Regulation of monocyte apoptosis and DNA extrusion in monocyte extracellular traps by PSGL-1: Relevance in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease characterized by severe organ damage and lacking curative treatment. While various immune cell types, especially dysfunctional B and T cells and neutrophils, have been related with disease pathogenesis, limited research has focused on the role of monocytes in SLE. Increased DNA extracellular traps, apoptosis and necrosis have been related to lupus pathogenesis. Our goal is to analyze the contribution of P-selectin glycoprotein ligand 1 (PSGL-1) in SLE monocytes to disease pathogenesis by investigating the control exerted by PSGL-1 on monocyte apoptosis and DNA extrusion in extracellular traps (METs). Monocytes from active disease patients (aSLE) exhibited reduced levels of PSGL-1. Importantly, lower PSGL-1 levels in SLE monocytes associated with several clinical characteristics, including anti-dsDNA autoantibodies, lupus anticoagulant, clinical lung involvement, and anemia. Monocytes from SLE patients showed higher susceptibility to apoptosis than healthy donors (HD) monocytes and PSGL-1/P-selectin interaction decreased secondary necrosis in HD but not in aSLE monocytes. Regarding METs, aSLE monocytes exhibited higher susceptibility to generate METs than HD monocytes. The interaction of HD monocytes with P-selectin induced Syk activation and reduced the levels of DNA extruded in METs. However, in aSLE monocytes, PSGL-1/P-selectin interaction did not activate Syk or reduce the amount of extruded DNA. Our data suggest a dysfunctional PSGL-1/P-selectin axis in aSLE monocytes, unable to reduce secondary necrosis or the amount of DNA released into the extracellular medium in METs, potentially contributing to lupus pathogenesis.

PSGL-1, ADAM8, and selectins as potential biomarkers in the diagnostic process of systemic lupus erythematosus and systemic sclerosis: an observational study.

Background: Early diagnosis and treatment of Systemic lupus erythematosus (SLE) and Systemic sclerosis (SSc) present significant challenges for clinicians. Although various studies have observed changes in serum levels of selectins between healthy donors and patients with autoimmune diseases, including SLE and SSc, their potential as biomarkers has not been thoroughly explored. We aimed to investigate serum profiles of PSGL-1 (sPSGL-1), ADAM8 (sADAM8) and P-, E- and L-selectins (sP-, sE- and sL-selectins) in defined SLE and SSc patient cohorts to identify disease-associated molecular patterns. Methods: We collected blood samples from 64 SLE patients, 58 SSc patients, and 81 healthy donors (HD). Levels of sPSGL-1, sADAM8 and selectins were analyzed by ELISA and leukocyte membrane expression of L-selectin and ADAM8 by flow cytometry. Results: Compared to HD, SLE and SSc patients exhibited elevated sE-selectin and reduced sL-selectin levels. Additionally, SLE patients exhibited elevated sPSGL-1 and sADAM8 levels. Compared to SSc, SLE patients had decreased sL-selectin and increased sADAM8 levels. Furthermore, L-selectin membrane expression was lower in SLE and SSc leukocytes than in HD leukocytes, and ADAM8 membrane expression was lower in SLE neutrophils compared to SSc neutrophils. These alterations associated with some clinical characteristics of each disease. Using logistic regression analysis, the sL-selectin/sADAM8 ratio in SLE, and a combination of sL-selectin/sE-selectin and sE-selectin/sPSGL-1 ratios in SSc were identified and cross-validated as potential serum markers to discriminate these patients from HD. Compared to available diagnostic biomarkers for each disease, both sL-selectin/sADAM8 ratio for SLE and combined ratios for SSc provided higher sensitivity (98% SLE and and 67% SSc correctly classified patients). Importantly, the sADAM8/% ADAM8(+) neutrophils ratio discriminated between SSc and SLE patients with the same sensitivity and specificity than current disease-specific biomarkers. Conclusion: SLE and SSc present specific profiles of sPSGL-1, sE-, sL-selectins, sADAM8 and neutrophil membrane expression which are potentially relevant to their pathogenesis and might aid in their early diagnosis.

