Neoantigen prediction and computational perspectives towards clinical benefit: recommendations from the ESMO Precision Medicine Working Group.

BACKGROUND: The use of next-generation sequencing technologies has enabled the rapid identification of non-synonymous somatic mutations in cancer cells. Neoantigens are mutated peptides derived from somatic mutations not present in normal tissues that may result in the presentation of tumour-specific peptides capable of eliciting antitumour T-cell responses. Personalised neoantigen-based cancer vaccines and adoptive T-cell therapies have been shown to prime host immunity against tumour cells and are under clinical trial development. However, the optimisation and standardisation of neoantigen identification, as well as its delivery as immunotherapy are needed to increase tumour-specific T-cell responses and, thus, the clinical efficacy of current cancer immunotherapies. METHODS: In this recommendation article, launched by the European Society for Medical Oncology (ESMO), we outline and discuss the available framework for neoantigen prediction and present a systematic review of the current scientific evidence. RESULTS: A number of computational pipelines for neoantigen prediction are available. Most of them provide peptide major histocompatibility complex (MHC) binding affinity predictions, but more recent approaches incorporate additional features like variant allele fraction, gene expression, and clonality of mutations. Neoantigens can be predicted in all cancer types with high and low tumour mutation burden, in part by exploiting tumour-specific aberrations derived from mutational frameshifts, splice variants, gene fusions, endogenous retroelements and other tumour-specific processes that could yield more potently immunogenic tumour neoantigens. Ongoing clinical trials will highlight those cancer types and combinations of immune therapies that would derive the most benefit from neoantigen-based immunotherapies. CONCLUSIONS: Improved identification, selection and prioritisation of tumour-specific neoantigens are needed to increase the scope of benefit from cancer vaccines and adoptive T-cell therapies. Novel pipelines are being developed to resolve the challenges posed by high-throughput sequencing and to predict immunogenic neoantigens.

[Ultrasound-guided trans-parietohepatic cholecystostomy in the critical patient: current indications]. / La colecistostomia trans-parietoepatica ecoguidata nel paziente critico: attuali indicazioni.

The appearance of acute cholecystitis can make to complicate a natural history of cholelitiasis or post-operating time of patients that have concomitant predisposition factors. The best therapy is the cholecystectomy but somewhere for the critical general conditions is too much dangerous to make a surgical procedure. However we need to stabilize patients conditions, also for a short time. Our experience suggest us that percutaneous transhepatic cholecystostomy is a simple method without any complications, efficacious to resolve the acute sepsis in patients with cholecystitis that not be able to tolerate a surgical procedure.

[Classification of varices with Doppler ultrasonography mapping. Clinical and hemodynamic correlation]. / Classificazione delle varici con mappaggio eco Doppler. Correlazione clinica ed emodinamica.

The authors report their experience concerning the outpatient echo Doppler mapping of peripheral venous circulation in the lower limbs. Out of a total of 240 patients examined, 190 showed monolateral venous insufficiency and 50 bilateral. These results were correlated to the age of the patients and showed that saphenous pathology tended to affect the entire vein in elderly patients, whereas only partial damage was observed in young patients. Varices seemed to appear with the first signs of the insufficiency of collateral veins before signs of saphenous insufficiency became manifest.

Amount and duration of alcohol intake as risk factors of symptomatic liver cirrhosis: a case-control study.

We carried out a hospital based case-control study involving 320 patients with symptomatic liver cirrhosis (LC) and 320 pair-matched control individuals, in order to estimate the dose-response relationship between both the daily amount and the duration of alcohol intake and the risk of LC. Lifetime alcohol consumption was measured by a standardized and reproducible questionnaire, and expressed as lifetime daily alcohol intake (LDAI) and duration of alcohol consumption (DAC). The odds ratio (OR) for LC was estimated by the conditional logistic regression. It increased from 1.0 for lifetime abstainers to 4.2 for LDAI of 225 g or more. Comparing durations of alcohol consumption of < or = 10 and > or = 30 years in the model, the ORs consistently decreased for all the LDAI categories: from 4.1 to 0.6 in the 25-50 g category; from 15.1 to 0.9 in the 75-100 g category; from 67.2 to 1.5 in the 125 g or more category. Our results suggest that the dose-dependent relationship between alcohol and LC may be mediated by the degree of individual susceptibility to the detrimental effect of alcohol to the liver.

