Single-cell ATAC and RNA sequencing reveal pre-existing and persistent cells associated with prostate cancer relapse.

Prostate cancer is heterogeneous and patients would benefit from methods that stratify those who are likely to respond to systemic therapy. Here, we employ single-cell assays for transposase-accessible chromatin (ATAC) and RNA sequencing in models of early treatment response and resistance to enzalutamide. In doing so, we identify pre-existing and treatment-persistent cell subpopulations that possess regenerative potential when subjected to treatment. We find distinct chromatin landscapes associated with enzalutamide treatment and resistance that are linked to alternative transcriptional programs. Transcriptional profiles characteristic of persistent cells are able to stratify the treatment response of patients. Ultimately, we show that defining changes in chromatin and gene expression in single-cell populations from pre-clinical models can reveal as yet unrecognized molecular predictors of treatment response. This suggests that the application of single-cell methods with high analytical resolution in pre-clinical models may powerfully inform clinical decision-making.

[Longitudinal bronchotomy on the pars membranacea for endoscopic foreign body extraction]. / Broncotomia longitudinale sulla pars membranacea per l'estrazione di un corpo estraneo endobronchiale.

Purpose of this paper is to report a case of foreign body obstructing the middle lobe bronchial origin, surgically removed through a longitudinal bronchotomy on the pars membranacea, reached by passing posteriorly to the lung. Anatomical considerations suggest that longitudinal bronchotomy on the pars membranacea of the main bronchial axis (stem bronchi, intermedius br (right), and lower lobar bronchus) gives direct visualization from inside also of foreign bodies sited into the lobar bronchi (sup, middle and lingula), allowing an easy bronchotomy repair, without lumen distortion or stenosis. Since the pulmonary artery is not in contact with this pars membranacea, broncho-arterial fistula in case of suture line dehiscence is quite improbable.

Interleukin-2 at different concentrations induces the growth of selective lymphoid cells.

The effects of low and high doses of recombinant interleukin-2 (rIL-2) on cultured peripheral blood mononucleated cells are reported with the aim to show the effects of this immunomodulator in different conditions. The proliferation of various cell types at different IL-2 concentrations was investigated and ultrastructural and enzymatic studies were performed. The data obtained indicate that grade and type of cell stimulation induced by IL-2 is correlated to the dose employed.

Effects of interleukin-2 for the treatment of malignant mesothelioma.

The hematological profile, and clinical parameters in mesothelioma affected patients undergoing a clinical trial to evaluate the efficacy of IL-2 treatment were investigated. Six patients were monitored for 6 weeks following therapy. Blood cell count, morphological and immunophenotypical analysis were performed, as well as clinical evaluations of the patients before and after therapy. Activation of the immune system (increase in lymphocytes, monocytes, eosinophils and HMS lymphocytes) induced by IL-2 was observed. The treatment was well tolerated: our patients had only mild adverse reactions controlled by symptomatic therapy. Eosinophilia represented the most evident negative effect. A slight decrease in CD4-positive subset of lymphocytes was observed after rIL-2 treatment. The therapy did not induce significant changes in the progression of the disease. In one patient necrosis at the tumoral site was observed after loco-regional rIL-2 administration.

Pulmonary sclerosing hemangioma in a woman with multiple uterine fibromas, cysts to thyroid and kidney.

Pulmonary sclerosing hemangioma (PSH) is a relatively rare, homogeneous solitary tumor. Its complex histology may suggest a number of malignancies. We report one case diagnosed in a 62-years-old woman with a tendency to benign tumors (uterine fibromas) and cystes (thyroide, kidney). Bronchoscopy was normal. Pulmonary scintiscan was not done. Chest X-ray showed a 2 x 2 cm solitary, dense, homogeneous, delimited nodule of the LLL. The intraoperatory diagnosis of adenocarcinoma led to lobectomy. The final diagnosis of PSH was done on fixed and stained samples. She is in good condition after 13 months of follow-up.

The role of functional respiratory tests in predicting pneumothorax during lung needle biopsy.

