Comblike dendrimers containing Tn antigen modulate natural killing and induce the production of Tn specific antibodies

Comblike glycodendrimers were prepared by the chemoselective ligation of cysteine-modified glycopeptides (1-7) with a 3-maleimidopropionate-modified linear synthetic carrier (8). Glycodendrimers bearing mono-, di-, or tri-Tn clusters (9-11) were tested as inhibitors using plant and mammalian lectins. In the former group, the Codium fragile lectin showed moderate discrimination among 9, 10, and 11. In the latter group, A and B isoforms of rat NKR-P1 lectin strongly discriminated between 9 and 10. 10 caused a 4-fold increase in killing of the NK resistant tumor cell lines at concentrations as low as 10(-8) M. Surprisingly, 11 interacted exclusively with the rat NKR-P1B isoform and inhibited efficiently natural killing in both rats and humans, even in the presence of the activating compounds 9 and 10. Dinitrophenol haptenization or influenza virus hemagglutinin T-cell epitope conjugation increased the immunogenicity of the parent compounds and resulted in the production of Tn specific antibodies.     

Plasma lipoproteins in transport of silibinin,an antioxidant flavonolignan from Silybum marianum

To assess the role of plasma lipoproteins in the transport of silibinin, an antioxidant flavonolignan, (125)I-labelled silibinin ((125)I-SB) administered perorally to the rat was used. The plasma (125)I-SB derived radioactivity was distributed among plasma lipoproteins according to their lipophilicity (TAG-rich lipoproteins 30-40% > LDL 15% > HDL 5%), and in the fraction of d > 1.215 containing albumin and other proteins a minority amount of radioactivity was found. Administration of (125)I-SB in a complex with phosphatidylcholine resulted in proportionally higher radioactivities in all fractions as well as in tissues. Dietary olive oil had a slightly decreasing effect on plasma concentrations of silibinin measured by HPLC as well as on (125)I-SB derived radioactivity in plasma and liver. In the TAG-rich lipoprotein fraction and HDL no effects of olive oil on the levels of (125)I-SB derived radioactivities were observed, however, at a 30 min interval the levels of (125)I-SB derived radioactivity in LDL and the heart were significantly decreased in the olive oil group. These results suggest that (i) silibinin is not resorbed by the chylomicron pathway, and (ii) the endogenous lipoprotein pathway VLDL --> LDL may play a role in the transport of silibinin from the liver to the extrahepatic tissues concurrently facilitating the lipoprotein antioxidant influence of silibinin.     

Characterization of a charybdotoxin-sensitive intermediate conductance Ca2+-activated K+ channel in porcine coronary endothelium: relevance to EDHF

1. This study characterizes the K(+) channel(s) underlying charybdotoxin-sensitive hyperpolarization of porcine coronary artery endothelium. 2. Two forms of current-voltage (I/V) relationship were evident in whole-cell patch-clamp recordings of freshly-isolated endothelial cells. In both cell types, iberiotoxin (100 nM) inhibited a current active only at potentials over +50 mV. In the presence of iberiotoxin, charybdotoxin (100 nM) produced a large inhibition in 38% of cells and altered the form of the I/V relationship. In the remaining cells, charybdotoxin also inhibited a current but did not alter the form. 3. Single-channel, outside-out patch recordings revealed a 17.1+/-0.4 pS conductance. Pipette solutions containing 100, 250 and 500 nM free Ca(2+) demonstrated that the open probability was increased by Ca(2+). This channel was blocked by charybdotoxin but not by iberiotoxin or apamin. 4. Hyperpolarizations of intact endothelium elicited by substance P (100 nM; 26.1+/-0.7 mV) were reduced by apamin (100 nM; 17.0+/-1.8 mV) whereas those to 1-ethyl-2-benzimidazolinone (1-EBIO, 600 microM, 21.0+/-0.3 mV) were unaffected (21.7+/-0.8 mV). Substance P, bradykinin (100 nM) and 1-EBIO evoked charybdotoxin-sensitive, iberiotoxin-insensitive whole-cell perforated-patch currents. 5 A porcine homologue of the intermediate-conductance Ca(2+)-activated K(+) channel (IK1) was identified in endothelial cells. 6. In conclusion, porcine coronary artery endothelial cells express an intermediate-conductance Ca(2+)-activated K(+) channel and the IK1 gene product. This channel is opened by activation of the EDHF pathway and likely mediates the charybdotoxin-sensitive component of the EDHF response.     

Intermetallic Phases Identification and Diffusion Simulation in Twin-Roll Cast Al-Fe Clad Sheet

Aluminium steel clad materials have high potential for industrial applications. Their mechanical properties are governed by an intermetallic layer, which forms upon heat treatment at the Al-Fe interface. Transmission electron microscopy was employed to identify the phases present at the interface by selective area electron diffraction and energy dispersive spectroscopy. Three phases were identified: orthorhombic Al₅Fe₂, monoclinic Al₁₃Fe₄ and cubic Al₁₉Fe₄MnSi₂. An effective interdiffusion coefficient dependent on concentration was determined according to the Boltzmann–Matano method. The highest value of the interdiffusion coefficient was reached at the composition of the intermetallic phases. Afterwards, the process of diffusion considering the evaluated interdiffusion coefficient was simulated using the finite element method. Results of the simulations revealed that growth of the intermetallic phases proceeds preferentially in the direction of aluminium.     

