CD70 (TNFSF7) is expressed at high prevalence in renal cell carcinomas and is rapidly internalised on antibody binding.

In order to identify potential markers of renal cancer, the plasma membrane protein content of renal cell carcinoma (RCC)-derived cell lines was annotated using a proteomics process. One unusual protein identified at high levels in A498 and 786-O cells was CD70 (TNFSF7), a type II transmembrane receptor normally expressed on a subset of B, T and NK cells, where it plays a costimulatory role in immune cell activation. Immunohistochemical analysis of CD70 expression in multiple carcinoma types demonstrated strong CD70 staining in RCC tissues. Metastatic tissues from eight of 11 patients with clear cell RCC were positive for CD70 expression. Immunocytochemical analysis demonstrated that binding of an anti-CD70 antibody to CD70 endogenously expressed on the surface of A498 and 786-O cell lines resulted in the rapid internalisation of the antibody-receptor complex. Coincubation of the internalising anti-CD70 antibody with a saporin-conjugated secondary antibody before addition to A498 cells resulted in 50% cell killing. These data indicate that CD70 represents a potential target antigen for toxin-conjugated therapeutic antibody treatment of RCC.

Proteomic analysis of the cell-surface membrane in chronic lymphocytic leukemia: identification of two novel proteins, BCNP1 and MIG2B.

B-cell-specific plasma-membrane proteins are potential targets for either small molecule or antibody-based therapies. We have sought to annotate proteins expressed at the cell surface membrane in patients with chronic lymphocytic leukemia (CLL) using plasma-membrane-based proteomic analysis to identify previously uncharacterized and potentially B-cell-specific proteins. Proteins from plasma-membrane fractions were separated on one-dimensional gels and trypsinized fractions subjected to high-throughput MALDI-TOF mass spectrometry. Using this method, many known B-cell surface antigens were detected, but also known proteins not previously described in this disease or in this cellular compartment, including cell surface receptors, membrane-associated enzymes and secreted proteins, and completely unknown proteins. To validate the method, we show that BLK, a B-cell-specific kinase, is located in the CLL-plasma-membrane fraction. We also describe two novel proteins (MIG2B and B-cell novel protein #1, BCNP1), which are expressed preferentially in B cells. MIG2B is in a highly conserved and defined gene family containing two plasma-membrane-binding ezrin/radixin/moesin domains and a pleckstrin homology domain; the Caenorhabditis elegans homolog (UNC-112) is a membrane-associated protein that colocalizes with integrin at cell-matrix adhesion complexes. BCNP1 is a completely unknown protein with three predicted transmembrane domains, with three alternatively spliced final exons. Proteomic analysis may thus define new potential therapeutic targets.

Cloning and expression profiling of Hpa2, a novel mammalian heparanase family member.

Heparan sulfate proteoglycans are important constituents of the extracellular matrix and basement membrane. Cleavage of heparan sulfate by heparanase, an endoglycosidase, is implicated in the extravasation of leukocytes and metastatic tumour cells, identifying this enzyme(s) as a target for anti-inflammatory and anti-metastatic therapies. The cloning of a cDNA encoding human heparanase (Hpa1) was reported recently, together with evidence indicating that the hpa1 gene is unique and unlikely to belong to a family of related genes. Here we report the cloning of a cDNA encoding a novel human protein, HPA2, with significant homology to Hpa1. Alternative splicing of the hpa2 transcript yields three different mRNAs, encoding putative proteins of 480, 534, and 592 amino acids. Sequence analyses predict that all three Hpa2 proteins are intracellular, membrane-bound enzymes. Hpa2 also shows a markedly different mRNA distribution to Hpa1 in both normal and cancer tissues. The difference in expression profiles and predicted cellular locations suggests that Hpa2 and Hpa1 proteins have distinct biological functions.

Evaluating architectural design review.

Architectural design review is a method of environmental management which is widely used by governmental agencies in both the United States and in Great Britain. Because design review is a governmental function, there is a major need to assess how well it works. Research covering over 29,000 respondents and 5,600 environmental scenes suggests that scientific protocols can be adapted to provide an accurate and efficient design review protocol. The protocol uses preference experiments to find the standardized mean difference [formula: see text] between a proposed project and a random sample of existing projects. Values of d will indicate whether the project will increase, maintain, or diminish the aesthetic merit of the sampled area. The protocol is illustrated by applying it to the case of design review for a single residence. Implications for further implementations are discussed.

