Pepsin in saliva as a biomarker for oropharyngeal reflux compared with 24-hour esophageal impedance/pH monitoring in pediatric patients.

BACKGROUND: Pepsin in saliva is a proposed biomarker for oropharyngeal reflux. Pepsin may be prevalent in saliva from subjects with gastro-esophageal reflux and may correlate with proximal reflux by intraluminal impedance/pH monitoring (MII/pH). METHODS: Patients (3 days to 17.6 years, n=90) undergoing 24-hour MII/pH monitoring and asymptomatic controls (2 months to 13.7 years, n=43) were included. Salivary pepsin was determined using a pepsin enzyme-linked immunosorbent assay. Eight saliva samples were collected from patients undergoing 24-hr MII/pH: (i) before catheter placement, (ii) before and 30 minutes after each of three meals, and (iii) upon awakening. One sample was collected from each control. KEY RESULTS: In MII/pH subjects, 85.6% (77/90) had at least one pepsin-positive sample compared with 9.3% (4/43) in controls. The range of pepsin observed in individual subjects varied widely over 24 hours. The average pepsin concentration in all samples obtained within 2 hours following the most recent reflux event was 30.7±135 ng/mL, decreasing to 16.5±39.1 ng/mL in samples collected more than 2 hours later. The frequency of pepsin-positive samples correlated significantly with symptom index (rS =0.332, P=.0014), proximal (rS =0.340, P=.0010), and distal (rS =0.272, P=.0095) MII events. CONCLUSIONS & INFERENCES: Concentration of salivary pepsin may not be an accurate measure of severity of reflux because of the wide range observed in individuals over 24 hours. Saliva samples must be obtained soon after a reflux event. Defining a regimen for optimal saliva collection may help to achieve the goal of using salivary pepsin as a biomarker for oropharyngeal reflux. CLINICAL TRIAL REGISTRY: NCT01091805.

The role of VEGF in the diabetic patients undergoing endovascular therapy of symptomatic aortic valve stenosis.

The aim of this study was to explore changes in plasma vascular endothelial growth factor (VEGF) in aged patients who undergone transcatheter aortic valve implantation or balloon angioplasty for the treatment of aortic stenosis. Plasma VEGF was measured in subjects with diabetes mellitus type 2 (DM) (n=21, age 79.2+/-1.6 years) and in non-diabetic subjects (non-DM) (n=23, age 84.4+/-0.7 years), using an ELISA kit. Before the procedure plasma levels of VEGF were significantly lower in DM than in non-DM patients (P<0.05). Plasma VEGF significantly increased in both groups (DM and non-DM) 24 h (387+/-64 vs. 440+/-30 pg/ml, P<0.05) and 72 h (323+/-69 vs. 489+/-47 pg/ml, P<0.05) after the endovascular procedure. However, the VEGF in DM patients was significantly lower compared to non-DM subjects up to one month after the endovascular procedure (283+/-47 vs. 386+/-38 pg/ml, P<0.05). We conclude that increased plasma VEGF in aged patients associates with atherosclerotic aortic valve stenosis. In spite of that plasma VEGF in DM was constantly significantly lower than in non diabetic patients, both before and after the endovascular procedure, possibly reflecting a disturbance of angiogenic/anti-angiogenic balance in diabetes.

Lipoprotein-associated phospholipase A2 mass level is increased in elderly subjects with type 2 diabetes mellitus.