Using torsional wave elastography to evaluate spring pot parameters in skin tumor mimicking phantoms.

Estimating the tissue parameters of skin tumors is crucial for diagnosis and effective therapy in dermatology and related fields. However, identifying the most sensitive biomarkers require an optimal rheological model for simulating skin behavior this remains an ongoing research endeavor. Additionally, the multi-layered structure of the skin introduces further complexity to this task. In order to surmount these challenges, an inverse problem methodology, in conjunction with signal analysis techniques, is being employed. In this study, a fractional rheological model is presented to enhance the precision of skin tissue parameter estimation from the acquired signal from torsional wave elastography technique (TWE) on skin tumor-mimicking phantoms for lab validation and the estimation of the thickness of the cancerous layer. An exhaustive analysis of the spring-pot model (SP) solved by the finite difference time domain (FDTD) is conducted. The results of experiments performed using a TWE probe designed and prototyped in the laboratory were validated against ultrafast imaging carried out by the Verasonics Research System. Twelve tissue-mimicking phantoms, which precisely simulated the characteristics of skin tissue, were prepared for our experimental setting. The experimental data from these bi-layer phantoms were measured using a TWE probe, and the parameters of the skin tissue were estimated using inverse problem-solving. The agreement between the two datasets was evaluated by comparing the experimental data obtained from the TWE technique with simulated data from the SP- FDTD model using Pearson correlation, dynamic time warping (DTW), and time-frequency representation. Our findings show that the SP-FDTD model and TWE are capable of determining the mechanical properties of both layers in a bilayer phantom, using a single signal and an inverse problem approach. The ultrafast imaging and the validation of TWE results further demonstrate the robustness and reliability of our technology for a realistic range of phantoms. This fusion of the SP-FDTD model and TWE, as well as inverse problem-solving methods has the potential to have a considerable impact on diagnoses and treatments in dermatology and related fields.

Reliability and robustness of a novel preclinical torsional wave-based device for stiffness evaluation.

In this work, we present a novel preclinical device utilizing Torsional Wave Elastography (TWE). It comprises a rotational actuator element and a piezoceramic receiver ring circumferentially aligned. Both allow the transmission of shear waves that interact with the tissue before being received. Our main objective is to demonstrate and characterize the reliability, robustness, and accuracy of the device for characterizing the stiffness of elastic materials and soft tissues. Experimental tests are performed using two sets of tissue mimicking phantoms. The first set consists of calibrated CIRS gels with known stiffness value, while the second test uses non-calibrated manufactured phantoms. Our experimental observations show that the proposed device consistently and repeatably quantifies the stiffness of elastic materials with high accuracy. Furthermore, comparison with established techniques demonstrates a very high correlation (> 95%), supporting the potential medical application of this technology. The results obtained pave the way for a cross-sectional study aiming to investigate the correlation between gestational age and cervical elastic properties during pregnancy.

Healthy human skin Kelvin-Voigt fractional and spring-pot biomarkers reconstruction using torsional wave elastography.

