Higher Vessel Density Research Articles

Prognostic significance of lymphangiogenesis in pharyngolaryngeal carcinoma patients.

BackgroundLymphatic vessel spread is considered a major route for head and neck squamous cell carcinoma metastasis. Formation of new lymphatic vessels could facilitate the process, raising the malignant potential of these tumours. Recent identification of lymphatic markers allows the study of the lymphangiogenesis phenomenon. We searched for molecular events involved in the lymphangiogenic process that could have prognostic value in laryngeal/pharyngeal carcinoma patients.Methods104 paraffin-embedded pharyngeal/laryngeal tumour samples were studied. Immunohistochemical analysis of podoplanin and double immunofluorescence analysis of Ki-67 and D2-40 were performed. Lymph vessel density (inside the tumour mass, at its periphery or considered as a whole) and the presence of tumour emboli inside lymphatics were recorded. The proliferative state of endothelial lymphatic cells was evaluated.ResultsLymphatic vessels were detected inside the tumour mass (75%) and in the surrounding tissue (80%); some of them in a proliferative state. Tumour emboli were detected in a high proportion of the cases (45%). Lymphatic vessel density was higher in the pharyngeal cases (p = 0.0029), in greater size (p = 0.039), more advanced stage primary tumours (p = 0.006) and in carcinomas of patients with affected nodes (p = 0.019). The presence of tumour emboli and a high global vessel density were indicators of poor prognosis (recorded as death from tumour) in the laryngeal group (p = 0.015 and p = 0.027, respectively), but notably not in the pharyngeal one. Interestingly, high global vessel density showed a negative prognostic value among pathologically staged N0 laryngeal carcinomas (p = 0.03).ConclusionsThe lymphangiogenic process correlated with aggressive tumour features (pN category, tumour size, tumour stage), but might play different roles in tumours arising from different anatomic sites.Our results suggest that detection of tumour emboli and assessment of global vessel density using the D2-40 antibody, may be useful in the clinical practice, as predictors of reduced survival among pN0 laryngeal carcinoma patients.

Improved heart function follows enhanced inflammatory cell recruitment and angiogenesis in 11βHSD1-deficient mice post-MI

Mice unable to locally regenerate corticosterone due to deficiency of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) have enhanced angiogenesis during acute myocardial infarct healing. The present study investigates the hypotheses that in these mice (i) inflammation and angiogenic signalling are promoted and (ii) longer-term remodelling and function are improved. Myocardial infarction (MI) was induced by coronary artery ligation in 11βHSD1(-/-) and wild-type (C57BL/6) mice. Studies were terminated 2, 4, 7, and 28 days post-surgery. Increased vessel density (CD31 immunoreactivity) on the infarct border was confirmed 7 days after MI in 11βHSD1(-/-) hearts (P < 0.05) and was accompanied by improved ejection fraction (ultrasound) compared with C57BL/6. During wound healing, recruitment of neutrophils (at 2 days after MI) and macrophages (from 4 days after MI) and expression of monocyte-chemoattractant protein-1 was increased in 11βHSD1(-/-) compared with C57BL/6 hearts (P < 0.05). Recruitment of alternatively activated YM1-positive macrophages was particularly enhanced in the period preceding increased vessel density and was accompanied by increased expression of pro-angiogenic IL-8. By 28 days post-MI, when the infarct scar had matured, higher vessel density was maintained in 11βHSD1(-/-) hearts and vessels were smooth-muscle coated. Infarct scars were thicker (P < 0.001) in 11βHSD1(-/-) compared with C57BL/6 hearts and ejection fraction was higher (P < 0.05). Increased vessel density in healing infarcts of mice deficient in 11(-/-)HSD1 follows recruitment of pro-reparative macrophages and increased pro-angiogenic signalling. Mature infarcts show less thinning and cardiac function is improved relative to wild-type mice, suggesting that 11βHSD1 may be a novel therapeutic target after MI.

