Comparative roadmaps of reprogramming and oncogenic transformation identify Bcl11b and Atoh8 as broad regulators of cellular plasticity.

Coordinated changes of cellular plasticity and identity are critical for pluripotent reprogramming and oncogenic transformation. However, the sequences of events that orchestrate these intermingled modifications have never been comparatively dissected. Here, we deconvolute the cellular trajectories of reprogramming (via Oct4/Sox2/Klf4/c-Myc) and transformation (via Ras/c-Myc) at the single-cell resolution and reveal how the two processes intersect before they bifurcate. This approach led us to identify the transcription factor Bcl11b as a broad-range regulator of cell fate changes, as well as a pertinent marker to capture early cellular intermediates that emerge simultaneously during reprogramming and transformation. Multiomics characterization of these intermediates unveiled a c-Myc/Atoh8/Sfrp1 regulatory axis that constrains reprogramming, transformation and transdifferentiation. Mechanistically, we found that Atoh8 restrains cellular plasticity, independent of cellular identity, by binding a specific enhancer network. This study provides insights into the partitioned control of cellular plasticity and identity for both regenerative and cancer biology.

Micrographic surgery (Tubingen torte technique) for the treatment of an invasive dermatofibrosarcoma protuberans with muscular involvement.

Dermatofibrosarcoma protuberans (DFSP) is an uncommon skin tumour characterized by a slow, infiltrative growth and marked tendency towards local recurrences. Wide surgical excision of 3-5 cm from the margins including the fascia is the recommended treatment. Mohs Micrographic Surgery has been shown to afford lower rates of recurrence. The Mohs-Tübingen technique is a variant indicated for very large excisions that allows a complete eradication of the tumour, preserving healthy tissues. We report the case of a 45 year-old man affected by DFSP of the right shoulder deeply infiltrating the muscles, referred to us for a recurrence after a large excision. We submitted the patient to Mohs-Tübingen surgery in collaboration with an orthopaedic surgeon due to the presence of muscular involvement of DFSP. Two surgical operations were necessary to obtain negative histology as the tumor was deeply infiltrating the prescapular muscles. After three years of follow-up, the patient did not have any recurrence and the normal mobility of the shoulder was preserved. Precocious diagnosis and adequate therapy are necessary for DFSP as not only the margins, but also the deep invasion of the tumor have to be carefully controlled.

Completion pneumonectomy for non-small cell lung cancer: experience with 59 cases.

OBJECTIVE: The objective of this study was to assess the results of completion pneumonectomy performed for non-small cell lung cancer, classified as second primary or recurrence/metastasis. METHODS: From 1982 to 2000, 59 patients underwent completion pneumonectomy for lung cancer, classified second primary or recurrence/metastasis according to a modified form of Martini's criteria, after a mean interval from first resection of 60 months for second primary lung cancers and 19 months for recurrences/metastases. RESULTS: Operative mortality was 3.4% and complications occurred in 30% of patients. Five-year survival rate for completely resected patients was 25% (median 20 months). No significant difference in long-term survival was detected between second primary and recurrent tumors; survival was not adversely affected by a resection interval of less than 2 years or less than 12 months. CONCLUSIONS: Completion pneumonectomy for non-small cell lung cancer is a safe surgical procedure in experienced hands; long-term survival is acceptable and the best results are obtained for stage I lung cancer. Distinction between second primary lung cancer and recurrence failed to demonstrate a prognostic value.

Sleeve lobectomy for non-small cell lung cancer and carcinoids: results in 160 cases.

OBJECTIVE: To assess operative mortality (OM), morbidity and long-term results of sleeve lobectomies performed for non-small cell lung cancer (NSCLC) and carcinoids during a 35-year period. METHODS: A retrospective review of patients who underwent a sleeve lobectomy for NSCLC and carcinoids was undertaken, univariate and multivariate analyses of factors influencing early mortality in NSCLC were performed and for this purpose the series was split into an early and a contemporary phase, the Kaplan-Meier method was used to calculate the cumulative survival rate, and statistical significance was calculated with the log-rank test. Causes of death were evaluated in relation to the stage of the disease. RESULTS: OM for NSCLC was 14.6% in the early phase and 6% in the contemporary one; late stenosis occurred in 7.7% of NSCLC patients in the early phase and in 2% in the contemporary one. No OM or late stenosis occurred in carcinoid patients. Three, 5 and 10-year survival rates excluding carcinoids were 77, 62 and 31% for stage I(A-B), 45, 34 and 27% for stage II(A-B), 33, 22 and 0% for stage III(A-B). The 10-year survival rate for carcinoids was 100%. There was no significant difference in long-term survival between stages II and III, while the difference between stage I and stages II and III was significant (P<0.001). When survival was analyzed in relation to nodal status, 3, 5 and 10-year survival rates were 71, 57 and 33% for N0 disease, 42, 33 and 22% for N1 disease, and 34 and 19% with the last observation at 82 months of 19% for N2 disease; there was no significant difference in survival between N1 and N2 disease. A second primary lung cancer occurred in six patients (3.7%) who underwent resection. Late mortality was not related to cancer in most stage I patients while in stages II and III patients it was related to local and distant recurrences. CONCLUSIONS: Sleeve lobectomy is a valid alternative to pneumonectomy: careful patient selection and surgical technique make it possible to achieve a mortality rate comparable to or lower than that for pneumonectomy along with a better quality of life. In addition, it allows further lung resection, if necessary.