This paper presents a novel method for reconstructing skin parameters using Probabilistic Inverse Problem (PIP) techniques and Torsional Wave Elastography (TWE) rheological modeling. A comprehensive examination was conducted to compare and analyze the theoretical, time-of-flight (TOF), and full-signal waveform (FSW) approaches. The objective was the identification of the most effective method for the estimation of mechanical parameters. Initially, the most appropriate rheological model for the simulation of skin tissue behavior was determined through the application and comparison of two models, spring pot (SP) and Kevin Voigt fractional derivative (KVFD). A numerical model was developed using the chosen rheological models. The collection of experimental data from 15 volunteers utilizing a TWE sensor was crucial for obtaining significant information for the reconstruction process. The study sample consisted of five male and ten female subjects ranging in age from 25 to 60 years. The procedure was performed on the ventral forearm region of the participants. The process of reconstructing skin tissue parameters was carried out using PIP techniques. The experimental findings were compared with the numerical results. The three methods considered (theoretical, TOF, FSW) have been used. The efficacy of TOF and FSW was then compared with theoretical method. The findings of the study demonstrate that the FSW and TOF techniques successfully reconstructed the parameters of the skin tissue in all of the models. The SP model's the skin tissue η values ranged from 8 to 12 P a · s , as indicated by the TOF reconstruction parameters. η values found by the KVFD model ranged from 4.1 to 9.3 P a · s . The µ values generated by the KVFD model range between 0.61 and 96.86 kPa. However, FSW parameters reveal that skin tissue η values for the SP model ranged from 7.8 to 12 P a · s . The KVFD model determined η values between 6.3 and 9.5 P a · s . The KVFD model presents µ values ranging between 26.02 and 122.19 kPa. It is shown that the rheological model that best describes the nature of the skin is the SP model and its simplicity as it requires only two parameters, in contrast to the three parameters required by the KVFD model. Therefore, this work provides a valuable addition to the area of dermatology, with possible implications for clinical practice.

Nonlinear fourth-order elastic characterization of the cornea using torsional wave elastography.

Measuring the mechanical nonlinear properties of the cornea remains challenging due to the lack of consensus in the methodology and in the models that effectively predict its behaviour. This study proposed developing a procedure to reconstruct nonlinear fourth-order elastic properties of the cornea based on a mathematical model derived from the theory of Hamilton et al. and using the torsional wave elastography (TWE) technique. In order to validate its diagnostic capability of simulated pathological conditions, two different groups were studied, non-treated cornea samples (n=7), and ammonium hydroxide ([Formula: see text]) treated samples (n=7). All the samples were measured in-plane by a torsional wave device by increasing IOP from 5 to 25 mmHg with 5 mmHg steps. The results show a nonlinear variation of the shear wave speed with the IOP, with higher values for higher IOPs. Moreover, the shear wave speed values of the control group were higher than those of the treated group. The study also revealed significant differences between the control and treated groups for the Lamé parameter [Formula: see text] (25.9-6.52 kPa), third-order elastic constant A (215.09-44.85 kPa), and fourth-order elastic constant D (523.5-129.63 kPa), with p-values of 0.010, 0.024, and 0.032, respectively. These findings demonstrate that the proposed procedure can distinguish between healthy and damaged corneas, making it a promising technique for detecting diseases associated with IOP alteration, such as corneal burns, glaucoma, or ocular hypertension.

Optical micro-elastography with magnetic excitation for high frequency rheological characterization of soft media.

The propagation of shear waves in elastography at high frequency (>3 kHz) in viscoelastic media has not been extensively studied due to the high attenuation and technical limitations of current techniques. An optical micro-elastography (OME) technique using magnetic excitation for generating and tracking high frequency shear waves with enough spatial and temporal resolution was proposed. Ultrasonics shear waves (above 20 kHz) were generated and observed in polyacrylamide samples. A cutoff frequency, from where the waves no longer propagate, was observed to vary depending on the mechanical properties of the samples. The ability of the Kelvin-Voigt (KV) model to explain the high cutoff frequency was investigated. Two alternative measurement techniques, Dynamic Mechanical Analysis (DMA) and Shear Wave Elastography (SWE), were used to complete the whole frequency range of the velocity dispersion curve while avoid capturing guided waves in the low frequency range (<3 kHz). The combination of the three measurement techniques provided rheology information from quasi-static to ultrasonic frequency range. A key observation was that the full frequency range of the dispersion curve was necessary if one wanted to infer accurate physical parameters from the rheological model. By comparing the low frequency range with the high frequency range, the relative errors for the viscosity parameter could reach 60 % and they could be higher with higher dispersive behavior. The high cutoff frequency may be predicted in materials that follow a KV model over their entire measurable frequency range. The mechanical characterization of cell culture media could benefit from the proposed OME technique.

