Prediction of tumor-reactive T cell receptors from scRNA-seq data for personalized T cell therapy.

The identification of patient-derived, tumor-reactive T cell receptors (TCRs) as a basis for personalized transgenic T cell therapies remains a time- and cost-intensive endeavor. Current approaches to identify tumor-reactive TCRs analyze tumor mutations to predict T cell activating (neo)antigens and use these to either enrich tumor infiltrating lymphocyte (TIL) cultures or validate individual TCRs for transgenic autologous therapies. Here we combined high-throughput TCR cloning and reactivity validation to train predicTCR, a machine learning classifier that identifies individual tumor-reactive TILs in an antigen-agnostic manner based on single-TIL RNA sequencing. PredicTCR identifies tumor-reactive TCRs in TILs from diverse cancers better than previous gene set enrichment-based approaches, increasing specificity and sensitivity (geometric mean) from 0.38 to 0.74. By predicting tumor-reactive TCRs in a matter of days, TCR clonotypes can be prioritized to accelerate the manufacture of personalized T cell therapies.

Role of hyperbaric oxygen exposure in reduction of lipid peroxidation and in multiple organ failure induced by zymosan administration in the rat.

The aim of the present study was to evaluate the effects of hyperbaric oxygen (HBO) therapy on multiple organ failure induced by zymosan. Administration of zymosan (500 mg/kg) in the rat induced neutrophil infiltration in the lung, liver, and intestine as evaluated by increase in myeloperoxidase (MPO) activity. Therefore, lipid peroxidation was significantly increased in zymosan-treated rats. This inflammatory process coincided with the damage of lung, liver, and small intestine. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to nitrotyrosine in the lung, liver, and small intestine of zymosan-shocked rats. HBO (2 absolute Atmosphere) exposure attenuates the increase in the tissue levels of MPO and malondialdehyde (MDA) caused by zymosan in the lung, liver, and intestine. In addition, HBO (2 absolute Atmosphere) was effective in preventing the development of lung, liver, and intestine injury. Taken together, the present results demonstrate that HBO may also be an efficacious treatment in multiple organ failure induced by zymosan.

Cardiac effects of formoterol and salmeterol in patients suffering from COPD with preexisting cardiac arrhythmias and hypoxemia.

OBJECTIVE: There are several reports of documented adverse cardiac effects during treatment with beta-agonists. Since one should be aware that this may be a problem in patients with preexisting cardiac disorders, we have conducted a randomized, single-blind, balanced, crossover, placebo-controlled study to assess the cardiac effects of two single doses of formoterol (12 microg and 24 microg) and one single dose of salmeterol (50 microg) in 12 patients suffering from COPD with preexisting cardiac arrhythmias and hypoxemia (PaO2<60 mm Hg). DESIGN: Each patient was evaluated at a screening visit that included spirometry, blood gas analysis, plasma potassium measurement, and 12-lead ECG. In following nonconsecutive days, all patients underwent Holter monitoring 24 h during each of the four treatments. Holter monitoring was started soon before drug administration in the morning. Plasma potassium level was measured before drug inhalation, at 2-h intervals for 6 h, and at 9, 12, and 24 h following administration. None of our patients took rescue medication during the 24-h period. RESULTS: Holter monitoring showed a heart rate higher after formoterol, 24 microg, than after formoterol, 12 microg, and salmeterol, 50 microg, and supraventricular or ventricular premature beats more often after formoterol, 24 microg. Formoterol, 24 microg, significantly reduced plasma potassium level for 9 h when compared with placebo, whereas formoterol, 12 microg, was different after 2 h and salmeterol, 50 microg, from 4 to 6 h. CONCLUSIONS: The results of this study suggest that if a COPD patient is suffering from preexisting cardiac arrhythmias and hypoxemia, long-acting beta-agonists may have adverse effects on the myocardium, although the recommended single dose of salmeterol and formoterol, 12 microg, allows a higher safety margin than formoterol, 24 microg.

