Sidelobe Suppression Techniques for Near-Field Multistatic SAR.

Multirotor Unmanned Air Systems (UAS) represent a significant improvement in capability for Synthetic Aperture Radar (SAR) imaging when compared to traditional, fixed-wing, platforms. In particular, a swarm of UAS can generate significant measurement diversity through variation of spatial and frequency collections across an array of sensors. In such imaging schemes, the image formation step is challenging due to strong extended sidelobe; however, were this to be effectively managed, a dramatic increase in image quality is theoretically possible. Since 2015, QinetiQ have developed the RIBI system, which uses multiple UAS to perform short-range multistatic collections, and this requires novel near-field processing to mitigate the high sidelobes observed and form actionable imagery. This paper applies a number of algorithms to assess image reconstruction of simulated near-field multistatic SAR with an aim to suppress sidelobes observed in the RIBI system, investigating techniques including traditional SAR processing, regularised linear regression, compressive sensing. In these simulations presented, Elastic net, Orthogonal Matched Pursuit, and Iterative Hard Thresholding all show the ability to suppress sidelobes while preserving accuracy of scatterer RCS. This has also lead to a novel processing approach for reconstructing SAR images based on the observed Elastic net and Iterative Hard Thresholding performance, mitigating weaknesses to generate an improved combined approach. The relative strengths and weaknesses of the algorithms are discussed, as well as their application to more complex real-world imagery.

Preparation for the next major incident: are we ready? Comparing major trauma centres and other hospitals.

OBJECTIVES: A major incident is any emergency requiring special arrangements by the emergency services. All hospitals are required by law to keep a major incident plan (MIP) detailing the response to such events. In 2006 and 2019, we assessed the preparedness and knowledge of key individuals in hospitals across England and found a substantial gap in responding to the MIP. In this report, we compare responses from doctors at major trauma centres (MTCs) and other hospitals (non-MTCs). METHODS: We identified trusts in England that received over 30 000 patients through the ED in the fourth quarter of 2016/2017. We contacted the on-call anaesthetic, emergency, general surgery and trauma and orthopaedic registrar at each location and asked three questions assessing their confidence in using their hospital's MIP: (1) Have you read your hospital's MIP? (2) Do you know where you can access your hospital's MIP guidelines? (3) Do you know what role you would play if an MIP came into effect while you are on call?We compared data from MTCs and non-MTCs using multinomial mixed proportional odds models. RESULTS: There was a modest difference between responses from individuals at MTCs and non-MTCs for question 2 (OR=2.43, CI=1.03 to 5.73, p=0.04) but no evidence of a difference between question 1 (OR=1.41, CI=0.55 to 3.63, p=0.47) and question 3 (OR=1.78, CI=0.86 to 3.69, p=0.12). Emergency medicine and anaesthetic registrars showed significantly higher preparedness and knowledge across all domains. No evidence of a systematic difference in specialty response by MTC or otherwise was identified. CONCLUSIONS: Confidence in using MIPs among specialty registrars in England remains low. Doctors at MTCs tended to be better prepared and more knowledgeable, but this effect was only marginally significant. We make several recommendations to improve education on major incidents.

Preparation for the next major incident: are we ready? A 12-year update.

OBJECTIVES: A major incident is any emergency that requires special arrangements by the emergency services and generally involves a large number of people. Recent such events in England have included the Manchester Arena bombing and the Grenfell Tower disaster. Hospitals are required by law to keep a major incident plan (MIP) outlining the response to such an event. In a survey conducted in 2006 we found a substantial knowledge gap among key individuals that would be expected to respond to the enactment of an MIP. We set out to repeat this survey study and assess any improvement since our original report. METHODS: We identified NHS trusts in England that received more than 30 000 patients through the emergency department in the fourth quarter of the 2016/2017 period. We contacted the on-call anaesthetic, emergency, general surgery, and trauma and orthopaedic registrar at each location and asked each individual to answer a short verbal survey assessing their confidence in using their hospital's MIP. RESULTS: Of those eligible for the study, 62% were able to be contacted and consented to the study. In total 50% of respondents had read all or part of their hospital's MIP, 46.8% were confident that they knew where their plan was stored, and 36% knew the role they would play if a plan came into effect. These results show less confidence among middle-grade doctors compared with 2006. CONCLUSIONS: Confidence in using MIPs among specialty registrars in England is still low. In light of this, we make a number of recommendations designed to improve the education of hospital doctors in reacting to major incidents.