Stromal PDGFRβ Expression in Prostate Tumors and Non-Malignant Prostate Tissue Predicts Prostate Cancer Survival

BackgroundThe identification of new prognostic markers for prostate cancer is highly warranted, since it is difficult to identify patients requiring curative treatment. Data from both experimental models and clinical samples have identified important functions of PDGFRβ on pericytes and fibroblasts in the tumor stroma.Methodology/Principal FindingsIn this study the prognostic significance of PDGFRβ in prostate cancer stroma, and in matched non-malignant tissue, was evaluated with immunohistochemistry. PDGFRβ expression was analyzed in normal and tumor stroma from more than 300 prostate cancer patients. High PDGFRβ expression in tumor stroma was associated with large tumor size, advanced stage, high Gleason score and high vessel density. Perivascular PDGFRβ staining in tumors was also correlated with high Gleason score. Correlations were also observed between PDGFRβ status in tumor stroma and non-malignant stroma. Similarly, high PDGFRβ expression in adjacent non-malignant tissue stroma correlated with large tumor size, advanced stage, high Gleason score and proliferation in non-malignant epithelium. Interestingly, high levels of PDGFRβ in the stroma of tumor and non-malignant tissue were associated with shorter cancer specific survival in prostate cancer patients.Conclusions/SignificanceThe study revealed a number of novel associations between stromal PDGFRβ expression in prostate tumors and several important clinical characteristics, including survival.

Expression of VEGF‐C and VEGF‐D as Significant Markers for Assessment of Lymphangiogenesis and Lymph Node Metastasis in Non‐Small Cell Lung Cancer

Vascular endothelial growth factor (VEGF)-C and VEGF-D induce lymphangiogenesis through activation of VEGF receptor 3 (VEGFR-3) and have been implicated in tumor spread to the lymphatic system. Lymph node dissemination critically determines clinical outcome and therapeutic options of patients with non-small cell lung cancer (NSCLC). However, the relationship of VEGF-C, VEGF-D, and lymph node metastasis in cancers, including NSCLC, is still controversial. To evaluate the relationship between lymphangiogenesis and lymph node metastasis, the expression of VEGF-C and VEGF-D in NSCLC tumors were detected by immunohistochemistry and quantitative real-time polymerase chain reaction (QRT-PCR). QRT-PCR revealed that in marginal region VEGF-C and VEGF-D mRNA was significantly higher than in tumor center, and VEGF-D mRNA was also higher than that in peritumoral lung tissue. Immunohistochemically, we observed the same heterogeneous expression of VEGF-C and VEGF-D proteins. The group with high expression of VEGF-C and VEGF-D in marginal region had a higher incidence of lymph node metastasis compared with the group with low expression. Furthermore, the group with high expression of VEGF-D in marginal region had a higher incidence of lymphatic invasion. The group with high peritumoral lymphatic vessel density (LVD) had higher expression of VEGF-C and VEGF-D mRNA compared with the group with low peritumoral LVD. Our studies suggested that the expression of VEGF-C and VEGF-D at invasive edge was significantly associated with lymph node metastasis or lymphatic invasion in patients with NSCLC and may be involved in regulation of lymphangiogenesis and lymph node metastasis in NSCLC.

High density of peritumoral lymphatic vessels is a potential prognostic marker of endometrial carcinoma: a clinical immunohistochemical method study

BackgroundThe lymphatic system is a major route for cancer cell dissemination and also a potential target for antitumor therapy. To investigate whether increased lymphatic vessel density (LVD) is a prognostic factor for nodal metastasis and survival, we studied peritumoral LVD (P-LVD) and intratumoral LVD (I-LVD) in samples from 102 patients with endometrial carcinoma;MethodsEndometrial carcinoma tissues were analyzed for lymphatic vessels by immunohistochemical staining with an antibody against LYVE-1. Univariate analysis was performed with Kaplan-Meier life-table curves to estimate survival, and was compared using the log rank test. Prognostic models used multivariate Cox regression analysis for multivariate analyses of survival;ResultsOur study showed that P-LVD, but not I-LVD, was significantly correlated with lymph vascular space invasion (LVSI), lymph node metastasis, tumor stage, and CD44 expression in endometrial carcinoma. Moreover, P-LVD was an independent prognostic factor for progression-free survival and overall survival of endometrial carcinoma;ConclusionsP-LVD may serve as a prognostic factor for endometrial carcinoma. The peritumoral lymphatics might play an important role in lymphatic vessel metastasis.