OBJECTIVE. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is extensively expressed by advanced atherosclerotic lesions and may play a role in plaque instability. We selected a group of elderly subjects that underwent transcatheter aortic valve implantation (TAVI) or balloon angioplasty (BA) and separated them into two groups, diabetic and nondiabetic, to compare the level of Lp-PLA2 mass between them. METHODS. 44 patients aged 79.6 ± 5.6 years with symptomatic severe aortic valve stenosis underwent TAVI (n = 35) or BA (n = 9). 21 subjects had confirmed type 2 diabetes mellitus. Lp-PLA2 mass was measured using an enzyme-linked immunosorbent assay kit (USCN Life Science, China) before and 3 days after the procedure. RESULTS. Lp-PLA2 mass was significantly elevated in this population (1296 ± 358 ng/mL before TAVI; 1413 ± 268 ng/mL before BA) and further increased after TAVI (1604 ± 437 ng/mL, P < 0.01) or BA (1808 ± 303 ng/mL, P < 0.01). Lp-PLA2 mass was significantly increased on the diabetic group before these interventions. CONCLUSION. Lp-PLA2 may be a novel biomarker for the presence of rupture-prone atherosclerotic lesions in elderly patients. Levels of Lp-PLA2 in diabetic patients may accompany the higher amount of small dense LDL particles seen in these subjects.

[Serious sepsis treatment in intensive care departments in the Czech Republic -  EPOSS Project pilot results]. / Lécba tezké sepse na pracovistích intenzivní péce v Ceské republice -  pilotní výsledky projektu EPOSS.

INTRODUCTION: Severe sepsis is still associated with significant morbidity and mortality, which is however different, as well as its management, depending on the region. What is the situation in the Czech Republic and what is the character of patients with severe sepsis is currently not known. The aim of the project is to describe the processes of care, outcome and characteristics of patients with severe sepsis admitted to the intensive care department of the Czech Republic. METHODS: This is a multicentre and observational project with retrospective enrollment of patients who meet the criteria for severe sepsis before or within 24 hours after admission to selected intensive care units (ICU EPOSS). RESULTS: 394 patients were analyzed. Median age at admission was 66 (56- 76) years, males predominated (58.9%) and the median APACHE II score on admission was 25 (19- 32). Patients were predominantly medical (56.9%) and most were secondary admitted from other ICU (53.6%). Meeting the criteria of severe sepsis was most frequently within the period (± 4 hours) of admission the EPOSS ICU (77.6%). Median total fluid intake during the first 24 hours was 6,680 (4,840- 9,450) ml. Most patients required mechanical ventilation (58.4%). Compliance with the resuscitation bundle of severe sepsis in our group was very good and was associated with lower mortality of patients. Most frequently, the EPOSS ICU length of stay (LOS) was 7 (3- 15) days and median hospital LOS was 13 (8- 28) days. Hospital mortality in our cohort was 35.8%. CONCLUSION: Introducing the project, which in its first stage obtained valuable and internationally comparable data about patients with severe sepsis admitted to the involved ICU in the Czech Republic.

Real-time digital heterodyne interferometer for high resolution plasma density measurements at ISTTOK.

With the implementation of alternating discharges (ac) at the ISTTOK tokamak, the typical duration of the discharges increased from 35 to 250 ms. This time increase created the need for a real-time electron density measurement in order to control the plasma fueling. The diagnostic chosen for the real-time calculation was the microwave interferometer. The ISTTOK microwave interferometer is a heterodyne system with quadrature detection and a probing frequency of 100 GHz (lambda(0)=3 mm). In this paper, a low-cost approach for real-time diagnostic using a digital signal programmable intelligent computer embedded system is presented, which allows the measurement of the phase with a 1% fringe accuracy in less than 6 micros. The system increases its accuracy by digitally correcting the offsets of the input signals and making use of a judicious lookup table optimized to improve the nonlinear behavior of the transfer curve. The electron density is determined at a rate of 82 kHz (limited by the analog to digital converter), and the data are transmitted for each millisecond although this last parameter could be much lower (around 12 micros--each value calculated is transmitted). In the future, this same system is expected to control plasma actuators, such as the piezoelectric valve of the hydrogen injection system responsible for the plasma fueling.

[Oxidative stress and its effects on insulin resistance and pancreatic beta-cells dysfunction: relationship with type 2 diabetes mellitus complications]. / Stress oxidativo e seus efeitos na insulino-resistência e disfunção das células Beta-pancreáticas: relação com as complicações da diabetes mellitus tipo 2.