Micro-CT scouting for transmission electron microscopy of human tissue specimens.

Transmission electron microscopy (TEM) provides sub-nanometre-scale details in volumetric samples. Samples such as pathology tissue specimens are often stained with a metal element to enhance contrast, which makes them opaque to optical microscopes. As a result, it can be a lengthy procedure to find the region of interest inside a sample through sectioning. We describe micro-CT scouting for TEM that allows noninvasive identification of regions of interest within a block sample to guide the sectioning step. In a tissue pathology study, a bench-top micro-CT scanner with 10 µm resolution was used to determine the location of patches of the mucous membrane in osmium-stained human nasal scraping samples. Once the regions of interest were located, the sample block was sectioned to expose that location, followed by ultra-thin sectioning and TEM to inspect the internal structure of the cilia of the membrane epithelial cells with nanometre resolution. This method substantially reduced the time and labour of the search process from typically 20 sections for light microscopy to three sections with no added sample preparation.

Comparison between a fixed-dose contrast protocol and a weight-based contrast dosing protocol in abdominal CT.

AIM: To compare a fixed-dose intravenous iodinated contrast medium protocol with weight-based dosing protocols for abdominal computed tomography (CT). MATERIALS AND METHODS: Fifty patients were scanned using a fixed-dose protocol, 50 patients were scanned using a full-dose weight-based contrast dosing protocol, and 13 patients were scanned using a reduced dose weight-based protocol. Radiodensity was measured at the portal vein, aorta, spleen, and liver. These values were plotted against contrast medium dose per unit weight. Images from all patients were anonymised and presented to two independent consultants who subjectively assessed contrast enhancement using a five-point Likert scale. RESULTS: Using a fixed-dose protocol, there was a statistically significant negative correlation and trend between patient weight and radiodensity at the portal vein, aorta, spleen, and liver. Using a full-dose weight-based contrast dosing protocol, there was no longer a statistically significant correlation or trend implying a more consistent degree of enhancement over a spectrum of patient weights. In addition, when the full-dose weight-based contrast dosing protocol was used, there was a statistically significant increase in the number of scans subjectively assessed as having ideal enhancement and a statistically significant decrease in the number of scans felt to have excessive enhancement when compared to a fixed-dose protocol. The weight-based dosing protocol used less contrast medium than the fixed-dose protocol and there was no evidence of contrast-induced acute kidney injury (CIAKI) in any of the patients that received a greater dose than that which they would have received using a fixed-dose protocol. The reduced-dose weight-based protocol showed less objective enhancement of the portal vein, abdominal aorta, spleen, and liver compared to the full-dose protocol and a reduction in the number of scans perceived as showing ideal enhancement. There was, however, no increase in the number of scans with poor or non-diagnostic enhancement. CONCLUSION: Weight-based contrast medium dosing has been shown to objectively provide more consistent vessel and solid-organ enhancement and subjectively improve image quality across a spectrum of weights. Depending on mean patient mass, it has also been shown to reduce overall contrast medium dose, and there is no evidence of CIAKI in patients that receive larger doses. This study also postulates that a standardised approach to contrast medium dose reduction in patients with renal impairment may be a viable strategy.

Xenon treatment protects against cold ischemia associated delayed graft function and prolongs graft survival in rats.

Prolonged hypothermic storage causes ischemia-reperfusion injury (IRI) in the renal graft, which is considered to contribute to the occurrence of the delayed graft function (DGF) and chronic graft failure. Strategies are required to protect the graft and to prolong renal graft survival. We demonstrated that xenon exposure to human proximal tubular cells (HK-2) led to activation of range of protective proteins. Xenon treatment prior to or after hypothermia-hypoxia challenge stabilized the HK-2 cellular structure, diminished cytoplasmic translocation of high-mobility group box (HMGB) 1 and suppressed NF-κB activation. In the syngeneic Lewis-to-Lewis rat model of kidney transplantation, xenon exposure to donors before graft retrieval or to recipients after engraftment decreased caspase-3 expression, localized HMGB-1 within nuclei and prevented TLR-4/NF-κB activation in tubular cells; serum pro-inflammatory cytokines IL-1ß, IL-6 and TNF-α were reduced and renal function was preserved. Xenon treatment of graft donors or of recipients prolonged renal graft survival following IRI in both Lewis-to-Lewis isografts and Fischer-to-Lewis allografts. Xenon induced cell survival or graft functional recovery was abolished by HIF-1α siRNA. Our data suggest that xenon treatment attenuates DGF and enhances graft survival. This approach could be translated into clinical practice leading to a considerable improvement in long-term graft survival.

