Extra Regions Research Articles

Correction of exon 2, exon 2–9 and exons 8–9 duplications in DMD patient myogenic cells by a single CRISPR/Cas9 system

Duchenne Muscular dystrophy (DMD), a yet-incurable X-linked recessive disorder that results in muscle wasting and loss of ambulation is due to mutations in the dystrophin gene. Exonic duplications of dystrophin gene are a common type of mutations found in DMD patients. In this study, we utilized a single guide RNA CRISPR strategy targeting intronic regions to delete the extra duplicated regions in patient myogenic cells carrying duplication of exon 2, exons 2–9, and exons 8–9 in the DMD gene. Immunostaining on CRISPR-corrected derived myotubes demonstrated the rescue of dystrophin protein. Subsequent RNA sequencing of the DMD cells indicated rescue of genes of dystrophin related pathways. Examination of predicted close-match off-targets evidenced no aberrant gene editing at these loci. Here, we further demonstrate the efficiency of a single guide CRISPR strategy capable of deleting multi-exon duplications in the DMD gene without significant off target effect. Our study contributes valuable insights into the safety and efficacy of using single guide CRISPR strategy as a potential therapeutic approach for DMD patients with duplications of variable size.

Mixing behavior and mass transfer of liquid–liquid two-phase flow in an annular microchannel with helical wires

Combining the advantages of high efficiency, low-pressure drop, and large throughput, the pore array-enhanced tube-in-tube microchannel (PA-TMC) is a promising microreactor for industrial applications. However, most of the mass transfer takes place in the upstream pore region, while the contribution of the downstream annulus is limited. In this work, helical wires were introduced into the annulus by adhering to the outer surface of the inner tube. Mixing behavior and mass transfer of liquid–liquid two-phase flow in PA-TMC with different helical wires have been systematically studied by a combination of experiments and volume of fluid (VOF) method. The introduction of helical wires improves the overall volumetric mass transfer coefficient KLa by up to 133% and the mass transfer efficiency E by up to 117%. The simulation results show that the helical wire brings extra phase mixing regions and increases the specific interface area, while accelerating the fluid flow and expanding the area of enhanced turbulent dissipation rate. Influences of helical wires in various configurations are compared by the comprehensive index I concerning the pressure drop and mass transfer performance simultaneously and a new correlation between KLa and specific energy consumption φ is proposed. This research deepens the understanding of the mixing behavior and mass transfer in the PA-TMCs and provides practical experience for the process intensification of microchannel reactors.

A developed fatigue analysis approach for composite wind turbine blade adhesive joints using finite-element submodeling technique

The intricate shapes and complex structures of composite wind turbine blades pose significant challenges in conducting local-fatigue simulation analyses. Moreover, adhesive joints, commonly found in composite wind turbine blades, are often neglected in current finite-element methods, leading to a dearth of empirical evidence to substantiate design enhancements for these blades. To tackle these issues, this study introduces a fatigue analysis methodology based on a finite-element submodeling approach to investigate the local fatigue behavior of composite wind turbine blades, with particular emphasis on the adhesive joints. The research begins with a comprehensive fatigue simulation analysis of composite wind turbine blades to establish boundary conditions for subsequent submodeling. A specialized submodel focusing on the trailing edge, a common adhesive joint, is constructed with the inclusion of extra adhesive regions for a more accurate depiction of their actual configuration. This submodel is subsequently utilized to investigate the localized fatigue characteristics unique to the trailing edge. Moreover, an exploration into the influence of the submodeling technique and its parameters on the predicted outcomes is carried out to enhance the applicability of the proposed methodology. These results significantly advance the understanding of local static and fatigue properties in large-scale structures through finite element analysis.

Time-series visual explainability for Alzheimer’s disease progression detection for smart healthcare

Artificial intelligence (AI)-based diagnostic systems provide less error-prone and safer support to clinicians, enhancing the medical decision-making process. This study presents a smart and reliable healthcare framework for detecting Alzheimer's disease (AD) progression. Early detection of AD before the onset of clinical symptoms is the most crucial step in starting timely treatment. To predict the conversion of cognitively normal patients to those with AD, three-dimensional (3D) magnetic resonance imaging (MRI) whole-brain neuroimaging methods have been extensively studied. However, depending on the 3D volume, this method is computationally expensive. To solve this problem, we used an approximate rank pooling method originally designed for video action recognition with a 3D MRI volume to obtain a compressed representation of multiple two-dimensional (2D) MRI slices. This study proposes a hybrid multimodal CNN-Bi-LSTM deep model for AD progression detection, in which the resulting dynamic 2D images are fused with cognitive features. Moreover, a novel explainable AI approach is proposed to provide visual explanations using the resulting longitudinal 2D dynamic images. Temporal explanations were provided by visualizing the affected brain regions captured using longitudinal 2D MRIs. By utilizing a sample of 1,692 subjects with multimodal data from the Alzheimer’s Disease Neuroimaging Initiative dataset, our method was assessed using a 10-fold cross-validation process. The model achieved an area under the receiver operating characteristics curve (AUC) of 94% using longitudinal 2D three-time-step dynamic image data. The fusion of 2D dynamic images with cognitive features enhanced the performance by 2% in terms of the AUC. Patients who gradually develop AD, show changes in various brain regions. For such patients, our system highlights the critical role of the hippocampus, medial amygdala, caudal hippocampus, and lateral amygdala at the initial time steps. In the late stages of AD, the system detects abnormalities in extra brain regions such as the medial temporal gyrus, superior temporal gyrus, fusiform gyrus, and caudal hippocampus; indicating that patients have completely progressed to AD.