Type 2 diabetes mellitus, the most prevalent and serious metabolic disease worldwide, is believed to result from the interaction between genetical and lifestyle factors. In genetically predisposed people, the combination of a hypercaloric ingestion and reduced physical activity is responsible for the appearance of insulin resistance. This state can be overcomed, until a certain point, with increments of insulin secretion (hyperinsulinemia). However, an insufficient compensation leads to a state of glucose intolerance, which can evolve to diabetes, according to actual knowledge. The noxious effects of the hyperglycemia, allied with the possible increase of free fatty acids, are mediated by highly reactive molecules, oxygen and nitrogen free radicals species (ROS and RNS). Recent data suggests that these reactive species are signalling molecules and are involved in the regulation of the cellular function, being its increased production or reduced elimination a cause of oxidative stress. Indeed, those free radicals act directly through oxidative damage on macromolecules (proteins, lipids, DNA) or indirectly, activating single transduction pathways sensible to stress mechanisms. In this review, we will consider the pathways recognized as the more significant in stress mechanisms, namely: NF-kB, JNK/SAPK, p38 MAPK, PKC, AGE/RAGE, hexosamines and poliol. These signalling cascades are believed to be responsible for the insulin resistance and reduced insulin secretion, therefore the use of innocuous antioxidant substances such as vitamin C, E and the a-lipoic acid, is seen as a possible step for type 2 diabetic complications management. We will also discuss acetylsalicylic acid potentialities in the above-mentioned pathologies.

Oxidative stress after acute and sub-chronic malathion intoxication in Wistar rats.

Malathion is an insecticide of the group of organophosphate pesticides (OPs), which shows strong insecticidal effects. However, it possesses mutagenic and carcinogenic properties and shows organ-specific toxicity in relation to the heart, kidney and other vertebrate organs. The exact mechanism of the genotoxic effects of malathion is not yet known. Free radical damage is an important direct or indirect factor in several pathological and toxicological processes, including malathion poisoning. The aim of the present study was the evaluation of oxidative damage in different tissues of Wistar rats, administered intra peritoneally at doses of 25, 50, 100 and 150mgmalathion/kg, after acute and sub-chronic malathion exposure. Oxidative stress evaluation was based on lipid peroxidation by levels of thiobarbituric acid reactive substances (TBARS), protein oxidation by levels of carbonyl groups, and also on the activities of superoxide dismutase and catalase, two antioxidant enzymes that detoxity superoxide radical (O(2)(-)) and hydrogen peroxide, respectively. The results showed that the most sensitive targets of oxidative damage were kidney, lung and diaphragm after acute treatment, and liver, quadriceps and serum after sub-chronic treatment. Also, in general, increased lipid peroxidation measured as TBARS levels seems to be a better biomarker of oxidative stress compared to the contents of protein carbonyls after acute and sub-chronic malathion treatments. The present findings reinforce the concept that oxidative stress and particularly lipoperoxidation, are involved in OPs toxicity.

Apolipoprotein (a) fragments in relation to human carotid plaque instability.