Vulvar and vestibulovaginal hemangiosarcoma in a cow: morphological and histopathological observations.

Background: Hemangiosarcoma is defined as the malignant mesenchymal neoplasm of endothelial cells. It is a common tumor affecting dogs and is very rare in cattle. Case description: A seven-year-old three months pregnant female Holstein Friesian cross-breed cow was presented with a history of a proliferating irregular dark red friable mass in the vulvar region for the past month. Findings/treatment and outcome: The surface of the mass had diffuse ecchymotic hemorrhages. Histopathological examination of the tissue biopsy specimen revealed unencapsulated, infiltrating neoplasm composed of numerous vascular channels with irregular borders surrounded by endothelial cells of variable sizes and shapes supported by a scanty fibrovascular stroma. Based on morphological and histopathological findings, the case was diagnosed as vulvar and vestibulovaginal hemangiosarcoma. The animal was culled due to the poor prognosis. Conclusion: To the best of our knowledge, this is the first report of vulvar and vestibulovaginal hemangiosarcoma in a cow.

Satisfaction and genital perception after orchiectomy for prostate cancer: does the technique matter? A randomized trial.

OBJECTIVES: Bilateral extracapsular or total orchiectomy (BEO) for prostate cancer is presumed to have psychological consequences after the surgery due to perception of an empty scrotum. Bilateral subcapsular orchiectomy (BSO) was designed to preserve perception of palpable testes. We compared the patients' satisfaction and genital perception following BEO and BSO. MATERIALS AND METHODS: Prostate cancer patients eligible for androgen deprivation therapy who opted for orchiectomy were enrolled in prospective randomized study. Patients with bleeding disorder or uncorrected coagulopathy, poor performance score, and psychiatric problems were excluded. Outlook to life and own health in-general, overall satisfaction to the procedure and genital perception was evaluated using modified Fugl-Meyer questionnaire (FMQ) which was administered before and after 3 months of the surgery. Patients were randomized to BEO and BSO groups at the time of surgery using block randomization. Primary outcome was to compare the genital perception of testicular loss and patients' satisfaction to BSO and BEO. Secondary outcomes included testosterone and PSA control, operative time, and complications. RESULTS: Total 35 patients were enrolled in each group which was comparable. There was no difference in PSA control at 3 months. Mean operative time and blood loss were significantly lesser in BEO group. FMQ score at 3 months did not show significant difference. Majority of the patients in both groups were satisfied with procedure and the aesthetic value of scrotum after surgery. However, 84% in BSO group did not feel that testes were removed on self-examination, as compared to 28% in BEO group. Majority patients in both groups did not report physical or psychological discomfort from change in scrotal content. CONCLUSIONS: Results showed that patients' satisfaction and genital perception following BSO and BEO were similar. Feeling of remaining intrascrotal contents after BSO did not had added psychological advantage in terms of perception of genitalia.

Comparative assessment of faecal diagnostics for detection and predictive modelling of endemic Fasciola hepatica infection in sheep and cattle on Australian farms.

The diagnosis, monitoring and flukicide efficacy testing of fasciolosis on-farm is reliant on non-terminal methods. The coproantigen ELISA (cELISA) has been recommended for diagnosis of fasciolosis and associated flukicide efficacy testing as an alternative to fluke egg counts for monitoring parasitism. Recently experimental multi-age infections have suggested that the reliability of efficacy results can be improved by a second cELISA testing at 6 weeks post-treatment (wpt) in addition to the generally accepted 1 wpt. A field study was conducted to determine the suitability of faecal fluke egg counts (FFEC) and cELISA as diagnostic, drug efficacy testing and epidemiological tools on Australian sheep and cattle farms. Faecal samples from sheep and/or cattle on three endemic farms were taken at monthly intervals for 12 months and examined by both methods. Normal farm management was maintained during the study period and opportunistic efficacy testing, in line with each farm's normal flukicide management was undertaken. Additionally, the suitability of the Ollerenshaw Index as a predictive model for fasciolosis under Australian conditions was examined. While both diagnostics demonstrated their value in the farm environment, the current data demonstrate a distinct and significant increase in diagnostic sensitivity for epidemiological studies by using the two tests in parallel. The agreement between the two diagnostics was found to be higher in cattle, despite the poor sensitivity of FFEC in this species. Similar levels of agreement between the two tests were demonstrated at both sheep properties, regardless of the marked difference in the intensity of F. hepatica challenge. Linear regression models demonstrated the results of the two diagnostics utilized in parallel were explained substantially (R2 = 0.91) as were series data (R2 = 0.88) when the respective models were fitted. In contrast, the fitted models for FFEC (R2 = 0.54) and cELISA (R2 = 0.58) were poor explanations for test outcomes. The outcomes of these models support previous findings that suggest that the two diagnostic tests are best utilized together, particularly in parallel. The application of the Ollerenshaw Index to Australian conditions requires further investigation.

