In vitro expansion of tumour cells derived from blood and tumour tissue is useful to redefine personalized treatment in non-small cell lung cancer patients.

The clinical development of locally and advanced non-small cell lung cancer (NSCLC) suffers from a lack of biomarkers as a guide in the selection of optimal prognostic prediction. Circulating Tumour Cells (CTCs) are correlated to prognosis and show efficacy in cancer monitoring in patients. However, their enumeration alone might be inadequate; it might also be critical to understand the viability, the apoptotic state and the kinetics of these cells. Here, we report what we believe to be a new and selective approach to visually detect tumour specific CTCs. Firstly, using labelled human lung cancer cells, we detected a specific density interval in which NSCL-CTCs were concentrated. Secondly, to better characterize CTCs in respect to their heterogeneous composition and tumour reference, blood and tumour biopsy were performed on specimens taken from the same patient. The approach consisted in comparing phenotype profile of CTCs, and their progenitor Tumour Stem Cells, (TSCs). Moreover, NSCL-CTCs were cultivated in short-time human cultures to provide response to drug sensitivity. Our bimodal approach allowed to reveal two items. Firstly, that one part of a tumour, proximal to the bronchial structure, displays a predominance of CD133+. Secondly, specific NSCL-CTCs Epithelial Cell Adhesion Molecule (EpCAM)+CD29+ can be used as a negative prognostic factor as well the high expression of CTCs EpCAM+. These data were confirmed by drug-sensitivity tests, in vitro, and by the survival curves, in vivo.

Protein-carbohydrate complex reveals circulating metastatic cells in a microfluidic assay.

Advances in carbohydrate sequencing technologies reveal the tremendous complexity of the glycome and the role that glycomics might have to bring insight into the biological functions. Carbohydrate-protein interactions, in particular, are known to be crucial to most mammalian physiological processes as mediators of cell adhesion and metastasis, signal transducers, and organizers of protein interactions. An assay is developed here to mimic the multivalency of biological complexes that selectively and sensitively detect carbohydrate-protein interactions. The binding of ß-galactosides and galectin-3--a protein that is correlated to the progress of tumor and metastasis--is examined. The efficiency of the assay is related to the expression of the receptor while anchoring to the interaction's strength. Comparative binding experiments reveal molecular binding preferences. This study establishes that the assay is robust to isolate metastatic cells from colon affected patients and paves the way to personalized medicine.

Surface properties and implantation site affect the capsular fibrotic overgrowth.

Transplantation of encapsulated pancreatic islets is a promising approach for the treatment of type 1 diabetes. Large-scale application of this technique, however, is hampered by insufficient biocompatibility of the capsules. In this study, we have evaluated the biocompatibility of a new synthetic material with six different chemical groups on their surface (amino, carboxy-sulfate, carboxylate, hydroxylate, sulfate, and PMMA) used for the fabrication of the microcapsules. Eight Lewis rats were inoculated with a suspension of empty capsules made for each candidate material in the retroperitoneal ileopsoas muscle and renal subcapsular space. Four weeks later kidney and muscle containing the capsules were explanted, paraffin embedded, sectioned and stained with Sirius Red and Masson's Trichrome for histological analysis. The amount of fibrosis was also ultrastructurally evaluated with scanning electron microscopy. The samples were then subjected to digitalized quantitative analysis using specific software to determine the degree of fibrotic overgrowth. The quantification of collagen deposition, calculated in proximity of the microcapsules, was expressed as a percentage of the total area and can be considered a good index for the biocompatibility, an essential prerequisite for functional pancreatic islet transplantation. The results show that subcapsular renal space is the best implantation site and the positive surface charge induces a more intense collagen synthesis.

Effect of Intralipid on some immunological parameters and leukocyte functions in patients with esophageal and gastric cancer.