PURPOSE: An elevated plasma level of lipoprotein (a) is an independent risk factor for atherothrombotic cardiovascular disease by yet undefined mechanisms. We have previously reported that matrix metalloproteinases cleave apolipoprotein (a) into 2 main fragments, F1 and F2, the latter (the C-terminal domain) exhibiting in vitro a high-affinity binding to extracellular matrix components, including fibrin(ogen). We therefore tested the hypothesis that the lipoprotein (a) matrix metalloproteinase-derived F2 is localized in potentially or morphologically unstable human carotid plaque at regions of increased matrix metalloproteinase activity. METHODS: Carotid plaques removed after endarterectomy (n = 18) were evaluated for structural features indicative of instability (thin fibrous cap, inflammation, and proximity of the necrotic core to the lumen); each plaque was classified as unstable (n = 10) or stable (n = 8). Western blot analysis was performed to quantitate apolipoprotein (a) and its fragments F1 and F2 in plaque extracts. Immunohistochemical staining was used to localize apolipoprotein (a) and its fragments within the atherosclerotic plaque. In situ zymography was used to determine regions of gelatinase (matrix metalloproteinase 2 and matrix metalloproteinase 9) activity. RESULTS: Western blot analyses demonstrated a 2.5-fold higher density of F2 in unstable plaques than in stable plaques (3.07 +/- 1.9 vs 1.18 +/- 0.8; P <.05). In morphologically unstable plaques, there was preferential distribution of F2 within regions of fibrous cap inflammation and/or foam cell accumulation and within abluminal necrotic cores. In morphologically stable plaques, however, localization was predominantly found in the medial smooth muscle cells. Regions of enhanced matrix metalloproteinase 2 and matrix metalloproteinase 9 activity co-localized with the transmural distribution of F2 within the plaque. CONCLUSIONS: These findings suggest that F2 in regions of increased matrix metalloproteinase activity is a potential mechanism for superimposed thrombotic events in morphologically unstable human carotid plaques. The relationship between plasma lipoprotein (a) levels and accumulation of F2 and the potential correlation of F2 to human plaque disruption and thrombosis warrant further study.

Increased flow and shear stress enhance in vivo transforming growth factor-beta1 after experimental arterial injury.

We have previously demonstrated that high-flow (HF) conditions inhibit experimental intimal hyperplasia. We hypothesized that such flow conditions may alter transforming growth factor-beta1 (TGF-beta1) after mural injury. The right common carotid artery (CCA) was balloon-injured in 54 New Zealand White male rabbits. Flow was thereafter preserved (normal flow [NF]), reduced by partial outflow occlusion (low flow [LF]), or increased by ligation of the left CCA (HF). Four sham-operated animals served as uninjured controls. Mean blood flow and pressure in the right CCA were measured before and after flow modulation and before euthanasia (3, 7, and 14 days). TGF-beta1 mRNA and protein levels in the right CCA were determined by Northern and ELISA analyses at each time point. At 7 and 14 days, intimal hyperplasia was quantified, and the transmural localization of TGF-beta1 was determined by immunohistochemical analysis. Mean flow was reduced from 22+/-1 to 10+/-3 mL/min in the LF group and increased to 34+/-2 mL/min in the HF group (P<0.001). Blood pressure was not different among the flow groups for all time points. Wall shear stress was markedly decreased in the LF group to 14+/-4 dyne/cm(2) and increased in the HF group to 63+/-6 dyne/cm(2) at 7 days compared with values in uninjured controls (39+/-2 dyne/cm(2), P<0.001) and the NF group (44+/-7 dyne/cm(2), P<0.001). At 14 days, wall shear stress was similar among the flow groups. The intima-to-media ratio was 5- and 2-fold greater in the LF group than in the HF and NF groups at 14 days. mRNA levels for TGF-beta1 and its active ligand were increased in the HF group by at least 2- and 3-fold, respectively, at 3 and 7 days compared with levels in uninjured controls and the LF group (P<0.05) but were not different among the flow groups at 14 days. TGF-beta1 preferentially localized in the abluminal vascular smooth muscle cells of the HF arterial segments. Flow- and shear-mediated release of TGF-beta1 may therefore play a role in abrogating the proliferative and migratory response of vascular smooth muscle cells in the early stages after mural injury.

Gene therapy enhances the antiproliferative effect of radiation in intimal hyperplasia.