Sex, complexity, and preferences for residential facades.

Analysis of variance was used to find out how much variance in preference for residential facades could be attributed to places or people. In accordance with previous findings based on other stimuli, places accounted for more preference variance than did people (16% vs 0%). More focused results were obtained for the subjective factor of sex and the objective design features of number of turns in a silhouette and the percentage of a facade covered by small visual elements. Preferences across sex correlated at .98, indicating that this demographic distinction had a very small effect. The objective factor of visual complexity had a multiple correlation of .90 with preference, indicating a strong effect. These results indicate that design features were much more effective than subjective factors in predicting preferences for environments. Moreover, this finding was reproducible over different samples of people and places.

Effect sizes of mountains and molehills.

Decisions about environmental aesthetics require valid, understandable measurements of visual impacts. This study reports well known scientific measurements (standardized mean contrasts) for well known images (mountains and molehills). The scientific measurements of visual preferences for mountains and molehills were 1.17 and -0.05, respectively.

Complexity of architectural silhouettes: from vague impressions to definite design features.

Nine 2-dimensional shapes were created that varied in four geometric factors: number of vertexes, symmetry, variation of lengths of line segments, and variation of angles. The shapes were evaluated in a paired-comparison experiment on the criterion of perceived complexity. Analysis of variance suggested that the most important predictor of shape complexity was the number of vertexes. P2/A (perimeter squared divided by the area) was also strongly correlated with judged complexity. Symmetry was a significant but weak predictor of complexity.

Of time and preference: temporal stability of environmental preferences.

Time is a central issue in discussions about art and in Berlyne's aesthetic theory. This article reports on the temporal stabilities of preferences for a novel and controversial building at three times after construction (2 years, 18 years, 23 years), and public preferences for 20 ordinary and noncontroversial buildings at three times over nine years. In all there were 5543 respondents. Analyses suggested that the initial response to the novel building was stable over the next 23 years, and the public responses for the 20 nonnovel buildings were stable over nine years. Implications for research are discussed.

A human cDNA sequence with homology to non-mammalian lysophosphatidic acid acyltransferases.

A novel human homologue of Escherichia coli, yeast and plant 1-acylglycerol-3-phosphate acyltransferase has been isolated from U937 cell cDNA. Expression of the cloned sequence in 1-acylglycerol-3-phosphate acyltransferase-deficient E. coli resulted in increased incorporation of oleic acid into cellular phospholipids. Membranes made from COS7 cells transfected with the cDNA exhibited higher acyltransferase activity towards a range of donor fatty acyl-CoAs and lysophosphatidic acid. Northern-blot analysis of the cDNA sequence indicated high levels of expression in immune cells and epithelium. Rapid amplification of cDNA ends revealed differentially expressed splice variants, which suggests regulation of the enzyme by alternative splicing. This cDNA therefore represents the first described sequence of a mammalian gene homologous to non-mammalian lysophosphatidic acid acyltransferases.

Analysis of proviral integration in human mammary epithelial cell lines immortalized by retroviral infection with a temperature-sensitive SV40 T-antigen construct.

A panel of eight conditionally immortal lines derived by infection of human breast epithelial cells with an amphotropic retrovirus transducing a ts mutant of SV40 large T-antigen was analyzed with respect to individual retroviral integration patterns. Each line contained multiple integration sites which were clonal and stable over extended passage. Similar integration patterns were observed between individual lines arising separately from the same stock of pre-immortal cells, suggesting a common progenitor. Retroviral integration analysis of pre-immortal cells at different stages of pre-crisis growth showed changes indicative of a progressive transition from polyclonality to clonality as the cells approached crisis. Each of the immortal lines contained a sub-set of the integration sites of their pre-immortal progenitors, with individual combinations and copy numbers of sites. Since all the cell lines appeared to originate from single foci in separate flasks, it is likely that each set arose from a common clone of pre-immortal cells as the result of separate genetic events. There was no evidence from this analysis to suggest that specific integration sites played any part either in the selection of pre-crisis clones or in the subsequent establishment of immortal lines.

