Investigations of impact biomechanics for penetrating ballistic cases.

This study aims to investigate the penetration of a projectile into a surrogate human tissue numerically, using Finite Element (FE) simulation. 20% Balistic Gelatin material (BG) is simulated with an elasto-plastic hydrodynamic constitutive law, and then impacted by steel spheres at different velocities. The results from the FE simulations are compared with existing experimental data and other analytical equations from the literature. To our knowledge, this is the first study to investigate a projectile penetration by numerical simulation, and then compare the results with analytical and experimental data from previous studies. This developed model gives encouraging results for further investigations of penetrating impact of projectile in the human body.

[Effectiveness of measures to promote and control on the appropriateness of preventive measures taken on farms--health surveillance, surveys on damages associated with risk]. / Efficacia delle azioni di promozione e controllo sull'appropriatezza delle misure di prevenzione adottate nelle aziende agricole--la sorveglianza sanitaria, le indagini sui danni associati al rischio.

The agricultural activities have several issues in the management of safety and health of workers. The study of two ASL of Central Italy (VT and RMH) intended to check the risk conditions in order to highlight most critical points and define a prevention and surveillance plan. We moved in these directions: verification of workplaces and work practices; examination of machineries and equipment; active search of occupational diseases. We analyzed some peculiar aspects of the health surveillance of 75 workers such as risk from sun exposure, significantly underestimated by employers and competent doctors, despite sun exposure diseases are included in the list for which reporting is mandatory. Our study shows that a targeted campaign of prevention and control can lead to an improvement in safety management, on the other hand shows the necessity to bring occupational health physician to assess and manage also less valuated risks as the sun exposure.

Rabbit tissue distribution of 3H-ouabain by digital radioautography.

3H-ouabain is useful to evaluate the tissue localization of Na,K-ATPase. In this work we determined the distribution of 3H-ouabain in rabbit tissue by digital radioautography. Using this method, we were able to obtain a comparison of various organs in a relatively short time (6.5 days), while with traditional radioautography, only the kidney was detectable after seven months of film exposure. The kidney has the highest intensity (concentration of 3H-ouabain), followed by the heart, liver, muscle, lung, spleen and finally brain, which was almost undetectable. In kidney the activity was higher in the cortex, while the other tissues displayed a more uniform intensity, suggesting that Na,K-ATPase was evenly distributed.

[A case of pseudophlebitis of the great saphenous vein: focal nodular myositis of the gracilis muscle]. / A propos d'un cas de pseudophlébite de la grande veine saphène: une myosite nodulaire focale du muscle gracilis.

A case of focal myositis in a healthy 68-year-old woman is described. The patient was admitted for evaluation of a painful soft-tissue mass localised on the medial side of the left thigh, initially misdiagnosed as thrombophlebitis of the v. saphena magna. Laboratory data were normal, in particular sedimentation rate and muscle enzyme levels. After exclusion of venous thrombosis, the mass localised in the left m. gracilis was surgically removed. Histologic examination of the biopsy specimen showed muscle cell necrosis and severe inflammation, with lymphocytic infiltration leading to the diagnosis of focal myositis. This is a rare benign inflammatory pseudotumour of skeletal muscle. The aetiology and pathogenesis of the disease remain unclear. It is most commonly seen in the lower extremities and may mimic thrombophlebitis or soft-tissue neoplasm. Ultrasound and magnetic-resonance scans are helpful, but definitive diagnosis is obtained only by histology. Because recurrent lesions in other skeletal muscles are possible, and a third of patients develop polymyositis, a follow-up of several years is recommended.

The low M(r) protein-tyrosine phosphatase is involved in Rho-mediated cytoskeleton rearrangement after integrin and platelet-derived growth factor stimulation.

The low molecular weight protein-tyrosine phosphatase (LMW-PTP) is an enzyme that is involved in the early events of platelet-derived growth factor (PDGF) receptor signal transduction. In fact, LMW-PTP is able to specifically bind and dephosphorylate activated PDGF receptor, thus modulating PDGF-induced mitogenesis. In particular, LMW-PTP is involved in pathways that regulate the transcription of the immediately early genes myc and fos in response to growth factor stimulation. Recently, we have found that LMW-PTP exists constitutively in cytosolic and cytoskeleton-associated localization and that, after PDGF stimulation, c-Src is able to bind and phosphorylate LMW-PTP only in the cytoskeleton-associated fraction. As a consequence of its phosphorylation, LMW-PTP increases its catalytic activity about 20-fold. In this study, our interest was to investigate the role of LMW-PTP phosphorylation in cellular response to PDGF stimulation. To address this issue, we have transfected in NIH-3T3 cells a mutant form of LMW-PTP in which the c-Src phosphorylation sites (Tyr(131) and Tyr(132)) were mutated to alanine. We have established that LMW-PTP phosphorylation by c-Src after PDGF treatment strongly influences both cell adhesion and migration. In addition, we have discovered a new LMW-PTP substrate localized in the cytoskeleton that becomes tyrosine-phosphorylated after PDGF treatment: p190Rho-GAP. Hence, LMW-PTP plays multiple roles in PDGF receptor-mediated mitogenesis, since it can bind and dephosphorylate PDGF receptor, and, at the same time, the cytoskeleton-associated LMW-PTP, through the regulation of the p190Rho-GAP phosphorylation state, controls the cytoskeleton rearrangement in response to PDGF stimulation.