BACKGROUND: Although ionizing radiation (IR) has been demonstrated to attenuate vessel wall restenosis and intimal hyperplasia (IH), dose-related mural injury and atrophy are possible deleterious side effects. We tested the hypothesis that a radiosensitizing strategy may improve IR-induced inhibition of in vivo vascular smooth muscle cells (VSMCs) without influencing apoptotic cell death. METHODS: In 28 New Zealand White rabbits, the right common carotid artery (CCA) was injured and subjected to low-flow conditions to promote IH. The CCA was transfected with an adenoviral vector incorporating the cytosine deaminase (CD) gene (1 x 10(9) PFU/ml). 5-Fluorocytosine (5-FC), a prodrug that is converted to the radiosensitizing agent 5-fluorouracil (5-FU) by CD, was thereafter administered intravenously. The CCA was exposed to 5 Gy IR at 24 h. Intimal/medial (I/M) area and thickness ratios were determined in the harvested CCAs at 14 days. VSMC proliferative and apoptotic indices were assessed with immunohistochemistry. RESULTS: A 50% reduction in I/M area was found in rabbits treated with IR and IR + CD/5-FC (0.19 +/- 0.03 and 0.18 +/- 0.02) when compared with untreated controls (UC) (0.37 +/- 0.06) (P = 0.005). This finding was substantiated by attenuation of I/M thickness in the IR groups [0.47 +/- 0.13 (IR), 0.41 +/- 0.11 (IR + CD/5-FC), 0.61 +/- 0.17 (UC)] (P = 0.007). The number of proliferating VSMCs was notably smaller when IR was combined with CD/5-FC (4.17 +/- 1.16 vs 2.97 +/- 1.09 log transformed cells/mm(2), P < 0.07). Apoptosis was similar in all groups. CONCLUSIONS: Both IR alone and IR combined with a radiosensitizing agent are effective in attenuating experimental IH. However, combination therapy is synergistic and achieves greater inhibition of VSMC proliferation and may involve selective killing of radioresistant S-phase VSMCs. IR + CD/5-FC represents a novel therapeutic strategy that offers potential for long-term control of IH.

Biomechanical factors as regulators of biological responses to vascular grafts.

Biomechanical forces have been implicated in the induction and progression of intimal hyperplastic thickening in vein, prosthetic, and endovascular bypass grafts. Graft implantation imposes significant alterations is shear and tensile forces. Such physical forces play an important role in modulating those cellular and molecular events that underlie regulation of vascular healing and adaptation. Characterization of such hemodynamic variables that induce perpetual medial vascular smooth muscle cell proliferation and migration will help in identification of those grafts at risk for occlusion and limited long-term patency and in design of therapeutic strategies that attenuate progressive intimal hyperplasia.

Irradiation for the treatment of intimal hyperplasia.

Intimal hyperplasia represents a serious complication limiting the long-term benefits of vascular interventions such as balloon angioplasty and stent placement. Although pharmacological interventions have attempted to curtail restenosis, they have not been shown to be effective to date. Radiotherapy is one alternative that has shown promise as an inhibitor of intimal hyperplasia in several animal models. Irradiation causes cell death by producing irreparable damage to DNA. This is believed to be the mechanism of inhibition of VSMC proliferation. Delivery of irradiation can be either intraluminal via an angiographically directed catheter or extraluminal using an external radiation source such as an x-ray device. Intraluminal irradiation has generally utilized either gamma or beta-emitting sources. Both have been effective in producing a dose response, although some studies advocate the use of beta-type irradiation as a safer, more efficient means of delivery. Extraluminal irradiation also has been an effective inhibitor of intimal hyperplasia. Studies suggest that this form of irradiation provides a more even-dose distribution to vessel walls than an intravascular delivery system. The use of radiotherapy has more recently been extended to clinical trials, and initial studies have shown promising results. The success of irradiation must be balanced with its potential complications including radiation-induced arteritis, coronary artery stenosis, and secondary development of malignancy. Although these have been associated with irradiation, the dose used in these cases was often considerably higher than those used in the treatment of intimal hyperplasia. Finally, with the advent of gene therapy, irradiation may provide an additional means of supplementing this new type of therapy through radiation-inducible gene therapy.

Management of recurrent carotid stenosis: should asymptomatic lesions be treated surgically?

