Analysis of leucocyte antibodies, cytokines, lysophospholipids and cell microparticles in blood components implicated in post-transfusion reactions with dyspnoea.

BACKGROUND AND OBJECTIVES: Post-transfusion reactions with dyspnoea (PTR) are major causes of morbidity and death after blood transfusion. Transfusion-related acute lung injury (TRALI) and transfusion-associated circulatory overload (TACO) are most dangerous, while transfusion-associated dyspnoea (TAD) is a milder respiratory distress. We investigated blood components for immune and non-immune factors implicated in PTR. MATERIAL AND METHODS: We analysed 464 blood components (RBCs, PLTs, L-PLTs, FFP) transfused to 271 patients with PTR. Blood components were evaluated for 1/antileucocyte antibodies, 2/cytokines: IL-1ß, IL-6, IL-8, TNF-α, sCD40L, 3/lysophosphatidylcholines (LysoPCs), 4/microparticles (MPs) shed from plateletes (PMPs), erythrocytes (EMPs) and leucocytes (LMPs). RESULTS: Anti-HLA class I/II antibodies or granulocyte-reactive anti-HLA antibodies were detected in 18.2% of blood components (RBC and FFP) transfused to TRALI and in 0.5% of FFP transfused to TAD cases. Cytokines and LysoPCs concentrations in blood components transfused to PTR patients did not exceed those in blood components transfused to patients with no PTR. Only EMPs percentage in RBCs transfused to patients with TRALI was significantly higher (P < 0.05) than in RBCs transfused to patients with no PTR. CONCLUSION: Immune character of PTR was confirmed mainly in 1/5 TRALI cases. Among non-immune factors, only MPs released from stored RBCs are suggested as potential mediators of TRALI. Our results require further observations in a more numerous and better defined group of patients.

Micro-computed tomography study of the abnormal osseous extensions of sella turcica.

The paper presents anatomical considerations on the abnormal ossification, which occurred around the dorsum of the sella turcica in the human skull of the female individual. Probably the morphological alterations of the sellar region were associated with extensive heterotopic ossification of the dura mater attached to the dorsum of the sella turcica and the posterior clinoid processes. The analysis of gray values of the voxels representing the areas of abnormal ossification indicated on variation in bone density in the entire sample. We have established that the highest mineralisation of bony tissue occurred in the marginal parts of the osseous extensions deriving from the posterior clinoid processes. The ossified parts of dura mater attached to the posterior clinoid processes showed significantly higher content of the hydroxyapatite (1.9 g/cm3) than the dorsum of the sella turcica (1.0 g/cm3).

Three dimensional visualisation and morphometry of bone samples studied in microcomputed tomography (micro-CT).

This article highlights the utility of micro-computed tomography (micro-CT) for characterisingmicroscale bone morphology. For this purpose we tested selected samplesof the human bones (Wormian bone, rib, lumbar vertebra) to reconstruct externaland internal morphological features. Selected bony samples were investigatedusing a micro-CT scanner (Skyscan 1172, N.V., Aartselaar, Belgium). The imageresolution of scans varied from 5 to 27 µm/pixel depending on the bone sample.We used CTvox software (by Skyscan) to perform volume rendering of the samples.Further, 3-dimensional geometrical models were reconstructed using theCTvol application. Such models enabled graphical distinction between osseouscomponents of various morphology and were used to visualise the Haversian canalsystem inside the compact bone of the rib. Applying a modified transfer functionfor volume rendering we presented the overall morphology of the Wormian boneand small vascular channels penetrating its interior. As an example of quantitativeanalysis based on micro-CT scans we compared the trabecular structure of thelumbar vertebrae with CTAn software. Significant differences in percent bonevolume (BV/TV) were determined. Micro-CT was found to be a very accurate andhelpful method to study small anatomical structures of the bones in micro scale.

Topography and morphometry of the subarcuate canal.

The current study shows in a close-up view anatomical relationship between the subarcuate canal and the osseous labyrinth. For this purpose we used micro-computed tomography which allowed performing three-dimensional reconstruction of the subarcuate canal and gave adequate data for estimation its diameter across its course. The diameter of the middle part (the most uniform) of the subarcuate canal varied from 0.28 mm to 0.46 mm. Hence, we calculated the centre of mass for each cross-section of the separated subarcuate canal. This procedure helped us to visualise trajectory of the subarcuate canal and its spatial orientation within the petrous bone. From our data we concluded that subarcuate canals revealed not well defined trajectories and their spatial orientation varied across the studied temporal bones.