Gemfibrozil in CAPD patients.

Gemfibrozil was given at a daily oral dose of 600 mg to 28 CAPD patients with serum triglyceride levels higher than 200 mg/dl, after 2 months of low fat low calorie diet. Eleven patients dropped out from the study due to spontaneous withdrawal of the drug or to intercurrent hospitalization. The 17 patients who completed the study showed a significant decrease in serum total (434.6 +/- 207.9 to 237.5 +/- 81.6 mg/dl, p 0.003), VLDL (268.8 +/- 17.8 to 109.2 +/- 66.9 mg/dl, p less than 0.005) and LDL (152.9 +/- 34.6 to 119.2 +/- 30.7 mg/dl, p = 0.004) triglyceride. Serum VLDL cholesterol increased from 108.7 +/- 54.1 to 59.9 +/- 29.0 mg/dl, p = 0.003. Serum HDL cholesterol decreased from 25.4 +/- 4.8 to 35.3 +/- 6.3 mg/dl (p less than 0.001). Apo A1 increased from 117.8 +/- 16.7 to 130.8 +/- 16.7 mg/dl (p less than 0.001) and Apo B increased from 173 +/- 26.7 to 137.8 +/- 21.9 mg/dl (p less than 0.001). No clinical or laboratory side effect was observed in the patients who completed the study. Therefore dropouts were due to the poor patients' compliance rather than to the severity of side effects.

[Action and tolerability of delayed-action buflomedil in obliterating arteriopathies of the lower limbs. A multicenter study]. / Attività e tollerabilità di buflomedil ritardo nelle arteriopatie obliteranti degli arti inferiori. Studio policentrico.

A new buflomedil retard preparation has been evaluated in 318 patients, suffering from obliterating arterial disease of the lower limbs, for 90 days at once daily 600 mg doses. After 30 days of treatment a significant (p less than 0.001) improvement of paresthesia, hypothermia and "claudicatio intermittens" has been found. The same trend has been found after 90 days with improvement of walking distance (+ 81.1%), walking time (+ 62.1%), walking speed (+ 15.0%) as well as a significant decrease of functional recovery time (-24.6%). The AA. stress the good tolerability and compliance of this new retard preparation.

Phosphodiesterase isozyme inhibition, activation of the cAMP system, and positive inotropy mediated by milrinone in isolated guinea pig cardiac muscle.

The purpose of the present study was to examine the interrelationships among phosphodiesterase (PDE) isozyme inhibition, cAMP formation, activation of cAMP-dependent protein kinase (cAPK), and positive inotropy in isolated guinea pig cardiac muscle mediated by the cardiotonic/vasodilator agent, milrinone. Milrinone was a potent and selective inhibitor of the "low Km" cAMP PDE isozyme (peak III) isolated by diethylaminoethyl ether cellulose chromatography, with IC50 values of 0.7 microM for peak III PDE and 100 microM for peak I PDE. In isolated papillary muscles frozen at peak inotropic responses, positive and significant correlations were evident between isometric force development as a function of cAMP content (r = 0.72, p less than 0.05) or cAPK activity ratio, an index of activation of cAPK (r = 0.79, p less than 0.001), for concentrations of milrinone from 0.1-1000 microM. Similar correlations were evident in muscles frozen at peak inotropic responses for the beta-adrenoreceptor agonist isoproterenol (r = 0.96, p less than 0.001; r = 0.98, p less than 0.001, respectively), but not for ouabain or Bay K-8644. The temporal sequence of these events was also quantitated for concentrations of milrinone (100 microM) and isoproterenol (3 nM) that produced approximately a 100% increase in isometric force. Whereas early time interval of force development (30 s, 1 min, isoproterenol; 30 s milrinone) were not accompanied by significant increases in either cAMP content or cAPK activity ratio, peak increases in force development for both isoproterenol (2 min) and milrinone (1 min) were related to peak increases in cAPK activity ratios. In summary, these results show that significant increases in cAMP content or cAPK activation are correlated with positive inotropy in isolated guinea pig papillary muscles with milrinone. These correlations occur at concentrations of milrinone that inhibit cardiac PDE isozymes and are similar to the known cAMP-dependent cardiostimulant isoproterenol. These data support the hypothesis that selective PDE isozyme inhibition is a mechanism by which milrinone effects positive inotropy.

