Assessment of vascularity in histological sections: effects of methodology and value as an index of angiogenesis in breast tumours

The aims of this study were to (a) determine how the quantification of blood vessels in histological sections (vascularity) is affected by the methodology used and (b) assess the value of vascularity as an index of angiogenesis by comparing tumour and normal breast tissue. Archival specimens of breast, lung and oral carcinoma, oral dysplasia and normal breast tissue were used to test the effects of the following experimental variables on vascularity: pretreatment of the sections (enzymatic digestion, heating), endothelial markers (von Willebrand factor and CD31 antibodies), method of quantification (highest microvascular density, average microvascular density and microvascular volume) and interobserver variations. All the variables examined significantly affected the estimated vascularity; this depended on the type of tissue and method used. The pretreatment of the sections before staining was the most important variable, altering the vascularity ranking of the tumours. Vascularity in breast tumours was similar to that of the normal breast intralobular stroma, suggesting that an area of high microvascular density in the tumour does not necessarily represent tumour-induced angiogenesis. Contradictory results have been published regarding the value of vascularity as a tumour prognostic factor. Our results suggest that statistically significant differences in vascularity values are most likely to arise from failure to optimize the staining protocol and from the method used to assess vascularity. © 1998 Chapman & Hall     

Factors influencing the neovascularization of experimental tumours

When experimental tumours are inoculated into a host animal, the tumour growth depends, among other things, on its vascular supply. This vascular supply has been shown to be initiated by substances released by the tumour tissue, and vascular sprouting towards implanted tumour substances has been extensively demonstrated in nonvascular tissue. Most tissues, however, already contain a vascular supply sufficient for their own needs. In such conditions, the host vascular system is probably incorporated into the tumour without much vascular sprouting. It is well known that, as a tumour grows larger, the center tends to become ischaemic and necrotic. It is not clear why the tumour vascularity does not respond to this development with reactive vascular proliferation, but increased interstitial tissue pressure and impaired fluid transport may be implicated.     

Changes in oviducal vascularity during the reproductive cycle of three oviparous lizards (Eumeces obsoletus, Sceloporus undulatus and Crotaphytus collaris)

Histologically derived estimates and ink suspension vascular casts were used to examine oviducal vascular changes. Vascularity peaked during gravidity and was correlated with maximal plasma progesterone concentrations. The vascular increase in the oviducal tissue was attributed exclusively to increased capillary densities. The greatest change occurred in the anterior uterus where incubation and egg shell secretion occur. Similar patterns of change in vascularity occurred in the infundibulum, although not as extreme as that seen in the anterior uterus, whereas no significant alterations were noted in the posterior uterus. These modifications mimic the pattern of vascular change occurring in viviparous lizards during simple placentation. We suggest that major changes in uterine vascularity may not be required for the evolution of simple chorioallantoic placentae in lizards.     

Spatial and temporal variations of Penilia avirostris and Evadne tergestina (Crustacea,Branchiopoda) in a Tropical Bay,Brazil

We analysed monthly samples collected in Guanabara Bay, with a conical net of 200 m mesh during 1985. The bay was divided into three areas: an outer region (area A), influenced by oceanic waters; an inner region (area C), influenced by fluvial inflow; and a transition region (area B) with intermediate features. Penilia avirostris and Evadne tergestina were observed in the three areas, with greatest densities, however, in the outermost region, which had the highest salinities and lowest temperatures. Penilia avirostris was more abundant in summer (March), a period with the greatest relative densities of nanoplankton. Evadne tergestina was also abundant in summer, but its peak fell in November, a period with a relative increase in microphytoplankton density in the bay. The two species disappeared in winter: Penilia avirostris was absent from May to August, whereas Evadne tergestina disappeared in August and September.     