Inhibitory effect of full-length human endostatin on in vitro angiogenesis.

Endostatin, a C-terminal product of collagen XVIII, is a very powerful angiogenesis inhibitor. In vivo experiments in mice indicate that endostatin dramatically reduces tumor mass without causing the onset of any resistance to the treatment. Recently, a 12-aa shorter human endostatin has been purified from plasma, but is ineffective in in vitro angiogenesis assays. Here we report that the full-length human recombinant endostatin has a potent inhibitory activity in in vitro angiogenesis assays. Two powerful angiogenic factors were used to stimulate endothelial cells: FGF-2 and VEGF-165. Endostatin prevented cell growth both in the basal condition and after stimulation with FGF-2 or VEGF-165. Migration of microvascular endothelial cells toward FGF-2 or VEGF-165 was impaired, both when cells were pretreated with the inhibitor and when endostatin was added together with the growth factors. Furthermore, experiments of inhibition of proliferation performed on nonmicroendothelial cells showed that endostatin was ineffective. This study indicates that human endostatin is a potent angiogenesis inhibitor and suggests its use in human anticancer therapy.

The low Mr phosphotyrosine protein phosphatase behaves differently when phosphorylated at Tyr131 or Tyr132 by Src kinase.

The low molecular weight phosphotyrosine protein phosphatase (LMW-PTP) is phosphorylated by Src and Src-related kinases both in vitro and in vivo; in Jurkat cells, and in NIH-3T3 cells, it becomes tyrosine-phosphorylated upon stimulation by PDGF. In this study we show that pp60Src phosphorylates in vitro the enzyme at two tyrosine residues, Tyr131 and Tyr132, previously indicated as the main phosphorylation sites of the enzyme, whereas phosphorylation by the PDGF-R kinase is much less effective and not specific. The effects of LMW-PTP phosphorylation at each tyrosine residue were investigated by using Tyr131 and Tyr132 mutants. We found that the phosphorylation at either residue has differing effects on the enzyme behaviour: Tyr131 phosphorylation is followed by a strong (about 25-fold) increase of the enzyme specific activity, whereas phosphorylation at Tyr132 leads to Grb2 recruitment. These differing effects are discussed on the light of the enzyme structure.

Secretory polymorphism and serous cutaneous gland heterogeneity in Bufo granulosus (Amphibia, Anura).

Three types of secretory products (a, b and c) in the poison glands of the Argentine toad Bufo granulosus have been detected under light microscope. The type a secretory product consists of granules of homogeneous density, type b of vesicles with a translucent compartment and type c of granules of varying density. Subsequent transmission electron microscope analysis disclosed obvious similarities in the secretory pathways of type a and c granules; the differences detected under light microscope are due to the functional phases observed. On the contrary, production of type b secretory vesicles involves a distinctive pathway. Therefore, two classes of glands (I and II) have been identified. Glands of the first class are typical of bufonid toads and produce granules provided with repeating substructure; glands of the second class, which manufacture a lucent product, are unusual in the family Bufonidae. Ultrastructural differences, consistent with the two gland classes, have also been described in the myoepithelia. The myocytes ensheathing class I secretory units possess striking cytoskeletal specializations, whereas those of class II glands are rich in sarcoplasmic reticulum. The distinctive ultrastructural traits detected in these myoepithelial cells have been compared with the results of previous studies on the dimorphic serous glands of Bombina. Findings point to the use of pharmacological treatment on the skin of anurans with different classes of serous glands to elicit differential secretory discharge.

Low molecular weight protein-tyrosine phosphatase tyrosine phosphorylation by c-Src during platelet-derived growth factor-induced mitogenesis correlates with its subcellular targeting.