This study has been undertaken to investigate if the intravenous (i.v.) infusion of fat emulsions may be associated with impairment of some immunological functions thus increasing the risk of septic complications. Fifteen malnourished patients with advanced gastric or esophageal cancer received for 2 weeks preoperatively and 1 week after surgery an isocaloric and isonitrogenous TPN treatment with Intralipid (group A: n=8) or glucose alone (group B: n=7) as energy substrate. Cluster analysis of 11 nutritional parameters and some tests of the humoral and cellular immunity (IgG, IgM, C3c, Factor B; polymorphonuclear (PMN) cells, total lymphocytes, T and B lymphocyte counts; 'in vitro' PMN chemotaxis, adherence to nylon fibers, phagocytosis of latex particles) were sequentially determined. The incidence and severity of post-operative infections were investigated and a 'sepsis score' was calculated for each patient. Pre- and postoperative TPN were not associated with an improvement of the nutritional status. The humoral and cellular immune parameters showed the same behaviour in patients receiving Intralipid and in controls. The chemotactic activity of PMN cells was constantly normal, granulocyte adherence fluctuated below the normality range in controls, whereas phagocytosis of latex was similar in both groups. Post-operative infectious episodes were less severe in patients receiving Intralipid. Our results do not confirm that Intralipid adversely affects some aspects of the humoral and cellular immune response.

Radioimmunoassisted follow-up and surgery vs traditional examinations and surgery after radical excision of colorectal cancer.

The authors report their experience in the radioimmunodetection of recurrent colorectal cancer in comparison with traditional examinations. 485 patients were studied after radical surgery for colorectal cancer: 168 (Group 1) were studied with a radioimmunoassisted follow-up plan including: Immunoscintigraphy (IS), serum markers assays, Radioimmunoguided Endoscopy (RIGE), Intraoperative Radioimmuno-localization (IORIL) in case of reoperation; 317 (Group 2) underwent a protocol with traditional examinations. In 24 patients of Group 1 IS was performed even at the time of their admission for primary cancer. 7 patients underwent RIGE for primary cancer and 16 in the follow-up. IORIL was performed in 12 patients with primary cancer and in 16 in the course of reoperation. The radioimmunodiagnostic methods were performed after a single administration of the radiolabeled MAb (111In F(ab')2 a-CEA and 111In B72.3). The radioimmunoassisted follow-up plan detected a greater number of recurrences than traditional examinations (27% vs 13%). The rates of radical reoperation in Group 1 and 2 were 61% and 37% respectively. 33% of the patients of Group 2 were alive 24 months after radical reoperation vs 62.5% of the patients of Group 1. Immunoscintigraphy demonstrated a good sensitivity and specificity, mainly in the detection of pelvic recurrences (sens. 92%, spec. 84%). The radioimmunoassisted follow-up plan was well accepted by the patients. RIGE led to the detection of 3 periluminal recurrences of rectal cancer that traditional investigations failed to demonstrate and in 5 cases influenced the patients management. IORIL detected minimal tumor foci (2 mm.) where pre and intraoperative study were negative, while the histopathologic examination gave evidence of tumor. Considering that the costs of the radioimmunodiagnostic methods and of traditional examinations are very similar, we can conclude that the radioimmunoassisted follow-up plan has a favorable cost/benefit rate and a remarkable impact on the treatment of patients with colorectal cancer.

Evaluation of possible causes of delayed hypersensitivity impairment in cancer patients.

The separate roles of malnutrition, advanced age, and stage of tumor growth as causes of impairment of delayed hypersensitivity response (DHR) was studied in 111 patients with solid tumors and in 56 nonneoplastic control patients matched for age, anatomical site of disease, degree of illness, and nutritional status. Pretreatment DHR to recall antigens (tuberculin, Candida, streptokinase-streptodornase, trichophyton) and to dinitrochlorobenzene in cancer patients with 9% anergic, 43% hypoergic, and 48% normoergic; the distribution of DHR in controls was not significantly different. In cancer patients, the serum albumin level showed an inverse correlation with the stage of tumor (p less than 0.01) and a positive correlation with the DHR (p less than 0.001); the serum albumin level was also in the controls positively correlated with the DHR (p less than 0.01), indicating that malnutrition in neoplastic or benign disease may cause depression of DHR. In well-nourished controls, age was inversely correlated with DHR (p less than 0.05), showing that aging itself may be another relevant cause of depression of DHR. The results of this study indicate that DHR in patients with solid tumors is similar to the DHR of nonneoplastic patients if matched for age, sex, and nutritional status. DHR impairment in cancer patients appears to be caused mainly by aging and by malnutrition due to the advanced progression of cancer.