Postnatal development of vascularity in the inferior colliculus of the young rat

The inferior colliculus in the rat midbrain is an auditory relay center whose functional maturation occurs postnatally. We examined by morphometry the vascularity and the nuclear profile density of the inferior colliculus in normal young rats at different ages (before and after the onset of auditory input). We also compared this region with a frontal region of the cerebral cortex in 24-day-old rats. The inferior colliculus from aldehyde-perfused Sprague-Dawley rats aged 5, 9, 14, and 24 days was analyzed by light microscopy of semithin plastic sections. The central region (mostly the central nucleus) was sampled at 5 levels representing its entire rostrocaudal extent. Patent-blood-vessel profiles were counted and classified according to their size and profile orientation. Counts of nuclear profiles in the same sections were also made. In the inferior colliculus of rats between 5 and 24 days of age, the small (less than 10-microns diameter) cross-sectioned vessel profiles increased over 6-fold in number per unit area. Correspondingly the vascular volume density, estimated by differential point counting, increased between these ages. However, there was a decrease in the number of neuronal and glial nuclear profiles per unit area, probably because of growth in the volume of the neuronal perikarya and processes, along with cell emigration reported to occur at early postnatal ages. This study has shown that an increase in vascularity in the central region of the rat inferior colliculus continues for up to 2 weeks after the onset of hearing.(ABSTRACT TRUNCATED AT 250 WORDS)     

Blood flow dynamics after photodynamic therapy with verteporfin in the RIF-1 tumor

In the present study, the effects of photodynamic therapy (PDT) with verteporfin on tumor blood flow and tumor regrowth were compared as verteporfin distributed in different compartments within the RIF-1 tumor. Tissue distribution of verteporfin was examined by fluorescence microscopy, and blood flow measurements were taken with a laser Doppler system. It was found that, at 15 min after drug administration, when verteporfin was mainly confined within the vasculature, PDT induced a complete arrest of blood flow by 6 h after treatment. PDT treatment at a longer drug-light interval (3 h), which allowed the drug to diffuse to the tumor interstitium, caused significantly less flow decrease, only to 50% of the initial flow in 6 h. A histological study and Hoechst 33342 staining of functional tumor vasculature confirmed the primary vascular damage and the decrease in tumor perfusion. The regrowth rate of tumors treated with 15-min interval PDT was 64% of that of the control group. However, when tumors were treated with 3-h interval PDT, the regrowth rate was not significantly different from that of the control, indicating that only the 15-min interval PDT caused serious damage to the tumor vascular bed. These results support the hypothesis that temporal pharmacokinetic changes in the distribution of the photosensitizer between the tumor parenchyma and blood vessels can significantly alter the tumor target of PDT.     

Neoadjuvant cisplatin for head and neck cancer: Simulation of a novel schedule for improved therapeutic ratio

Current clinical studies support the role of neoadjuvant cisplatin administration prior to curative radiotherapy or radio-chemotherapy for advanced head and neck cancer. Nevertheless, based on locoregional control rates the studies indicate that there is need to redesign cisplatin-based schedules for induction chemotherapy, thus the ideal treatment regimen is yet to be established. While the pharmacokinetics/dynamics of daily cisplatin regimens correspond better with the cell cycle properties of head and neck cancers, weekly regimens are more commonly employed in clinics due to lower complications. Yet, the high rates of adverse events induced by current cisplatin schedules often represent a limiting factor in the overall success of the treatment.The aim of the present paper was to model the pharmacodynamic properties of cisplatin and to simulate and compare various neoadjuvant treatment regimens in regards to their effect on tumour control. Treatment simulation was undertaken on a virtual squamous cell carcinoma of the head and neck, previously grown by computer-based probabilistic methods.The model suggests that a novel cisplatin treatment, given every three days is comparable, in regards to tumour control, with the daily administration and more effective than the weekly regimen in neoadjuvant settings. Endpoints were assessed in terms of cell population regrowth after treatment cessation followed by two weeks of unperturbed growth. Simulation of two weeks low-dose daily cisplatin followed by two weeks ‘free growth’ lead to 15% population regrowth, while weekly high-dose cisplatin over three weeks, followed by two weeks unperturbed growth resulted in 52% tumour cell regrowth. The proposed novel schedule of low-dose third-daily cisplatin gives closer tumour regrowth to daily administration (27% versus 15%) than to the weekly regimen (52%) and also similar cell distribution along the cell cycle as the daily one, suggesting therefore comparable response to subsequent treatment.The advantage of using a third-daily drug regimen would be a decrease in normal tissue complication rates compared to daily administration and possibly an increase in tumour control when compared to the ‘conventional’ weekly cisplatin delivery.     