Postconstruction validation of photomontage simulations.

Validation of simulation media is of concern to many environmental planners and researchers. Using results from an actual architectural competition, this article correlated preferences obtained from various preconstruction media with preferences obtained from as-built photographs. The photomontage medium performed the best with a correlation of .94.

Expression of p53 in premalignant and malignant squamous epithelium.

Using three antibodies (JG8, CM-1 and 1081) directed to the p53 protein, strong positivity was found in 16/47 (34.0%) of mucosal squamous cell carcinomas of the head and neck and in two squamous carcinoma cell lines (LICR-LON- HN5 and HN6Rr). The presence of the mutant p53 was confirmed in the cell lines as substitutions in exon 7 (codon 238, TGT greater than AGT) and exon 5 (codon 152, CCG greater than CTG) respectively. Positive staining was seen only in the undifferentiated cells and progressively lost as the cells keratinized, both in the tumour specimens and in the cell lines. Similar results were seen in areas of dysplasia, well removed from the site of the primary tumour. Staining of epidermal lesions showed positivity in 2/12 (16.6%) cases of Bowen's disease, 0/12 (0.0%) cases of solar keratosis, 0/10 (0.0%) basal cell carcinomas and in 3/20 (15.0%) squamous cell carcinomas. These results are discussed in relation to the multifocal origin of squamous cell carcinomas, the role of p53 mutations in squamous cell carcinomas from different sites and the significance of the 'basal' distribution of p53 as a normal growth regulator. The possible significance of the distribution of p53 in squamous epithelium as it relates to papilloma virus infection is also considered.

Hepatitis B virus-associated hepatocellular carcinoma in African patients.

We have examined several tumors from South African patients with hepatitis B virus (HBV)-associated hepatocellular carcinoma for the presence of integrated viral DNA. In contrast with our findings in patients from Taiwan, few copies of the viral genome were integrated in each tumor. Furthermore, Southern hybridization showed similarities in integration patterns between different tumors. We are presently constructing genomic libraries from selected single-copy tumors in order to make a more detailed analysis of HBV DNA integration at the molecular level.

A new method for teaching architectural studios: use of objective feedback to help design students refine their capacity for self-evaluation.

The literature suggests that there is concern about the traditional method of teaching architecture. This article describes an alternate method of teaching design studios in which scientific assessments are used instead of the traditional architectural injuries. Analysis suggests that the scientific method of teaching design leads to improvements in the students' abilities to predict how other students would respond to their projects.

Mapping of two novel transcripts of bovine papillomavirus type 4.

A cDNA library was synthesized using RNA from bovine papillomavirus type 4 (BPV-4)-induced papillomas. The major viral transcript was characterized by sequencing of its cDNA, primer extension mapping and S1 nuclease protection studies. The transcript initiates at multiple sites between nucleotide (nt) 777 and nt 902, contains a splice junction between nt 1016 and nt 3376 and terminates at nt 4034, using the early polyadenylation signal at nt 4009. Sequencing of viral cDNAs and mRNA revealed deviations from the reported genomic sequence between nt 3412 and nt 3460. These resulted in a temporary frameshift, abolished the translation termination codon of the E5a open reading frame (ORF), and introduced a new termination codon in the E4 ORF. The new uninterrupted E5 ORF was found to have greater homology to the E4 ORFs of other papillomaviruses than to their E5 ORFs. The E5 ORF of BPV-4 is joined to a five codon ORF in the leader exon of the major transcript, in the same way as the E4 ORF in the major transcripts of BPV-1 and human papillomavirus type 11. On this basis, it has been redesignated E4, and the viral genomic sequence revised. Two novel transcriptional promoters of BPV-4 were defined: a putative controller of the multiple major RNA start sites around nt 870 and a minor TATA box at nt 691. In addition, minor early region transcripts were mapped which initiate between nt 3071 and nt 3152. None of these transcripts utilizes the splice site at nt 3376. These messages may express E2-encoded functions.