Opium may affect coronary artery disease by inducing inflammation but not through the expression of CD9, CD36, and CD68.

The molecular mechanisms of opium action with regard to coronary artery disease (CAD) have not yet been determined. The aim of this study was to evaluate the effect of opium on the expression of scavenger receptors including CD36, CD68, and CD9 tetraspanin in monocytes and the plasma levels of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), malondialdehyde (MDA), and nitric oxide metabolites (NOx) in CAD patients with and without opium addiction. This case-control study was conducted on three groups: (1) opium-addicted CAD patients (CAD + OA, n = 30); (2) CAD patients with no opium addiction (CAD, n = 30); and (3) individuals without CAD and opium addiction as the control group (Ctrl, n = 17). The protein and mRNA levels of CD9, CD36, and CD68 were evaluated by the flow cytometry and quantitative polymerase chain reaction (RT-qPCR) methods, respectively. The consumption of atorvastatin, aspirin, and glyceryl trinitrate was found be higher in the CAD groups compared with the control group. The plasma level of TNF-α was significantly higher in the CAD + OA group than in the CAD and Ctrl groups (p = 0.001 and p = 0.005, respectively). MDA levels significantly increased in CAD and CAD + OA patients in comparison with the Ctrl group (p = 0.010 and p = 0.002, respectively). No significant differences were found in CD9, CD36, CD68, IFN-γ, and NOx between the three groups. The findings demonstrated that opium did not have a significant effect on the expression of CD36, CD68, and CD9 at gene and protein levels, but it might be involved in the development of CAD by inducing inflammation through other mechanisms.

Band 3 Protein: An Effective Interrogation Tool of Storage Lesions in RBC Units.

The present study aims to investigate the changes in different parameters related to the storage time of red blood cell (RBC) units. Microscopic, flow cytometric, and electrophoretic assessments were employed every few days for 60 days to investigate the alterations in morphology, size, phosphatidylserine (PS) externalization, and membrane proteins over time. Morphological transformation from discocytes to spherocytes progressed as the storage time increased, which was accompanied by an increment of cellular size. However, this storage period did not result in the externalization of significant amounts of PS (p > 0.05). Mean Fluorescence Intensity (MFI) values increased by 11% to 23% between days 21 and 35 compared to the day 1 sample (p < 0.001). By day 60, the MFI decreased to about 70% of the day 1 sample. The analysis of membrane proteins' distribution showed a significant drop in band 3 expression after 35 days (p < 0.05 and 0.001 on days 42 and 60, respectively); however, no significant change was observed up to five weeks (p > 0.05). The inconsistency observed between Eosin-5-Maleimide (5-EMA) binding and the relative band 3 content could be due to additional accessibility of 5-EMA to hidden domains of other membrane proteins on RBCs as a result of increased mean corpuscular volume (MCV) and changes in morphology. Overall, our present study represents a step-wise and time-dependent series of events that progressively affects stored RBCs.

Opium may affect coronary artery disease by inducing inflammation but not through the expression of CD9, CD36, and CD68.

The molecular mechanisms of opium with regard to coronary artery disease (CAD) have not yet been determined. The aim of the present study was to evaluate the effect of opium on the expression of scavenger receptors including CD36, CD68, and CD9 tetraspanin in monocytes and the plasma levels of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), malondialdehyde (MDA), and nitric oxide metabolites (NOx) in patients with CAD with and without opium addiction. This case-control study was conducted in three groups: (1) opium-addicted patients with CAD (CAD+OA, n=30); (2) patients with CAD with no opium addiction (CAD, n=30); and (3) individuals without CAD and opium addiction as the control group (Ctrl, n=17). Protein and messenger RNA (mRNA) levels of CD9, CD36, and CD68 were evaluated by flow cytometry and reverse transcription-quantitative PCR methods, respectively. Consumption of atorvastatin, aspirin, and glyceryl trinitrate was found to be higher in the CAD groups compared with the control group. The plasma level of TNF-α was significantly higher in the CAD+OA group than in the CAD and Ctrl groups (p=0.001 and p=0.005, respectively). MDA levels significantly increased in the CAD and CAD+OA groups in comparison with the Ctrl group (p=0.010 and p=0.002, respectively). No significant differences were found in CD9, CD36, CD68, IFN-γ, and NOx between the three groups. The findings demonstrated that opium did not have a significant effect on the expression of CD36, CD68, and CD9 at the gene and protein levels, but it might be involved in the development of CAD by inducing inflammation through other mechanisms.