Single up-conversion nanocrystal as a local temperature probe of electrically heated silver nanowire.

Luminescence thermometry is a powerful technique for monitoring temperature in a sensitive, remote (through light), and minimally invasive manner. Up to now, many macroscopic and microscopic luminescence temperature probes exploiting different temperature sensing schemes have been investigated, with the majority of the studies using aggregates of nanothermometers. This work presents isolated single up-converting NaYF4:Er3+/Yb3+ nanocrystals as functional temperature indicators operating in a standard confocal microscopy configuration. More specifically, the nanocrystals were used to monitor the temperature of a single silver nanowire, whose temperature was controlled electrically via the Joule process. We demonstrate that individual nanocrystals placed near the nanowire can precisely determine the temperature distribution in its surroundings. These results, which combine nanoscopic heat generation with temperature readout using isolated nanocrystals, represent an essential step for the application of isolated single nanoprobes for luminescence thermometry at the nanoscale.

The involvement of NMDA and AMPA receptors in the mechanism of antidepressant-like action of zinc in the forced swim test.

Antidepressant-like activity of zinc in the forced swim test (FST) was demonstrated previously. Enhancement of such activity by joint administration of zinc and antidepressants was also shown. However, mechanisms involved in this activity have not yet been established. The present study examined the involvement of the NMDA and AMPA receptors in zinc activity in the FST in mice and rats. Additionally, the influence of zinc on both glutamate and aspartate release in the rat brain was also determined. Zinc-induced antidepressant-like activity in the FST in both mice and rats was antagonized by N-methyl-D-aspartic acid (NMDA, 75 mg/kg, i.p.) administration. Moreover, low and ineffective doses of NMDA antagonists (CGP 37849, L-701,324, D-cycloserine, and MK-801) administered together with ineffective doses of zinc exhibit a significant reduction of immobility time in the FST. Additionally, we have demonstrated the reduction of immobility time by AMPA receptor potentiator, CX 614. The antidepressant-like activity of both CX 614 and zinc in the FST was abolished by NBQX (an antagonist of AMPA receptor, 10 mg/kg, i.p.), while the combined treatment of sub-effective doses of zinc and CX 614 significantly reduces the immobility time in the FST. The present study also demonstrated that zinc administration potentiated a veratridine-evoked glutamate and aspartate release in the rat's prefrontal cortex and hippocampus. The present study further suggests the antidepressant properties of zinc and indicates the involvement of the NMDA and AMPA glutamatergic receptors in this activity.

Microwave-assisted synthesis and characterization of novel chitosan-based biomaterials for pelvic organ prolapse treatment.

Pelvic organ disorders affect up to one in four women in the United States. The prevalence of pelvic organ prolapse (POP) is increasing with each year, particularly in the setting of prolonged life expectancy and an aging population. Current treatment approaches, including polypropylene monofilaments are associated with numerous painful and worrisome side-effects. Therefore, scientists are looking for new solutions. A promising alternative to the current treatment is tissue engineering, which can be utilized to re-create support to the vagina and pelvic organs. Tissue engineering requires the use of three-dimensional scaffolds, derived from biocompatible materials. Chitosan is a natural polymer, obtained from shellfish exoskeletons. It is known for its biodegradability, lack of cytotoxicity and non-pyrogenicity. Due to the presence of free hydroxyl and amino groups, it may undergo various modifications. In this paper, we describe a new type of chitosan-based biomaterials, which can be used as a new alternative scaffold that may provide support to prolapse organs. The chitosan scaffold was obtained under microwave radiation using multifunctional amino and organic acids. We discuss the scaffold's characteristics, with an emphasis on its chemical structure and morphology. Fourier transform infrared spectroscopy (FT-IR) analysis confirmed cross-linking processes with preservation of free amino groups. Moreover, mechanical durability, the stability and swelling ability of the scaffolds in a simulated body fluid were investigated. All of the prepared scaffolds demonstrated very good antioxidant activity and biodegradability. Importantly, the biocompatibility of chitosan scaffolds was examined on human vaginal VK2/E6E7 cell line. No evidence of toxicity was documented, and the cells maintained their presence on the studied materials. These results allude to the lack of toxicity of the scaffolds, and indicate that chitosan-based scaffold should be further investigated in in vivo studies as they may be a promising alternative treatment to pelvic organ prolapse.

Towards electrophysiological correlates of auditory perception of temporal order.