Inhibition of the low Km cyclic AMP phosphodiesterase and activation of the cyclic AMP system in vascular smooth muscle by milrinone.

The purposes of the present study were to quantitate the effects of the cardiotonic/vasodilator milrinone on phosphodiesterase (PDE) isozymes isolated from vascular (aortic) smooth muscle from several species, and to quantitate changes in cellular cyclic AMP (cAMP) content, activation of cAMP protein kinase (cAPK) and vasorelaxation by milrinone in isolated guinea pig aortic smooth muscle. With PDE isozymes isolated from rat (Wistar-Kyoto or spontaneously hypertensive rats), guinea pig, monkey or canine aortic smooth muscle, milrinone is a potent (IC50 = 0.16-0.90 microM) and selective (100 times peak III relative to peak I) peak III inhibitor. The potency of milrinone and other vascular peak III PDE inhibitors parallels their potency as vasorelaxants in isolated guinea pig aortic rings (r = 0.86; P less than .01). Vasorelaxation of phenylephrine-contracted (3 microM) guinea pig aortic rings is accompanied by significant increases in cAMP content or cAPK activation with concentrations of milrinone greater than or equal to 10 microM. Temporally, significant increases in cAMP content accompany significant vasorelaxation; however, activation of cAPK is not significantly increased until at least 1 to 2 min after addition of milrinone. Similar concentration and temporal relationships are seen with the cAMP-related vasorelaxants papaverine and forskolin. As with milrinone, a temporal dissociation between increases in cAMP content and increases in cAPK activity ratio is evident.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes induced by gemfibrozil on lipidic, coagulative and fibrinolytic pattern in patients with type IV hyperlipoproteinemia.

A short term open study (90 days) was carried out with Gemfibrozil in patients (n = 10) suffering from type IV hyperlipoproteinemia. For the whole observation period (120 days) the patients followed a standard diet. Every 30 days the following parameters were checked: total cholesterol, serum triglycerides, HDL-C and its fractions (HDL2-C and HDL3-C), apolipoprotein A1 and B, glucose, fibrinogen, plasminogen, euglobulin lysis time, antithrombin III, alpha-2-antiplasmin and PTT. The administration of the drug caused a reduction of serum triglycerides by 39.5%, an increase of HDL-C by 16.2% together with a significant increase of cholesterol bound to HDL2 (+27.6%). A significant increase was also noticed for the mean levels of apolipoprotein A1 (+19.8%), confirming thus the antidislipidemic effect of the drug. Significant reductions were also found in the mean levels of fibrinogen and alpha-2-antiplasmin together with a return to normality of mean values of antithrombin III and of the euglobulin lysis time. The effect on the lipid, haemocoagulative and fibrinolytic parameters shows that the administration of Gemfibrozil causes favourable changes both on the hyperlipoproteinemic pattern and on the thrombophilic state present in these patients.

Protein kinase C activity and reactivity to phorbol ester in vascular smooth muscle from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY).