Placental growth throughout the last two thirds of pregnancy in sheep: vascular development and angiogenic factor expression

Morphometric methodologies were developed and applied to investigate the patterns of vascular development in maternal (caruncular; CAR) and fetal (cotyledonary; COT) sheep placentas throughout the last two thirds of gestation. We also examined the expression levels of the major angiogenic factors and their receptors in CAR and COT sheep placentas. Although the vascularity of the CAR tissues increased continuously from Day 50 through Day 140 of pregnancy, those of the COT tissues increased at about twice the instantaneous rate (i.e., the proportionate increase/day) of the CAR. For CAR, vascularity increased 2-fold from Day 50 through Day 140 via relatively small increases in capillary number and 2- to 3-fold increases in capillary diameter. For COT, the increased vascularity resulted from a 12-fold increase in capillary number associated with a concomitant 2-fold decrease in capillary diameter. This large increase in fetal placental capillary number, which was due to increased branching, resulted in 6-fold increases in total capillary cross-sectional area and total capillary surface, per unit of COT tissue. Different patterns of expression of the mRNAs for angiogenic factors and their receptors were observed for CAR and COT. The dilation-like angiogenesis of CAR was correlated with the expression of vascular endothelial growth factor receptor-1 (FLT1), angiopoietin-2 (ANGPT2), and soluble guanylate cyclase (GUCY1B3) mRNAs. The branching-like angiogenesis of COT was correlated with the expression of vascular endothelial growth factor (VEGF), FLT1, angiopoietin-1 (ANGPT1), ANGPT2, and FGF2 mRNAs. Monitoring the expression of angiogenic factors and correlating the levels with quantitative measures of vascularity enable one to model angiogenesis in a spatiotemporal fashion.     

Photoradiation therapy of cutaneous angiosarcomas in mice

Cutaneous angiosarcoma is a relatively rare but devastating malignant vascular tumor. It has a high incidence of recurrence following conventional therapeutic modalities applied either singly or in combination. The increased vascularity of cutaneous angiosarcomas, facilitating selective uptake and retention of a photosensitizing agent, such as hematoporphyrin derivative (HPD), suggests that these tumors would respond well to photoradiation therapy. To study the feasibility of this treatment modality, transplantable hemangiosarcomas were implanted in B6C3F1 female mice. Within 2.5 to 3.5 hours after intraperitoneal administration of HPD, fluorescence was recorded in the tumor as compared with surrounding normal skin. When these photosensitized tumors were exposed to 70 J/cm2 of laser energy from an argon-pumped dye laser at 630 nm, the tumors showed marked necrosis within 24 hours. In another series, the tumors were initially photosensitized with HPD for 3 hours and then treated with laser energy ranging from 0 to 96 J/cm2. A dual labeling procedure demonstrated a dose-related decrease in DNA synthesis rate in tumors that were exposed to 0 to 30 J/cm2 at 24 hours after treatment. Furthermore, tumor tissue exposed to laser energy in excess of 30 J/cm2 showed no significant cellular DNA synthesis. These data, supported by histologic evidence of tissue destruction, suggest that photoradiation therapy has a great potential as a therapeutic modality for cutaneous angiosarcomas.     