Slot blotting and flow cytometry: two efficient assays for platelet antibody screening among patients with platelet refractoriness.

BACKGROUND AND OBJECTIVES: Frequent platelet transfusion may lead to the formation of alloantibodies and immune-mediated platelet destruction. Currently, identifying economic and effective screening methods is necessary for the management of platelet transfusion while different tests were recommended. The present study aims to challenge the performance of slot blotting (SB) and flow cytometry (FC) assays in detecting immune platelet refractoriness. MATERIALS AND METHODS: Sera from 118 patients who received blood components and were clinically suspected of platelet refractoriness were enrolled. Platelet-reactive antibodies were explored in parallel by SB, FC and monoclonal antibody-specific immobilization of platelet antigens (MAIPA) techniques. In a further study, chloroquine-treated platelets were incubated with MAIPA-positive serum, and then, the results of the SB and FC techniques were compared. RESULTS: Using MAIPA as a reference, antibodies were detected in 51 sera, with specificity for human leucocyte antigens (HLA), human platelet antigens (HPA) or both HLA/HPA, in 27, 18 and 6 patients, respectively. The sensitivity and specificity of SB and FC were 86·3%, 88·1%, 82·4% and 95·5%, respectively. The Spearman correlation revealed significant (P < 0·001) correlations between FC (r = 0·763) and SB (r = 0·738) with MAIPA. In respect to HPA antibody detection, SB had 83·3% sensitivity and 92·6% specificity compared to 91·7% and 96·3% for FC while both approaches are acceptable (P < 0·001, r = 0·69; P < 0·001, r = 0·773) and can be recommended. CONCLUSIONS: The present study acknowledges that among the used methods, the flow cytometry's performance is the most appropriate, but slot blotting, with acceptable sensitivity, can be used as an acceptable and convenient procedure for platelet antibody screening.

Variable Presentation of Hereditary Spherocytosis in an Iranian Family.

Hereditary spherocytosis (HS), a familial defect involving red blood cell (RBC) membrane proteins, is associated with reduced deformability, increased fragility, and progressive destruction of spherical cells. The present study focuses on three subjects of a family showing a history of repeated episodes of lethargy and pallor of unknown etiology. All patients displayed reticulocytosis and spherocytosis and one of them had anemia and splenomegaly. The patients underwent screening tests to rule in/out possible underlying disorders, and deficiency/dysfunction of RBC membrane proteins was suspected. Definitive diagnosis can be made on the basis of membrane protein analysis by quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Interestingly, all patients showed marked decrease in the protein 4.2 expression and therefore, HS was confirmed. This case report highlights the simultaneous occurrence of protein 4.2-dependent "typical" and "atypical" HS in a family and serves as a reminder to clinicians to consider RBC membrane disorders in patients presenting with suspicious and unexplained clinical signs.

Evaluation of MDR1 and MRPA genes expression in different types of dry cutaneous leishmaniasis.