In 243 of 945 patients having lung percutaneous needle biopsy, we retrospectively evaluated 10 independent variables that potentially influence the incidence of pneumothorax (PNX): age, sex, lesion size, number of needle passes, needle intrapulmonary route, distance of the lesion from the chest wall, FEV1, FVC, residual volume, and total lung capacity percent predicted. The subjects considered (mean age, 61 years; 192 men and 51 women) performed respiratory function tests within 1 year of the procedure. The sample was also subdivided into four functional groups: restricted patients, obstructed ones, obstructed with alveolar hyperinflation patients, and normal subjects. The variables significantly correlated to PNX were the length of the needle intraparenchymal route (p<0.05) and the distance of the lesion from chest wall greater than 0 cm (p<0.01). None of the functional parameters was determinant in predicting PNX occurrence. Bronchial obstruction was not significantly associated with a higher risk of PNX. Nevertheless, when alveolar hyperinflation was associated with bronchial obstruction, the risk increased significantly (odds ratio, 2.38).

Surveillance transbronchial biopsy in the diagnosis of acute lung rejection in heart and lung and lung transplant recipients.

From March 1991 to December 1993, 30 patients underwent transbronchial biopsy (TBB) after lung transplantation (16 with a heart lung transplant, 7 with a single lung transplant, and 7 with a double lung transplant). The now standard TBB technique was used. Initially, TBB was performed only when clinically indicated, i.e. when there were sound reasons to suspect the existence of acute rejection (AR) or pulmonary infection. From 1992, all the patients were entered into a prospective study, the protocol of which called for serial "surveillance" TBB to be performed, in addition to those for clinical indications, 15 days, 2, 3, 6, 9 and 12 months after the transplant, and then annually. One hundred and twenty nine transbronchial biopsies were performed in 2.5 yrs. Of the 121 successful TBBs, 54 (45%) were positive, i.e. showed signs of acute rejection. Sixty six of 129 (51%) of the TBBs were performed because of clinical indications, 45 of them (68%) within the first 3 months following the transplant. The other 63 were surveillance biopsies. About two thirds of the clinically indicated TBBs and more than a quarter of the surveillance TBBs, yielding adequate samples, were positive for AR > or = A2 (mild rejection). The sensitivity and specificity of the method in detecting AR were 91 and 100%, respectively. The overall incidence of complications was 10.8% (pneumothorax in 9% of cases). There were no deaths correlated to the procedure. Our results confirm the decisive role of TBB in the diagnosis of acute lung rejection. The high incidence of mild acute rejection, and the occasional finding of moderate acute rejection in stable asymptomatic patients, support the use of surveillance TBB in the first 6 months.

Coexpression of aspartic proteinases and human leukocyte antigen-DR in human transplanted lung.

Aspartic proteinases have recently been shown to be implicated in antigen processing. We explored the expression of two aspartic proteinases, cathepsins E and D, and of human leukocyte antigen-DR (HLA-DR) molecules in a consecutive series of 80 transbronchial biopsies from transplanted lungs. For controls, we studied five normal donor lungs (not suitable for transplantation on account of thoracic trauma) and macroscopically normal areas of three cancer-affected lungs. Two of the five unsuitable donor lungs showed minimal inflammatory changes. Macroscopically normal samples from the three cancerous lungs showed mild and focal inflammatory infiltrates. In histologically normal lungs, HLA-DR expression was limited to professional antigen-presenting cells. Macroscopically normal lung samples with minimal inflammatory changes from both donor and cancer lungs showed variable HLA-DR expression by alveolar and bronchial epithelial cells and by endothelial cells. All transplanted lung biopsies showed HLA-DR expression by epithelial (alveolar and bronchial) and endothelial cells, with a trend for increased positivity in acute rejection. Cathepsin E was restricted to Clara and to rare bronchus-associated lymphoid tissue-related epithelial cells in histologically normal lung samples, whereas minimal de novo cathepsin E expression by rare alveolar pneumocytes was noted in control lung samples exhibiting minimal inflammatory changes. In all transplanted lung biopsies, cathepsin E was diffusely expressed de novo by hyperplastic alveolar epithelial cells, regardless of the presence or degree of rejection. Cathepsin D was expressed only by alveolar macrophages and by ciliated bronchial cells of normal, minimally inflamed, and transplanted lungs. In transplanted lung, Clara cells and several hyperplastic alveolar pneumocytes coexpressed HLA-DR and cathepsin E, whereas all alveolar macrophages and a few ciliated cells coexpressed cathepsin D and HLA-DR. The present investigation suggests that the de novo expression of cathepsin E and HLA-DR by hyperplastic alveolar pneumocytes of transplanted lung may be crucial for antigen processing and presentation to recipient competent T cells, and thus for the triggering of the immune-inflammatory cascade that leads to rejection.