Low P-Selectin Glycoprotein Ligand-1 Expression in Neutrophils Associates with Disease Activity and Deregulated NET Formation in Systemic Lupus Erythematosus.

Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by the generation of anti-DNA autoantibodies due to exposure of immune cells to excessive amounts of extracellular DNA. Lack of P-selectin in mice induces the development of a lupus-like syndrome and patients with cutaneous lupus have reduced P-selectin expression in skin vessels. Using flow cytometry we analyzed in healthy donors and patients the expression of P-selectin Glycoprotein Ligand-1 (PSGL-1) in circulating neutrophils and the implication of PSGL-1/P-selectin interaction in neutrophil extracellular traps (NETs) generation. We found a statistical significance that neutrophils from active SLE patients have a reduced expression of PSGL-1 and low levels of PSGL-1 in neutrophils from SLE patients associated with the presence of anti-dsDNA antibodies, clinical lung involvement, Raynaud's phenomenon, and positive lupus anticoagulant. PSGL-1 is present along the DNA in the NET. In healthy donors, neutrophil interaction with immobilized P-selectin triggers Syk activation, increases the NETs percentage and reduces the amount of DNA extruded in the NETs. In active SLE patients, neutrophil interaction with P-selectin does not activate Syk or reduce the amount of DNA extruded in the NETs, that might contribute to increase the extracellular level of DNA and hence, to disease pathogenesis.

Damage Detection Using Ultrasonic Techniques in Concrete-Filled Steel Tubes (CFSTs) Columns.

Concrete-filled steel tubes (CFSTs) are structural elements that, as a consequence of an incorrect elaboration, can exhibit internal defects that cannot be visualized, being usually air voids. In this work, the detection of internal damage in CFST samples elaborated with a percentage of contained air voids in concrete, was carried out by performing a complete ultrasound scan using an immersion tank. The analysis of the ultrasound signals shows the differences presented in the amplitude of the fundamental frequency of the signal, and in the Broadband Ultrasound Attenuation (BUA), in comparison with a sample without defects. The main contribution of this study is the application of the BUA technique in CFST samples for the location of air voids. The results present a linear relationship between BUA averages over the window of the CFSTs and the percentage of air voids contained (Pearson's correlation coefficient r = 0.9873), the higher percentage of air voids, the higher values of BUA. The BUA algorithm could be applied effectively to distinguish areas with defects inside the CFSTs. Similar to the BUA results, the analysis in the frequency domain using the FFT and the STFT was sensitive in the detection of internal damage (Pearson's correlation coefficient r = -0.9799, and r = -0.9672, respectively). The results establish an improvement in the evaluation of CFST elements for the detection of internal defects.

Targeted nanotherapy with everolimus reduces inflammation and fibrosis in scleroderma-related interstitial lung disease developed by PSGL-1 deficient mice.

BACKGROUND AND PURPOSE: Interstitial lung disease (ILD) is the main cause of mortality in systemic sclerosis (SSc), and current therapies available are of low efficacy or high toxicity. Thus, the identification of innovative less toxic and high efficacy therapeutic approaches to ILD treatment is an urgent need. The interaction of P-selectin glycoprotein ligand-1 (PSGL-1) with P-selectin initiates leukocyte extravasation and deletion of the corresponding gene (Selplg) induces a SSc-like syndrome with high incidence of ILD in aged mice. EXPERIMENTAL APPROACH: Aged PSGL-1 KO (Selplg-/- ) mice were used to assess the therapeutic effects of nanotherapy with everolimus, included in liposomes decorated with high MW hyaluronic acid (LipHA+Ev) and administered intratracheally to specifically target CD44-expressing lung cells. KEY RESULTS: PSGL-1 KO mice had increased numbers of CD45+ and CD45- cells, including alveolar and interstitial macrophages, eosinophils, granulocytes and NK cells, and myofibroblasts in bronchoalveolar lavage (BAL). CD45+ and CD45- cells expressing pro-inflammatory and pro-fibrotic cytokines were also increased. Lungs from PSGL-1 KO mice showed increased immune cell infiltration and apoptosis and exacerbated interstitial and peribronchial fibrosis. Targeted nanotherapy with LipHA+Ev decreased the myofibroblasts in BAL, cells producing proinflammatory and profibrotic cytokines, and the degree of lung inflammation at histology. LipHA+Ev treatment also decreased the severity of peribronchial and interstitial lung fibrosis, from moderate to mild levels. CONCLUSIONS AND IMPLICATIONS: In PSGL-1 KO mice, targeted nanotherapy with LipHA+Ev was an effective treatment for SSc-ILD, reducing the number of inflammatory and fibrotic cells in BAL and reducing inflammation and fibrosis in lungs.

