Biomedical subjects
Publications and source records attributed to M Rusca.
Persistent pleural space must be considered a possible complication after lung resection surgical treatment. Although in most cases the evolution is favourable, the authors suggest the use of the "pleural tent" technique, that compared to others thoracoplasty procedures offers some advantages. A short historical review of the thoracoplasty techniques is presented together with a detailed description of the above-mentioned technique.
The lack of an ideal heart-lung preservation solution is one of the principal factor that limits the wide spread of transplantation. The aim of this work was to investigate the efficacy of Haemaccel (HM) on isolated human pulmonary artery endothelial cells comparing its effects with those of University of Wisconsin (UWS). Subcultures of HPAEC were inoculated at the density of 5,000 cells per cm2 in 9 cm2 well-plates. Cells were incubated with HM and UWS for 6 hrs at 10 degrees C. Cellular viability was analysed by the total protein content (cytotoxicity index) and by the rate of protein synthesis (metabolic index). The results showed that HM and UWS did no show a significant differences in the toxicity when compared with the control; on the contrary, HM seems to determine a less inhibitory effect on cellular metabolism permitting a more rapid cellular metabolic recovery than UWS. Thus, HM appears to be more suitable for the preservation of isolated HPAEC than UWS.
The preoperative diagnosis of the involvement of the N2 lymph nodes is very important in patients with NSCLC for the most appropriate treatment. The classical diagnostic techniques for mediastinal exploration, mediastinoscopy and left anterior mediastinotomy, have been recently integrated by videothoracoscopy. FromJanuary 1993 to April 1994 186 patients with NSCLC suitable for surgery, were observed in our Department. 18 patients (9%) had CT evidence of N2 disease. In 10 cases the sites of the nodal enlargement were the right paratracheal station (#2 according to Naruke) and the right tracheobronchial station (#4), in 4 the subcarinal station (#7), in 2 the subaortic (#5) and in the remaining 2 cases the paraaortic station (#6). 14 mediastinoscopies were performed to investigate the stations 2,4 and 7,2 videothoracoscopies for station 5 and 2 left anterior mediastinotomies for station 6. The histological diagnosis was obtained in all cases without intraoperative or postoperative complications. Because the sequence chemotherapy-surgery seems to obtain the best results in the treatment of N2 disease the preoperative diagnosis of nodal involvement is of outstanding importance. Until recently mediastinoscopy and anterior left mediastinotomy were considered the standard techniques to explore mediastinum, now also videothoracoscopy has been introduced. In our experience, the integration of all the above techniques allowed a complete study of each suspect N2 site. Particularly the videothoracoscopy is very useful to safely biopsy under direct vision the aortic window lymph nodes.
Pulmonary endothelial cells are known to be fundamental for lung preservation and one of the most serious limiting factors observed during transplantation is the stress to which these cells are subjected. On this premise, strenuous efforts should be made to select and employ the preservation solution best able to prolong ischemia time and thus prevent cytotoxic effects. The aim of this study was to identify the solution with minimum toxicity on endothelial cells. For this purpose, we analysed the noxious effect of solutions such as Euro Collins (ECS), Belzer (UWS) and Low-potassium Dextran (LPD) on endothelial cells after 12 hours of incubation at 10 degrees C. For each solution, we examined the modifications produced on the nuclei, mitochondria and cellular wall of human pulmonary-artery endothelial cells by transmission electron microscopy and recorded the results on an ultrastructural grading scale. As regards morphological alterations incompatible with cell life, the most cytotoxic solution proved to be ECS. UWS and LPD, on the contrary, appeared to preserve cells relatively well, and no perceptible difference was observed between the two solutions. In conclusion, it is interesting to note that although ECS is widely used for lung preservation, the results of our study indicate that a 12 hours at 10 degrees C in this solution may exert adverse effect on pulmonary endothelial cells.