Correlation between ultrastructural and histochemical parameters in lymphokine-activated killer (LAK) cells reacting with target cells in vitro.

Ultrastructural and fluorescence data allowed us to study the most important moments of the interaction between lymphokine-activated killer (LAK) cells against target cells (Chang) in vitro. The LAK cells, maintained at low doses of recombinant interleukin-2, were able to recognize, bind and destroy the tumoral cells. Before the attack, the LAK cells were characterized by a cytoplasm with a high ribosomes content; after the identification and the interaction cell-cell, a degeneration of the tumoral cell was observed. These observations allowed us to suppose that the interaction between the two types of cells may be mediated by a receptoral membrane system without the action of lytic enzymes.

Therapeutic embolization of bronchial artery: a successful treatment in 209 cases of relapse hemoptysis.

Still today bronchial artery embolization (BAE) is an important procedure in the management of hemoptysis, as an alternative to, or in association with, surgical or medical therapy. From 1974 to 1990, BAE was performed in 209 patients who suffered from hemoptysis with different etiopathogeneses and severity. Bronchiectasis were the most frequent indication (46%) followed by tuberculosis (31%), cystic fibrosis (16%), lung cancer (4%), and aspergillosis (3%). BAEs were performed by means of polyvinyl alcohol particles and absorbable gelatin sponge. In a few cases complete occlusion of the main truncus of particularly large bronchial arteries was obtained by use of Gianturco steel coin (5 mm o) technique. In the last seven years, together with the traditional angiographic techniques, selective digital angiography (SDA) was performed, above all in preliminary control phases, to evaluate occlusion during embolization. SDA reduced catheterization time and the mean quantity of contrast administered, decreasing side effects; 98% of hemoptysis was controlled in the first twenty-four hours, the other 2% in the following forty eight hours; 16% relapses occurred within the first year; 27 patients needed reembolization (15 patients twice, 11 patients three times, 1 patient five times). No complications were seen. If diagnosis, therapeutic indications, operative technique, and equipment selection are adequate, BAE has a high reliability in patients affected by relapsing hemoptysis, which is difficult to resolve.

Pneumocystis carinii pneumonia in heart transplant recipients.

Seven cases of Pneumocystis carinii pneumonia (PCP) (two in 1988, three in 1989, one in 1990 and one in 1991) have been observed in a group of 241 heart transplant recipients transplanted in Pavia, Italy, from November 1985 through December 1991. Median time to onset of symptoms was 100 days after transplantation (range 59-333 days). Diagnosis was achieved in all patients by cytological examination of bronchoalveolar lavage (BAL) fluid and/or transbronchial biopsy. Clinical and roentgenographic features were remarkably similar in all PCP-affected heart transplant recipients. A dry, persistent hacking cough associated with dyspnoea was consistently observed. Fever ranged from 37.6 to 39.4 degrees C, median leukocyte count and median arterial oxygen saturation (SaO2) values were 7,300/mm3 (range 3,000-16,000/mm3) and 61% (range 49.3-93%), respectively. Median CD4+ count at the onset of symptoms was 211/mm3 (range 28-739/mm3). The only patient experiencing a recurrence of PCP had a CD4+ cell count of 28/mm3 at the end of treatment with trimethoprim-sulfamethoxazole (TMP-SMX). In all patients human cytomegalovirus was isolated from BAL fluids; however, treatment with TMP-SMX alone (20 mg/kg/day of TMP) was consistently followed by a complete recovery.

Report of one case of familial sarcoidosis.