OBJECTIVE: The resistance to antimony-containing glucantime is a major obstacle to successful treatment, especially in endemic areas. Looking the molecular mechanisms involved in this drug resistance will help in choosing the best treatment. The aim of this study was to evaluate the expression of multidrug-resistance 1 (MDR1) and multidrug-resistance protein A (MRPA) genes in acute, chronic non-lupoid, and chronic lupoid forms of dry type cutaneous leishmaniasis (DTCL). RESULTS: MDR1 gene was over-expressed as 14.4- and 1.56-folds in the chronic lupoid and acute forms compared with the chronic non-lupoid form, respectively. Results comparison showed P < 0.05 between the chronic non-lupoid and acute groups, P < 0.01 between acute and chronic lupoid groups, and P < 0.001 between the chronic non-lupoid and chronic lupoid groups. MRPA gene was over-expressed as 266 and 17.7-fold in the chronic lupoid and chronic non-lupoid forms compared with the acute form, respectively. Statistical analysis showed P < 0.01 between the chronic non-lupoid and chronic lupoid groups, P < 0.05 between acute and chronic non-lupoid groups, and P < 0.001 between the acute and chronic lupoid groups.

Attenuating the ear canal feedback pressure of a laser-driven hearing aid.

Microphone placement behind the pinna, which minimizes feedback but also reduces perception of the high-frequency pinna cues needed for sound localization, is one reason why hearing-aid users often complain of poor sound quality and difficulty understanding speech in noisy situations. In this paper, two strategies are investigated for minimizing the feedback pressure (thereby increasing the maximum stable gain, MSG) of a wide-bandwidth light-activated contact hearing aid (CHA) to facilitate microphone placement in the ear canal (EC): (1) changing the location of the drive force and its direction at the umbo, and (2) placing an acoustic damper within the EC to reduce the feedback pressure at the microphone location. The MSG and equivalent pressure output (EPO) are calculated in a 3D finite element model of a human middle ear based on micro computed tomography (micro-CT) images. The model calculations indicate that changing the umbo-force direction can decrease feedback pressure, but at the expense of decreased EPO. However the model shows improvements in MSG without sacrificing EPO when an acoustic damper is placed in the EC. This was verified through benchtop experimentation and in human cadaver temporal bones. The results pave the path towards a wide-bandwidth hearing aid that incorporates an EC-microphone design.

In-plane and out-of-plane motions of the human tympanic membrane.

Computer-controlled digital holographic techniques are developed and used to measure shape and four-dimensional nano-scale displacements of the surface of the tympanic membrane (TM) in cadaveric human ears in response to tonal sounds. The combination of these measurements (shape and sound-induced motions) allows the calculation of the out-of-plane (perpendicular to the surface) and in-plane (tangential) motion components at over 1,000,000 points on the TM surface with a high-degree of accuracy and sensitivity. A general conclusion is that the in-plane motion components are 10-20 dB smaller than the out-of-plane motions. These conditions are most often compromised with higher-frequency sound stimuli where the overall displacements are smaller, or the spatial density of holographic fringes is higher, both of which increase the uncertainty of the measurements. The results are consistent with the TM acting as a Kirchhoff-Love's thin shell dominated by out-of-plane motion with little in-plane motion, at least with stimulus frequencies up to 8 kHz.

Simultaneous full-field 3-D vibrometry of the human eardrum using spatial-bandwidth multiplexed holography.

Holographic interferometric methods typically require the use of three sensitivity vectors in order to obtain three-dimensional (3-D) information. Methods based on multiple directions of illumination have limited applications when studying biological tissues that have temporally varying responses such as the tympanic membrane (TM). Therefore, to measure 3-D displacements in such applications, the measurements along all the sensitivity vectors have to be done simultaneously. We propose a multiple-illumination directions approach to measure 3-D displacements from a single-shot hologram that contains displacement information from three sensitivity vectors. The hologram of an object of interest is simultaneously recorded with three incoherently superimposed pairs of reference and object beams. The incident off-axis angles of the reference beams are adjusted such that the frequency components of the multiplexed hologram are completely separate. Because of the differences in the directions and wavelengths of the reference beams, the positions of each reconstructed image corresponding to each sensitivity vector are different. We implemented a registration algorithm to accurately translate individual components of the hologram into a single global coordinate system to calculate 3-D displacements. The results include magnitudes and phases of 3-D sound-induced motions of a human cadaveric TM at several excitation frequencies showing modal and traveling wave motions on its surface.