Postero-lateral thoracotomy is the standard access in thoracic surgery, and can be carried out dividing or preserving the serratus anterior muscle. This last method, besides its advantages, is still a matter of discussion, in fact traction has been claimed to cause severe muscular damage equal to section. We have studied 20 patients (16 males and 4 females, mean age 63 years), who underwent postero-lateral thoracotomy sparing the serratus anterior, after 6 months from surgery, both clinically and by means of electromyography (EMG), to evaluate the functional status of the spared muscles. In 4 cases (20%) there wasn't any neurogenic damage nor clinical evidence of winging scapula; 3 cases (15%) had medium neurogenic damage. The remaining 13 (65%) cases had a medium neurogenic damage and only one patient showed a winging scapula, neverthless being able to lift the arm to shoulder level. Our data confirmed that retraction during surgery does not damage the serratus anterior, leaving a functionally valid muscle.
Primary pulmonary lymphomas are uncommon tumours arising from mucosa associated lymphoid tissue (MALT). The authors report a case diagnosed by transbronchial biopsy and treated by surgery only. They discuss the distinct clinicalpathological features and debate the different ways of treating this condition.
This investigation was designed to show an original methodology for the assessment of lung preservation and to analyze the efficacy of a low potassium polygelin solution (haemaccel [HM]) on isolated human pulmonary artery endothelial cells. The effects of HM were compared with those of low potassium dextran (LPD), Belzer (University of Wisconsin [UWS]), and Euro-Collins solutions. The viability of the endothelial cultures was assessed by means of both total protein content and recovery of metabolic cellular function expressed as the protein synthesis rate after 6 hr and 16 hr of incubation at 10 degrees C. Our results failed to show any significant difference in the total protein content for HM, LPD, and UWS, both after 6 and 16 hr of incubation; however, the Euro-Collins-preserved sample revealed a significant drop in this parameter as early as 6 hr after the start. This finding was regarded as a clear indication of cellular cytotoxicity. In contrast, the metabolism recovery capacity of the cells varied significantly between HM and UWS at 6 hr and among HM, LPD, and UWS at 16 hr; at 6 hr, however, no significant difference was observed between HM and LPD. In conclusion, HM appears to exert a more significant effect on human pulmonary artery endothelial cell metabolism recovery than do the other fluids, thus suggesting its suitability as a long-term pulmonary perfusate.
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UNLABELLED: The attempt to synthesise efficacious solution to prolong lung preservation is, at present one of the most interesting challenges in transplant research. Recently, several issues emphasise the central role of ionic composition of lung-flush solutions, underlining, however, that colloid-free solutions are clearly detrimental. We have been studying a complex extracellular type solution (SPAL UP) synthesised to minimise the pathological events that occur during both preservation and reperfusion period. We report the results of toxicity of SPAL UP on normal human fibroblasts obtained from foetal lung (WI-38). WI-38 cells were seeded at 1.4 x 10(4)/cm2 in disposable plastic 12-well plates. After 3 days, cells were incubated in SPAL UP, Beltzer (UWS), Low Potassium Dextran (LPD) and Eurocollins (ECS) solutions for 6 hours at 10 degrees C. Cellular viability was evaluated by the rate of protein synthesis exploiting the incorporation of 35S-Methionine (2 microCi/ml) in growth medium with 10 mM unlabelled Methionine during 30 minutes incubation at 37 degrees C. The results were expressed as nmol. 35S-Methionine/mg of proteins/minute, and presented as means +/- SD of data of three (n = 3) well for each solution studied. RESULTS: the viability at time 0 before incubation (considered as control) was 1.65 +/- 0.1; after hypothermic preservation the data were respectively as follow: SPAL UP 0.51 +/- 0.09; UW 0.24 +/- 0.02; ECS 0.19 +/- 0.01; LDP 0.19 +/- 0.05. CONCLUSIONS: in this "in vitro" model SPAL UP solution provides a significantly (p < 0.05) better cell preservation than regular UW, ECS and LPD solutions.