Familial sarcoidosis is a quite unusual occurrence. We describe two affected sisters, in which the clinical features of the disease were fairly similar. Interestingly, the two patients and two unaffected siblings share an HLA haplotype [A10 (26); CW7; B16 (38); DRW6; DQW1] rare in Caucasians.

Detection of malignant pleural effusions by tumor marker evaluation.

Cytologic examination and determination of tumor markers (PHI, LDH, alpha-1-glycoprotein, alpha-2-HS-glycoprotein, beta 2-microglobulin, ferritin [corrected], sialic acid, IgE, fetoprotein, CEA, beta HCG and beta 1-SP-glycoprotein) were carried out in pleural fluid samples obtained from 70 patients with suspected neoplasia. Tumor markers were also determined in sera. The protein content of all pleural effusions was greater than or equal to 3 g/dl. Patients were grouped according to diagnosis as follows: (a) 42 with neoplastic diseases (7 mesotheliomas and 19 lung, 4 ovarian, 3 breast and 8 miscellaneous cancers), (b) 22 with benign inflammations and (c) 6 with congestive effusions. Of the parameters examined, only CEA and beta-HCG [corrected] gave information that the effusion was probably malignant. Using 6 ng/ml as cut-off for CEA and 10 mIU/ml for beta HCG, the sensitivity was 57.1% and 45.2%, respectively, specificity was 92.8% for both parameters and test efficiency 0.75 and 0.69, respectively. When CEA and beta HCG were considered together sensitivity increased to 73.8% and efficiency to 0.78. CEA and/or beta HCG were positive in the pleural effusions of 19 of the 20 malignant pleural effusions, all with a negative cytologic examination, which subsequently became positive in 8. Because of their high specificity, these two parameters are a useful tool and can be routinely measured to evaluate pleural effusions of dubious origin, even if CEA and beta HCG cannot, on [corrected] their own, define the primary malignancy.

[Serum and bronchial concentrations of amoxicillin administered with a bronchial fluidizer]. / Osservazioni sperimentali sulle concentrazioni seriche e bronchiali dell'amoxicillina somministrata in associazone ad un fluidificante bronchiale.

The authors compared the serum and bronchial concentration of amoxycillin administered alone and in association with carboxymethylcysteine. The determinations were carried out in 10 patients affected with exacerbated acute and chronic bronchopneumopathies, treated first with amoxycillin alone (15 g/day in 3 administrations) and then with amoxycillin at the same dosage and carboxymethylcysteine (450 mg/day in 3 administrations). The bronchial secretions were collected during bronchoscopy performed 2 hours after the last administration of antibiotic. The bronchial secretion values of amoxycillin administered alone varied from 0.92 mcg/ml to 1.88 mcg/ml with a mean value of 1.44 mcg/ml. The percentage ratio between levels in bronchial secretion and levels in the serum varied from 12.7 to 36.1 with a mean value of 23.2. The administration of the amoxycillin-fluidizing agent association determined a statistically significant increase of the antibiotic levels in the bronchial secretions, varying from 1.26 mcg/ml to 6.39 mcg/ml, with a percentage ratio from 19.6 to 103.0.

[Immunological monitoring of patients operated on for epidermoid bronchogenic carcinoma]. / Monitoraggio immunologico di pazienti operati per carcinoma broncogeno epidermoidale.

Sequential monitoring of non specific immunity has been carried out in 16 patients with squamous cell bronchogenic carcinoma undergoing radical surgery. At the time of diagnosis total lymphocyte counts, T-lymphocyte counts and the blastogenic response of lymphocytes to PHA were only slightly depressed; on the contrary C3c and C4 serum levels were significantly increased (P less than 0.01). Sequential determinations of the immunologic profile at monthly intervals up to the 12th postoperative month showed a transient increase of total lymphocyte and T-lymphocyte levels; the lymphocyte blastogenic response to PHA and also C3c and C4 serum levels progressively decreased. Delayed hypersensitivity response to cutaneous antigens, neutrophil chemotaxis, serum levels of IgG, IgA, IgM and B-lymphocyte counts were constantly found within normal range.