Comparative assessment of faecal diagnostics for detection and predictive modelling of endemic Fasciola hepatica infection in sheep and cattle on Australian farms.

The diagnosis, monitoring and flukicide efficacy testing of fasciolosis on-farm is reliant on non-terminal methods. The coproantigen ELISA (cELISA) has been recommended for diagnosis of fasciolosis and associated flukicide efficacy testing as an alternative to fluke egg counts for monitoring parasitism. Recently experimental multi-age infections have suggested that the reliability of efficacy results can be improved by a second cELISA testing at 6 weeks post-treatment (wpt) in addition to the generally accepted 1 wpt. A field study was conducted to determine the suitability of faecal fluke egg counts (FFEC) and cELISA as diagnostic, drug efficacy testing and epidemiological tools on Australian sheep and cattle farms. Faecal samples from sheep and/or cattle on three endemic farms were taken at monthly intervals for 12 months and examined by both methods. Normal farm management was maintained during the study period and opportunistic efficacy testing, in line with each farm's normal flukicide management was undertaken. Additionally, the suitability of the Ollerenshaw Index as a predictive model for fasciolosis under Australian conditions was examined. While both diagnostics demonstrated their value in the farm environment, the current data demonstrate a distinct and significant increase in diagnostic sensitivity for epidemiological studies by using the two tests in parallel. The agreement between the two diagnostics was found to be higher in cattle, despite the poor sensitivity of FFEC in this species. Similar levels of agreement between the two tests were demonstrated at both sheep properties, regardless of the marked difference in the intensity of F. hepatica challenge. Linear regression models demonstrated the results of the two diagnostics utilized in parallel were explained substantially (R2 = 0.91) as were series data (R2 = 0.88) when the respective models were fitted. In contrast, the fitted models for FFEC (R2 = 0.54) and cELISA (R2 = 0.58) were poor explanations for test outcomes. The outcomes of these models support previous findings that suggest that the two diagnostic tests are best utilized together, particularly in parallel. The application of the Ollerenshaw Index to Australian conditions requires further investigation.

A rapid method for labelling CD4+ T cells with ultrasmall paramagnetic iron oxide nanoparticles for magnetic resonance imaging that preserves proliferative, regulatory and migratory behaviour in vitro.

A number of techniques have been developed to track the migration of T cells in vivo, but they all suffer significant shortcomings, including the examination of selected organs rather than the organism as a whole--thus precluding longitudinal studies--or limitations imposed by poor spatial resolution and the application of ionizing radiation. By conjugating the HIV tat peptide to ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles in a reaction yielding a mean valence of 45, a magnetic resonance (MR) contrast agent was synthesised that allowed T cells to be efficiently labelled within just 5 min. The USPIO nanoparticles were incorporated into both the cytoplasm and nucleus of labelled cells, which retained normal in vitro proliferative responses to a polyclonal stimulus; suppressive responses mediated by labelled CD4(+) CD25(+) regulatory T cells; chemotactic responses to the chemokine CXCL-12; and transmigration of an activated endothelial monolayer. We believe that this rapid, efficient and essentially non-toxic approach to labelling both murine and human T cells for MRI holds considerable promise, paving the way for the wider immunological application of this exciting technology.

Source of oncofetal ED-B-containing fibronectin: implications of production by both tumor and endothelial cells.