Deladenus siricidicola,bedding (Neotylenchidae) parasitism evaluation in adult Sirex noctilio, Fabricius, 1793 (Hymenoptera: Siricidae)

This work aimed to evaluate the Deladenus siricidicola, Bedding (Neotylenchidae)parasitism in adults Sirex noctilio, Fabricius, 1793 (Hymenoptera: Siricidae). Timber was sampled by cutting out pieces of 0.80 m in length in Sirex noctilio attacked and Deladenus siricidicola inoculated Pinus taeda. Longs were 15-20 cm in diameter, according to the tree age. Samples were packed in gauze-cages, for daily observations, till S. noctilio adults emergence. The emerged insects were transported, in plastic containers to the laboratory, where they were sectioned and dissected under stereoscopic microscopy to observ the nematode occurrence. From the initially proposed ten units, nine of them were evaluated in a total of 1,810 emerged adult insects, being 1,441 males and 369 females. Nematode parasitism was shown in 267 males and 74 females, in a total of 341 infected insects (18.84 percent)

Biochemical and microscopic evidence for the internalization of drug-containing mast cell granules by macrophages and smooth muscle cells.

During mast cell degranulation the soluble component of the granule is released into extracellular fluid, whereas two neutral proteases and heparin proteoglycans form the extracellular granule remnants. These structures are negatively charged and bind with high affinity LDL and other basic molecules. In this study we show that granule remnants expelled into extracellular fluid are able to bind the aminoglycoside antibiotic gentamicin and the anticancer agent doxorubicin in a dose-dependent manner. In addition, granule remnants loaded with the two basic substances are subsequently phagocytosed by macrophages. Indeed, when cells are incubated for 24 h with 1 mg/ml gentamicin, the intracellular concentration of the drug, which in basal conditions is extremely low, increases significantly in the presence of degranulating mast cells (from 5.1 +/- 1.0 to 25.4 +/- 2.5 microg/mg protein) and a good correlation between histamine release and gentamicin uptake is evident. The antineoplastic agent doxorubicin can penetrate cells by passive diffusion; however, when mast cells are added to macrophage monolayer, incubated for 30 min with 50 microM of the antineoplastic agent, a significant increase in intracellular doxorubicin concentration is observed (from 3.5 +/- 0.2 to 4.7 +/- 0.2 microg/mg protein). Internalization of granule remnants carrying gentamicin or doxorubicin is also evident in smooth muscle cells of the synthetic phenotype. In particular, when smooth muscle cells are incubated for 24 h with 1 mg/ml gentamicin, addition of isolated granules increases the uptake from 2.4 +/- 0.2 to 4.8 +/- 0.4 microg/mg protein. Similar results are obtained in smooth muscle cells incubated for 4 h with doxorubicin 50 microM (from 3.3 +/- 0.2 to 4.8 +/- 0.5 microg/mg protein). Data are confirmed by microscopic experiments by means of fluorescence microscopy and electron microscopic studies. The study demonstrates that basic substances can enter phagocytic cells when loaded to granule remnants. The phenomenon can be of particular interest for substances like the aminoglycosides that do not cross biological membranes; indeed, the storage of these antibiotics in phagocytic cells could have important consequences on their antibacterial activity in vivo. Macrophages and smooth muscle cells can also act as a reservoir for doxorubicin. High concentrations of the antineoplastic agent in these cells could be responsible for toxicity, as well as play an important role in the transport of the drug to tumor cells.

Deladenus siricidicola, bedding (Neotylenchidae) parasitism evaluation in adult Sirex noctilio, Fabricius, 1793 (Hymenoptera: Siricidae).