PURPOSE: The purpose of this study was to determine factors that may influence patient selection for surgery in recurrent carotid stenosis (RCS) and to contrast the results of primary and secondary carotid endarterectomy (CENDX) with regard to operative morbidity and stroke prevention. METHODS: Forty-eight patients who underwent CENDX for RCS (RCS-OP group) were compared with a contemporaneous group of 40 patients who on at least one post-CENDX duplex ultrasonography study had a greater than 50% stenosis but did not undergo operation (RCS-NO-OP group). This latter group was drawn from 1053 follow-up duplex studies in 348 patients who underwent primary CENDX between the years 1983 and 1993. Each of these two groups was compared with a metanalysis of six key series derived from the literature. RESULTS: No significant differences were seen in the demographics or the incidence of risk factors between the two groups except for a higher incidence of coronary artery disease (p < 0.03) and peripheral vascular disease (p < 0.001) in the RCS-OP group. The operation-specific stroke rate was 2.1%, and the 30-day mortality was also 2.1%. Symptomatic RCS was the indication in 56% of cases. Important anatomic differences were found between groups. The duplex/arteriographic degree of stenosis was greater than 90% in 75% of the patients in the RCS-OP group, whereas only 10% of the patients in the RCS-NO-OP group had greater than 80% stenosis, most being in the 50% to 80% range. An unexpected finding was the sudden progression to occlusion in 10 (25%) of 40 in the RCS-NO-OP group, with 2 (5%) of 10 of the occlusions presenting as unheralded strokes. Overall, a stroke without an antecedent transient ischemic attack occurred in 3 (7.5%) of 40 of patients in the RCS-NO-OP group, all in patients with greater than 75% stenosis on their last documented scan preceding the stroke. CONCLUSION: Given the relatively low stroke rate with surgery in the RCS-OP group (2.1%) and the higher incidence of unheralded strokes (7.5%) in the RCS-NO-OP group, a more aggressive approach may be warranted in patients with asymptomatic high-grade (> 75%) RCS, a strategy not unlike that adopted for primary CENDX.

Automatic vessel segmentation and quantification of the rat aortic ring assay of angiogenesis.

BACKGROUND: Pharmacologic control of angiogenesis is a promising new approach to the treatment of a variety of pathologic conditions including cancer. The recently developed in vitro rat aortic ring model provides a simple, reproducible assay for discovering angiogenic agonists and antagonists. However, quantification of results in this assay is time consuming, tedious, and subjective, because it involves visual inspection of images and manually counting the newly formed microvessels extending from the cultured aortic ring. This report describes an automated image analysis-based procedure for quantification of this assay that overcomes these difficulties. EXPERIMENTAL DESIGN: The designed image processing algorithm segments the vessels from gray scale images. A high-pass filter is used, and the results are separated into nonvascular and vessel compartments based on object size and shape. Quantification relies on identification of vessels intersecting a closed transect set a fixed distance from the aortic ring. The number and the total area of these vessels are determined. The entire operation has been automated and packaged in an application called Vessels. RESULTS: The correlation between computer-determined vessel area/vessel number and visual microvessel count is high (r2 = 0.91 and r2 = 0.86, respectively). CONCLUSION: Vessels offers high-speed, fully automatic batch processing including production of a hard copy for documentation. The application runs on the Apple family of computers. On a Quadra 800, the application can process approximately 30 images/hour, which is approximately 2.5 times faster than manual quantification of this assay.

Heritable alterations at the adenine phosphoribosyltransferase (APRT) locus in human lymphoblastoid cell lines.