Successful bronchial revascularization in experimental single lung transplantation.

It is the purpose of this paper to report our experience with bronchial artery revascularization in an experimental model of single lung transplantation in swine. Thirty-three large white pigs weighing 20-40 kg underwent left lung allotransplantation. In 24 animals, bronchial artery revascularization was attempted by anastomizing the aortic patch containing the bronchial artery orifice with the recipient descending aorta. Eight survivors were put to death on postoperative days 11-15; five animals were put to death or died on postoperative days 2-9; the other animals died intra-operatively or within a few hours. Preservation of left bronchial vascularization was achieved in all cases attempted, as documented by post-mortem injection of dye (methylene blue) or contrast medium. Five of the 8 animals surviving for 11-15 days showed diffuse graft hepatization, associated with diffuse vascular thrombosis. Whether this was caused by damage to the endothelium due to poor graft preservation or by rejection was unclear. In animals surviving for 11-15 days without gross lung pathology, the anastomosis and bronchial mucosa were completely normal; in contrast, bronchial ischaemic changes were found in nonrevascularized animals and in survivors with graft hepatization. Our experience confirms that re-anastomosis of the bronchial arteries can prevent bronchial healing problems in single lung transplantation. The pig is an ideal model for these experiments since the bronchial arteries have a constant common aortic origin, allowing easy identification and preservation of left bronchial vascularization.

Potentials of liposomes in diagnosis and treatment of pulmonary metastases: an experimental study in the rat.

OBJECTIVE: The ultimate goal of the therapy of lung metastases is to destroy all malignant cells while sparing normal ones. Liposomes represent a novel approach for the selective transport of tracers and therapeutic agents to cancer cells because of their flexibility, low toxicity, wide range of possible variants, simplicity to make, and because agents can be entrapped in them in their native states in large amounts. We have studied the biodistribution of "Stealth" liposomes in the experimental model of lung metastases in the rat. METHODS: The secondaries were induced by i.v. injection 20. 10(6) cancer cells (DHD/K12/TRb line) in BD-IX rats. The study of the liposome biodistribution in the rat was carried out by the use of unilamellar liposomes with homogeneous size distribution (0.1 microns), the liposomes were labeled with Cholesteryl-Bodipy. The rats were sacrificed at scheduled times after the injection; blood, urine, metastatic and healthy lung, colon, liver and spleen were analysed by a microcytofluorimetric examination. RESULTS: Liposomes prolonged the circulation time of Cholesteryl-Bodipy. Only spleen and lung metastases exhibited an accretion of fluorescent liposomes. CONCLUSIONS: The biodistribution of such formulation of liposomes in rats with lung metastases, may be of considerable importance in diagnosis and therapy of the secondaries, for increasing the concentration of tracers and therapeutic agents in tumor tissue while minimizing the likelihood of aspecific distribution and toxicity to non target tissue.

How to study boron biodistribution in liver metastases from colorectal cancer.

The purpose of this study was to evaluate boron distribution for a safe and effective BNCT (Boron Neutron Capture Therapy) of liver metastases. Samples both from healthy and tumour liver parenchyma were analysed, after i.v. boron administration, by: alpha particles counting under neutron irradiation; morphological analysis by standard haematoxylin-eosin staining; neutron autoradiography. Our method was unaffected by the cytological heterogeneity inside tumour nodules; it demonstrated selective boron distribution in tumour tissue and predicted estimated mean therapeutic doses in tumour and safety doses in healthy tissue. The time interval for efficient BNCT was 2 to 4 hours after i.v. boron administration.

Sequential evaluation of immunoreactivity in patients with melanoma undergoing surgery and adjuvant therapy.