Protein kinase C (PKC) activity in aortic and renal arterial smooth muscle from SHR (20-23 wk male; mean arterial pressure = 178 mm Hg) and WKY (age/sex matched; mean arterial pressure = 126 mm Hg) was quantitated. Activity was greatest in the particulate fractions relative to the soluble fractions in all sources. The only difference between SHR and WKY was in the soluble fraction from SHR renal arteries, which had 2 fold more activity (255 pmol/mg/min) when compared with WKY (136 pmol/mg/min). This difference was not apparently related to force modulation, since the magnitude of isometric force development in renal arteries in response to phorbol 12,13-dibutyrate was not different between SHR and WKY. The magnitude of force developed in response to phorbol 12,13-dibutyrate and PKC activity in the particulate fraction was greatest in aorta vs. renal arteries in both WKY and SHR. These results suggest that regional vascular differences in the amount of PKC activity may exist which are not apparently related to a disease state (i.e., hypertension). These differences may be related to differential sensitivity to phorbol ester-mediated contractions in isolated smooth muscle.

Evaluation of a sustained-release buccal nitroglycerin preparation in stable angina pectoris.

The efficacy of a 2.5 mg sustained-release buccal nitroglycerin preparation given 8-hourly in effort-induced stable angina pectoris was investigated by means of graded exercise testing in 15 patients. An initial double-blind crossover study, compared with a placebo (duration six days) was followed by a further 15-day open treatment period on the active drug. Exercise testing was carried out 2 and 7 h after tablet administration at the conclusion of active treatment in the double-blind phase, and 2 h after tablet administration at the conclusion of the open treatment phase. Workload and exercise duration were significantly increased and mean electrocardiographic ST segment depression and ST segment recovery time significantly reduced by buccal nitroglycerin in the initial phase of the study, and these improvements were maintained over the subsequent 15-day assessment period. Systolic blood pressure at rest was significantly decreased by active treatment; other haemodynamic parameters remained unchanged. The study demonstrated the efficacy of sustained-release buccal nitroglycerin in effort-induced stable angina pectoris, and an absence of (+) tolerance to the therapeutic effects of this mode of nitrate administration.

Mechanical anterior resection for carcinoma of the midrectum: long-term results.

Long term results are reported on 108 patients treated by curative mechanical anterior resection for midrectum carcinoma. The functional results were good but there was a large number of local recurrences for carcinoma at Dukes C stage (28% of the total cases). Oncological results suggest that greater importance should be given to early diagnosis and to pathogenesis and risk factors for recurrence.

Identification and characterization of substance P receptors on LRM55 glial cells.

Substance P (SP) receptors were described by the specific binding of [3H]SP to several neuronal and glial cell lines. The neuronal cell lines N18 and NG108-15 were found to contain few if any SP receptors (less than 5 fmol/mg of protein). The glial cell line LRM55 contained large numbers (Bmax = 707 fmol/mg of protein) of a single class of SP binding sites (Kd = 276 pM). [3H]SP binding could be inhibited by a number of c-terminal SP fragments and the tachykinins physalaemin, eledoisin and kassinin. The binding kinetics and pharmacology of these receptors are similar to those the authors have previously described in the brain. Activation of SP receptors was shown to inhibit cyclic AMP-dependent, beta adrenergic-stimulated taurine release from LRM55 glial cells. SP inhibition must occur by mechanisms affecting taurine release after adenylate cyclase activation, inasmuch as SP has no significant effect on beta adrenergic-stimulated increases or basal levels of intracellular cyclic AMP.

Activation of beta-adrenergic receptors stimulates release of an inhibitory transmitter from astrocytes.

Activation of beta-adrenergic receptors on astrocytes in primary cell culture results in the release of taurine, an inhibitory transmitter. Taurine release occurs via a cyclic AMP-mediated intracellular pathway, because (a) taurine release and intracellular cyclic AMP accumulation have similar pharmacologies and time courses of activation and (b) N6,O2'-dibutyryl cyclic AMP stimulates release with a time course similar to that observed with the beta-adrenergic agonist isoproterenol. These results describe a previously unrecognized physiological function of astrocytes in the CNS-receptor-mediated release of the neuroactive amino acid taurine. This observation indicates that astrocytes may function as local regulators of neuronal activity.

Beta-receptor-stimulated and cyclic adenosine 3',5'-monophosphate-mediated taurine release from LRM55 glial cells.