Human lymphoblastoid cell lines derived from WI-L2 exhibit unexpected frequencies of diaminopurine (DAP) resistant mutants. The background mutant fractions of 10(-7) to 10(-8) in untreated cultures are much lower than the frequencies expected for loss of a heterozygous autosomal locus (10(-5) to 10(-6), yet much higher than expected for a homozygous locus (10(-10) to 10(-12). We used aminopterin, adenine and thymidine (AAT) to select DAP-sensitive (DAPS) revertants from one resistant line. The background frequency of DAPR in these revertant cell lines ranged from 3.5 to 6.5 x 10(-4), approximately the square root of 10(-7). Thus these data suggest that both alleles of aprt are inactivated at similarly high frequencies. They also indicate that the DAPS revertants were heterozygotes (aprt +/-) or hemizygotes (aprt +/0) and that WI-L2 was homozygous (aprt+/+). Mutational dose-response studies with X-rays, ethyl methanesulfonate (EMS), and ICR-191 were conducted in 4 of these revertant cell lines. EMS and ICR-191, which induce mainly point mutations, did not induce an increase in mutant fraction. A dose of 200 cGy X-rays, however, induced a frequency of 10(-3). Treatment of DAPR cells with 5-azacytidine induced a significant increase in reversion to DAPS. Southern blot analysis of the aprt gene after digestion with MspI or HpaII also suggests that differential methylation changes may play a major role in the generation of DAP sensitivity and resistance.

Different patterns of inhibition of adrenaline-induced platelet aggregation and kinetics in vivo by acetylsalicylic acid and indobufen.

The inhibitory effect of 50 mg/kg lysyl acetylsalicylic acid (ASA) intravenously injected 24 and 4 h before 80 micrograms/kg adrenaline, 40 mg/kg indobufen injected 15 min before, or a combination of ASA plus indobufen on the platelet aggregation and kinetics was evaluated in anaesthetized dogs previously injected with 111indium- (111In-) labelled platelets using gamma-camera dynamics studies. The highest degree of inhibition, indicated by the lowest platelet aggregation ratio decrease and the most significant 111In-labelled platelet mobilization from hepatic and splenic stores with a corresponding increase of labelled and unlabelled platelet counts in blood, was obtained in ASA-treated dogs. Such changes were less marked when ASA plus indobufen was injected. In indobufen-treated dogs a similar mobilization of 111In-labelled platelets with an increase in circulating platelet numbers was evident only after the second adrenaline injection. It is concluded that platelet mobilization is mainly dependent on the production of an ASA-sensitive substance.

Acute effects of adrenaline on platelet aggregation and kinetics in vivo.

The platelet (Plt) contribution to cardiovascular events triggered by adrenergic stress is supported by some experimental and necropsy data, but a cause and effect link and mechanism have not yet been clearly demonstrated. Adrenergic stress was simulated by three successive adrenaline (A) injections (80 micrograms/kg) in anesthetized dogs previously infused with indium111 labelled Plt (111In-Plt) for a gamma-camera study. Adrenaline induced an acute and significant decrease in the Plt aggregation ratio (PAR) and a parallel decrease in the Plt count as well as in the circulating 111In-Plt, mirroring a significant and persistent Plt sequestration mainly in the liver and spleen. The long-lasting and significant decrease in the circulating Plt count observed 15 min after each A injection can be explained by a persistent retention of Plt. Further studies are in progress in order to disclose if this corresponds to platelet microaggregate embolization in microvessels of the organs so far analysed. If this postulate can be confirmed the hypothesis of an adrenergic stress triggering of ischemic events will be justified.

3-(1-Indolinyl)benzylamines: a new class of analgesic agents.

An extensive series of 3-(1-indolinyl)benzylamines and related compounds was synthesized and tested for analgesic activity. After a detailed study of structure-activity relationships, 3-(1-indolinyl)benzylamine (2b) was selected for further investigation as the most interesting member of this novel class of compounds. It was active in both the phenylquinone writhing and tail-flick assays for analgesic activity. No motor deficits were observed in the rotorod test, and 2b was found to be free of any other effects on the central nervous system. The compound did not bind to opiate receptors, since it was inactive in inhibiting the stereospecific binding of [3H]naloxone in rat brain homogenates. Thus, 3-(1-indolinyl)benzylamine represents a novel analgesic with an unusual chemical structure and biological profile.