The low molecular weight phosphotyrosine phosphatase (LMW-PTP) is an enzyme that is involved in the early events of platelet-derived growth factor (PDGF) receptor signal transduction. Our previous results have shown that LMW-PTP is able to specifically bind and dephosphorylate activated PDGF receptor, thus modulating PDGF-induced mitogenesis. In particular LMW-PTP is involved in pathways that regulate the transcription of the immediately early genes myc and fos in response to growth factor stimulation. In this study we have established that, in nontransformed NIH3T3 cells, LMW-PTP exists constitutively in cytosolic and cytoskeleton-associated localization and that, after PDGF stimulation, c-Src is able to bind and to phosphorylate LMW-PTP only in the cytoskeleton-associated fraction. As a consequence of its tyrosine phosphorylation, LMW-PTP significantly increases its catalytic activity. After PDGF stimulation these two LMW-PTP pools act on distinct substrates, contributing in different manners to the PDGF receptor signaling. The cytoplasmic LMW-PTP fraction exerts its well known action on activated PDGF receptor. On the other hand we have now demonstrated that the cytoskeleton-associated LMW-PTP acts specifically on a few not yet identified proteins that become tyrosine-phosphorylated in response to the PDGF receptor activation. Finally, these two LMW-PTP pools markedly differ in the timing of the processes in which they are involved. The cytoplasmic LMW-PTP pool exerts its action within a few minutes from PDGF receptor activation (short term action), while tyrosine phosphorylation of cytoskeleton-associated LMW-PTP lasts for more than 40 min (long term action). In conclusion LMW-PTP is a striking example of an enzyme that exerts different functions and undergoes different regulation in consequence of its subcellular localization.

Detection of perfusion defects during coronary occlusion and myocardial reperfusion after thrombolysis by intravenous administration of the echo-enhancing agent BR1.

The purpose of this study was to detect myocardial perfusion defects as a result of coronary occlusion and myocardial reperfusion after thrombolysis with intravenous (i.v.) administration of the echo contrast agent BR1 (Bracco Research, Switzerland), which consists of microbubbles (median diameter 2.5 microm) containing sulfur exafluoride in a phospholipidic shell. To generate a coronary thrombosis, a copper coil was advanced into the left circumflex coronary artery in eight anesthetized dogs with opened chest cavities. Coronary occlusion occurred 18 +/- 10 minutes after the insertion of the coil and was documented both by an electromagnetic flow meter (as zero blood flow) and by radiolabeled microspheres (as myocardial perfusion defect). After 2 hours of occlusion, streptokinase was infused i.v.; reperfusion was documented by both the flow-meter and microspheres. Left ventricular cavity enhancement was apparent after all contrast injections. Peak cavity intensity did not increase with dose and was not affected by signal processing (suggesting signal saturation), whereas the duration of contrast effect significantly increased with the dose (from 26 +/- 16 to 147 +/- 74 seconds). Myocardial contrast intensity also increased after contrast (from 15 +/- 12 to 21 +/- 18 gray level/pixel, p < 0.001). Contrast echo detected myocardial perfusion defects (corresponding to 17% +/- 11% of LV cross-sectional area) in all the injections performed during coronary occlusion and detected myocardial reperfusion with a sensitivity of 50% versus microspheres. The extent of perfusion defects by contrast echo showed a good correlation with microspheres (r = 0.73). Myocardial reperfusion was not detected by changes in heart rate, aortic pressure, pulmonary arterial pressure, cardiac output, left ventricular fractional area change, or wall-motion score index. Hemodynamic parameters were not affected by contrast injections. Thus, the i.v. administration of BR1 allows us to accurately detect myocardial perfusion defects during coronary occlusion and, to a lesser extent, myocardial reperfusion after thrombolysis.

Acylphosphatase is involved in differentiation of K562 cells.

The level of both isoforms of acylphosphatase was evaluated in the human erythroleukemia K562 cell line during differentiation. K562 cells were treated with PMA, which induces megakaryocytic differentiation, and with aphidicolin or hemin, which stimulate erythrocytic differentiation. While the MT isoform showed an average 10-fold increase independently of the differentiating agent used, only hemin treatment caused a similar increase of the CT isoform, suggesting a different role of the two isoforms in the cell. Treatment with either hemin or aphidicolin of K562 cells overexpressing the two acylphosphatase isoforms suggested the possibility that acylphosphatases play a role in the onset of differentiation.

Characterization of a novel nucleolytic activity of acylphosphatases.