Down-regulation of artery in moderately differentiated hepatocellular carcinoma related to tumor development

Hepatocellular carcinoma develops in a multistep process. Previous studies have revealed changes in blood supply in hepatocellular carcinoma during its carcinogenesis. However, little is known about the relationship between tumor vasculature and the biological behavior of moderately differentiated hepatocellular carcinoma which demonstrates varied degrees of biological behavior. We immunohistochemically assessed intratumoral arterial vessel density (by high-molecular-weight caldesmon and calponin) and microvessel density (by CD34) in 123 cases of moderately differentiated hepatocellular carcinomas, and compared these densities with clinicopathological findings. Arterial vessel density and microvessel density of 19 well-differentiated and 37 poorly differentiated hepatocellular carcinomas were also evaluated. The arterial vessel density of moderately differentiated hepatocellular carcinomas with capsule formation, infiltration to the capsule, portal venous invasion, and high Ki-67 labeling index was lower than that of moderately differentiated hepatocellular carcinomas without these pathological findings (high-molecular-weight caldesmon: P < .0001, P = .0074, P = .0009, P = .0244, calponin: P < .0001, P = .0695, P = .0033, and P = .0155, respectively). The low arterial vessel density group (<10) of moderately differentiated hepatocellular carcinomas tended to show poorer overall survival than the high arterial vessel density group (>or=10) (high-molecular-weight caldesmon: P = .0347, calponin: P = .0404). The arterial vessel density and microvessel density of moderately differentiated hepatocellular carcinomas were significantly higher than those of well-differentiated hepatocellular carcinomas (high-molecular-weight caldesmon: P = .022, calponin: P = .027, CD34: P = .036) and poorly differentiated hepatocellular carcinomas (high-molecular-weight caldesmon, calponin and CD34: P < .0001). The moderately differentiated hepatocellular carcinomas with lower arterial vessel density had more malignant potential than those with higher arterial vessel density. The changes of arterial vessel density in moderately differentiated hepatocellular carcinomas were suggested.

Role of lymphangiogenesis in lung cancer.

Lung cancer represents one of the most frequent causes of death due to neoplastic disease in Poland and around the world. The high mortality which accompany neoplastic diseases used to be ascribed mainly to dissemination of cancerous cells. Studies on animal models suggest that tumour lymphangiogenesis represents the principal factor in the process of metastases formation. Lymphangiogenesis involves a process of formation of new lymphatic vessels from already existing lymphatic capillaries. Lymphangiogenesis is stimulated by vascular endothelial growth factors (VEGF) and other, recently reported factors, such as, e.g., cyclooxygenase 2, fibroblast growth factor 2, angiopoetin-1 and the insulin-resembling growth factor. In lymphangiogenesis a key role is played by neutropilin 2 or podoplanin and this promoted development of studies on lymphangiogenesis. Activation of VEGF-C/VEGF-D/VEGFR-3 axis increases motility and invasiveness of neoplastic cells, promotes development of metastases in several types of tumours such as, e.g., lung cancer, mammary carcinoma, cancers of the neck, prostate and large intestine. In recent years lymphangiogenesis provided topic of many studies. A positive correlation was detected between expressions of VEGF-C/D and VEGFR-3 in non-small cell lung cancer. In patients with lung cancer with high expression of VEGF-C a markedly abbreviated survival was noted. Positive correlation was detected between expression of VEGF-C and VEGF-D on one hand and expression of LYVE-1 on the other in sentinel lymph nodes with metastases of neoplastic cells in patients with non-small cell lung cancer. Also, high density of lymphatic vessels and high density of intraneoplastic microvessels proved to be independent poor prognostic indices in patients with non-small cell lung cancer. Extensive hope is linked to studies on inhibitors of lymphangiogenesis, which may improve results of treatment also in tumour patients.