The immunologic profile of 15 patients undergoing surgery and adjuvant chemoimmunotherapy for cutaneous melanoma was studied for a mean period of 18 months. In vivo cellular immunity was assayed by evaluation of delayed hypersensitivity response (DHR) to primary antigen and a panel of recall antigens. In vitro cellular immunity was evaluated to means of total and T-lymphocyte counts in peripheral blood and by the lymphocyte blastogenic response to phytohemagglutinin stimulation. Humoral immunity was assayed by determining the serum levels of IgG, IgA and IgM and of complement components C3c, C4 and Factor B. Phagocytic activity was studied by testing leukocyte chemotaxis, neutrophil phagocytosis and leukocyte random migration. The in vitro parameters were determined preoperatively at diagnosis, 6 times during the first 2 postoperative weeks, and then every month during adjuvant therapy. No correlation was found between DHR and clinico-pathologic stage of tumor, or with subsequent clinical course. Significant depression of total lymphocyte and T-lymphocyte count and blastogenic response of lymphocytes was found at diagnosis. The lymphocyte response to PHA decreased significantly in the early postoperative period but returned to preoperative levels one week after surgery. Periodic fluctuations of lymphocyte blastogenic response and progressive decrease of total lymphocyte counts and T-lymphocyte counts were observed during the 18-month follow-up. No significant alterations of immunoglobulin levels were recorded at diagnosis or during the postoperative period. Complement levels were within normal values preoperatively; in the early postoperative period a transient increase of C3c, C4 and Factor B was recorded, then complement levels progressively decreased. Parameters of phagocytic activity were normal at diagnosis and fluctuated within the normal range throughout the whole period of study.

Boron-loaded liposomes in the treatment of hepatic metastases: preliminary investigation by autoradiography analysis.

Boronophenylalanine (BPA)-loaded conventional and stabilized liposomes were prepared by the reversed phase evaporation method to treat liver metastases by boron neutron capture therapy. Conventional vesicles were composed of phosphatidylcholine and cholesterol, molar ratio 1:1. To obtain stealth liposomes, GM1 or PEG were included in the lipidic bilayer at a concentration of 6.67 or 5 mol%, respectively. Large unilamellar vesicles were formulated encapsulating BPA in the liposome aqueous compartment as a complex with fructose; BPA free base also was embedded into the lipidic bilayer. In vivo experiments were carried out after intravenous injection of liposome suspensions in BD-IX strain rats in which liver metastases had been induced. Alpha particle spectroscopy associated with histological analysis was performed to visualize boron spatial distribution in liver. Simultaneously, tissue boron concentrations were determined using inductively coupled plasma-mass spectroscopy. Results showed that PEG-modified liposomes accumulated boron in therapeutic concentrations (> 30 micrograms boron/g tissue) in metastatic tissue. The PEG-liposomes could be further explored in enhancing boron delivery to tumor cells.

[Glucose-1-phosphate associated with total parenteral feeding. Study of various metabolic effects and leukocyte functions]. / Il glucosio-1-fosfato in associazione alla nutrizione parenterale totale. Studio di alcuni effetti metabolici e di alcune funzioni leucocitarie.

The effects of glucose-1-phosphate (250-500 ml given by i.v. infusion over a period of 3-4 h for 3-5 days) on the quantity of exogenous insulin administered and on leukocyte adhesion and chemotaxis were studied in 4 groups of post-operative patients submitted to TPN (total parenteral nutrition), TPN + glucose-1-phosphate, standard infusion therapy and standard infusion therapy + glucose-1-phosphate respectively. With the use of glucose-1-phosphate the quantity of insulin administered could be significantly reduced (P less than 0.01) thereby an increased utilization of glucose, induced by the phosphorylated saccharide, was confirmed. Moreover, glucose-1-phosphate, through its action on intracellular sugar metabolism, increased leukocyte adhesion during TPN. This phenomenon might have a favourable effect on some of the functions carried out by these cells on the inflammatory response.

