Tumour development is dependent upon the formation of an adequate blood supply through angiogenesis. Vascular endothelial growth factor (VEGF) is one of the most potent and specific pro-angiogenic factors associated with tumour development. Vascular endothelial growth factor is elevated in dogs with a variety of neoplastic tumours and has been linked to an increased risk for metastasis and a poorer prognosis in several tumours. Spirocerca lupi (S. lupi) is a nematode of canids which infests the oesophagus where it forms a nodule. The oesophageal nodule can develop into a neoplastic tumour namely osteosarcoma, fibrosarcoma or anaplastic sarcoma. The pathogenesis of the neoplastic transformation is poorly understood. Diagnosis of neoplastic transformation can be challenging and is based on endoscopy-guided biopsies which are invasive, expensive and may yield non diagnostic samples. The aim of this prospective study was to determine if serum and plasma VEGF levels could be used to distinguish between neoplastic and non-neoplastic spirocercosis. Twenty four dogs were enrolled in the study, 9 with non-neoplastic, 9 with neoplastic spirocercosis, and 6 control dogs. Plasma and serum samples for VEGF analysis were collected at diagnosis. Measurement of VEGF was done using a canine VEGF Quantikine ELISA kit. Statistical analysis to compare the means of the VEGF concentrations between the groups was performed using the Kruskal-Wallis followed by the Dunn’s test. Significance was set at p<0.05 The median plasma VEGF concentration of the dogs with neoplastic spirocercosis 629pg/ml (range 282 – 2366) was higher than the median plasma VEGF concentrations of both the non-neoplastic 0pg/ml (range 0 – 716) and controls 0pg/ml (range 0 – 0) (p<0.001). The median serum VEGF concentration of the neoplastic dogs 69pg/ml (range 0 – 212) was higher than the serum VEGF concentrations in the non-neoplastic 0pg/ml (range 0 – 44.13) and control 0pg/ml (range 0 – 39.4) (p=0.001). Plasma VEGF at a cut off value of 250pg/ml was determined to have a sensitivity of 100%, specificity of 77.8%, a PPV of 81.8% and a NPV of 100% for determining neoplastic transformation. Serum VEGF at a cut off value of 25pg/ml was determined to have a sensitivity of 88.9%, specificity of 100%, a PPV of 100% and a NPV of 90% for determining neoplastic transformation. Both plasma and serum VEGF concentrations can be used to differentiate between non-neoplastic and neoplastic spirocercosis. Plasma VEGF concentrations were higher than serum VEGF concentrations, contrary to what is reported in literature. Both plasma and serum VEGF concentrations can, therefore, potentially be used for diagnosis of neoplastic vs. non-neoplastic cases in canine spirocercosis. There is a need to perform more studies to determine cut-off concentrations that would maximize the sensitivity and specificity for determining neoplastic transformation in canine spirocercosis as well as to determine the role of VEGF in the pathogenesis of the neoplastic transformation. Copyright / Dissertation (MMedVet)--University of Pretoria, 2012. / Companion Animal Clinical Studies / unrestricted