Recent studies have postulated that the temporal order (TO) of two successive events can be correctly identified if they are separated by an inter-stimulus interval (ISI) of at least 30 ms duration. Using Auditory Evoked Potentials, we tested 21 students for the cortical activation associated with TO detection of two successively presented tones in either 'easy' (ISI=60 ms) or 'difficult' (ISI=10 ms) conditions. The amplitude of P2 component was related to difficulty of TO perception and was significantly higher in 'difficult' than 'easy' condition. Moreover, in 'difficult' condition the correlation analyses revealed a negative association at both Fz and Cz electrodes between P2 amplitudes and the correctness level. Correct responses in this condition were accompanied by lower P2 amplitudes than the incorrect ones.

Influence of band 3 protein absence and skeletal structures on amphiphile- and Ca(2+)-induced shape alterations in erythrocytes: a study with lamprey (Lampetra fluviatilis), trout (Onchorhynchus mykiss) and human erythrocytes.

Amphiphiles which induce either spiculated (echinocytic) or invaginated (stomatocytic) shapes in human erythrocytes, and ionophore A23187 plus Ca(2+), were studied for their capacity to induce shape alterations, vesiculation and hemolysis in the morphologically and structurally different lamprey and trout erythrocytes. Both qualitative and quantitative differences were found. Amphiphiles induced no gross morphological changes in the non-axisymmetric stomatocyte-like lamprey erythrocyte or in the flat ellipsoidal trout erythrocyte, besides a rounding up at higher amphiphile concentrations. No shapes with large broad spicula were seen. Nevertheless, some of the 'echinocytogenic' amphiphiles induced plasma membrane protrusions in lamprey and trout erythrocytes, from where exovesicles were shed. In trout erythrocytes, occurrence of corrugations at the cell rim preceded protrusion formation. Other 'echinocytogenic' amphiphiles induced invaginations in lamprey erythrocytes. The 'stomatocytogenic' amphiphiles induced invaginations in both lamprey and trout erythrocytes. Surprisingly, in trout erythrocytes, some protrusions also occurred. Some of the amphiphiles hemolyzed lamprey, trout and human erythrocytes at a significantly different concentration/membrane area. Ionophore A23187 plus Ca(2+) induced membrane protrusions and sphering in human and trout erythrocytes; however, the lamprey erythrocyte remained unperturbed. The shape alterations in lamprey erythrocytes, we suggest, are characterized by weak membrane skeleton-lipid bilayer interactions, due to band 3 protein and ankyrin deficiency. In trout erythrocyte, the marginal band of microtubules appears to strongly influence cell shape. Furthermore, the presence of intermediate filaments and nuclei, additionally affecting the cell membrane shear elasticity, apparently influences cell shape changes in lamprey and trout erythrocytes. The different types of shape alterations induced by certain amphiphiles in the cell types indicates that their plasma membrane phospholipid composition differs.

Cognitive impairment in HIV infection.

HIV-infected subjects at various stages of illness but without opportunistic cerebral disease were evaluated using a comprehensive, cognitively-based neuropsychological protocol and measures of levels of depression and anxiety. The data indicated a prominent attentional disorder among impaired subjects; however, language, visual-spatial and memory functioning were not deficient. There was also evidence suggesting executive function deficit. Depression contributed a small additional component in differentiating the groups. These findings help to specify the nature of the cognitive disturbance associated with HIV encephalopathy and are consistent with the pathological effects of primary infection of the brain by HIV. In addition, they provide a specific basis for ameliorative treatment with psychostimulant medication.

Coupling of beta and gamma activity in corticothalamic system of cats attending to visual stimuli.

We have previously shown that local field potentials (LFPs) recorded in the lateral geniculate nucleus (LGN) and primary visual cortex (VCx) of the cat contain significantly more beta frequency activity when the animal attends to visual than to auditory stimuli. In the present study we utilised data from the same experiments to calculate the cross-correlation between envelopes of filtered beta (16-24Hz) and gamma (30-45Hz) oscillatory signals recorded at the same sites. Correlation values obtained from visual trials were significantly higher than those calculated for the auditory task. This observation was typical in those LGN and VCx recording sites which corresponded to the central representation of the visual field. The cross-correlation function peaked within a 20 ms time window from the centre of the cross-correlogram. These findings support the hypothesis that beta activity provides an excitatory background for the appearance of oscillations in the gamma band.

Spontaneous variability reveals principal components in cortical evoked potentials.