Regional changes of brain serotonin and its metabolite 5-hydroxyindolacetic Acid and development of immunosuppression in submissive mice

The production of submissive behavior in C57BL/6J mice during 10 or 20 days of social confrontations resulted in increases in serotonin (5-HT) content in the amygdala, hippocampus, nucleus caudatus, Al1, A10, A9, and hypothalamus. The level of 5-hydroxyindolacetic acid (5-HIAA) was higher in most structures after 20 daily encounters compared to animals tested for 10 days. The ratio 5-HIAA/5-HT was increased in the nucleus raphe, accumbens, A9, and hypothalamus in mice displaying submission during 10 and 20 confrontations. The experience of defeats during 10 days accompanied with 5-HT system activation in a number of brain structures (nucleus raphe, accumbens, and A9) produced immunosuppression. With increasing number of confrontations the ratio 5-HIAA/5-HT was decreased in the same structures and a tendency to the immune response elevation appeared.     

Clinical and laboratory prognostic factors in patients with metastatic renal cell carcinoma treated with sunitinib and sorafenib after progression on cytokines

ObjectivesThe aim of this retrospective study was to analyze prognostic factors in patients with metastatic renal cell carcinoma treated with tyrosine kinase inhibitors (TKIs) sunitinib or sorafenib after progression on cytokine therapy.Materials and methodsA national database of patients treated with targeted agents was used as the data source. A total of 319 patients treated with sunitinib (n = 181) or sorafenib (n = 138) after progression on cytokine therapy were analyzed.ResultsPrognostic factors significantly associated with poor overall survival in a multivariable Cox model included the time from diagnosis to the start of treatment with TKIs<1 year, increased neutrophil counts, increased lactate dehydrogenase, and Eastern Oncology Cooperative Group performance status 2 or higher. The parameters showing statistically significant association with progression-free survival included time from diagnosis to the beginning of treatment with TKI<1 year, increased lactate dehydrogenase, and Eastern Oncology Cooperative Group performance status 2 or higher. We have also validated the International Metastatic Renal Cell Carcinoma Database Consortium prognostic model in our cohort of patients.ConclusionWe demonstrate that the International Database Consortium prognostic model performs well for European patients treated with TKIs, including sunitinib or sorafenib, after progression on cytokines and suggest that a reduction from original 6 down to 4 parameters is possible.     

Ultrasound imaging of the infraorbital foramen and simulation of the ultrasound-guided infraorbital nerve block using a skull model

Purpose

Infraorbital nerve block is used for intraoperative and postoperative analgesia in nasal and oral surgery procedures, as well as in the chronic pain settings. Ultrasound guidance has not been described in the literature. The aim of the study was to assess the quality of ultrasound imaging of the infraorbital foramen and develop an “in-plane” technique of the block using a skull model.

Methods

The infraorbital foramina were assessed on five skull models immersed in the water bath. Ultrasound-guided simulation of an in-plane infraorbital nerve block was then performed. Slightly curved needle was placed close to the foramina and its visibility was recorded. Success rate and time to locate infraorbital foramina, success rate and time to insert the needle close to the foramina under ultrasound and correlation between the ultrasound and caliper measurements were evaluated and recorded. Data for the left and right foramen were compared.

Results

The infraorbital foramina were successfully located using ultrasound in all 20 cases. Simulation of infraorbital nerve block was also successful in all measurements. The time difference between locating or simulating blockade of the left and right infraorbital foramina was not statistically significant. Correlation between ultrasound measurement and direct measurement using a caliper was satisfactory for the distances between the inferior orbital rim and the inferior margin of the infraorbital foramen but poor for the distances between the lower rim of the orbit and the superior margin of the foramen.

Conclusions

This experimental study suggests that the infraorbital foramen is easily located using ultrasound and an “in-plane” ultrasound-guided technique for infraorbital nerve blockade is feasible on the model.     

Effects of Austenitization Temperature and Pre-Deformation on CCT Diagrams of 23MnNiCrMo5-3 Steel

The combined effect of deformation temperature and strain value on the continuous cooling transformation (CCT) diagram of low-alloy steel with 0.23% C, 1.17% Mn, 0.79% Ni, 0.44% Cr, and 0.22% Mo was studied. The deformation temperature (identical to the austenitization temperature) was in the range suitable for the wire rolling mill. The applied compressive deformation corresponded to the true strain values in an unusually wide range. Based on the dilatometric tests and metallographic analyses, a total of five different CCT diagrams were constructed. Pre-deformation corresponding to the true strain of 0.35 or even 1.0 had no clear effect on the austenite decomposition kinetics at the austenitization temperature of 880 °C. During the long-lasting cooling, recrystallization and probably coarsening of the new austenitic grains occurred, which almost eliminated the influence of pre-deformation on the temperatures of the diffusion-controlled phase transformations. Decreasing the deformation temperature to 830 °C led to the significant acceleration of the austenite → ferrite and austenite → pearlite transformations due to the applied strain of 1.0 only in the region of the cooling rate between 3 and 35 °C·s⁻¹. The kinetics of the bainitic or martensitic transformation remained practically unaffected by the pre-deformation. The acceleration of the diffusion-controlled phase transformations resulted from the formation of an austenitic microstructure with a mean grain size of about 4 µm. As the analysis of the stress–strain curves showed, the grain refinement was carried out by dynamic and metadynamic recrystallization. At low cooling rates, the effect of plastic deformation on the kinetics of phase transformations was indistinct.