[Controlled study of unspecific cellular immunity in cancer patients]. / Studio controllato dell'immunità cellulare aspecifica in pazienti neoplastici.

Cellular immunity has been studied in 92 patients with solid tumors undergoing surgery, in order to evaluate immunocompetence at the time of diagnosis and to assess the prognostic value of parameters of cellular immunity. The results show that total lymphocyte counts, T-lymphocyte counts and lymphocyte blastogenic responses are moderately depressed at diagnosis in the cancer patients as compared to age matched controls. These parameters of cell mediated immunity "in vitro" seem to be of limited prognostic value, since no correlation was found with the clinical course during the first 6 postoperative months. Depression of delayed hypersensitivity response to cutaneous antigens appeared to be an index of poor prognosis.

[Cervical thymic cysts. Considerations on a case]. / Le cisti timiche cervicali. Considerazioni su di un caso.

The Authors report a case of cervical thymic cyst. The only distinguishing feature is the presence of thymic tissue, containing Hassall's corpuscles, in the wall. From review of literature rarity of this swelling is manifest, and so is difficulty of preoperative diagnosis. As far as pathogenesis is concerned, long consideration has failed to resolve the question of their developmental or degenerative origin. Treatment consists of excision of the cyst and its accompanying tract. Prognosis is usually good.

[Simple ulcer of the colon. Considerations on a case]. / L'ucera simplex del colon. Considerazioni su di un caso.

A case of "simple ulcer" of the colon is reported. The lesion is uncommon and not well known. As far as pathogenesis is concerned, the Authors emphasize the role of peptic, neurogenic, circulatory, bacterial, toxic and mechanical factors. The following clinical picture was observed: sudden and acute abdominat pain due to colonic perforation with pneumoperitoneum. As already described by literature preoperative correct diagnosis was not possible; this is usually due to the fact that the ulcer tends to perforate before other symptoms arise. During surgery suture of the ulcer, temporary colostomy and Mikulicz's drainage were performed; after 4 months colostomy was closed.

[Possible causal factors in postoperative immunodepression]. / Studio di alcuni possibili fattori causali dell"immunodepressione postoperatoria.

The relationships between postoperative immunodepression and duration of general anesthesia, volume of blood transfusions, cortico-steroid administrations and amount of surgical trauma have ben studied, in cancer patients undergoing radical operations. Postoperative depression of cellular immunity, assayed by the lymphocyte blastogenic response to phytohemagglutinin, was significantly correlated with the amount of surgical trauma and did not correlate with blood transfusions and corticosteroid administrations, nor with the duration of anesthesia.

[Postoperative infections. A prospective analysis of 1396 cases]. / Le infezioni postoperatorie. Analisi prospettica di 1396 casi.

Postoperative infections are the most frequent complications in surgery and are the commonest cause of the lengthening of hospital stay. The purpose of this study is to prospectively evaluate the incidence and predisposing factors of postoperative infections in 1396 surgical patients admitted to our Institute from 1984 to 1988. Patients undergoing minor surgical procedures (wound less than 2 cm) were excluded from the study. Patients were evaluated daily during hospital stay for onset of infections and results recorded on data sheet. Hemocultures in septic patients and samples of exudate at site of infection were taken whenever possible for aerobic and anaerobic cultures. 368 patients (26.36%) had at least one postoperative septic complication; (79 of them [5.65%] had two or more infections). The following infections were recorded: wound infections: 148 (10.60%); respiratory tract infections: 144 (10.31%); urinary tract infections 125 (8.95%); miscellaneous infections 11 (0.78%); thrombophlebitis 23 (1.64%); FUO 10 (0.71%). The most important predisposing factor for wound infection was endogenous contamination (wound infections: 18/499 [3.60%] in clean, 42/594 [7.67%] in potentially contaminated, 57/217 [26.26%] in contaminated and 31/86 [36.04%] in dirty operations). The duration of the anaesthesia was found to correlate with an increased incidence of respiratory tract infections (4.49% anaesthesia less than 60 min; 7.21% anaesthesia greater than 60 less than 120 min; 15.31% greater than 120 min anaesthesia). Urinary infections were more frequent when the patients where catheterized at least once in the postoperative period (24.86% vs 3.2%).