COMPARISON OF TETRAPLOID AND SINGLE GENE-INDUCED GIGAS VARIANTS IN CHICKPEA (CICER ARIETINUM). II. MORPHOLOGICAL AND ANATOMICAL LEAF CHARACTERS

Two genomic variants of a chickpea (Cicer arietinum L.) parental line have been developed which exhibit gigas characters. The two genotypes were the result of a single-gene mutation (gigas) and induced tetraploidy of a single parental line. The two genotypes plus parental strain were investigated to determine the similarity-of-effect of polyploidy and this single-gene mutation on leaf anatomy and morphology. Leaves consisted of two rows of alternatively arranged leaflets. Both the tetraploid and parental lines had the same mean number of leaflets per leaf while the gigas plants had fewer, but mean total leaf surface area was greater in the gigas plants. Quantitative comparison of mesophyll and vascular tissue and air space volume density (V_v) showed that leaves of the tetraploid plants had the greatest mesophyll cell density (V_vm) and least air space density. Mesophyll cell density was equal in the parental and single-gene mutant while parental leaves had the greatest vascular tissue density. The greater mesophyll cell density values of the polyploid were due to larger mean mesophyll cell size, not to greater cell numbers per unit area. Leaf models based on tissue density and leaflet size showed tetraploid plants had the greatest productivity potential per unit of leaflet surface area. However, if models were based on a whole leaf, gigas plants had the greatest productivity potential since they had larger total leaf area. The effectiveness of using structural models to predict physiological potential in plant tissues will be tested in future studies.     

The role of mechanical host-tumour interactions in the collapse of tumour blood vessels and tumour growth dynamics

A mathematical model of residual stress evolution in a growing vascular tumour is presented, in an attempt to elucidate the poorly understood phenomenon of vascular collapse. Whereas earlier studies in this area have neglected the effects of mechanical interactions between the tumour and the surrounding host tissue, the significance of these interactions for the long-term development of a tumour is now considered. The model predicts tumour stress distributions which reflect the distinctive patterns of vascular collapse reported in experimental studies. Moreover, while neglecting mechanical host/tumour interactions results in the eventual complete regression of the tumour to its avascular dormant size in the event of vascular collapse, this new model points to the possibility of oscillations in the tumour's size in the long term.     

Cellular abnormalities of blood vessels as targets in cancer

Tumor blood vessels have multiple structural and functional abnormalities. They are unusually dynamic, and naturally undergo sprouting, proliferation, remodeling or regression. The vessels are irregularly shaped, tortuous, and lack the normal hierarchical arrangement of arterioles, capillaries and venules. Endothelial cells in tumors have abnormalities in gene expression, require growth factors for survival and have defective barrier function to plasma proteins. Pericytes on tumor vessels are also abnormal. Aberrant endothelial cells and pericytes generate defective basement membrane. Angiogenesis inhibitors can stop the growth of tumor vessels, prune existing vessels and normalize surviving vessels. Loss of endothelial cells is not necessarily accompanied by simultaneous loss of pericytes and surrounding basement membrane, which together can then provide a scaffold for regrowth of tumor vessels. Rapid vascular regrowth reflects the ongoing drive for angiogenesis and bizarre microenvironment in tumors that promote vascular abnormalities and thereby create therapeutic targets.     

Vascularity at sites of gastro-jejunal anastomoses.

Recent interest in vascularity in relation to peptic ulceration poses the question of relationship between three entities: (1) site of gastro-jejunal (post-anastomotic) ulcer, (2) site of the anastomotic line and (3) site of junction and possible vascular anastomosis between gastric and jejunal circulations in the region of the anastomosis. Vascular injection studies in dogs show that the main gastric and jejunal circulations run up to, but do not communicate across, the surgical anastomosis, there being a line of least vascularity at the anastomosis. Since anastomotic ulcers occur distal to the anastomotic line, this supports the view that such ulcers are situated wholly in an area of jejunal circulation.     