This work aimed to evaluate the Deladenus siricidicola, Bedding (Neotylenchidae) parasitism in adults Sirex noctilio, Fabricius, 1793 (Hymenoptera: Siricidae). Timber was sampled by cutting out pieces of 0.80 m in length in Sirex noctilio attacked and Deladenus siricidicola inoculated Pinus taeda. Longs were 15-20 cm in diameter, according to the tree age. Samples were packed in gauze-cages, for daily observations, till S. noctilio adults emergence. The emerged insects were transported, in plastic containers to the laboratory, where they were sectioned and dissected under stereoscopic microscopy to observ the nematode occurrence. From the initially proposed ten units, nine of them were evaluated in a total of 1,810 emerged adult insects, being 1,441 males and 369 females. Nematode parasitism was shown in 267 males and 74 females, in a total of 341 infected insects (18.84%).

Endocytosis of gentamicin in a proximal tubular renal cell line.

The mechanisms by which aminoglycosides are accumulated in renal proximal tubular cells remain unclear. Adsorptive mediated endocytosis, via a common pathway for cationic proteins, or receptor endocytosis, mediated by the glycoprotein 330/megalin, have been proposed to be involved in gentamicin transport in renal cells. We used the LLC-PK1 cell line, derived from the pig proximal tubule, to explore further the regulation of gentamicin endocytosis in these cells and to determine the role of clathrin mediated endocytosis and G proteins in this function. Gentamicin endocytosis was strictly temperature dependent, whereas total uptake (endocytosis plus binding) did not significantly differ at 4 or 37 degrees C. Substances that suppress receptor mediated, clathrin dependent endocytosis, such as monensin, phenylarsine oxide and dansylcadaverine, or inhibit caveolae mediated endocytosis, such as nystatin, did not affect gentamicin entrance in LLC-PK1 cells. Among substances that disrupt the actin cytoskeleton, only cytochalasin D, that is active also on fluid phase endocytosis, significantly reduced the intracellular concentrations of the aminoglycoside. Other maneuvers that perturb clathrin dependent endocytosis without affecting clathrin independent pathway, such as acidification of cytosol or incubation in hypertonic medium, were also without effect. Mastoparan, a well known stimulator of heterotrimeric G proteins, strongly increased endocytosis of gentamicin, and the same effect was evident with two other G protein stimulators, aluminum fluoride and fluoride alone; however the effect seems not to be mediated by an activation of adenylyl cyclase. In conclusion, gentamicin endocytosis in LLC-PK1 cells is probably clathrin independent, limited by cytochalasin D, which interacts with cytoskeleton, and increased by substances like mastoparan and aluminum fluoride, which activate heterotrimeric G proteins.

[Lung resection after pneumonectomy for bronchogenic carcinoma]. / Resezioni polmonari dopo pneumonectomia per carcinoma broncogeno.

METHODS: Four patients who previously underwent pneumonectomy for lung cancer, underwent a new contralateral resection for a metachronous cancer. Functional evaluation of these patients was not different from the standard one and included ECG, spirometry, blood gas analysis and a careful clinical evaluation. A femoro-femoral bypass was used in 2 patients to work on a collapsed lung, High Frequency Jet Ventilation was used in 1 while the last patient was hyperoxygenated and then ventilation was discontinued for the time required for the resections. RESULTS: Each method was free of complications. Operative mortality was nil. Two patients are alive free of disease at 66 and 14 months, two died for metastatic disease at 4 (N2 paratracheal nodes) and 12 months. CONCLUSIONS: Patients with a single lung should not be denied a further resection on contralateral lung. A routine evaluation of cardiopulmonary function may be sufficient but a more sophisticated evaluation including exercise oxygen consumption test, cardiac catheterization, lung perfusion scan, should be used when there is some doubt on the possibility of the patients to sustain the resection. Due to the high operative risk of this kind of patients mediastinoscopy should be performed even if preoperative CT scan of the chest shows minimally enlarged mediastinal nodes.

Lung resection on single residual lung after pneumonectomy for bronchogenic carcinoma.

Seven patients who had undergone a pneumonectomy for lung cancer developed a second tumor in the remaining lung after a mean time of 28.5 months and underwent a further resection. Preoperative evaluation was based on standard functional tests and on the "stair climbing test". Three patients were operated on using an extracorporeal oxygenator to work on a collapsed lung, three using standard anesthesiologic techniques, and one using high-frequency jet ventilation. There was no operative mortality. Complications occurred in two patients, requiring a temporary tracheostomy in one case. No patient required home oxygen supplementation. Four patients died of metastatic disease after 4, 8, 10, and 12 months, while two patients are alive and free of disease after 83 and 9 months, one is alive and free of symptoms but with a local recurrence after 29 months. Lung resection for bronchogenic carcinoma on a single lung can be safely performed provided that careful clinical judgment is used; long-term survival can be achieved with the resection of the new tumor.