Three-dimensional vibrometry of the human eardrum with stroboscopic lensless digital holography.

The eardrum or tympanic membrane (TM) transforms acoustic energy at the ear canal into mechanical motions of the ossicles. The acousto-mechanical transformer behavior of the TM is determined by its shape, three-dimensional (3-D) motion, and mechanical properties. We have developed an optoelectronic holographic system to measure the shape and 3-D sound-induced displacements of the TM. The shape of the TM is measured with dual-wavelength holographic contouring using a tunable near IR laser source with a central wavelength of 780 nm. 3-D components of sound-induced displacements of the TM are measured with the method of multiple sensitivity vectors using stroboscopic holographic interferometry. To accurately obtain sensitivity vectors, a new technique is developed and used in which the sensitivity vectors are obtained from the images of a specular sphere that is being illuminated from different directions. Shape and 3-D acoustically induced displacement components of cadaveric human TMs at several excitation frequencies are measured at more than one million points on its surface. A numerical rotation matrix is used to rotate the original Euclidean coordinate of the measuring system in order to obtain in-plane and out-of-plane motion components. Results show that in-plane components of motion are much smaller (<20%) than the out-of-plane motions' components.

Measurements of three-dimensional shape and sound-induced motion of the chinchilla tympanic membrane.

UNLABELLED: Opto-electronic computer holographic measurements were made of the tympanic membrane (TM) in cadaveric chinchillas. Measurements with two laser wavelengths were used to compute the 3D-shape of the TM. Single laser wavelength measurements locked to eight distinct phases of a tonal stimulus were used to determine the magnitude and the relative phase of the surface displacements. These measurements were made at over 250,000 points on the TM surface. The measured motions contained spatial phase variations consistent with relatively low-order (large spatial frequency) modal motions and smaller magnitude higher-order (smaller spatial frequency) motions that appear to travel, but may also be explained by losses within the membrane. The measurement of shape and thin shell theory allowed us to separate the measured motions into those components orthogonal to the plane of the tympanic ring, and those components within the plane of the tympanic ring based on the 3D-shape. The predicted in-plane motion components are generally smaller than the out-of-plane perpendicular component of motion. Since the derivation of in-plane and out-of plane depended primarily on the membrane shape, the relative sizes of the predicted motion components did not vary with frequency. SUMMARY: A new method for simultaneously measuring the shape and sound-induced motion of the tympanic membrane is utilized to estimate the 3D motion on the membrane surface. This article is part of a special issue entitled "MEMRO 2012".

Digital holographic measurements of shape and 3D sound-induced displacements of Tympanic Membrane.

Acoustically-induced vibrations of the Tympanic Membrane (TM) play a primary role in the hearing process, in that these motions are the initial mechanical response of the ear to airborne sound. Characterization of the shape and 3D displacement patterns of the TM is a crucial step to a better understanding of the complicated mechanics of sound reception by the ear. In this paper, shape and sound-induced 3D displacements of the TM in cadaveric chinchillas are measured by a lensless Dual-Wavelength Digital Holography system (DWDHS). The DWDHS consists of Laser Delivery (LD), Optical Head (OH), and Computing Platform (CP) subsystems. Shape measurements are performed in double-exposure mode and with the use of two wavelengths of a tunable laser while nanometer-scale displacements are measured along a single sensitivity direction and with a constant wavelength. In order to extract the three principal components of displacement in full-field-of-view, and taking into consideration the anatomical dimensions of the TM, we combine principles of thin-shell theory together with both, displacement measurements along the single sensitivity vector and TM surface shape. To computationally test this approach, Finite Element Methods (FEM) are applied to the study of artificial geometries.