Study on the characteristics of contrast-enhanced ultrasound and its utility in assessing the microvessel density in ovarian tumors or tumor-like lesions

Angiogenesis is a critical factor in tumor growth and metastasis, and microvessel density (MVD) was an important parameter for assessing vessels in tumors. However, radiologic assessment of tumor vascularity is not yet well established. In our study, we aimed at investigating the efficacy of contrast-enhanced ultrasonography (CEUS) in exploring the vascularity of the ovarian tumors or tumor-like lesions to assess the relationship between the parameters of the peak intensity (PI) and area under curve (AUC) on CEUS and MVD in ovarian masses. Compared to the contrast-enhanced ultrasound technique, conventional ultrasound shows limitation in differentiating benign and malignant ovarian tumors. The former is promising in improving the sensitivity of detecting small vessels and blood flow in ovarian tumors. Our results showed clear differences in enhancement patterns between benign and malignant ovary tumors or tumor-like lesions. The PI and AUC in the malignant tumors were significantly higher than those in the benign tumors or tumor-like lesions (p=0.001 and =0.01, respectively). The MVD was 43.1 ± 20.4 in the benign tumors or tumor-like lesions and was 65.3 ± 22.3 in the malignant ones (p= 0.01). In both the benign and malignant groups, the PI and AUC were correlated significantly with the MVD (r=0.595, p = 0.001; r =0.533, p = 0.003, respectively). The PI and AUC in CEUS can reflect the MVD in ovarin tumors. The PI and AUC of the ovarian masses in the contrast transvaginal sonography show significant correlation with the angiogenesis and may help in assessing tumor vascularity in ovarian masses.     

The nutrition of the human meniscus: A computational analysis investigating the effect of vascular recession on tissue homeostasis

The meniscus is essential to the functioning of the knee, offering load support, congruency, lubrication, and protection to the underlying cartilage. Meniscus degeneration affects ∼35% of the population, and potentially leads to knee osteoarthritis. The etiology of meniscal degeneration remains to be elucidated, although many factors have been considered. However, the role of nutritional supply to meniscus cells in the pathogenesis of meniscus degeneration has been so far overlooked. Nutrients are delivered to meniscal cells through the surrounding synovial fluid and the blood vessels present in the outer region of the meniscus. During maturation, vascularization progressively recedes up to the outer 10% of the tissue, leaving the majority avascular. It has been hypothesized that vascular recession might significantly reduce the nutrient supply to cells, thus contributing to meniscus degeneration. The objective of this study was to evaluate the effect of vascular recession on nutrient levels available to meniscus cells. This was done by developing a novel computational model for meniscus homeostasis based on mixture theory. It was found that transvascular transport of nutrients in the vascularized region of the meniscus contributes to more than 40% of the glucose content in the core of the tissue. However, vascular recession does not significantly alter nutrient levels in the meniscus, reducing at most 5% of the nutrient content in the central portion of the tissue. Therefore, our analysis suggests that reduced vascularity is not likely a primary initiating source in tissue degeneration. However, it does feasibly play a key role in inability for self-repair, as seen clinically.     

Macrofungal diversity and community ecology in mature and regrowth wet eucalypt forest in Tasmania: A multivariate study

Abstract The biodiversity of macrofungi in mature and young regrowth Tasmanian wet forests is described at the species level and at the community level. The macrofungal communities studied were much more species‐rich than their vascular plant counterparts, with the total number of macrofungal taxa outnumbering vascular plants by four to one. This ratio applied in both mature and young regrowth forest sites. Some 242 taxa of macrofungi were recorded, of which 132 were identified to species level, the remainder to species groups or higher taxa. Distinct communities could be discerned from multivariate analysis (ordination and classification) of vascular plant and macrofungal data from the mature and regrowth sites. The two vascular plant communities had different fire histories, and this difference is also assumed to account for the separation of the macrofungal communities of the two forest types. There was generally a high level of congruence between the vascular plant and the macrofungal communities. However, one young regrowth site, which was relatively close to the mature sites in the ordination space for the analysis of vascular plants, was distant from the mature forest sites for the analysis of macrofungi. Another regrowth site, which had experienced wildfire rather than silvicultural regeneration, clustered with mature sites for some analyses of the macrofungal assemblage. Variation in the macrofungal communities was correlated with a different set of the measured environmental variables than was variation in the vascular plant communities. Mature and young regrowth forests were found to have distinctly different macrofungal floras, with approximately 40% of the taxa in each forest type being restricted to that type of site. Suitable indicator taxa (restricted or preferential to particular forest types) for use in further studies are suggested.     