Endobronchial extramedullary plasmacytoma. Report of one case.

A patient underwent a subtotal resection of the tracheobronchial carina for an obstructing endobronchial lesion. Preoperative biopsies of the lesion were not diagnostic. After resection, the histological examination of the specimen removed demonstrated an extramedullary plasmacytoma infiltrating the bronchial wall. Immunohistochemical studies showed monoclonality for kappa light chains. The postoperative course was uneventful and the screening for multiple myeloma was negative. No adjuvant treatment was given and the patient is currently alive and free of disease 63 months after the resection. Primary endobronchial plasmacytoma is a very rare disease: it is unclear which is the best treatment for endobronchial plasmacytoma. However, complete surgical resection has allowed a long-term survival, free of disease.

Prevention of atrial tachyarrhythmias after non-cardiac thoracic surgery by infusion of magnesium sulfate.

The possible role of magnesium sulfate (MgSO4) infusion in the prevention of atrial tachyarrhythmias after non-cardiac thoracic surgery was evaluated through a prospective study of two hundred patients who underwent non-cardiac thoracic surgery. The patients (who fulfilled the following requirements among others: no myocardial infarction in the previous six months, normal renal function, no use of digitalis or antiarrhythmic drugs, not undergone emergency operations or video-assisted thoracic surgery), were randomly assigned to receive MgSO4 infusion in all circumstances (Mg group), or either no treatment or, if aged over 70 or in cases of pneumonectomy or an intrapericardial procedure, application of digoxin starting on the day of operation (control group). 95 patients were enrolled in the Mg group and 105 in the control group. 93 patients in the Mg group and 101 in the control group were evaluated. Post-operative atrial tachyarrhythmias, mainly atrial fibrillation, occurred in 10 patients (10.7%) in the Mg group and in 27 (26.7%) patients in the control group (chi 2 = 7.009, df = 1. p = 0.008). It is concluded that infusion of MgSO4 is an effective means of reducing the incidence of atrial tachyarrhythmias after non-cardiac thoracic surgery.

Penetration of vancomycin into human lung tissue.

Vancomycin penetration into lung tissue was evaluated in thirty patients following the administration of 1 g of vancomycin as a 1 h i.v. infusion. Mean concentrations (range) of vancomycin in lung tissue were 9.6 (6.3-12.1) mg/kg at 1h, 5.7 (4.7-7.4) mg/kg at 2 h, 4.2 (0.8-6.5) mg/kg at 3-4 h, 2.4 (1.4-4.7) mg/kg at 6 h, and 2.8 (0.9-7.8) mg/kg at 12 h after the end of infusion. Ratios of lung tissue to serum concentration ranged 0.24 to 0.41 at 1 and 12 h, respectively. One of six patients observed at 6 h, and 3 of 7 patients at 12 h did not have detectable levels of vancomycin in lung tissue. A 1 h iv infusion of a 1 g dose of vancomycin does not achieve sustained lung concentrations above the MIC for susceptible staphylococci over a dosing interval of 12 h. Therefore, a more appropriate modality of administration, such as continuous infusion, should be considered.

Optimised method for determination of ticlopidine in serum by high performance liquid chromatography.

In this study we verified whether high performance liquid chromatography with UV detection (HPLC-UV) could be optimised and hence become a useful method for measuring ticlopidine concentrations in patient blood. The extraction step was improved by adding isoamylic alcohol in the extraction mixture, resulting in a better recovery. Moreover, the extraction efficiency was consistently ameliorated by evaporation to dryness of the organic extract; we could therefore utilise a lower (213 nm) wavelength, corresponding to the maximum of absorption, achieving a better sensitivity. With these technical improvements, the limit of quantitation is 0.02 microgram/ml, and the limit of detection is 0.01 microgram/ml, hence comparable to levels obtained with gas-liquid chromatography with nitrogen detection (GC-NPD). In addition, HPLC method, if compared with GC-NPD, is simpler and can be easily used for routine determinations of a lot of serum samples.

Left pneumonectomy for a second primary tumor after right upper lobectomy for superior sulcus tumor.

We report on a patient who underwent a left pneumonectomy for a second primary lung cancer after a right upper lobectomy and upper chest wall resection for a superior sulcus tumor. Although the postoperative course was complicated by sputum retention which required a temporary tracheostomy the outcome was good and the patient is now living without supplementary oxygen supply. Left pneumonectomy may be considered in patients with previous contralateral lobectomy when the preoperative evaluation of the patient shows an acceptable predicted postoperative pulmonary function. A long-term cure and a good quality of life must, however, be offered.