UNLABELLED: The preservation of the lung for transplantation for a long period is still a problem not solved. Euro-Collins (EC) and Belzer (UW) solution are the most widely used. The aim of this work is to analyse the direct influence of both EC and UW solutions on endothelial cells of human pulmonary artery by means of an ultrastructural analysis. The arteries were obtained from 3 patients that underwent pneumonectomy and prepared with the no touch technique. The arteries were divided in 15 specimens and preserved in EC and UW solution at 4 degrees C for 6 and 10 hours. The specimens were fixed in osmic acid veronal buffer 1% and embedded in Durcupan. Ultrastructural examination was done with transmission electron microscopy (TEM) and the influence of the solutions was evaluated using a grading scale with scores ranging from 0 to 4 that express the damages of the cellular wall, mitochondria and nuclei. The data are expressed as mean +/- standard deviation (n = 5). Student's t-test was used for statistical comparison between the solutions. RESULTS: after 6 hours of preservation in EC and UW the scores were 5.2 +/- 0.45 and 4.8 +/- 0.84 (p = 0.373) while after 10 hours were respectively 8.2 +/- 0.84 and 6.8 +/- 0.84 (p = 0.029). In conclusion our experimental model suggests that there are no significant differences between EC and UW after hypothermic 6 hours preservation while endothelial cells are better preserved after 10 hours in UW solution.
Chylopericardium as an isolated complication after open heart surgery is a rare event. We present here a case of late postoperative chylopericardium and comment on the clinical course, the aetiopathology and treatment.
Tracheobronchial disruption is one of the less common injury associated with blunt thoracic trauma and the diagnostic evaluation is often misleading. Early diagnosis and repair of these injuries are essential to minimize the risk of patient's death and his future respiratory performances. Mechanisms of injury, clinical presentation, diagnosis and management of 5 our patients are reviewed. The good results achieved encourage an aggressive approach in these disruptions that can be life threatening.
The authors report their standard protocol for single lung transplant in New Zealand rabbit, the anaesthesiological approach consisted of a premedication with Ketamine 35 mg/kg i.m., Atropine sulphate 0.25 mg/kg s.c. and Fentanil 0.1 mg/i.m.; a jugular vein was cannulated and the anaesthesia was carried out administering Ketamine 10-15 mg/kg every 20 min., plus Fentanil 0.1 mg every 30 min. The rabbit was intubated through a tracheostomy, curarized and room air was administered using an appropriate ventilator device. The heart-lung en bloc removal from the donor was performed through a midline sternotomy. The left lung was then separated leaving an adequate cuff of left atrium inclusive of right pulmonary veins which were ligated using a 7-0 thread the graft prepared in this way was then preserved by putting it into the perfusion solutions. The recipient rabbit, after anaesthesia, underwent left pneumonectomy through a left toracotomy the bronchial anastomoses was first performed, followed respectively by the arterial and the atrial sutures. The controlateral pulmonary artery was then clamped and sequential arterial samples were taken from descending aorta previously cannulated to obtain arterial gases tests. The recipient rabbit has been followed up this way no more than 2 hour time, than it was sacrificed.
Several and different solutions have been used for lung preservation but, at present, fluids and solutions are quite alike. In the last years extracellular type solutions have been progressively tested in experimental researches and have shown a better protection vs intracellular one. In this research we have studied the effect of a low-potassium solution normally used as plasma expander (Haemmaccel, HM) on isolated foetal human fibroblasts (WI-38). HM has been compared with Belzer solution (UWS) after 16 hrs incubation at 10 degrees C and low-potassium solutions with dextran 2% and 5% after 6 hrs and 16 hrs incubation at 10 degrees C. Wi-38 cells have been seeded at the density of 9x10(4)/cm2 onto plastic well plates. Cellular viability was measuring using the rate of protein synthesis through the incorporation of 3H leucine in growth medium during a 1 hr incubation at 37 degrees C. The results were expressed as nmol 3 H leu/mg of proteins/minute and presented as means +/- SD; the comparison has been performed by the one way variance analysis test. After 16 hrs incubation at 10 degrees C HM preserves WI-38 cells significantly better than UWS. Comparing HM with LPD 2% and 5% a significant difference both at 6 hrs and at 16 hrs was observed. Our preliminary in vitro results confirm that low-potassium solutions are less toxic on isolated lung cells than intracellular one. Polygelin contained in HM could be considered a suitable colloidal substance that determine a better preservation if compared with Low-Potassium Dextran solutions.
