Biology of liver metastases.

A primary goal of cancer research is an increased understanding of the molecular mechanisms mediating the process of cancer metastasis. Analyses of colon cancer cells (the seeds) and the microenvironment (the soil) have increased our understanding of the biologic mechanisms mediating metastasis formation. Insight into the molecular mechanisms regulating the pathobiology of colon cancer metastasis, as well as a better understanding of the interaction between the metastatic cell and the host environment (including the vasculature), should provide a foundation for new therapeutic approaches. To the clinician, it is readily apparent that by the time metastases form, most steps in the metastatic cascade have completed. Therefore, therapy to down-regulate or interrupt the last stages of metastasis, proliferation and angiogenesis as well as mechanisms to disrupt cell survival signals seems the most promising areas of investigation.

Epidermal growth factor receptor blockade with C225 plus gemcitabine results in regression of human pancreatic carcinoma growing orthotopically in nude mice by antiangiogenic mechanisms.

Both epidermal growth factor receptor (EGF-R) signaling mechanisms and angiogenesis have been evaluated as independent targets for therapy of human pancreatic carcinoma, but a link between the two processes has been identified only recently. This study evaluated whether EGF-R blockade therapy with anti-EGF-R antibody C225 inhibits pancreatic carcinoma growth and metastasis in an orthotopic nude mouse model via tumor-mediated angiogenesis and whether gemcitabine potentiates this effect. In vitro treatment of human pancreatic carcinoma L3.6pl cells with C225 inhibited EGF-R autophosphorylation, producing a maximum of 20% cytostasis. Treatment with C225 plus gemcitabine resulted in additive cytotoxic effects that increased with increasing gemcitabine concentrations. Dose-dependent decreases in expression of the angiogenic factors vascular endothelial growth factor and interleukin 8 (but not basic fibroblast growth factor) were observed in the C225-treated cells (mRNA and protein levels). In L3.6pl tumors established in the pancreas of nude mice, systemic therapy with C225 alone and C225 in combination with gemcitabine resulted in growth inhibition, tumor regression, and abrogation of metastasis; median tumor volume was reduced from 538 to 0.3 and to 0 mm3, respectively. Gemcitabine treatment alone reduced median tumor volume from 538 to 152 mm3. Liver metastases were present in 50% of the controls, 30% of the gemcitabine-treated animals, and 20% of C225-treated animals. No macroscopically visible liver metastases were observed in the combination treatment group. As early as 11 days after C225 treatment, the median percentage of proliferating cell nuclear antigen-positive cells was substantially reduced compared with gemcitabine treatment alone (26% versus 73%, respectively) versus controls (92%), correlating with in vivo blockade of EGF-R activation. Similarly after 11 days treatment, production of vascular endothelial growth factor and interleukin 8 was significantly lower in C225 and C225 plus gemcitabine-treated tumors versus gemcitabine-treated and control tumors. Significant differences in microvessel density were observed 18 days after C225 or combination treatments (but not gemcitabine alone) in direct correlation with the difference in percentage of apoptotic endothelial cells, as visualized by double immunofluorescence microscopy. These experiments indicate that therapeutic strategies targeting EGF-R have a significant antitumor effect on human L3.6pl pancreatic carcinoma growing in nude mice which is mediated in part by inhibition of tumor-induced angiogenesis, leading to tumor cell apoptosis and regression. Furthermore, this effect is potentiated in combination with gemcitabine.

Zenker's diverticulum in the elderly: a neurologic etiology?