Completion pneumonectomy: experience with 47 cases.

From 1982 through March 1994, fourty-seven patients underwent completion pneumonectomy for a reappearing lung cancer, lung metastases, late complications, benign lung diseases, and early complication of bronchial or pulmonary artery sleeve resections. Intraoperative bleeding was higher than in standard pneumonectomy; there was one intraoperative mortality (2.3%). Operative mortality was 14.9% overall but was 3.6% in completion pneumonectomy for lung cancer, 20% for late complications and benign disease, and 57% for the treatment of early complications of sleeve resections. Three and five-year survival in patients with lung cancer who survived the operation was 43.8% and 28.7% according to the Kaplan-Meier method; no significant difference in long-term survival was present between patients with a second primary lung cancer or recurrence. Completion pneumonectomy is indicated in reappearing lung cancer and should be considered in benign disease when a less invasive procedure is not available. Completion pneumonectomy for the treatment of early complications of bronchial or pulmonary artery sleeve resections has a very high mortality but no alternative is available.

[Pulmonary lymphangioleiomyomatosis. Case report]. / La linfangioleiomiomatosi (LAM) polmonare. Presentazione di un caso clinico.

Pulmonary lymphangioleyomiomatosis (LAM) is a rare disease that affects only women. About one hundred cases are reported in the literature. This disease progresses to respiratory failure and its course can be slowed by hormonal therapy. A new case is reported; this woman was admitted to our Division for a right hydropneumothorax and after pleural drainage fluid demonstrated to be a chylous effusion; after a thoraco-abdominal CT scan the diagnosis of LAM was suspected. She was operated on after a few days because of continuous air and chyle leakage. A closure of lung leakage was performed with a stapler device that allowed the removal of a small specimen of lung for histopathological examination and a parietal pleurectomy was added with the aim of obtaining a pleural space obliteration. The histopathological examination revealed changes consistent with LAM and also positive was the hormonal steroid receptors assay. Adhesion of the lung to the chest wall was obtained but an antero-basal pouch persisted with continuous chylous leakage. A transposition of omentum into the chest was made with the aim of obliterating the pouch and favour chyle reabsorption and no drainage was left. Another CT scan demonstrated complete obliteration of the pouch and no fluid accumulation; the procedure proved to be successful and the patient was discharged with hormonal therapy.

Chylothorax after pleuro-pulmonary surgery: a rare but unavoidable complication.

The incidence of chylothorax after pleuro-pulmonary operations as well as its treatment is reported. Of 1744 operations performed postoperative chylothorax developed in 13 (0.74%). It resulted in two cases from the transection of the thoracic duct, in six from the transection of the so-called minor lymph channels, which drain lymph from mediastinal nodes straight into the thoracic duct or drain lung segments into the thoracic duct via the pulmonary ligament; the transection of these channels occurred during mediastinal lymphadenectomy or during the section of the pulmonary ligament. In 5 patients the site of leakage was not determined as reoperation was not required. Conservative treatment with low-fat diet and medium-chain triglycerides and/or total parenteral nutrition was attempted in all but one patient but was successful only in 5 cases whose mean losses were 292 ml/day. Seven patients were reoperated after a mean of 11 days; their mean losses were 930 ml/day. One patient was reoperated on the third postoperative day without attempting conservative treatment; his mean loss was 850 ml/day. Lymphadenectomy seems to be an important risk factor for postoperative chylothorax. Chyle leakage around 500 ml/day or higher that tends to decrease below 500 ml/day after a few days of dietary manipulation usually stops within 10-20 days, while leakage over 500 ml/day that does not tend to decrease below 500 ml/day seldom if ever stops without surgery, so that a more aggressive attitude is justified.

Lung resections concomitant to coronary artery bypass grafting.

From 1980 through 1993 ten patients underwent concomitant coronary artery bypass grafting and lung resection via median sternotomy. In eight patients a lung malignancy was resected, of which one was a small cell lung cancer. The lung resection was carried out before cardiopulmonary bypass in eight patients and during cardiopulmonary bypass in two. Coronary artery bypass grafting was performed using saphenous vein in eight patients; internal mammary artery was used as arterial conduit in two patients. There was one postoperative death while postoperative complications during hospital stay occurred in two patients. Pulmonary bleeding did not occur in any patient in whom lung resection was performed either before or during cardiopulmonary bypass. Both the patients who had internal mammary artery grafting experienced complications related to an associated lobectomy. A staged procedure is advisable if internal mammary artery has to be used and a lobectomy is required. The long-term survival in the patients with lung cancer was less than expected but the number of patients is too small to draw definite conclusions.