Microvascular system of the lumbar dorsal root ganglia in rats. Part I: a 3D analysis with scanning electron microscopy of vascular corrosion casts

So far, the morphological features of the vascular system supplying the dorsal root ganglion (DRG) have been inferred only from microangiograms. However, in the past most of these studies lacked 3D observations. This study presents the details of the microvasculature of the lumbar DRG visualized by scanning electron microscopy of vascular corrosion casts. Wistar rats were anesthetized with intraperitoneal sodium pentobarbital. After thoracotomy, the vascular system was perfused with heparinized saline, and Mercox resin was injected into the thoracic aorta. After polymerization of the resin, the vascular casts were macerated with potassium hydroxide, washed with water, and dried. The casts were examined with a scanning electron microscope. The vascular cast of the DRG was observed to have a higher density of vessels than the nerve root. Bifurcation or anastomoses of capillaries took place at approximately right angles, in a T-shaped pattern. Within the DRG, both the arterial supply and the capillary network contained blood flow control structures (ring-shaped constrictions in the cast probably representing a vascular sphincter in the microvessel). Three types of vessels could be distinguished: tortuous, straight, and bead-like capillaries. The dilations, bulges, and tortuousness of capillaries could serve the function of locally increasing the capillary surface area in a sensory neuron. The results of this study suggest a causal relationship between the metabolic demands of local neuronal activity and both the density of the capillary network and the placement of the blood flow control structures.

Accelerated Angiogenic Induction and Vascular Integration in a Novel Synthetic Scaffolding Matrix for Tissue Replacement

Reduced or delayed neovascularization is a major obstacle with regard to tissue-engineered constructs. The aim of this study was to evaluate the early microvascular response to a novel degradable ε-caprolactone terpolymer matrix. ε-caprolactone terpolymer matrices (Suprathel Plus®; Institute of Textile and Process Engineering, Denkendorf, Germany) were implanted into dorsal skinfold chambers of balb/c mice (n=10). Microcirculatory changes were observed by intravital fluorescence microscopy. Scaffolding matrices from PEGT/PBT copolymer were used as controls (n=10). The formation of de novo vascular networks within both scaffolding matrices was noted throughout the experiment. A vascular ingrowth of perfused microvessels into the matrices up to 600 μm apart from the edge was noted within 10 days of implantation. The earliest signs of neoangiogenesis were visible in ε-caprolactone terpolymer matrices on day 1. In both scaffolds the new developed vessels extended centripetally from the border of the matrices towards the center and anastomosed to form a perfused microvascular network. There was significantly earlier onset of vascularization, increased vascularized area and higher vessel density in ε-caprolactone terpolymer matrices compared to PEGT/PBT copolymer matrices were observed. The scaffolding matrix from ε-caprolactone terpolymer allowed for an earlier and more intense induction of angiogenesis and displayed the tendency to vascularize more rapidly within a shorter period of time after transplantation compared to PEGT/PBT copolymer scaffolds, thus indicating its potential application for tissue engineering purposes.

Morphometric analysis of CD34‐positive vessels in salivary gland adenoid cystic and mucoepidermoid carcinomas

Carcinomas of the salivary glands are uncommon and morphologically a diverse group of malignancies. To evaluate the prognostic value of CD34 immunostaining of the vessels in adenoid cystic carcinoma (AdCC) and mucoepidermoid carcinoma (MEC), an automated image analysis method was used. In a nationwide study, covering salivary gland cancer (SGC) patients in Finland 1991-1996, 37 AdCC and 18 MEC patients (M 25, F 30, age 25-90, mean 63) were included. In addition to clinical characteristics the size, shape, staining intensity and vessel density in CD34 immunostained histologic samples were measured. Altogether 4433 vessels were measured from AdCC and 2615 from MEC tumor. Of the total tumor vessels measured, 2651 were from patients who deceased with disease (Group I) and 4397 were from specimens derived from those who did not die of disease (Group II) during the 10-year follow-up. The staining intensity was significantly higher in MEC than in AdCC tumor (P = 0.0005). In MEC, the Group I patients had a higher staining intensity among high-grade patients compared with patients with low grade disease, whereas the tumors in Group II had a lower staining intensity among the high-grade compared with the low grade tumors (P = 0.018). A higher vessel density was found in patients with MEC in group II compared with group I (P = 0.017). The staining intensity of CD34 positive vessels in MEC was higher than in AdCC. In MEC, higher staining intensity of vessels in high-grade tumors and lower vessel density in all MEC patients, predicted poor survival.