A novel enzymatic activity on nucleic acids was discovered in both muscle type (MT) and erythrocyte or common type (CT) isoforms of acylphosphatase, an enzyme that was previously known as a hydrolase (E.C.3.6.1.7). Both deoxyribonucleic and ribonucleic hydrolitic activity were assayed on a variety of substrates. Our results demonstrate that acylphosphatase possesses both Mg++ dependent deoxyribonuclease and ribonuclease activities, at pH ranging from 5.0 to 6.8. Furthermore, we present evidences, for both isoenzymatic forms, of the coexistence of exonucleolytic and endonucleolytic activities on DNA.

Low M(r) phosphotyrosine protein phosphatase interacts with the PDGF receptor directly via its catalytic site.

Many proteins bind to the activated platelet derived growth factor receptor (PDGF-R) either directly or by means of adapter molecules. Up to now all these proteins were shown to transmit and amplify the signal started with PDGF-R stimulation. In a recent study our group had demonstrated that low M(r) phosphotyrosine protein phosphatase (LMW-PTP) specifically interacts with PDGF-R in NIH3T3 cells. In the present study we have attempted to clarify the modality of interaction, both in vivo and in vitro, of these two proteins, using a catalytically inactive LMW-PTP mutant. Our results indicate that LMW-PTP and PDGF-R interact directly, without the necessity of any adapter protein. This interaction leads to PDGF-R dephosphorylation and, presumably, interrupts one or more of the mitogenic pathways that originate from receptor activation.

The new nitroderivative ITF 296 improves collateral blood flow in a canine model of coronary thrombosis.

The actions of ITF 296 and isosorbide dinitrate (ISDN), 20, 70, and 200 g/kg/min, on myocardial transmural blood flow distribution during acute thrombotic occlusion of the left circumflex coronary artery (LCX) have been evaluated in seven and three anesthetized open-chest dogs, respectively, and compared with four animals receiving vehicle. Occlusion of LCX was achieved in 14 +/- 2 min by the insertion of a copper coil. This caused transmural myocardial ischemia in the LCX area, while leaving blood flow in the left anterior descending coronary artery (control area) unaffected. Infusion of ITF 296 (200 g/kg/min) increased transmural coronary flow in the border zone and in the control area without affecting blood flow in the central ischemic area. ISDN, given in the same dose, reduced systemic blood pressure but did not affect LCX blood flow. In three dogs with residual perfusion in the LCX central area ITF 296 also increased blood flow. These results confirm that ITF 296 promotes an increase of flow to the border zone, thus possibly reducing the area of myocardial infarction.

Intracoronary air-filled albumin microspheres for myocardial blood flow measurement.

OBJECTIVES: The aim of this study was to explore the possibility of quantifying coronary blood flow by myocardial contrast echocardiography with air-filled serum albumin microspheres (Albunex). BACKGROUND: Air-filled albumin microspheres have been proposed as an intravascular tracer for the study of myocardial perfusion by contrast echocardiography. METHODS: In six anesthetized open chest dogs, the left circumflex coronary artery was cannulated and perfused by a roller pump with blood from the femoral artery. Both air-filled albumin microspheres (0.4 ml, 2 x 10(8) spheres/ml) and technetium-99m-labeled albumin were injected as a bolus into the coronary cannula at baseline and after treatment with dipyridamole (0.56 mg/kg body weight intravenously for 4 min). Two-dimensional echographic images of the left ventricular short axis were digitized to generate myocardial time-intensity curves; myocardial radioactivity was measured by an external detector to generate radionuclide time-activity curves. RESULTS: After dipyridamole, left circumflex coronary artery blood flow (as measured by both the pump and an electromagnetic flow meter) significantly increased (from 1.06 +/- 0.28 to 3.61 +/- 1.43 ml/min per g of myocardium). Peak intensity and rise time of contrast echo curves were able to differentiate baseline myocardial perfusion from coronary hyperemia but did not show any significant correlation with coronary blood flow. A weak inverse correlation with coronary blood flow was provided by myocardial mean transit time of air-filled albumin microspheres (r = 0.33). Conversely, a close inverse correlation with coronary blood flow was obtained by myocardial mean transit time of technetium-99m-labeled albumin (r = 0.95). Myocardial transit time of air-filled albumin microspheres (1.95 +/- 0.60 s) was also markedly shorter than that of labeled albumin (5.35 +/- 3.43 s, p < 0.001) and the measurements were less reproducible. CONCLUSIONS: In this experimental study, coronary blood flow was not adequately quantified by myocardial contrast echocardiography with intracoronary injection of air-filled albumin microspheres.

Myocardial washout of sonicated iopamidol does not reflect the transmural distribution of coronary blood flow.