[Hypotension during acute myocardial infarct. Different pattern of parameters of renal function in relation to the localization of necrosis]. / Ipotensione durante IMA. Diverso contegno di alcuni parametri di funzione renale in rapporto alla sede di necrosi.

BACKGROUND: The influence on renal function caused by acute myocardial infarction complicated by hypotension has been studied. Particularly, a possible difference in renal functional pattern related to different localization and/or extension of necrotic myocardial area was investigated. METHODS: The study has been performed in 12 cases of acute myocardial infarction, involving anteroseptal (7) or inferodorsal (5) myocardial wall. The score of parietal asynergy resulted 13.28 in anterior vs 7.6 in inferior localization (p < 0.001). RESULTS: The decrease in systolic pressure, conversely, resulted not significantly different in the two groups. The renal function was assessed by measuring the urinary output (V) and creatinine clearance (CrCl). During the first 24 hours, both parameters resulted more preserved in inferior than in anterior AMI (V-552 vs 214 ml; p < 0.005; CrCl 70.6 vs 33.42; p < 0.001). Thus 1) the pressure decrease resulted unrelated to the size of asynergy and 2) the markers of renal function, although the decrease in brachial pressure has been identical in the two groups, have been shown to decrease more profoundly in the anterior infarction. DISCUSSION AND CONCLUSIONS: This may depends upon the reflex interruption of efferent nervous sympathetic drive toward the renal arteriolar bed, occurring in inferior infarction, despite of the systemic hypotension, and able to exercising a beneficial influence on the glomerular filtration rate (GFR). This reflex modulation has been demonstrated as effective in the inferior, but not in the anterior localization of infarction, this different haemodynamic pattern being able to explain the more pronounced decrease in GFR and diuresis in anterior than inferior AMI, as observed in our study.

[The dobutamine-dopamine combination versus amrinone in congestive heart failure with a marked edematogenic sign complicated by functional kidney failure. A comparison between 2 different models of inotropic stimulation and diuresis potentiation]. / Associazione dobutamina-dopamina versus amrinone nello scompenso cardiaco congestizio a marcata impronta edemigena complicato da insufficienza renale funzionale. Confronto tra due diversi modelli di stimolazione inotropa e di potenziamento della diuresi.

BACKGROUND: We evaluated the diuretic output in patients with decompensated chronic heart failure (CHF), previously treated by i.v. infusion with dobutamine and dopamine (dob-dop) or with amrinone (amr). Our target was to identify the possible discrepancies in urinary output perhaps linked to the different type of inotropic stimulation in the two subsets. METHODS: Adjunctive therapy with dob-dop or amr was chosen because the administration of diuretics only, without cardiac support, as tested in previous hospitalizations, had been demonstrated to produce unfavourable results, mainly expressed by finding of a low output syndrome in 50% of cases or more. The administration of i.v. infusion was maintained during 17 hours (1000 min approximatively), and included infusion in separate pumps of the two amines, dobutamine at dose of 5 micrograms/kg/min and dopamine at dose of 2.8 micrograms/kg/min or, alternatively, i.v. infusion of amr, administered at dose of 7 micrograms/kg/min. Infusion volumes were similar in the two subsets. The two subsets were homogeneous relatively to renal impairment, i.e. to the parameters (urinary Na, U/P creatinine, U/P urea, urinary osmolality) we fixed as markers idoneous to demonstrate the occurrence of organic renal damage (acute tubular necrosis). RESULTS: The diuresis was recovered in all 24 patients, and the urine volume resulted more pronounced in the subset attributed to the dob-dop at both the 8th and the 17th hour readings. We found no harmful alterations in HR and AP, whereas renal function parameters have been shown to enhance in both the dob-dop and amr arms. The diuretic effectiveness of the SIEV obtained by catecholamine implementation exercised a synergistic, favourable effect on diuresis, renal flow, glomerular filtration rate, and sodium post-proximal delivery. Amr resulted less effective then dob-dop simultaneous administration relatively to the diuretic effect. No remarkable differences were found in the two subsets as regards the heart rate, whereas a decrease in arterial pressure was found after amr. A persistent shift towards a condition of chronic renal failure, was identified in 4/24 patients, the two groups despite of the prolonged treatment at optimized doses: no remarkable side effects were reported. CONCLUSIONS: Thus, the selective effect upon renal hemodynamics, as exercised by dob-dop infusion low doses of dop, together with the enhanced renal output due to dob, has been shown to be more effective than amr influence: thus, the catecholamine therapeutical approach has been demonstrated to possess the best effectiveness in excitation of diuresis, among the CHF oliguric patients.