A safe hydraulic architecture as wood anatomical explanation for the difference in distribution of the mangroves Avicennia and Rhizophora

Summary Wood anatomical characteristics are an important source of information about how trees are coping with the conflicting requirements of optimal hydraulic conductivity and safety. In this study we compared the vessel characteristics of Avicennia marina and Rhizophora mucronata to contribute to a better understanding of the difference in distribution of these two mangrove species. Density, grouping and diameters of vessels together with vessel element length of A. marina trees growing on seven different study sites in Gazi Bay (Kenya) were measured and compared with the vessel characteristics of R. mucronata of the same research area. Furthermore, the relation of environmental factors related to the water relations of mangrove trees to the wood anatomy of both species was tested. Vessel density of both species and vessel grouping of A. marina are higher with higher salinity. In addition, A. marina was shown to have a higher vessel density, a higher vessel grouping, smaller vessel diameters and shorter vessel element lengths as compared to R. mucronata. As the vessel characteristics of A. marina are related to protection against cavitation or the effects of cavitation, we can infer that this species has a water transport system which can be considered safer under water limiting conditions than that of R. mucronata. The contrast in the safety of the water transport system between the two mangrove species studied is reflected in their distribution at local scale, over the land‐sea ecotone, and it may also explain the wide latitudinal distribution of A. marina as compared to R mucronata. This conclusion emphasizes the importance of the anatomy of the water transport system for survival under diverse environmental conditions. In general it is shown that dominant species of a single ecosystem can deal with the stressful conditions they live in by contrasting ecological strategies.

Expression of cytokines and chemokines and microvasculature alterations of the tongue from patients with chronic Chagas’ disease

Chagas' disease is caused by the protozoan Trypanosoma cruzi and continues to be a significant public health problem, since 10 million people are still infected in Latin America. The purpose of this study was to analyze the microvasculature alterations as well the expression of cytokines and chemokines in the tongues from patients with chronic Chagas' disease (CC; n = 18), comparatively with a non-chagasic group (NC; n = 22). We observed several vascular alterations in the tongue of CC such as a greater vascular diameter, increased vascular wall area, high density of the blood vessels, and increased thickening of the capillary basement membrane. The expression of cytokines interferon gamma and tumor necrosis factor alpha and chemokine macrophage inflammatory protein 1alpha were significantly down-regulated in the tongue of CC group. These results demonstrated that, in the tongue of chagasic patients, a microvascular abnormality and immunological impairment occurs, probably due to chronic inflammation evoked by T. cruzi antigens.

Leaf tissue water relations and hydraulic properties of sclerophyllous vegetation on white sands of the upper Rio Negro in the Amazon region

Abstract:The objective of this study was to explore the leaf tissue water relations in terminal branches, as well as the relations between xylem structure and function of five sclerophyllous species coexisting on white sands within the Amazon region. In these species, which possess costly leaves and thrive in an extremely nutrient-poor habitat, the preservation of leaf survival would be of comparable importance to the preservation of xylem vessels. Three trees per species were tagged in the field for all measurements. Minimum leaf water potential (Ψ) was −1.53 ± 0.61 and −0.94 ± 0.10 MPa during rainless and rainy days, respectively. The Ψ for turgor loss averaged −1.92 ± 0.05 MPa. Therefore, minimum Ψ was maintained within a safety range above the critical value for turgor loss. Xylem (Kx) and leaf (Kl) specific conductivity averaged 1.4 ± 0.22 and 0.00033 ± 0.000045 kg m−1 s−1 MPa−1, respectively. Water supply was favoured in species with higher vessel density, and all species depended on relatively less abundant larger vessels for water transport. This would be advantageous because leaves were unable to develop very negative water potentials in order to maintain transpiration. High transpiration rates may be restricted to a few hours daily so as to prevent cavitation of widest vessels.