It has been shown in previous studies that myocardial contrast echocardiography provides quantitative information on coronary blood flow. However, the ability of contrast echo to assess the transmural (endo/epicardial) distribution of blood flow is still debated. To test this hypothesis, the left circumflex coronary arteries of six anaesthetized open-chested dogs were cannulated and perfused with blood from the femoral artery. At different rates of coronary blood flow, during adenosine-induced coronary vasodilation, sonicated iopamidol and radionuclide labelled microspheres were injected into the coronary cannula, immediately proximal to a mixing chamber. Two-D echo images were digitized and myocardial time-intensity curves were obtained for the endocardial, mid- and epicardial layers. A good correlation existed between contrast washout of the entire ventricular wall and coronary flow (r = 0.85). However, the washout rate from the endo-, mid- and epicardial layers showed weak correlations with corresponding regional blood flows measured by microspheres (r = 0.56, 0.71 and 0.58, respectively). No significant relationship was found between the endo/epicardial washout ratio and the corresponding flow ratio by microspheres. Thus, measurement of the transmural distribution of coronary blood flow by myocardial contrast echocardiography remains an elusive goal.

Flow quantitation by contrast echocardiography. Effects of intervening tissue and of the angle of incidence between flow and ultrasonic beam.

The combination of a standardized echographic contrast agent with the analysis of the ultrasonic radio frequency (RF) signal allowed in vitro flow quantitation in a circulation model. The purpose of this study was to investigate both the effects of biological tissues, intervening between probe and insonated structure, and the effects of the angle of incidence between flow and ultrasonic beam on RF flow quantitation. Thus, the contrast agent SHU 454 was intravenously injected (0.4 ml) as a bolus into a circulation model, at variable flow rates, while keeping the pressure and volume of the vessel constant. Injections were performed with saline interposed between probe and vessel and after the addition of the subcutaneous tissue of a pig; injections were also performed using the probe normal to the flow and with an angle of incidence of 45 degrees. Echographic data were recorded by a mechanical sector scanner, capable of sampling the RF signal from a region of interest positioned in the center of the vein. Contrast echo time-intensity curves were generated. As expected, both peak intensity and the area under the curves decreased with intervening tissue (-58 and -70% of baseline values, respectively, p < 0.001). Surprisingly, mean transit time also decreased with intervening tissue (from 1.12 +/- 0.25 seconds with saline, to 0.92 +/- 0.13 seconds with tissue, p < 0.001), thus producing a systematic overestimation of flow (21% on the average). To compensate for signal attenuation, contrast injections were repeated in the presence of tissue after increasing the electronic signal amplification (10 dB), and transit time did not significantly differ from control.(ABSTRACT TRUNCATED AT 250 WORDS)

Flow quantitation by radio frequency analysis of contrast echocardiography.

Contrast echocardiography has the potential for measuring cardiac output and regional blood flow. However, accurate quantitation is limited both by the use of non-standard contrast agents and by the electronic signal distortion inherent to the echocardiographic instruments. Thus, the aim of this study is to quantify flow by combining a stable contrast agent and a modified echo equipment, able to sample the radio frequency (RF) signal from a region of interest (ROI) in the echo image. The contrast agent SHU-454 (0.8 ml) was bolus injected into an in vitro calf vein, at 23 flow rates (ranging from 376 to 3620 ml/min) but constant volume and pressure. The ROI was placed in the centre of the vein, the RF signal was processed in real time and transferred to a personal computer to generate time-intensity curves. In the absence of recirculation, contrast washout slope and mean transit time (MTT) of curves (1.11-8.52 seconds) yielded excellent correlations with flow: r = 0.93 and 0.95, respectively. To compare the accuracy of RF analysis with that of conventional image processing as to flow quantitation, conventional images were collected in the same flow model by two different scanners: a) the mechanical sector scanner used for RF analysis, and b) a conventional electronic sector scanner. These images were digitized off-line, mean videodensity inside an identical ROI was measured and time-intensity curves were built. MTT by RF was shorter than by videodensitometric analysis of the images generated by the same scanner (p < 0.001). In contrast, MTT by RF was longer than by the conventional scanner (p < 0.001). Significant differences in MTT were also found with changes in the gain setting controls of the conventional scanner. To study the stability of the contrast effect, 6 contrast injections (20 ml) were performed at a constant flow rate during recirculation: the spontaneous decay in RF signal intensity (t1/2 = 64 +/- 8 seconds) was too long to affect MTT significantly. In conclusion, the combination of a stable contrast agent and a modified echocardiographic instrument provides accurate quantitation of flow in an in vitro model; RF analysis is more accurate than conventional processing as to flow quantitation by contrast echocardiography.