Effect of hyperbaric oxygen therapy in experimental subcutaneous and pulmonary infections due to Pseudomonas aeruginosa.

About 80% of nosocomial infections are caused by aerobic bacteria. Pseudomonas aeruginosa is a Gram-negative bacterium belonging to the Pseudomonadaceae family; P. aeruginosa is responsible for 6-22% of all hospital infections. The aim of this study was to evaluate the efficacy of hyperbaric oxygen (HBO2) therapy (2 atm abs x 55 min.day-1) alone for 8 days and combined with antibiotic chemotherapy (amikacin 15 mg.kg-1.day-1 for 8 days by intraperitoneal route) in rats infected subcutaneously and via the pulmonary route. In the rats infected by P. aeruginosa, HBO2 induced a reduction in mortality and morbidity with bacteria eradication in blood culture, bronchial aspirate, and skin biopsies when compared to control. These effects were increased by the use of amikacin, an antibiotic used for the treatment of sensitive Gram-negative bacteria.

[Colorectal obstruction caused by cancer]. / L'occlusione retto-colica da cancro.

The authors report their experience, matured in 10 years, in the management of cancer of the rectum and large bowel complicated by obstruction; 425 patients out of 493 total cases of colorectal cancer presented an intestinal obstruction and underwent emergency surgery. In 386 cases it was possible to perform resection (91%). The surgical treatment of the patients with right colon obstruction usually consisted of a right hemicolectomy. The surgical treatment of left colon obstruction is still controversial; in the experience of the authors it was accomplished by Hartmann operation mainly until 1985; in recent years the authors have introduced a procedure of intraoperative anterograde irrigation of the colon and they have performed resection-anastomosis in a single stage successfully but only in selected patients. In 4 out of twelve cases of cancer of the splenic flexure a subtotal colectomy was performed with one stage ileo-sigmoid anastomosis crowned with success. The authors examine the operative and postoperative mortality of the patients with colorectal cancer and point out that 5 years survival is worse in patients with colorectal obstruction compared to elective operations (24% versus 41%).

[The therapy of arterial hypertension: a comparison between ACE inhibitors and angiotensin-II-receptor antagonists]. / Terapia dell'ipertensione arteriosa: confronto tra ACE-inibitori e antagonisti recettoriali dell'angiotensina II.

The efficaciousness of ACE inhibitors in arterial blood hypertension is well known. These drugs decreased the incidence of hypertension and myocardial infarction in population. However, they increase tissue levels of some kinines, that may be responsible of some adverse reactions (cough, etc.). Angiotensin-receptor antagonists can minimize the adverse reactions due to kinine accumulation and may increase the safety of the antihypertensive drug-treatment. Pharmacological and clinical aspects of angiotensin-receptor antagonists are discussed.

Effects of hyperbaric oxygen exposure on a zymosan-induced shock model.