Using principal component analysis, we studied trial to trial, spontaneous variability of evoked potentials (EPs) recorded from rat barrel cortex after whisker stimulation. This method allowed for extraction of two distinct components of EP which overlapped in the time domain. Our results are consonant with the previously described depth distribution of current sources and the extracted components can be therefore attributed to activities of two pyramidal cell classes: supra- and infragranular. Qualitatively similar results were found in both anaesthetized and alert animals.

Transient changes of electrical activity in the rat barrel cortex during conditioning.

To reveal the dynamics of neurophysiological changes in the rat barrel cortex induced by conditioned stimulation we recorded the local micro-electroencephalographic (EEG) activity and evoked potentials (EPs) in barrel cortex to stimulation of a single vibrissa before and after pairing it with a mild electric shock applied to the rat's tail. Following the introduction of the reinforcing stimulus, the amplitude of the first negative component of evoked potentials in the cortex on the conditioned side grew in relation to the same component of control potentials, evoked by stimulation of the opposite symmetrical vibrissa. This change was accompanied by a latent decrease in spectral power of the EEG within the alpha and beta frequency bands in both hemispheres. The observed changes in both of these electrical manifestations of enhanced neuronal activity reverted after two (EP) or three (EEG) days of conditioning. These results are discussed in relation to the putative activity of neuromodulatory systems.

Identification of principal components in cortical evoked potentials by brief surface cooling.

OBJECTIVES: The evoked potential recorded by a single electrode in rat's barrel cortex after whisker stimulation was shown to be composed of two main principal components shifted in time by about 3 ms. The purpose of this study was to verify the hypothesis that these components represent activity of supra- and infragranular pyramidal cell classes. RESULTS: Our results show that a brief cooling pulse applied to the cortical surface abolishes the shorter latency component, which may therefore be attributed to the response of supragranular pyramidal cells. CONCLUSIONS: The longer latency principal component, which disappears only with strong cooling pulses, is proposed to represent postsynaptic activity of infragranular pyramidal neurons.

Effect of repeated treatment with mirtazapine on the central alpha1-adrenergic receptors.

Mirtazapine (MIR) is an antidepressant which enhances noradrenergic and serotonergic 5-HT1A neurotransmission via antagomism of central alpha2-adrenergic autoreceptors and heteroreceptors. The drugs does not inhibit noradrenaline and serotonin reuptake but blocks the 5-HT, and 5-HT3 receptors and has high affinity only for central and peripheral histamine H1 receptors. The present study was aimed at determining whether repeated MIR treatment induced adaptive changes in the alpha1-adrenergic receptors, similar to those reported by us early for tricyclic antidepressants, The experiments were carried out on male mice and rats. MIR was administered at a dose of 10 mg/kg once or repeatedly (twice daily for 14 days). The obtained results showed that MIR administrated repeatedly potentiated the methoxamine- induced exploratory hyperactivity in rats and clonidine-induced aggressiveness in mice, those effects being mediated by alpha1-adrenergic receptors. MIR given repeatedly (but not acutely) increased the binding (Bmax ) of [3H]prazosin to alpha1-adrenergic receptors in cerebral cortex, however, the ability of the alpha1-adrenoceptor agonist phenylephrine to compete for the these sites was not significantly changed. The above results indicate that repeated MIR administration increases the responsiveness of alpha1-adrenergic system (behavioural and biochemical changes), as tricyclics do. However, the question whether the increased functional responsiveness found in the present study is important for the clinical antidepressant efficacy, remains open.

Electron paramagnetic resonance study of erythrocyte membrane fluidity in renal transplant recipients.

Renal transplantation (KTx) patients receiving calcineurin inhibitors-cyclosporine (CsA) or tacrolimus (TAC)-are known to be at risk for the development of posttransplant hemolytic uremic syndrome. The syndrome has been reported to occur more often with CsA than TAC treatment. It has also been noted that CsA affects erythrocyte membrane fluidity. The aim of this study was to investigate whether the impairment of erythrocyte membrane fluidity is similar among patients under treatment with different calcineurin inhibitors (CsA or TAC). Venous blood was collected from 29 KTx patients and from nine healthy volunteers. To investigate the fluidity of intact erythrocyte membranes spin label electron paramagnetic resonance spectroscopy was applied. Comparison of values for controls versus CsA-treated patients demonstrated a significant decrease in membrane viscosity in the hydrophobic region of the lipid bilayer among CsA-treated patients, whereas there was no significant difference between control and patients treated with TAC. There was no significant difference in the molecular order close to the polar heads of lipid molecules among all groups. The observed changes in erythrocyte membrane fluidity among CsA-treated patients and the lack of this phenomenon in the TAC group may correlate with more frequent prevalence of hemolytic anemia among CsA than TAC-treated patients.