Lun Samantha Wei Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 142-174). / Abstracts in English and Chinese. / Contents --- p.ii / Acknowledgements --- p.vii / Abstract --- p.viii / 摘要 --- p.xi / Chapter Section 1. --- Introduction / Chapter 1.1 --- General review of the epididymis --- p.1 / Chapter 1.1.1 --- Structure of the epididymis --- p.1 / Chapter 1.1.2 --- Function of the epididymis --- p.3 / Chapter 1.1.3 --- Regulation of the epididymal function --- p.5 / Chapter 1.2 --- Vascular endothelial growth factor (VEGF) --- p.8 / Chapter 1.2.1 --- VEGF peptides --- p.8 / Chapter 1.2.2 --- Biological activities of VEGF --- p.10 / Chapter 1.2.3 --- Hormonal regulation of VEGF --- p.11 / Chapter 1.3 --- VEGF receptors --- p.12 / Chapter 1.3.1 --- Flt-1 or VEGFR1 --- p.12 / Chapter 1.3.2 --- Flk-1 or VEGFR2 --- p.13 / Chapter 1.4 --- Caveolae --- p.15 / Chapter 1.4.1 --- Overview of caveolae --- p.15 / Chapter 1.4.2 --- Caveolins/caveolin-1 --- p.16 / Chapter 1.4.3 --- Caveolae and VEGF --- p.18 / Chapter 1.4.4 --- Caveolae and the epididymis --- p.20 / Chapter 1.5 --- VEGF/ VEGF receptors in the epididymis --- p.20 / Chapter 1.6 --- Aims of study --- p.22 / Chapter Section 2. --- Materials and Methods / Chapter 2.1 --- Materials --- p.24 / Chapter 2.2 --- Animal surgery --- p.35 / Chapter 2.2.1 --- Animals --- p.35 / Chapter 2.2.2 --- Castration and hemi-castration --- p.35 / Chapter 2.2.3 --- Efferent duct ligation (EDL) --- p.36 / Chapter 2.2.4 --- Tissue collection --- p.37 / Chapter 2.3 --- Epididymal cell culture --- p.38 / Chapter 2.4 --- Sample preparation --- p.40 / Chapter 2.4.1 --- Collection of epididymal plasma and sperm --- p.40 / Chapter 2.4.2 --- Purification of caveolae fraction --- p.41 / Chapter 2.5 --- Reverse-transcription polymerase chain reaction (RT-PCR) and semi-quantitative RT-PCR --- p.43 / Chapter 2.5.1 --- Preparation of RNA from epididymal tissues --- p.43 / Chapter 2.5.2 --- Quantitation of total RNA --- p.44 / Chapter 2.5.3 --- Reverse transcription (RT) and polymerase chain reaction (PCR) --- p.44 / Chapter 2.5.4 --- Purification and authenticity confirmation of PCR products --- p.50 / Chapter 2.6 --- Western immunoblotting --- p.53 / Chapter 2.6.1 --- Preparation of protein --- p.53 / Chapter 2.6.2 --- SDS-PAGE --- p.53 / Chapter 2.6.3 --- Western immunoblotting --- p.55 / Chapter 2.7 --- Immunohistochemistry --- p.56 / Chapter 2.7.1 --- Preparation of tissue sections --- p.56 / Chapter 2.7.2 --- Immunohistochemical staining of tissue sections --- p.57 / Chapter 2.7.3 --- Immunostaining of cultured cells --- p.59 / Chapter 2.8 --- Enzyme Linked Immunosorbant Assay (ELISA) --- p.59 / Chapter 2.9 --- Statistical analyses --- p.60 / Chapter Section 3. --- Results / Chapter 3.1 --- Expression and localization of VEGF in the rat epididymis --- p.62 / Chapter 3.1.1 --- RT-PCR of VEGF in the rat epididymis --- p.62 / Chapter 3.1.2 --- Western immunoblot of VEGF in the rat epididymis --- p.63 / Chapter 3.1.3 --- Developmental changes in VEGF expression in the rat epididymis --- p.66 / Chapter 3.1.4 --- Immunolocalization of VEGF in the rat epididymis --- p.66 / Chapter 3.1.5 --- Summary of the localization and expression of VEGF in the rat epididymis --- p.72 / Chapter 3.2 --- Expression and localization of VEGF receptors in the rat epididymis --- p.74 / Chapter 3.2.1 --- RT-PCR of Flt-1 and sFlt-1 in the rat epididymis --- p.74 / Chapter 3.2.2 --- Western immunoblot of Flt-1 in the rat epididymis --- p.74 / Chapter 3.2.3 --- Immunolocalization of Flt-1 in the rat epididymis --- p.75 / Chapter 3.2.4 --- RT-PCR of Flk-1 in the rat epididymis --- p.75 / Chapter 3.2.5 --- Western immunoblot of Flk-1 in the rat epididymis --- p.79 / Chapter 3.2.6 --- Immunolocalization of Flk-1 in the rat epididymis --- p.79 / Chapter 3.2.7 --- Summary on the localization and expression of VEGF receptors in the rat epididymis --- p.83 / Chapter 3.3 --- Detection of VEGF immunoreactivity in epididymal plasma and sperm lysate collected from cauda epididymidis --- p.83 / Chapter 3.4 --- Effect of