Vasomotion of Intraradicular Microvessels in Rat

This study is to investigate the changes of vasomotion of intraradicular microvessels in vivo. We have observed microvascular corrosion casts of the lumbar nerve root by scanning electron microscopy and used an immunohistochemical technique to investigate the presence and distribution of autonomic and sensory nerve in blood vessels of the nerve root. It is generally considered that the genesis of radiculopathy associated with the degenerative conditions of the spine may result from both mechanical compression and circulatory disturbance. However, the neurogenic control of intraradicular blood flow has received little attention in the past. For three-dimensional observation of intraradicular vessels, we used scanning electron microscopic examination of microvascular corrosion casts in ten Wister rats. To investigate the mechanism of vasomotion of the nerve root, we used immunohistochemical methods. The sections were incubated overnight with antisera to tyrosine hydroxylase, choline acetyl transferase, substance P, calcitonin-gene-related peptide, vasoactive intestinal peptide, somatostatin, neuropeptide Y, leucine-enkephalin, cholecystokinin octapeptide, brain-nitric oxide synthase, and endothelium-nitric oxide synthase. Abidin-biotin complex method was used as the immunohistochemical procedure and the sections were observed under the light microscope. The general view of whole vascular casts of the lumbar spinal cord and nerve roots showed a high density of vessels. Bifurcation or anastomoses of capillaries approximately took place at right angles in a T-shaped pattern and capillaries showed a lot of ring-like compressions. This ring-like compression on the cast may represent a vascular sphincter in the microvessels. This study also reveals the existence of perivascular adrenergic, cholinergic, peptidergic, and nitroxydergic innervation with a possible role in neurogenic regulation of nerve root circulation. Perivascular nerve plexuses around intraradicular microvessels suggest that the autonomic nerves play an important role in intraradicular circulation.

ANATOMY OF THE URETHRAL SUPPORTING LIGAMENTS DEFINED BY DISSECTION, HISTOLOGY, AND MAGNETIC RESONANCE IMAGING (MRI) OF FEMALE CADAVERS AND OF HEALTHY NULLIPAROUS WOMEN

sizes of individual lymphatic vessels were categorized as small ( 0.5mm). To compare the lymphatic density (counts/mm2), the number of lymphatic vessels of the five random areas was counted in each specimen by light microscopy at low power (x40). RESULTS: LYVE-1 expression and the lymphatic density of muscular propria were significantly higher than those of other layers, including epithelium, lamina propria, perivesical fat, and serosa (p < 0.0001, respectively). The size of lymphatic vessels in muscular propria was significantly smaller than in other layers (p < 0.0001). The greatest distribution of lymphatic vessels was identified in the border areas consisting of lamina propria and muscular propria, or muscular propria and adventitia, including perivesical fat and serosa. CONCLUSIONS: We found that high density of lymphatic vessels was identified in a border area consisting of lamina propria and muscularis propria. When biologically aggressive bladder cancer deeply or widely extends into the muscularis propria, cancer cells potentially invade the lymphatic vessels depending on its volume in this border area. A distribution of lymphatic vessels in normal bladder urothelium in this study possibly supports the clinical association of lymph node involvement with the depth of muscle invasion. LYVE-1 antibody was a useful specific marker for illustration of the lymphatic vessel in conventional paraffin-embedded specimens of the urinary bladder.