OBJECTIVE: To evaluate the effects of hyperbaric oxygen (HBO) therapy on zymosan-induced shock in rats. Zymosan, a cell wall component of the yeast Saccharomyces cerevisiae, induces inflammation by causing the production of various cytokines and pro-inflammatory mediators. The administration of zymosan to rats represents a new experimental shock model by inducing acute peritonitis, severe hypotension, and signs of systemic illness. However, it has been recently proposed that the zymosan-induced shock, like septic shock, may be mediated by overproduction of nitric oxide. DESIGN: Experimental study. SETTING: Institute of Pharmacology and Toxicology, 2nd University of Naples, Naples, Italy. SUBJECTS: Male rats were treated with zymosan (500 mg/kg) by intraperitoneal route, with HBO (2 Absolute Atmosphere) or with zymosan and HBO (2 Absolute Atmosphere). MEASUREMENTS AND MAIN RESULTS: Peritoneal exudate, plasma, and peritoneal nitric oxide metabolites (NOx) and zymosan determined a time-dependent increase in peritoneal and plasma NOx concentrations, and peritoneal leukocytes were determined. Moreover, symptomatology was observed. The administration of zymosan caused the appearance of a severe illness in the rats characterized by ruffled fur, lethargy, conjunctivitis, diarrhea, and a significant loss of body weight. All zymosan-treated rats developed an acute peritonitis, producing turbid exudate. Zymosan determined a time-dependent increase in peritoneal, plasma NOx, and tumor necrosis factor (TNF)-alpha concentrations. Morbidity of zymosan shocked rats has been attenuated and no mortality was observed by treatment with HBO. These findings were associated with a significant reduction either of peritoneal leukocytes and exudate, or plasma and peritoneal NOx concentrations. Moreover, TNF-alpha levels were significantly reduced in animals shocked by zymosan and treated with HBO.

[Effects of hyperbaric oxygenation in skin and pulmonary infections caused by Pseudomonas aeruginosa]. / Effetti dell'ossigenoterapia iperbarica nelle infezioni sperimentali cutanee e polmonari da Pseudomonas aeruginosa.

About 80% of nosocomial infections are caused by aerobic bacteria. The Pseudomonas aeruginosa is a Gram-negative bacterium pertaining to the Pseudomonadaceae family. P. aeruginosa is responsible for 6-22% of all hospital infections. The aim of this paper is to evaluate the efficacy of both hyperbaric oxygen-therapy (HBO 2 Atm x 35 min/day) alone for 8 days and when associated to the chemoantibiotic therapy (amikacine 15 mg/kg/day for 8 days intraperitoneal), in rats infected through pulmonary and subcutaneous intake. In rats affected by P. aeruginosa, HBO induces a significant reduction in mortality and morbility with bacteria eradication in blood culture findings, bronchial aspirate and skin biopsies. These effects were increased by the use of amikacine which is an antibiotic used for the treatment of Gram-negative bacteria.

Effects of hyperbaric oxygen exposure on non-adrenergic non-cholinergic responses of rat trachea.

The effects of prolonged (20 day) hyperbaric exposure (HBO) to oxygen on non adrenergic non cholinergic (NANC) contractile and relaxant responses of rat trachea were examined. The electrical field stimulation (EFS) of rat tracheal rings was performed at 30 Hz and contractile and relaxant responses were assessed in the absence or in the presence of pretreatment with L-nitro-arginine-methyl-ester (L-NAME), an inhibitor of NO synthase, and L-Arginine (L-ARG), a precursor of NO synthesis, plus L-NAME. Our data demonstrated that L-NAME significantly (p < 0.05) enhanced the contractile responses induced by EFS (controls 30.6 +/- 0.99%; L-NAME 76.07 +/- 2.00%) and statistically (p < 0.05) reduced the relaxant component of EFS (controls 31.10 +/- 0.46; L-NAME 15.00 +/- 0.12); these effects were reversed when tissues were pretreated with L-ARG plus L-NAME, suggesting that NO plays a modulatory role in cholinergic neurotransmission and participates in EFS relaxant responses. Moreover, prolonged HBO exposure (20 days) at 202.6 and 303.9 kPa did not modify the contractile or relaxant responses induced by EFS, nor modify the L-NAME or L-ARG effects on EFS responses.