castration on VEGF and VEGF receptor expression in the rat epididymis --- p.84 / Chapter 3.4.1 --- Effect of castration with or without testosterone replacement on VEGF expression in the rat epididymis --- p.84 / Chapter 3.4.2 --- Effect of castration with or without testosterone replacement on Flt-1 expression in the rat epididymis --- p.94 / Chapter 3.4.3 --- Effect of castration with or without testosterone replacement on Flk-1 expression in the rat epididymis --- p.98 / Chapter 3.5 --- Effect of efferent duct ligation and hemicastration on VEGF peptide levels in the rat epididymis --- p.102 / Chapter 3.5.1 --- Effect of efferent duct ligation on VEGF expression in the rat epididymis --- p.102 / Chapter 3.5.2 --- Effect of hemi-castration on VEGF expression in the rat epididymis --- p.106 / Chapter 3.6 --- "Summary on the effects of castration, efferent duct ligation, and hemicastration on the epididymal weight, and VEGF/VEGF receptor expression in the rat epididymis" --- p.107 / Chapter 3.6.1 --- "Summary on the effects of castration, efferent duct ligation and hemicastration on the epididymal weight" --- p.107 / Chapter 3.6.2 --- "Summary on the effects of castration, efferent duct ligation and hemicastration on VEGF and VEGF receptor expression in the rat epididymis" --- p.112 / Chapter 3.7 --- Localization and expression of caveolin-1 and 226}0ؤ2 in the rat epididymis --- p.113 / Chapter 3.7.1 --- RT-PCR of caveolin-1 and caveolin-2 in the rat epididymis --- p.113 / Chapter 3.7.2 --- Western immunoblot of caveolin-1 and caveolin-2 in the rat epididymis --- p.114 / Chapter 3.7.3 --- Immunolocalization of caveolin-1 in the rat epididymis --- p.117 / Chapter 3.7.4 --- Summary on the localization and expression of caveolin-1 and -2 in the rat epididymis --- p.119 / Chapter 3.8 --- Co-localization of VEGF receptors with caveolae in the rat epididymis --- p.119 / Chapter Section 4. --- Discussion --- p.124 / Chapter 4.1 --- VEGF expression and localization --- p.124 / Chapter 4.2 --- VEGF receptors expression and localization --- p.129 / Chapter 4.3 --- Possible VEGF action in the rat epididymis --- p.133 / Chapter 4.4 --- Regulation of VEGF and its receptor expression by androgen and/or other testicular factors --- p.136 / References

Hintergrund: In dieser retrospektiven Studie soll der Einfluss von Rauchen, Alter und systemischer Medikation auf den Behandlungserfolg einer Anti-VEGF Therapie bei altersbedingter Makuladegeneration (AMD) über einen Zeitraum von 24 Monaten untersucht werden. Patienten und Methode: 100 Patienten mit choroidalen Neovaskularisationen bei AMD wurden in die Studie eingeschlossen. Best korrigierter Visus (BV), Anzahl der Injektionen in 24 Monaten sowie Rauchgewohnheiten und systemische Medikation wurden für die Analysen berücksichtigt. Ausschlusskriterium war ein BV < 0,1 zu Beginn der Behandlung. Resultate: 42 Raucher (inkl. 31 Exraucher) mit 23,5 Packyears (Py) im Median wurden identifiziert. Je mehr Py ein Patient rauchte, umso niedriger war sein BV nach der letzten Injektion (p = 0,009). Je mehr Zigaretten pro Tag ein Raucher rauchte, umso mehr Injektionen erhielt er in 24 Monaten (p = 0,0042). Bluthochdruckpatienten hatten einen niedrigeren BV nach der letzten Injektion (p = 0,045). Schlussfolgerungen Rauchen ist nicht nur ein Risikofaktor für die Entwicklung einer AMD, sondern auch für die Effektivität der Anti-VEGF Therapie. Auch unter sozioökonomischen Gesichtspunkten ist dies ein weiterer Grund, Patienten zur Aufgabe des Rauchens aufzufordern. / Background: To investigate the influence of smoking, age and systemic