Abstract 512: Infiltrating CD4 Positive T cells may Support an Angiogenic Switch in Human Coronary Atheroma

Background: Pathologic angiogenesis has been implicated in the development of high-risk plaques where nascent immature leaky vessels become the primary source of intraplaque hemorrhage contributing to necrotic core expansion. The stage of lesion progression, however, favoring an angiogenic response has not been identified. Methods and Results: Multiple coronary lesions collected from 73 sudden coronary death victims at autopsy were examined. Plaque morphologies included pathologic intimal thickening (PIT), fibroatheromas whose cores were in an early stage (eFA) or late stage (lFA) of necrosis, thin-cap fibroatheromas (TCFA) and ruptures (pR). Lesions were characterized by immunostaining using specific markers directed against endothelial cells (CD31/CD34), smooth muscle cells (α-actin), macrophages (CD68), and T-lymphocytes (CD45RO, CD4). Mean intraplaque vessel densities per mm 2 between lesions with PIT and eFA increased significantly (PIT= 15.84±16.73 vs. eFA= 42.07±40.70, p=0.0054) where maximal numbers of vessels were found in pR (66.57±33.42). Overall, intraplaque vessels associated with actin 3 SMCs, were generally &lt;10%, suggesting a more immature phenotype. Increased vessel densities coincided with greater numbers of inflammatory macrophages and T-cells. Notably, T-cell numbers in areas with high intraplaque vessel densities increased approximate 3-fold between lesions with PIT and eFA (PIT= 62.62±76.50 vs. eFA= 182.33±171.54, p=0.0043) with maximal numbers displayed in TCFAs and pRs, in particular at the intimal/medial border. Populations infiltrating T cells were predominantly CD4 3 and most of these appeared to be activated (human leukocyte antigen DR positive). Conclusion: Atherosclerotic coronary arteries express a latent proangiogenic phenotype coinciding with a localized increased in activated CD4 3 T-cells and early plaque necrosis.

Expression of thymosin β10 and its role in non–small cell lung cancer

The exact role of thymosin beta10 in lung cancer progression remains unclear. We investigated by immunohistochemistry the expression of thymosin beta10 protein in tumors and tumor-adjacent tissues from 69 patients with non-small cell lung cancer. The relationship of thymosin beta10 expression with vascular endothelial growth factor, vascular endothelial growth factor-C, microvessel density, and lymphatic vessel density was determined; clinicopathologic factors and surgical treatment outcome were also studied. The results showed that thymosin beta10 was mainly expressed in the cytoplasm of lung cancer cells, and the overexpression of thymosin beta10 was correlated with advanced clinical stage (P = .026), distant metastases (P = .016), lymph node metastases (P = .007), poor degree of differentiation (P = .03), and poor postoperative survival (P = .004). Furthermore, thymosin beta10 overexpression was associated with vascular endothelial growth factor (P = .004), vascular endothelial growth factor-C (P = .017), microvessel density (P = .000), and lymphatic vessel density (P = .002). The lowest survival rate was observed in the patients with high thymosin beta10, positive vascular endothelial growth factor, and high microvessel density (P = .007) or in the patients with high thymosin beta10, positive vascular endothelial growth factor-C, and high lymphatic vessel density (P = .005). These results suggest that thymosin beta10 might induce microvascular and lymphatic vessel formation by up-regulating vascular endothelial growth factor and vascular endothelial growth factor-C in lung cancer tissues, thus promoting the distant and lymph node metastases and being implicated in the progression of non-small cell lung cancer.

The presence of tumor-infiltrating FOXP3 + lymphocytes correlates with intratumoral angiogenesis in endometrial cancer