[Tumors of the breast in old age]. / I tumori della mammella nell'età geriatrica.

After reviewing the literature on this topic the authors studied all patients admitted to the Institute of Surgical Pathology I at the University of Pavia for breast cancer between 1-1-1974. An analysis of the findings revealed that: (a) Out of 993 patients with breast cancer, 204 (20.54%) belonged to the geriatric age group (> 65 years old) and of the latter 135 (66.17%) belonged to the 3rd age group and 69 (33.82%) belonged to the 4th age group (> 75 years old). (b) In the 3rd age group 25 (18.51%) were benign tumours and 110 (81.48%) were malignant, whereas in the 4th age group 4 (5.79%) were benign and 65 (94.20%) were malignant. (c) Familial patterns were also examined and were found to be positive in 47 (23.03%) cases in the 3rd age group and in 30 (14.71%) in the 4th age group. (d) In the physiological anamnesis it is interesting to note that there was 1 menarche before 12 (0.9%) in the 3rd age group; delayed menopause (after 50) in 66 cases (60%) in the 3 rd age group and in 29 cases (44.61%) in the 4 th age group. (e) In the pathological anamnesis of the patients in question it was observed that the most frequent site of previous tumours was the breast (72 cases, equivalent to 10.9% in the 3rd age group; 9 cases, 13.8% in the 4th age group). (f) In the next pathological anamnesis and at EOL it was observed that the tumours was most often found in the supero-external quadrant, in the right breast in the 3rd age group (66 cases, 48.89%) and in the left one in the 4th age group (42 cases, 60.87%). (g) Test included breast cancer scan, mammography, aspirated needle and biopsy where required. (h) Surgery in the form of Halsted's mastectomy was the preferred therapy and as early as possible. Some Authors use conservative therapy. (i) Histological tests were performed in all patients and revealed that the most frequent malignant tumour was ductal carcinoma. (j) Postoperative complications took the form of wound suppuration. (k) Only one tumour (carcinoma) was observed in male patient aged 83 years old, without metastasis. From the above findings it can be concluded that the most evident risk factor for breast cancer is the influence of the estrogen hormone.

[Leukocyte adhesiveness and chemotaxis during infusion of Intralipid]. / Studio della adesivita e chemiotassi leucocitaria durante infusione di Intralipid.

The effect of an intravenously administered lipid emulsion (Intralipid) on granulocyte functions has been studied. The study has been carried out in 20 healthy individuals, divided into two groups: 10 individuals have been infused with Intralipid 10% i.v.; the other 10 individuals (controls) have not been infused with any pharmacologically active solution. In all the subjects the following parameters have been studied: blood granulocyte count, granulocyte and leucocyte chemotaxis. The results showed a transient fall of granulocyte adherence during the intralipid infusion; no alterations of granulocyte count and leucocyte chemotaxis were found.

[A comparative study among different therapeutic approaches to portal hypertension and bleeding esophageal varices]. / Studio comparativo tra diversi approcci terapeutici all'ipertensione portale ed alle varici esofagee sanguinanti.

Recurrence of haemorrhage in patients with portal hypertension is the most feared life-threatening complication and the one which most often conditions patient survival. The present study compares the results obtained in two groups of patients treated by surgery and endoscopic sclerotherapy, respectively, and a control group treated with traditional medical therapy during bleeding episodes and subsequently given no further treatment. The patients in each group were subdivided into three different risk classes on the basis of the Child classification. Patients treated surgically mainly belonged to Child classes A and B, whereas those treated by endoscopic sclerotherapy belonged to Child class C. The long-term survival results show no significant differences between the two groups. On the other hand, both groups show better survival data than the untreated patients. On the basis of the results obtained, the authors believe that surgical treatment appears to most indicated in patients belonging to the lower risk classes (Child A and B), whereas endoscopic sclerotherapy is better suited to patients belonging to the higher risk class (Child C).