medication on the effectiveness of an anti-VEGF therapy in patients with exsudative age-related macular degeneration (AMD). Patients and methods: A total of 100 patients were included in the retrospective analysis. Data included best-corrected visual acuity (BCVA) and number of injections in 24 months. Subjects with BCVA < 0.1 at baseline were rejected. All patients were interviewed about smoking habits and current systemic medication. Results: The study comprised 42 smokers (including 31 past-smokers) with a median of 23.5 packyears (py). The more py a patient had smoked, the lower BCVA was after the last injection (p = 0.009). The more cigarettes per day a smoker had smoked the more injections he had received (p = 0.0042). Patients with arterial hypertension had a lower BCVA after the last injection (p = 0.045). Conclusions: Smoking is not only a risk factor for the development of AMD but also for the effectiveness of an anti-VEGF treatment. Also from a socio-economic point of view AMD patients should be instructed to quit smoking.

Hintergrund In einer retrospektiven Studie in der Augenklinik Würzburg wurde die Ranibizumabtherapie bei Patienten mit altersabhängiger Makuladegeneration (AMD) im klinischen Alltag ausgewertet. Methoden Patientenakten von Patienten mit AMD, die im Jahr 2007 mit der Ranibizumabtherapie begannen, wurden untersucht. Daten wurden bis zum Ende der Behandlung und/oder Nachbeobachtung bis 2009 gesammelt. Der primäre Endpunkt war das Verhältnis der Patienten, die weniger als 15 Buchstaben (bzw. 0,3 logMAR Einheiten) an Visus verloren zwischen Beginn und nach 12 Monaten. Ergebnisse 375 Patienten wurden einbezogen, nur 298 Patienten beendeten die Untersuchung nach einem Jahr. Nach 12 Monaten verloren 72% der Patienten weniger als 15 Buchstaben. Die Sehschärfe verbesserte sich bis 12 Wochen nach der ersten Injektion und verschlechterte sich danach wieder. Patienten mit mehr als 3 Injektionen profitierten mehr als Patienten mit weniger Injektionen. Durchschnittlich wurden 4,25 Injektionen innerhalb eines Jahres gegeben. Der durchschnittliche Rückgang der Netzhautdicke betrug 50 µm. Schlussfolgerung Intravitreale Injektionen von Ranibizumab in der Augenklinik Würzburg führten zu einer Visusverbesserung. Der Visusgewinn konnte nach 3 Monaten nicht gehalten werden. Bessere Reinjektionskriterien, mehr OCT Untersuchungen und besseres Nachsorgemanagement sollten entwickelt werden. / Background Real-life ranibizumab therapy used in patients with wet age related macular degeneration (AMD) was assessed in a retrospective study in Augenklinik Würzburg Methods Medical records of patients with AMD, who started ranibizumab treatment in 2007, were evaluated. Data were collected until the end of treatment and/or monitoring until 2009. The primary end point was the proportion of patients losing fewer than 15 letters (0,3 logMAR units) from baseline visual acuity at 12 months. Results 375 patients were included, only 298 patients finished one-year examination. At 12 months 72% of the patients lost fewer than 15 letters. Visual acuity improved until 12 weeks after first injection and was not maintained afterwards. Patients with more than 3 Injections profited more than patients with less injections. In average, there were 4,25 injections given in one year. The average decrease in retinal thickness was 50µm. Conclusions Intravitreal administration of ranibizumab in Augenklinik Würzburg improved vision acuity. The vision gain was not maintained after 3 months. Criteria for reinjection, more oct examinations and better control management must be developed.