Torsional wave elastography to assess the mechanical properties of the cornea.

Corneal mechanical changes are believed to occur before any visible structural alterations observed during routine clinical evaluation. This study proposed developing an elastography technique based on torsional waves (TWE) adapted to the specificities of the cornea. By measuring the displacements in the propagation plane perpendicular to the axis of the emitter, the effect of guided waves in plate-like media was proven negligible. Ex vivo experiments were carried out on porcine corneal samples considering a group of control and one group of alkali burn treatment ([Formula: see text]OH) that modified the mechanical properties. Phase speed was recovered as a function of intraocular pressure (IOP), and a Kelvin-Voigt rheological model was fitted to the dispersion curves to estimate viscoelastic parameters. A comparison with uniaxial tensile testing with thin-walled assumptions was also performed. Both shear elasticity and viscosity correlated positively with IOP, being the elasticity lower and the viscosity higher for the treated group. The viscoelastic parameters ranged from 21.33 to 63.17 kPa, and from 2.82 to 5.30 Pa s, for shear elasticity and viscosity, respectively. As far as the authors know, no other investigations have studied this mechanical plane under low strain ratios, typical of dynamic elastography in corneal tissue. TWE reflected mechanical properties changes after treatment, showing a high potential for clinical diagnosis due to its rapid performance time and paving the way for future in vivo studies.

Experimental evidence of shear waves in fractional viscoelastic rheological models.

Fractional viscoelastic rheological models, such as the Kelvin Voigt Fractional Derivative model, have been proposed in the literature for modelling shear wave propagation in soft tissue. In this article, our previously developed wave propagation model for transluminal propagation based on a Kelvin Voigt Fractional Derivative wave equation is experimentally validated. The transluminal procedure uses the transmission and detection of shear waves through the luminal wall. The model was compared against high-speed camera observations in translucent elastography phantoms with similar viscoelastic properties to prostate tissue. An ad hoc cross-correlation procedure was used to reconstruct the angular displacement from the high-speed camera observations. Rheometry and shear wave elastography were used for characterising the shear wave velocity dispersion curve for the phantoms. Fractional viscoelastic properties were derived after fitting the dispersion curve to its analytical expression. Propagation features and amplitude spectra from simulations and high-speed camera observations were compared. The obtained results indicate that the model replicates the experimental observations with acceptable accuracy. The model presented here provides a useful tool to model transluminal procedures based on wave propagation and its interaction with the mechanical properties of the tissue outside the lumen.

IGF-1 facilitates extinction of conditioned fear.

Insulin-like growth factor-1 (IGF-1) plays a key role in synaptic plasticity, spatial learning, and anxiety-like behavioral processes. While IGF-1 regulates neuronal firing and synaptic transmission in many areas of the central nervous system, its signaling and consequences on excitability, synaptic plasticity, and animal behavior dependent on the prefrontal cortex remain unexplored. Here, we show that IGF-1 induces a long-lasting depression of the medium and slow post-spike afterhyperpolarization (mAHP and sAHP), increasing the excitability of layer 5 pyramidal neurons of the rat infralimbic cortex. Besides, IGF-1 mediates a presynaptic long-term depression of both inhibitory and excitatory synaptic transmission in these neurons. The net effect of this IGF-1-mediated synaptic plasticity is a long-term potentiation of the postsynaptic potentials. Moreover, we demonstrate that IGF-1 favors the fear extinction memory. These results show novel functional consequences of IGF-1 signaling, revealing IGF-1 as a key element in the control of the fear extinction memory.