ED-B fibronectin (FN) is a FN isoform derived from alternative splicing of the primary transcript of a single gene. Its expression on tumor stroma and neoformed tumor vasculature and its absence, with few exceptions, in normal adult tissues imply a prognostic and diagnostic value for ED-B FN. We investigated the location and source of ED-B FN because this will be of importance both in understanding its role in tumor development and in designing strategies to target this molecule. We have confirmed that ED-B FN is expressed in the majority of breast and colorectal carcinoma tissue samples, with strong immunohistochemical staining around the tumor cells and in the tumor stroma. No staining of tumor neovasculature was seen. ED-B FN is produced by a range of tumor and endothelial (both primary and transformed) cell lines, as detected by reverse transcription-PCR, but is not expressed at the plasma membrane. Strong expression of human ED-B FN is seen in tumor xenografts. These data indicate that neoplastic cells can act as the source of ED-B FN in tumors. The lack of cell surface expression on tumor cell lines has clear implications for the design of therapeutic strategies which target this molecule.

Characterisation of a light chain loss variant of the BCL1 lymphoma.

Vaccination of BALB/c mice with idiotypic (id) IgM derived from the murine B cell lymphoma BCL1, protects the animals from challenge with tumour cells. Escape of the tumour cells from immune control is associated with the selection of variant cells which fail to express significant levels of id IgM on their cell surface. We have previously isolated one such variant, SNAG 1, and shown that, while it expresses less than 10% of the levels of surface IgM of the parental BCL1 lymphoma, it continues to synthesise id material which can be detected within the cell. In this report we present a detailed characterisation of this variant and show that the tumour cells no longer synthesise the lambda light chain. This failure to produce the light chain causes the mu heavy chains in SNAG 1 to remain marooned in the endoplasmic reticulum. The mu heavy chains in SNAG 1 have a normal mol. wt and isoelectric point, and so appear not to be mutated. This is unlike the vast majority of light chain loss variants, in which the heavy chains have been shown to contain deletions. Investigation of the mechanisms responsible for the loss of light chain synthesis demonstrated that, while mRNA for the light chain is present, and of a normal size, there was no production of light chain protein in a cell free system. This indicates that the failure to express light chain by SNAG 1 cells is due to an inability to translate the light chain mRNA into the detectable levels of lambda light chain protein.

Characterization of a new monoclonal anti-Fc gamma RII antibody, AT10, and its incorporation into a bispecific F(ab')2 derivative for recruitment of cytotoxic effectors.

Fc gamma RII (CDw32) on monocytes is capable of triggering both phagocytosis and lysis of chick red blood cells (CRBC) coated with antibody of the appropriate isotype. In this report we describe the production and characterization of a mouse monoclonal IgG1 antibody specific for Fc gamma RII and compare its activity in binding studies, tissue distribution and redirected cellular cytotoxicity (RCC), with the previously identified anti-Fc gamma RII antibodies KB61 and IV.3. Immunohistochemical and flow cytometry analyses demonstrated that AT10 binds very strongly to Fc gamma RII on normal monocytes, but only weakly to that expressed on lymphocytes. This pattern does not correspond to the staining seen with either KB61 or IV.3, and appears to give an intermediate profile. The binding constant (Ka) for the Fab' fragment of AT10 was calculated at 5.3 x 10(8) M-1, four times higher than that for KB61 (1.4 x 10(8) M-1). Bispecific F(ab')2 antibodies were constructed from Fab' fragments of AT10 or KB61 thioether-linked to Fab' from an anti-CRBC monoclonal antibody. These bispecific derivatives directed monocyte cytotoxicity against CRBC as efficiently as either a monoclonal or polyclonal anti-chick erythrocyte antibody. The bispecific F(ab')2 antibodies had a distinct advantage over the conventional reagents, in that they were not blocked in the presence of human Fc gamma at 3.5 mg/ml (a concentration comparable with that provided by IgG in serum). Therefore, bispecific derivatives constructed with the high affinity anti-Fc gamma RII antibody, AT10, may be used as therapeutic reagents for targeting tumour cell lysis in vivo.

Ileostomy obstruction by ingested apricot stone with clinical-radiological-pathological correlation.

Patients with stomas often present with bowel obstruction, often secondary to adhesions. This case describes the presentation, investigation and management of a 62-year-old woman with an end ileostomy, who presented to hospital with acute abdominal pain and subacute bowel obstruction. Further questioning revealed the recent ingestion of an apricot stone and this was identified by multimodality imaging as the cause of the luminal obstruction in the distal ileum, just proximal to the stoma. After a failed period of conservative management, examination under anaesthesia was performed and digital extraction attempted, but this was unsuccessful. Rather than surgical stoma revision, endoscopic removal was achieved. The patient improved and was discharged the following day. However, her small bowel obstruction relapsed within 48 h. She was readmitted and underwent stoma revision with no further problems.