Adrenergic stimulation of LRM55 glial cells results in the release of the neuroactive amino acid taurine. The present study characterizes the receptors involved in taurine release and shows that taurine release is mediated by cyclic adenosine 3',5'-monophosphate (cAMP). beta-Receptors in LRM55 cells were first characterized by [125I]iodohydroxybenzylpindolol binding. Binding was stereospecific and saturable with time and ligand concentration. Kinetic analysis of equilibrium binding at 37 degrees C revealed a single component of high affinity (Km = 113 pm; Bmax = 52.1 +/- 5.0 fmol/mg of protein). The pharmacologies of the stimulation of cAMP accumulation and taurine release were similar. The agonists isoproterenol (IPR), epinephrine (E) and norepinephrine (NE) showed a rank order of potency characteristic of a beta-adrenergic system (IPR greater than E greater than or equal to NE). The beta-antagonists alprenolol and propranolol inhibited the IPR stimulation of both processes; the alpha-antagonist phentolamine did not. The dependence of taurine release on cAMP was further suggested by the similarity of the two time courses and was demonstrated by the stimulation of taurine release by the cAMP analogue dibutyryl cAMP. Thus, one physiological response of glial cells to beta-adrenergic stimulation is the release of taurine. Receptor-activated release of taurine from glia represents a previously undescribed neuronal-glial interaction by which glia may actively regulate neuronal excitability.

Treatment of vasospastic angina pectoris at rest with nitroglycerin ointment: a short-term controlled study in the coronary care unit.

The effectiveness of nitroglycerin ointment in vasospastic angina pectoris at rest was evaluated in 10 patients selected for study. The study was performed after a 24 hour control period, and a randomized single-blind crossover experimental design was followed. Two percent nitroglycerin ointment (15 mg) or placebo ointment was administered every 6 hours for a period of 48 hours each; the first treatment period was followed by a second in which each preparation was used for a 24 hour period. All patients were hospitalized in the coronary care unit; an objective evaluation was carried out using a multichannel electrocardiographic recording to assure recognition of the painless ischemic episodes. Coronary angiography showed critical stenosis of one or two vessels in 9 of the 10 patients; spasm was demonstrated in 3. Results of the ergonovine test were positive in nine of nine patients. Nitroglycerin ointment produced a significant reduction in the mean daily number of episodes during the first (12.5 +/- 3.9 versus 0.5 +/- 0.4, p less than 0.02) as well as the second treatment period (10.6 +/- 3.8 versus 0.6 +/- 0.4, p less than 0.02). These results demonstrate that nitroglycerin ointment provides effective, long-lasting protection against angina due to coronary spasm.

[Sustained effect of nitroglycerin ointment on exercise tolerance in patients with effort angina (author's transl)]. / Efficacia prolungata dell'unguento di nitroglicerina nell'angina pectoris da sforzo.

The effect on exercise tolerance of nitroglycerin (NTG) ointment was studied by exercise test on bicycle ergometer in a selected group of patients with effort angina and angiographically proven critical coronary lesions. NTG ointment (15 mg) and placebo were administered in a double-blind cross-over randomized trial on two consecutive days; exercise test was carried out before the administration of NTG ointment and placebo, and repeated after one hour, 2 and 1/2 hours and 4 hours. Compared to control and placebo tests, NTG ointment produced a highly significant increase of duration of exercise and of total work load in all tests, as well as a significant decrease of maximal ST segment depression. After NTG ointment administration resting systolic blood pressure showed a significant decrease after one hour, 2 and 1/2 hours and 4 hours, resting heart rate a significant increase after 2 and 1/2 and 4 hours, and peak exercise heart rate a significant increase after 4 hours. Double product at rest was unaffected by NTG ointment, while at peak exercise it showed a significant increase only after 4 hours. Our data show that in patients with effort angina NTG ointment produces an important increase of exercise capacity which persists at least for 4 hours after the administration. Therefore NTG ointment seems to be of high value in the treatment of effort angina.