Experimental Evidence of Generation and Reception by a Transluminal Axisymmetric Shear Wave Elastography Prototype.

Experimental evidence on testing a non-ultrasonic-based probe for a new approach in transluminal elastography was presented. The proposed modality generated shear waves by inducing oscillatory rotation on the lumen wall. Detection of the propagated waves was achieved at a set of receivers in mechanical contact with the lumen wall. The excitation element of the probe was an electromagnetic rotational actuator whilst the sensing element was comprised by a uniform anglewise arrangement of four piezoelectric receivers. The prototype was tested in two soft-tissue-mimicking phantoms that contained lumenlike conduits and stiffer inclusions. The shear wave speed of the different components of the phantoms was characterized using shear wave elastography. These values were used to estimate the time-of-flight of the expected reflections. Ultrafast ultrasound imaging, based on Loupas' algorithm, was used to estimate the displacement field in transversal planes to the lumenlike conduit and to compare against the readouts from the transluminal transmission-reception tests. Experimental observations between ultrafast imaging and the transluminal probe were in good agreement, and reflections due to the stiffer inclusions were detected by the transluminal probe. The obtained experimental evidence provided proof-of-concept for the transluminal elastography probe and encouraged further exploration of clinical applications.

miR-320c Regulates SERPINA1 Expression and Is Induced in Patients With Pulmonary Disease.

INTRODUCTION: Alpha-1 antitrypsin deficiency (AATD) is a genetic condition resulting in lung and liver disease with a great clinical variability. MicroRNAs have been identified as disease modifiers; therefore miRNA deregulation could play an important role in disease heterogeneity. Members of miR-320 family are involved in regulating of multiple processes including inflammation, and have potential specific binding sites in the 3'UTR region of SERPINA1 gene. In this study we explore the involvement of miR-320c, a member of this family, in this disease. METHODS: Firstly in vitro studies were carried out to demonstrate regulation of SERPINA1 gene by miR-320. Furthermore, the expression of miR-320c was analyzed in the blood of 98 individuals with different AAT serum levels by using quantitative PCR and expression was correlated to clinical parameters of the patients. Finally, HL60 cells were used to analyze induction of miR-320c in inflammatory conditions. RESULTS: Overexpression of miR-320 members in human HepG2 cells led to inhibition of SERPINA1 expression. Analysis of miR-320c expression in patient's samples revealed significantly increased expression of miR-320c in individuals with pulmonary disease. Additionally, HL60 cells treated with the pro-inflammatory factor lipopolysaccharide (LPS) showed increase in miR-320c expression, suggesting that miR-320c responds to inflammation. CONCLUSION: Our findings demonstrate that miR-320c inhibits SERPINA1 expression in a hepatic cell line and its levels in blood are associated with lung disease in a cohort of patients with different AAT serum levels. These results suggest that miR-320c can play a role in AAT regulation and could be a biomarker of inflammatory processes in pulmonary diseases.

Relevance of PSGL-1 Expression in B Cell Development and Activation.