Though described in 1769, the etiology of Zenker's diverticulum remains unclear. Various primary esophageal motor disorders have been proposed, but no consistent manometric pattern or anatomic etiology has been uniformly recognized. An association with clinical neurologic disease at our institution prompted a review of 12 cases of Zenker's diverticulum in patients over 60 years of age, treated in the last 8 years. Nine patients (75%) underwent cricopharyngeus myotomy and diverticulectomy, with uniformly good results. Ten patients (83%) had an associated neurologic disorder, substantiated by cranial CT or MRI, in most cases. A wide range of neurologic problems were identified, but a strong trend toward brainstem or basilar lesions was present. As expected, the etiology of the neurologic abnormality in most patients in this group was cerebrovascular disease, but two patients had peripheral neuropathies. We suggest that the etiology of Zenker's diverticulum in the elderly may be neurologic in origin. Esophageal motor disorders, including incomplete upper esophageal sphincter opening and increased hypopharyngeal pressures, which may result in Zenker's diverticulum, may be a manifestation of central or peripheral neurologic disease in the elderly.

Molecular determinants of colon cancer metastasis.

Colon cancer metastasis is a tightly regulated process that requires a cancer cell to express genes that allow progression through various distinct steps. Aberrations in gene expression by cancer cells leads to transformation, growth, angiogenesis, invasion, dissemination and survival in the circulation, attachment in the organ of metastasis, and again invasion, growth, and angiogenesis. In addition to the genotype/phenotype of the tumor cell, for a tumor cell to become a clinically relevant metastasis, it must be able to respond appropriately to the environment. This includes being able to utilize growth factors and blood vessels from the organ of metastasis for the benefit of the tumor mass. Understanding the molecular and biologic mechanisms of colon cancer metastasis will allow the development of rationale therapeutic strategies that are more likely to impact the natural history of this disease than current therapies.

Effect of macrophage stimulation on collagen biosynthesis in the healing wound.

Immunomodulators that enhance macrophage function have been shown to be beneficial in a number of wound-healing models in humans and in experimental animals. The exact mechanism of this improved healing is unclear. To assess the role of collagen biosynthesis, the immunomodulator glucan phosphate was utilized in two murine models of wound healing, i.e., colon anastomosis and full-thickness skin incision. Tensile strength was evaluated using computer-assisted constant velocity tensiometry. Collagen biosynthesis was determined by assaying hydroxyproline content of wound hydrolysates by N-(9-fluorenyl)methoxycarbonyl/o-phthalaldehyde high-performance liquid chromatography. Experimental animals were treated with (1-3)-beta-D-glucan phosphate (250 mg/kg) intravenously 24 hours prior to colon anastomosis or skin incision. A second dose of glucan phosphate was given immediately postoperatively. Control animals received dextrose and water (5% w/v) intravenously. Tensile strength and hydroxyproline content were measured on postoperative Day 3. In the skin wound model, glucan phosphate treatment increased (P < 0.05) tensile strength by 42 per cent (342.5 +/- 12.2 vs 241.8 +/- 4.8 g), and hydroxyproline content was increased by 23.5 per cent (242.0 +/- 14.4 vs 196.8 +/- 10.5 pmol/microg; P < 0.05). In the glucan phosphate group, colon tensile strength was significantly (P < 0.05) increased by 34 per cent (34.2 +/- 2.3 g vs 45.8 +/- 2.1 g), and hydroxyproline content was increased by 7 per cent (47.45 +/- 3.31 vs 44.34 +/- 3.74 pmol/microg). These data indicate that macrophage modulation with glucan phosphate will increase tensile strength in experimental colon and skin wounds. In addition, we observed a positive correlation between glucan phosphate treatment, wound tensile strength, and collagen biosynthesis.

Receptor binding and internalization of a water-soluble (1-->3)-beta-D-glucan biologic response modifier in two monocyte/macrophage cell lines.