Currently, Eurocollins' (EC) solution (high-potassium concentration) is the most widely clinically used pulmonary perfusate. However, recently, experimental studies have reported an increase of the lung ischemic period using low-potassium solutions. The purpose of our study, is to investigate the influence of the EC ionic composition and the effect of hyperosmolarity due to the glucose concentration on isolated alveolar type II epithelial cells. Pneumocytes type II were isolated from pathogen free Wistar rats using the modified Dobbs' method. Cells were incubated for 6 hours at 4 degrees C in EC, Collins (CL) and Ringer Lactate (RL) solutions. After that, cellular viability was evaluated by analysis of the protein synthesis assay by measuring the 35 S methionine uptake during an incorporation period of one hour at 37 degrees C (picomol 35 S met/mg proteins/h). Mean +/- standard deviation and Student "t"-test were used for data presentation and results comparison. Cellular viability at time 0 (control) before cellular incubation was 3.93 +/- 0.38. After 6 hours at 4 degrees C the results were respectively as follows: EC = 2.16 +/- 0.13; CL = 2.63 +/- 0; RL = 3.21 +/- 0.04. Our results suggest that the low-potassium extracellular type solution (RL) shows a protection on isolated type II epithelial cells statistically significant (p < 0.05) if compared with EC solution. Moreover CL solution, that has the same ionic composition EC but without glucose, presents a less cytotoxic effects on incubated cells than EC, confirming a deleterious influence of solution hyperosmolarity.
Chest wall tumors are rare being less than 1% of all the neoplasms. While the treatment of benign tumor and of solitary metastatic is standardized, the therapy of the primary malignant is not the same for all the authors. From January 1980 to December 1993 46 patients affected by chest wall neoplasms, 29 men (63%) and 17 women (37%), mean age of 51 yrs. (range 31-77 yrs.), were treated surgically. For benign tumors only resection was performed while for malignant and metastatic tumors after the resection the reconstruction was performed respectively: in 11 cases with Marlex mesh, in 1 case with the omentum, in 2 cases with Marlex and Methyl Methacrylate. Muscle transposition was performed in 1 case with pectoralis major covering a Dura Mater prosthesis. We have no peroperative death and no complications in 37 (80%) patients. Minor complications in 9 cases; in 1 case the sternal prosthesis was removed for infection. The median follow up was of 40.50 months. Actually the 48% of patients with primary and metastatic tumor are alive. From our results we conclude that the wide radical resection with reconstruction when necessary is the treatment of choice for the tumors of the chest wall.
It has been shown that Verapamil (V) improves lung preservation, but it is not clear whether the effect is due to vasodilation, with a more homogeneous cooling effect during hypothermic preservation, or a direct cytoprotection. Attempting to verify this, we studied the cellular viability of rats' pneumocytes type II primary cultures separately incubated in the first experiment for 6 hours in Ringer Lactate (RL) +/- V (10 mg/l) at 25 degrees C and at 4 degrees C, and in the second experiment for the same period in RL, EuroCollins (EC) and Collins (CL) solution +/- V (10 mg/l) at 4 degrees C. We analyzed the influence of the drug using two methods: the total protein content in each culture and the metabolic function of the cells using the rate of protein synthesis by means of 35 S methionine uptake assay. All data showed that there were no significant statistical difference (Student t-test, p > 0.05) in the use of verapamil during alveolar type II cells either in normothermic or hypothermic preservation. Thus we conclude that the protection of this calcium channel blocker during lung preservation is probably due to the effect on pulmonary vessels rather than to a direct cytoprotective effect on the lung cells.
The alteration of calcium homeostasis is of outstanding importance for the cytotoxic reactions that place after ischemia, for this reason calcium channel blockers have been used with the purpose to protect the lung during transplantation. This work analyses the effect of Diltiazem at two different doses (10 mg/l and 50 mg/l) on Wistar rat alveolar type II cells, incubated for 8 hours at 37 degrees C and at 4 degrees in an electrolytic solution. Total protein content and the rate of protein synthesis derived from 35S Methionine uptake were used to evaluate cells viability. The data showed that Diltiazem did not improve cellular viability after warm and cold metabolic ischemia either using 10 mg/l or 50 mg/l, while at 4 degrees C a significantly cytotoxic effect (p < 0.05) was observed. At this temperature toxicity was independent on the dose used.