Objectives CD4 +CD25 + regulatory T-cells (Tregs), that express the transcription factor FOXP3, suppress effector T-cell populations and can enable tumour cells to evade the host immune response. In this study, we investigated the numbers of FOXP3 + Tregs in the normal and malignat endometrium and examined potential links with tumor angiogenesis. Methods Paraffin-embedded tissues from 79 patients with stage I endometrial adenocarcinoma and 12 samples from normal endometrium were analyzed using immunohistochemistry for the detection of FOXP3 + lymphocytes. The presence of FOXP3 + lymphocytic infiltration was correlated with the tumor vascular density, the hypoxia inducible factors HIF-1α and HIF-2α, VEGF, estrogen and progesterone receptor expression. Survival analysis was also performed. Results In normal endometrium, FOXP3 was expressed by stroma infiltrating lymphocytes, with a mean number 8 (range 5–11) lymphocytes per ×100 optical field. In tumors, 55/79 (69.6%) cases showed little FOXP3 + lymphocytic infiltration (0–2 per ×100 optical field). In the remaining 24/79 (30.4%) cases that were scored as positive the mean score ranged from 3–8 (median 5). Low numbers of FOXP3 + lymphocytes significantly correlated with tumoral ER negativity and low vascular density. Survival analysis showed no significant impact of FOXP3 lymphocytic infiltration, although there was a trend towards worse prognosis. Conclusions The correlation between the presence of FOXP3 + Tregs and high vessel density in endometrial adenocarcinomas suggests a link between immunity, intratumoral angiogenesis and poor prognosis. However, further studies are required as significantly fewer Tregs were detected in the tumor microenvironment compared to normal endometrium.

Vascular thickness and cytostatic-resistant bronchial carcinoma

A new way of treatment of malignant tumours was suggested by blocking angiogenesis and thus creating subpopulations of cells which, due to their decreased oxygen content, react differently to treatment. The relationship between angiogenesis (vessel density) and resistance to cytostatic drugs was investigated. Results were analysed retrospectively for 83 untreated patients (73 men, 10 women; mean age 60 [37-75] years) with non-small cell bronchial carcinoma. Vessel density and resistance proteins were determined immunohistochemically. Radioactive incorporation of nucleic acid precursors with and without doxorubicin was measured with the in vitro short-term test. There were significant differences between vessel density and various resistance proteins. With low vessel density (less than mean value) glutathione-S-transferase pi (P < 0.01), thymidylate synthase (P < 0.05) and metallothioneine (P < 0.05) were significantly increased. P-glycoprotein showed a corresponding relationship, but it was not statistically significant, while there was no relationship with topoisomerase II. Bronchial carcinoma with low vessel density was more frequently resistant in vitro to doxorubicin than carcinomas with higher vessel density (P < 0.05). Non-small cell bronchial carcinoma with low vessel density more frequently expresses resistance proteins and is more commonly resistant to doxorubicin. Blocking angiogenesis to improve the results of treatment is therefore problematic.

Chitosan Scaffold Enhances Angiogenesis within an in vitro Reconstructed Bladder Wall, an Animal Study

Background: The function of a reconstructed bladder dependson the contracting bladder wall. This can be obtainedwith sufficient angiogenesis. Polyglycolic acid (PGA) is usedas a cell vehicle for bladder reconstruction. Chitosan supportsadhesion and differentiation of endothelial cells. Theaim of the study was to compare PGA with PGA/chitosan‘sandwich’ grafts for bladder wall regeneration. Materialsand Methods: 3T3 fibroblasts were seeded on 6 PGA and 3chitosan scaffolds and incubated 3 days at 37ºC and 5% CO2before implantation. Three rats underwent bladder reconstructionwith PGA cell seeded grafts and 3 with PGA graftscovered with chitosan cell seeded grafts (‘sandwich’ graft).Three rats as controls were not operated on. After 6 monthsreconstructed tissue was stained with H&E. Vessels wereidentified by immunohistochemistry by CD31 and CD34staining. A 4-level scale was used, (+++) referred to evidentmarker presence (high density of vessels) and (-) as lack of amarker (no vessels). Results: No complications during the 6-month period were noted. PGA/chitosan grafts were evaluatedas (+) while PGA grafts were evaluated as (+) and (-) bythe use of the CD31 antibody. The control group was evaluatedas (+). PGA/chitosan grafts were evaluated as (++) and(+) while PGA grafts were evaluated as (++) and (-) by CD34staining. The control group was evaluated as (+) and (-). Conclusion:Chitosan improved PGA abilities as a cell matrix andenhanced angiogenesis within the in vitro reconstructedgraft.