Lipid-mediated enhancement of transfection by a nonviral integrin-targeting vector.

Nonviral vectors consisting of integrin-targeting peptide/DNA (ID) complexes have the potential for widespread application in gene therapy. The transfection efficiency of this vector, however, has been limited by endosomal degradation. We now report that lipofectin (L) incorporated into the ID complexes enhances integrin-mediated transfection, increasing luciferase expression by more than 100-fold. The transfection efficiency of Lipofectin/Integrin-binding peptide/DNA (LID) complexes, assessed by beta-galactosidase reporter gene expression and X-gal staining, was improved from 1% to 10% to over 50% for three different cell lines, and from 0% to approximately 25% in corneal endothelium in vitro. Transfection complexes have been optimized with respect to their transfection efficiency and we have investigated their structure, function, and mode of transfection. Both ID and LID complexes formed particles, unlike the fibrous network formed by lipofectin/DNA complexes (LD). Integrin-mediated transfection by LID complexes was demonstrated by the substantially lower transfection efficiency of LKD complexes in which the integrin-biding peptide was substituted for K16 (K). Furthermore, the transfection efficiency of complexes was shown to be dependent on the amount of integrin-targeting ligand in the complex. Finally, a 34% reduction in integrin-mediated transfection efficiency by LID complexes was achieved with a competing monoclonal antibody. The role of lipofectin in LID complexes appears, therefore, to be that of a co-factor, enhancing the efficiency of integrin-mediated transfection. The mechanism of enhancement is likely to involve a reduction in the extent of endosomal degradation of DNA.

Platelet alpha 2-adrenoreceptor binding in elderly depressed patients.

Specific binding of 3H-clonidine to platelet membranes was measured in depressed elderly patients and in an elderly control group. Maximum specific binding was significantly higher in depressed patients than in the control group, whereas the binding affinity was not significantly different.

Phenotypic and functional differences between human saphenous vein (HSVEC) and umbilical vein (HUVEC) endothelial cells.

The vascular endothelial cell (EC) plays an essential role in the pathogenesis of inflammation, transplant rejection and tumour metastasis. Most research on vascular ECs uses human umbilical vein endothelial cells (HUVECs). However, HUVECs are derived from immune-naive foetal tissue, and show significant functional differences from adult vascular endothelium. In this paper, we characterise an alternative model based on human saphenous vein ECs (HSVECs), describe their culture conditions and provide a detailed functional comparison with HUVECs. Compared with HUVECs, HSVECs show an increased sensitivity to ox-LDL and a reduced response to cytokines, as indicated by adhesion molecule expression as well as leukocyte adhesion and transmigration. With respect to their ability to present antigen, HSVECs have a higher level of HLA-DR, CD40 and ICOS-L following cytokine stimulation. In addition, HSVECs upregulate the costimulatory ligand CD80 (B7.1) following CD40 ligation, and support allogeneic T cell proliferation, while HUVECs fail to express CD80. Due to differential expression of adhesion molecules, poorly differentiated tumour cell lines also showed more adhesion to HSVECs than to HUVECs. These results indicate that HSVECs have advantages over HUVECs for studying adult vascular endothelial pathology in vitro.

In vivo selection of sFv from phage display libraries.

The development of phage display technology has facilitated the development of many new and sometimes novel antibody based reagents for scientific research. However, present methods for selection from phage-sFv display libraries are limited to selection against purified antigens or ex vivo cells of known origin and phenotype. Existing methods therefore preclude the isolation of sFv against unknown molecules in their natural environment, where expression is complex and subject to diverse control mechanisms. Since such a complex environment is difficult to mimic in vitro, the development of an in vivo selection procedure would greatly enhance the selection from phage display antibody libraries and lead to the development of reagents against cell surface molecules in their natural environment. This would be particularly advantageous for isolation of sFv against vascular endothelium which can readily change phenotype when cultured and is believed to express molecules in a tissue specific manner and in response to different stimuli. We describe here the development of an in vivo selection procedure in the mouse and demonstrate its potential for the selection of sFv from a phage-sFv library. The target antigen for one sFv is expressed solely on the thymic endothelium, while the second, a 165-170 kDa molecule in present on both thymic endothelium and the perivascular epithelium.