Transport of traffic-related aerosols in urban areas.

This study was undertaken to assess the influence of traffic on particulate air pollution in an urban area, and to characterise the short-range transport of the aerosols generated by traffic. The study was conducted in Kraków, a city located in southern Poland with a population of approximately 800,000. Aerosol samples were collected using automatic sampling equipment at five sites located at different distances from the main road in Kraków, ranging from 5 to 1500 m. The sampling set-up allowed standardisation of the results due to continuous determination of the meteorological parameters (temperature, atmospheric pressure, wind speed and direction, rainfall and humidity). Aerosol particles were separated according to aerodynamic diameter into two size fractions: > 1.9 microm (coarse fraction); and 1.9-72 microm (fine fraction). The concentrations of 27 elements were measured in both size fractions (Na, Mg, Al, Si, P, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Ga, Ge, As, Se, Br, Rb, Sr, Zr, Ba, Pb). The multielement analyses were performed by Particle-Induced X-ray Emission (PIXE) spectrometry. Traffic contribution to particulate air pollution was determined on the basis of 13 elements which were present above the detection limit in all samples (Mg, Al, Si, P, S, K, Ca, Ti, Mn, Fe, Cu, Zn, Pb). It was found that the traffic contribution in the coarse size fraction was approximately 80% up to 150 m from the road; it dropped abruptly by a factor of 2 over a distance of 150-200 m and declined further to 20% at 1500 m from the road. Traffic contribution for the fine particle concentrations of individual elements was 50-70% in the close vicinity of the road (5 m); then there was a decrease, followed by an increase at a greater distance from the road. Possible explanations for this behaviour of the fine particles are given.

Light level induced reorganization of cat's lateral geniculate nucleus receptive fields: a spatiotemporal study.

The spatiotemporal organization of receptive fields (RF) of the cat’s lateral geniculate cells was investigated under several scotopic and mesopic light adaptation levels, using stationary and moving light stimuli. A given adaptation level was reached by changing of either the diffuse background or the adapting central spot intensity. Both methods gave the same qualitative results. The RFs were prominently reorganized after dark adaptation. Under such conditions, the central response area was enlarged and the classic excitatory and inhibitory surrounds were hardly visible in most cases. A minimum light level was needed for developing classic surrounds. Both inhibitory and excitatory surrounds changed their spatial positions simultaneously at higher light adaptation levels. ON- and OFF-center cells changed their spatiotemporal organization similarly, thus indicating similar underlying mechanisms.

Two-unit recordings from the lateral geniculate nucleus of the cat. Some inhibitory interactions.

Pairs of units were recorded by single electrodes from cat’s LGN. Three types of pairs were distinguished. Two of them were classified as retinogeniculate pairs with positive cross-correlograms (ON center – ON center, and OFF center – OFF center); the third type consisted of negatively correlated pairs of LGN neurones. In each case the spatiotemporal organizations of both receptive fields (RFs) were analyzed at several levels of background luminance. An increase of background illumination induced more pronounced compression of the retinal than of the related geniculate ON-center RFs; a strong, central stimulation evoked oscillatory responses only in the LGN cells. The LGN neuron pairs showed a significant overlap of their RFs antagonistic areas. Potentiation of inhibition at the LGN level is discussed in relation to a proposed functional model.

Inhibitory mechanisms within the receptive fields of the lateral geniculate body of the cat.

The spatiotemporal organization of receptive fields (RF) of neurons in lateral geniculate body (LGB) was analyzed using several experimental methods. The stimulation of the RF by two separate spots showed largely extended inhibitory field overlapping also the central region. The increase of light-adaptation or barbiturate anesthesia levels enhanced the effectiveness of RF inhibitory surround in qualitatively different manner. The correlated neurons pairs were found, showing the reciprocal arrangement of inhibitory and excitatory areas of their RFs. It was possible to identify some of these pairs as consisting of relay cell and intergeniculate interneuron. The neurons from perigeniculate nucleus were also investigated. They had a synaptic input from the retina and were not activated antidromically from the visual cortex. Their RFs were large with non-concentric ON-OFF type of organization. Both types of geniculate interneurons were used to propose a functional model of LGB circuitry reviewing the data of all presented experiments.