PSGL-1 is expressed in all plasma cells, but only in a small percentage of circulating B cells. Patients with systemic sclerosis (SSc) show reduced expression of PSGL-1 in B cells and increased prevalence of pulmonary arterial hypertension. PSGL-1 deficiency leads to a SSc-like syndrome and SSc-associated pulmonary hypertension in female mice. In this work, the expression of PSGL-1 was assessed during murine B cell development in the bone marrow and in several peripheral and spleen B cell subsets. The impact of PSGL-1 absence on B cell biology was also evaluated. Interestingly, the percentage of PSGL-1 expressing cells and PSGL-1 expression levels decreased in the transition from common lymphoid progenitors to immature B cells. PSGL-1-/- mice showed reduced frequencies of peripheral B cells and reduced B cell lineage-committed precursors in the bone marrow. In the spleen of WT mice, the highest percentages of PSGL-1+ populations were shown by Breg (90%), B1a (34.7%), and B1b (19.1%), while only 2.5-8% of B2 cells expressed PSGL-1; however, within B2 cells, the class-switched subsets showed the highest percentages of PSGL-1+ cells. Interestingly, PSGL-1-/- mice had increased IgG+ and IgD+ subsets and decreased IgA+ population. Of note, the percentage of PSGL-1+ cells was increased in all the B cell subclasses studied in peritoneal fluid. Furthermore, PSGL-1 engagement during in vitro activation with anti-IgM and anti-CD40 antibodies of human peripheral B cells, blocked IL-10 expression by activated human B cells. Remarkably, PSGL-1 expression in circulating plasma cells was reduced in pulmonary arterial hypertension patients. In summary, although the expression of PSGL-1 in mature B cells is low, the lack of PSGL-1 compromises normal B cell development and it may also play a role in the maturation and activation of peripheral naïve B cells.

Hyperelastic Ex Vivo Cervical Tissue Mechanical Characterization.

This paper presents the results of the comparison between a proposed Fourth Order Elastic Constants (FOECs) nonlinear model defined in the sense of Landau's theory, and the two most contrasted hyperelastic models in the literature, Mooney-Rivlin, and Ogden models. A mechanical testing protocol is developed to investigate the large-strain response of ex vivo cervical tissue samples in uniaxial tension in its two principal anatomical locations, the epithelial and connective layers. The final aim of this work is to compare the reconstructed shear modulus of the epithelial and connective layers of cervical tissue. According to the obtained results, the nonlinear parameter A from the proposed FOEC model could be an important biomarker in cervical tissue diagnosis. In addition, the calculated shear modulus depended on the anatomical location of the cervical tissue (µepithelial = 1.29 ± 0.15 MPa, and µconnective = 3.60 ± 0.63 MPa).

miR-320c Regulates SERPINA1 Expression and Is Induced in Patients With Pulmonary Disease.

INTRODUCTION: Alpha-1 antitrypsin deficiency (AATD) is a genetic condition resulting in lung and liver disease with a great clinical variability. MicroRNAs have been identified as disease modifiers; therefore miRNA deregulation could play an important role in disease heterogeneity. Members of miR-320 family are involved in regulating of multiple processes including inflammation, and have potential specific binding sites in the 3'UTR region of SERPINA1 gene. In this study we explore the involvement of miR-320c, a member of this family, in this disease. METHODS: Firstly in vitro studies were carried out to demonstrate regulation of SERPINA1 gene by miR-320. Furthermore, the expression of miR-320c was analyzed in the blood of 98 individuals with different AAT serum levels by using quantitative PCR and expression was correlated to clinical parameters of the patients. Finally, HL60 cells were used to analyze induction of miR-320c in inflammatory conditions. RESULTS: Overexpression of miR-320 members in human HepG2 cells led to inhibition of SERPINA1 expression. Analysis of miR-320c expression in patient's samples revealed significantly increased expression of miR-320c in individuals with pulmonary disease. Additionally, HL60 cells treated with the pro-inflammatory factor lipopolysaccharide (LPS) showed increase in miR-320c expression, suggesting that miR-320c responds to inflammation. CONCLUSION: Our findings demonstrate that miR-320c inhibits SERPINA1 expression in a hepatic cell line and its levels in blood are associated with lung disease in a cohort of patients with different AAT serum levels. These results suggest that miR-320c can play a role in AAT regulation and could be a biomarker of inflammatory processes in pulmonary diseases.