Feasibility and safety of silicone rubber contrast-enhanced microcomputed tomography in evaluating the angioarchitecture of prostatectomy specimens

This ethics committee-approved pilot study was carried out with informed consent. A protocol was developed to assess the feasibility of in vitro Microfil injection of prostate cancer specimens followed by analysis with micro-computed tomography (microCT) to characterize the functional vascularity of prostatic tissue and evaluate its safety with respect to the preservation of a specimen for pathologic examination. The visible prostatic arteries of two surgically resected prostates frompatients with known prostate cancer (PCa) were injected with MicrofilMV-122 contrast medium immediately after removal. The specimens were scanned using microCT and were qualitatively examined using three-dimensional analysis software (MicroView; GE Healthcare Biosciences). The Microfil perfusion in the two samples was sufficient to view the functional vascularity arising from a major prostatic artery, up to a resolution of 17.626 μm without any indication of adverse effects due to Microfil injection. Malignant prostatic regions showed a greater vascular density on histology but decreased vascular perfusion compared with benign prostatic regions. The use of microCT on Microfil-injected prostates seems to be a feasible and specimen-preserving method for visualizing the three-dimensional vessel patterns present in resected human prostates.     

Surgical treatment of a giant tibial high-grade mixofibrosarcoma with preservation of limb function: a case report

Myxofibrosarcoma is one of the most common sarcomas in elderly patients showing a slight male prevalence. The tumor is mainly located in lower and upper extremities and rarely in trunk, neck and feet. We present a case of a 84-year-old man referred to our tumour center with a giant and neglected high-grade tibial myxofibrosarcoma in the anteromedial side of tibial mid-diaphysis. Large size lesions in association with older age may jeopardise the maintenance of limb vitality, vascularity and stability.Authors performed a complete tumour resection, followed by reconstruction of bone and soft tissue defects with cement, plate and a musculocutaneous gastrocnemius flap, in order to cover the underlying bone and promote uneventful healing and perfusion of the operated extremity.At 2 years postoperatively, limb salvage, good functional outcome and no tumour recurrence were reported while the patient was able to effectively perform the majority of the daily activities.     

ANOMALOUS GROWTH AND VEGETATIVE ANATOMY OF SIMMONDSIA CHINENSIS

The anatomy of the stem, root, and leaf of Simmondsia chinensis (Link) Schneider was investigated, as well as the mode of tissue formation in the stem. Perivascular tissue is present as part of the primary body; outermost cell layers of this tissue mature as a fibrous sheath. The first short-lived extrafascicular cambium is generated within the remaining parenchymatous perivascular tissue. Successive independent extrafascicular cambia, organized as complete rings or large arcs, arise within peripheral conjunctive parenchyma produced by previous cambia. Extrafascicular cambia produce secondary xylem centripetally and conjunctive tissue bands and strands of secondary phloem centrifugally. Conjunctive tissue initials produce raylike structures of conjunctive tissue; true vascular rays are absent. The phellogen is actually a region of transition where the peripheral conjunctive parenchyma of previous extrafascicular cambia undergoes further cellular subdivision; a true phellogen is lacking. Xylem bands do not represent annual or seasonal growth increments, and secondary growth in Simmondsia is an unequivocal example of the “concentric” anomaly.