Automated annotation and visualisation of high-resolution spatial proteomic mass spectrometry imaging data using HIT-MAP.

Spatial proteomics has the potential to significantly advance our understanding of biology, physiology and medicine. Matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) is a powerful tool in the spatial proteomics field, enabling direct detection and registration of protein abundance and distribution across tissues. MALDI-MSI preserves spatial distribution and histology allowing unbiased analysis of complex, heterogeneous tissues. However, MALDI-MSI faces the challenge of simultaneous peptide quantification and identification. To overcome this, we develop and validate HIT-MAP (High-resolution Informatics Toolbox in MALDI-MSI Proteomics), an open-source bioinformatics workflow using peptide mass fingerprint analysis and a dual scoring system to computationally assign peptide and protein annotations to high mass resolution MSI datasets and generate customisable spatial distribution maps. HIT-MAP will be a valuable resource for the spatial proteomics community for analysing newly generated and retrospective datasets, enabling robust peptide and protein annotation and visualisation in a wide array of normal and disease contexts.

Apoptosis in cells of bronchoalveolar lavage: a cellular reaction in patients who die with sepsis and respiratory failure.

OBJECTIVE: Apoptosis represents a physiologic clearance mechanism in human tissues. The role of apoptosis has not been examined in lung cell populations, such as alveolar macrophages of septic patients, an organ frequently insulted in these patients. This study was designed to examine the effect of sepsis on the apoptosis of alveolar macrophages. DESIGN: Prospective study. SETTING: Intensive care unit and surgical intensive care and trauma unit of a large university hospital in Athens, Greece. PATIENTS: Bronchoalveolar lavage was obtained from 20 consecutive patients who met the criteria for sepsis, admitted to two intensive care units. Bronchoalveolar lavage was obtained from nine volunteers without lung disease who served as controls. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The specimens were analyzed by using annexin V binding, terminal deoxynucleotidyl transfer-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL), DNA laddering, light microscopy, and immunohistochemistry. Spontaneous apoptosis of bronchoalveolar lavage cells and particularly of alveolar macrophages was significantly decreased in septic patients compared with nonseptic controls. This finding was confirmed by using morphologic criteria and the TUNEL method. Furthermore, gel electrophoresis of DNA obtained from bronchoalveolar cells revealed that DNA fragmentation was not necessarily associated with apoptotic cell death. The bcl-2 gene was minimally expressed in the control group. An inverse correlation was found between the percentage of apoptotic alveolar macrophages and the severity of sepsis. CONCLUSIONS: The prolonged survival of lung cells in septic patients and especially of alveolar macrophages may be attributable to the inhibition of apoptosis. This seems to represent an initial attempt of the host to increase the defense capacity to kill the invading microorganism, resulting in an unbalanced tissue load of cells and an uncontrolled release of toxic metabolites. Furthermore, the inhibition of apoptosis in septic patients may explain why lung function is impaired, leading to sepsis-induced acute respiratory distress syndrome and death.

Histocompatibility antigens and geographic tongue.

The HLA-A, B and HLA-DR antigens were investigated in 50 unrelated Greek persons with geographic tongue and in 380 healthy control persons. An increased incidence of DR5 and DRW6 antigens was observed in the blood of persons with geographic tongue. Ten (20%) of them had the DRW6 antigen, and it was significantly increased compared with 29 (7.6%) of the controls (p < 0.01, RR = 3.32). Twenty-seven (54%) of the experimental group showed DR5 antigen compared with 136 (35.7%) of the controls (p < 0.025, RR = 2.18). On the contrary, only 12% (5) of the experimental group had the B51 antigen in comparison with the controls (26.3%) (p < 0.05, RR = 0.37). A decrease of the DR2 antigen was also found in the persons with geographic tongue (24%) in comparison with the controls (39.2%) (p < 0.05, RR = 0.58).

Quantitative severity of stress thallium-201 myocardial perfusion single-photon emission computed tomography defects in one-vessel coronary artery disease.

The relation between the quantitative myocardial perfusion defect severity of exercise thallium-201 single-photon emission computed tomography (SPECT) and the quantitative degree of coronary stenosis was examined in 18 patients with 1-vessel disease (> or = 50% diameter stenosis), and abnormal thallium-201 SPECT. A total of 26 vessels were analyzed. Thallium-201 SPECT quantitative defect severity score was derived by summing the number of pixels in a coronary territory in which counts fell below the normal mean and multiplied by the number of SDs by which they fell below the normal mean. The thallium-201 defect severity score was significantly (p < 0.001) related to the maximal percent luminal diameter narrowing (r = 0.93), percent area narrowing (r = 0.89), absolute stenotic area (r = 0.79), and absolute stenotic diameter (r = 0.81). As expected, the strongest relation between thallium-201 defect severity and quantitative angiographic indexes was in the low and high ranges of coronary stenosis, with more variability and lower correlation coefficients (percent diameter: r = 0.75, p < 0.02, percent area stenosis: r = 0.63, p < 0.05) in the middle ranges (50 to 80% diameter stenosis). This observation is likely to be due to the complex flow characteristics across stenotic lesions. The findings suggest that in a select population, thallium-201 defect severity is potentially useful for noninvasive characterization of the functional severity of coronary artery stenosis and may complement coronary angiography in predicting functionally significant stenosis.

