis, and the therapeutic implications. Prior reports of patients with narrow complex tachycardia with VA block consist of isolated case reports. The differential diagnosis of this disorder includes: automatic junctional tachycardia, AV nodal reentry with final upper common pathway block, concealed nodofascicular (ventricular) pathway, and intra-Hissian reentry. Between June 1994 and January 1996, six patients with narrow complex tachycardia with episodes of ventriculoatrial block were referred for evaluation. All six patients underwent attempted radiofrequency ablation of the putative arrhythmic site. Three of six patients had evidence suggestive of a nodofascicular tract. Intermittent antegrade conduction over a left-sided nodofascicular tract was present in two patients and the diagnosis of a concealed nodofascicular was made in the third patient after ruling out other tachycardia mechanisms. Two patients had automatic junctional tachycardia, and one patient had atrioventricular nodal reentry with proximal common pathway block. Attempted ablation in the posterior and mid-septum was unsuccessful in patients with nodofascicular tachycardia. In contrast, those with atrioventricular nodal reentry and automatic junctional tachycardia readily responded to ablation. The presence of a nodofascicular tachycardia should be suspected if: (1) intermittent antegrade preexcitation is recorded, (2) the tachycardia can be initiated with a single atrial premature producing two ventricular complexes, and (3) a single ventricular extrastimulus initiates SVT without a retrograde His deflection. The presence of a nodofascicular pathway is common in patients with narrow complex tachycardia and VA block. Unlike AV nodal reentry and automatic junctional tachycardia, the response to ablation is poor.