Both white and brown adipose tissues exhibit extensive vascularity. Increased angiogenesis in brown adipose tissue (BAT) is crucial for brown fat activation and thermogenesis in animals during cold acclimation. BAT can be similarly activated by food intake to generate heat through cellular respiration, in a process known as diet induced thermogenesis. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that regulates both pathological and physiological angiogenesis and can stimulate cell proliferation, migration, survival and vessel permeability. However, VEGF has also been shown to affect an increasing number of non-vascular cells such as skeletal muscle and kidney podocytes. The expression and function of VEGF in white and brown adipocytes are not fully understood. We have previously shown that the expression of VEGF is concomitantly regulated with skeletal muscle differentiation. Here we show that VEGF is expressed in BAT and all major white adipose depots in mice. VEGF expression was increased during white and brown adipocyte differentiation and was regulated in cultured brown adipocytes by the \(PPAR\gamma\) agonist troglitazone and by \(PGC1\alpha\) in BAT in vivo. Systemic VEGF neutralization led to brown adipocyte apoptosis in vivo, loss of mitochondrial cristae and increased mitophagy and was associated with increased inflammation and fibrosis. VEGFR2 was expressed in both brown preadipocytes and adipocytes. Blockade of VEGF signaling using anti-VEGFR2 antibody DC101 increased brown adipocyte apoptosis in vitro. VEGF also functioned as a mitogen and survival factor for brown preadipocytes. VEGF 164 and VEGF 188, isoforms that can bind heparan sulfate proteoglycans, comprise >98% of total VEGF in BAT, subcutaneous and perigonadal fat depots. Embryos that lacked VEGF 164 and 188 displayed abnormal BAT development with fewer brown adipocytes, lower levels of mitochondrial uncoupling protein 1 and Cox IV. These results indicate a direct role for VEGF signaling in brown adipocytes and preadipocytes and suggest the importance of heparan sulfate binding VEGF isoforms in BAT development. Elucidation of the role of VEGF signaling in adipocytes is vital to understanding adipose tissue expansion and activation and may reveal novel therapeutic targets for the activation of brown fat in humans.

Acute Respiratory Distress Syndrome (ARDS) is the most extreme form of acute lung injury and continues to have a significant morbidity and mortality. Unfortunately, the mechanisms involved in the recovery and repair of the lung following ARDS remain poorly understood. An understanding of these is pivotal to improving outcome from acute lung injury. Several observational studies have suggested a potential relationship between Vascular Endothelial Growth Factor (VEGF) in the lung and the development/outcome of ARDS. In this thesis, three potential mechanisms underlying these observations have been explored: 1. What is the anatomical distribution of VEGF receptor and isoform expression in normal and ARDS lung? How does this change at early and later time points following acute lung injury? 2. Are human type 2 alveolar epithelial (ATII) cells a source of and target for VEGF? How does exposure to a pro-inflammatory milieu modify their expression of VEGF isoforms and receptors? 3. Is there a relationship between a functional VEGF polymorphism and susceptibility to developing and severity of ARDS? I have demonstrated VEGF receptor expression on both sides of the alveolarcapillary membrane with upregulation in later ARDS. All three principal isoforms (VEGF121, VEGF165 and VEGF189) are expressed in normal human lung with uniform downregulation of all three in early ARDS, which normalises with increasing time following injury. I have not found any evidence of VEGF isoform switching. I have also demonstrated human ATII cells are both a significant cellular source of and a target for VEGF (via VEGF receptor expression) confirming autocrine VEGF activity in the lung. VEGF is an ATII cell survival factor. ATII cells differentially respond to pro-inflammatory stimuli by increasing VEGF isoform but not receptor expression, which may serve as a regulatory control mechanism. Finally, I have demonstrated the VEGF 936 T allele increases susceptibility to and the severity of lung injury. The T allele is associated with an increase in plasma VEGF level in ARDS patients but intra-alveolar levels are unaffected.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Diss. (sammanfattning) Linköping : Linköpings universitet, 2006. / Härtill 4 uppsatser.

Zugl.: Giessen, Universiẗat, Diss., 2008.

Zugl.: Giessen, Universiẗat, Diss., 2008.