Glucan phosphate, a water-soluble, chemically defined (1-->3)-beta-D-glucan biologic response modifier, has been reported to exert antisepsis activity and accelerate wound healing. In this study we describe the specific binding of glucan phosphate to human and murine monocyte/macrophage cell lines, U937 and J774A.1, respectively. At 37 degrees C, equilibrium binding was rapidly achieved, i.e., within 1 min. In U937 cells, binding occurred with an affinity (Kd) of 37 microM and a Bmax of 65 x 106 binding sites/cell at 37 degrees C. In J774A.1 cells, glucan phosphate bound with an affinity (Kd) of 24 microM and a Bmax of 53 x 106 binding sites/cell at 37 degrees C. In both cases there was insignificant nonspecific binding. We further demonstrated that bound glucan phosphate cannot be displaced by a 50-fold excess of unlabeled ligand, suggesting internalization of glucan phosphate. Transmission electron microscopy showed significantly increased cytoplasmic vacuolization and significantly decreased mitotic activity in glucan phosphate-treated U937 cells compared with that in untreated cells. Pullulan, a random coil alpha-(1-->4)-(1-->6)-linked glucose polymer that served as a control, did not compete for the same binding site as glucan phosphate in either cell line, indicating the specificity of the binding site for (1-->3)-beta-D-glucans. We conclude that water-soluble pharmaceutical grade (1-->3)-beta-D-glucan phosphate specifically binds to and is internalized by U937 and J774A.1 cells.

Benefits of pulmonary artery catheter and transesophageal echocardiographic monitoring in laparoscopic cholecystectomy patients with cardiac disease.

BACKGROUND: Because the abdominal insufflation and desufflation associated with laparoscopic procedures may adversely effect a compromised myocardium, patients with significant cardiopulmonary disease should be closely monitored during these procedures. The utility of intraoperative pulmonary artery catheter (PAC) and transesophageal echocardiography (TEE) monitoring was studied in 10 patients with moderate to severe cardiopulmonary disease to identify patients at greatest risk for cardiovascular complications during laparoscopic cholecystectomy. METHODS: Ten patients were enrolled in this prospective study; 7 had suffered a previous myocardial infarction, 6 had undergone coronary artery bypass grafting, and 9 had disease classified as Goldman's class II or greater. The heart was monitored by TEE throughout the laparoscopic cholecystectomy by using real-time, two-dimensional mode to study the wall thickness and motion. Several PAC measurements were taken directly: cardiac output, systemic vascular resistance, pulmonary artery wedge pressure, and central venous pressure. Heart rate and blood pressure were also obtained at corresponding intervals. Cardiac index, stroke volume, and left and right ventricular stroke work were then calculated. RESULTS: TEE demonstrated no significant changes in ventricular wall motion throughout laparoscopy. In patients who had postoperative cardiovascular complications, significant changes in cardiac index, left ventricular stroke work, and stroke volume were seen after pneumoperitoneum release. Compared to that of patients who did not develop complications, the cardiac index in those with complications dropped 42% (3.10 +/- 0.72 versus 1.80 +/- 0.10 L/min per m2, respectively; P < 0.01); left ventricular stroke work dropped 64% (139.00 +/- 11.36 versus 50.38 +/- 10.55 g x min/beat, respectively; P < 0.01); and stroke volume dropped 51% (86.90 +/- 12.68 versus 42.50 +/- 5.08 mL/beat, respectively; P < 0.01). CONCLUSIONS: PCA monitoring in patients with compromised cardiac function is useful in identifying patients who may not tolerate hemodynamic changes after pneumoperitoneum release. Normalization of hemodynamic changes secondary to abdominal insufflation and desufflation in patients with compromised hearts may not occur in patients with compromised hearts may not occur for hours postoperatively. Abnormal hemodynamic changes occur within the first hour after desufflation in patients who later develop cardiovascular complications, which are heralded by significant drops in left ventricular stroke work, cardiac index, and stroke volume. TEE did not prove to be useful for intraoperative monitoring.

Rattlesnake bite of the face: case report and review of the literature.

Most poisonous snakebites with envenomation involve the extremities. A case where the bite occurred on the face is presented. Problems in management included marked coagulopathy and prolonged airway compromise, requiring 9 days of nasotracheal intubation. The bite site did not develop necrosis and required only local wound care.