HLA-associated non-responsiveness to hepatitis B vaccine.

HLA-A,B,DR antigens of two groups, one of normal individuals (N- = 31) and another of CRF (Chronic Renal Failure) patients (K- = 37), who did not develop anti-HBs protective antibodies after Hepatitis B (HB) vaccination, were compared, respectively, to the HLA antigens of two corresponding control groups (N+ = 52, K+ = 49), who responded to the vaccine. A statistically significant difference (Pc less than 0.02) in the frequency of HLA-DR3 was observed between responders and non-responders. An increased frequency of HLA-A1 and HLA-B8 in N- as well as of HLA-A1 and HLA-B35 in K- was also noticed, but this was not of statistical significance. As these antigens have been associated to both HBs antigenemia as well as chronic active hepatitis, we suggest that these genes or other genes in linkage to those may suppress the response to HBV vaccination while, in parallel, they may predispose to an autoimmune course of Hepatitis.

Prognostic implications of angiographically normal and insignificantly narrowed coronary arteries.

The clinical presentation and prognosis of 1,977 consecutive patients with normal coronary arteries or "insignificant" coronary artery disease (CAD) (no major epicardial artery with 75% or more luminal diameter narrowing) were examined. Compared with patients with significant CAD, these patients had a lower frequency of traditional cardiac risk factors and abnormalities on the rest and exercise electrocardiogram. Cardiac survival was 99% at 5 years of follow-up and 98% at 10 years for patients with normal or insignificantly narrowed coronary arteries. Patients with normal coronary arteries differed from those with insignificant CAD in their myocardial infarction free survival rate: 99% at 5 years and 98% at 10 years for patients with normal coronary arteries, compared with 97% at 5 years and 90% at 10 years for patients with insignificant CAD. A strong relation occurred between the amount of insignificant CAD and follow-up cardiac events (chi 2 = 21.5, p less than 0.0001). Cardiac risk factors were statistically related to the risk of follow-up cardiovascular events when considered alone (chi 2 = 4.93, p = 0.026), but this relation lost significance after adjusting for the effect of coronary anatomy. Patients in both groups continued to have cardiac symptoms that resulted in frequent hospitalizations, medication use and job disability. Almost 50% in any given year of follow-up could not perform activities of high metabolic equivalent requirement and 70% had continuing symptoms of chest discomfort. Although these patients are at low risk of death, many remain functionally impaired for years.

Effects of blockade of alpha 1- and alpha 2-adrenoceptors on vasoconstrictor responses to single and twin pulse stimulation in rat tail artery.

Perfused/superfused proximal segments of Sprague-Dawley rat tail artery were stimulated at supramaximal voltage with two 1 ms square wave pulses. The pulse interval was either 10 or 20 s. With either interval the response to each pulse was similar, amounting to about 20 mm Hg increase in perfusion pressure. Prazosin (0.1 nM) approximately halved the response to both pulses whereas idazoxan (30 nM) was without effect. With an interval of 10 s, cocaine (4 microM) greatly increased the response to the first but not to the second pulse; in the presence of cocaine, prazosin (1 nM) again reduced both responses whereas idazoxan (30 nM) reduced the response to the first pulse but increased the response to the second. With an interval of 20 s, cocaine increased the responses to both pulses to a similar degree; in the presence of cocaine, idazoxan reduced the responses to both pulses. The results suggest that in rat tail artery, inhibition of the neuronal uptake process is required if noradrenaline released after stimulation with a single pulse is to reach smooth muscle alpha 2-adrenoceptors, and if feedback inhibition is to persist for more than 10 s.

Evidence for autoinhibition of stimulation-induced noradrenaline release from vasa deferentia of the guinea-pig and rat.

Both phenoxybenzamine and idazoxan increased the efflux of radioactivity elicited by a train of stimulation (4 pulses at 5 Hz) in vasa deferentia preincubated with [3H]-noradrenaline. Phenoxybenzamine increased the release of radioactivity from vasa stimulated with a single pulse, whereas idazoxan did not. The contractile response in both guinea-pig and rat vasa was biphasic: phenoxybenzamine enhanced the initial twitch component and reduced the second component in guinea-pig vasa stimulated with a single pulse or a train of pulses. Idazoxan enhanced both phases of the response of guinea-pig vasa stimulated with a train of pulses but did not affect the response to stimulation with a single pulse. The effect of phenoxybenzamine in increasing the efflux of radioactivity produced by a single pulse of stimulation was abolished by cocaine, indicating that the increase in efflux was due to blockade of noradrenaline uptake. Contractile responses of guinea-pig vasa stimulated with a single pulse in the presence of cocaine were unaltered by phenoxybenzamine, whereas with a train of stimulation the twitch component was enhanced and the second phase was reduced. The effects of phenoxybenzamine or idazoxan on the efflux of radioactivity from rat vasa portions were qualitatively the same as were observed in whole vasa. The contractile response of the prostatic portion consisted of a rapid twitch with a single pulse of stimulation, but was biphasic with a train of stimulation; the response of the epididymal portion was biphasic with either a single pulse or a train of pulses. These results suggest that there is no inhibitory feedback modulation of noradrenaline release with a single pulse of stimulation in guinea-pig and rat vasa deferentia whereas, with a train of stimulation, there is autoinhibition of noradrenaline release.