Graph network measures reveal distinct white matter abnormalities in motor and extra-motor brain regions of two UMN-predominant ALS subtypes

BackgroundRoutine clinical magnetic resonance imaging (MRI) shows bilateral corticospinal tract (CST) hyperintensity in some patients with upper motor neuron (UMN)-predominant ALS (ALS-CST+) but not in others (ALS-CST–). Although, similar in their UMN features, the ALS-CST+ patient group is significantly younger in age, has faster disease progression and shorter survival than the ALS-CST– patient group. Reasons for the differences are unclear. MethodIn order to evaluate more objective MRI measures of these ALS subgroups, we used diffusion tensor images (DTI) obtained using single shot echo planar imaging sequence from 1.5 T Siemens MRI Scanner. We performed an exploratory whole brain white matter (WM) network analysis using graph theory approach on 45 ALS patients (ALS-CST+) (n = 21), and (ALS-CST–) (n = 24) and neurological controls (n = 14). ResultsSignificant (p < 0.05) differences in nodal degree measure between ALS patients and controls were observed in motor and extra motor regions, supplementary motor area, subcortical WM regions, cerebellum and vermis. Importantly, WM network abnormalities were significantly (p < 0.05) different between ALS-CST+ and ALS-CST– subgroups. Compared to neurologic controls, both ALS subgroups showed hubs in the right superior occipital gyrus and cuneus as well as significantly (p < 0.05) reduced small worldness supportive of WM network damage. ConclusionsSignificant differences between ALS-CST+ and ALS-CST– subgroups of WM network abnormalities, age of onset, symptom duration prior to MRI, and progression rate suggest these patients represent distinct clinical phenotypes and possibly pathophysiologic mechanisms of ALS.

SLICE: An algorithm for incorporating ultrasonography in the assessment of shocked or breathless patients.

SLICE is an algorithm for the integration of point-of-care ultrasound in the assessment and resuscitation of the shocked or breathless patient. It aims to determine the patient's fluid status, and identify reversible causes for the patient's clinical picture. SLICE stands for 'In a patient who is Shocked/Short of breath, scan the Lungs, IVC, Cardiac and Extra regions as indicated'. Its key advantages are that it explicitly guides resuscitative fluid management, can be performed rapidly and by clinicians with a broad range of sonographic experience, and can be used in a broad range of clinical scenarios. Its use has been successfully taught and implemented in routine clinical practice at our local institution.

Engineering Electromagnetic Field Distribution and Resonance Quality Factor Using Slotted Quasi-BIC Metasurfaces.

Dielectric metasurfaces governed by bound states in the continuum (BIC) are actively investigated for achieving high-quality factors and strong electromagnetic field enhancements. Traditional approaches reported for tuning the performance of quasi-BIC metasurfaces include tuning the resonator size, period, and structure symmetry. Here we propose and experimentally demonstrate an alternative approach through engineering slots within a zigzag array of elliptical silicon resonators. Through analytical theory, three-dimensional electromagnetic modeling, and infrared spectroscopy, we systematically investigate the spectral responses and field distributions of the slotted metasurface in the mid-IR. Our results show that by introducing slots, the electric field intensity enhancement near the apex and the quality factor of the quasi-BIC resonance are increased by a factor of 2.1 and 3.3, respectively, in comparison to the metasurface without slots. Furthermore, the slotted metasurface also provides extra regions of electromagnetic enhancement and confinement, which holds enormous potential in particle trapping, sensing, and emission enhancement.

Suspected Prostate Malignancy in Elderly Male Results in an Incidental Finding of Cellular Angiofibroma of the Male Pelvis

Cellular angiofibroma (CAF) is a rare benign mesenchymal tumor that affects both genders in the genitourinary region [1]. Nucci et al. first described this entity in the vulva in female in 1997 [2]. Cellular angiofibroma is generally a slow-growing and form asymptomatic mass that primarily arises in the vulvar-vaginal region in female and the inguinal region in males [3]. Although rare, cases have been reported in the pelvic and extra pelvic regions [4]. It mainly affects women mostly during the fifth decade of life [1]. “CAF” is the applied terminology of this tumor in accordance with World Health Organization [5]. It can easily be misdiagnosed as spontaneous perineal herniation in clinical practice [6]. Detailed pathological examination and correlation with immunohistochemistry workup are imperative to distinguish it from various soft tissue tumors which affect the pelvic regions. Differential diagnosis of this tumor includes both benign and malignant lesions such as spindle cell lipoma, solitary fibrous tumor, angiomyofibroblastoma, leiomyoma, and aggressive angiomyxoma [7]. Treatment with simple local excision is generally curable due to low local recurrence and no chance of metastasizing [8]. Cellular angiofibroma has a good prognosis [9]. However, preoperative distinction of cellular angiofibroma from malignant tumors can be challenging [10]. Periurethral CAF is rare with only two cases reported in the periurethral region [11, 12]. The current study presents a third rare case of cellular angiofibroma affecting the periurethral region in 69-year-old male that measured approximately 7 cm. This case was suspected a prostate malignancy with possible bladder invasion on the radiology imaging.

Cryo-EM structures of PI3Kα reveal conformational changes during inhibition and activation

Phosphoinositide 3-kinases (PI3Ks) are lipid kinases essential for growth and metabolism. Their aberrant activation is associated with many types of cancers. Here we used single-particle cryoelectron microscopy (cryo-EM) to determine three distinct conformations of full-length PI3Kα (p110α-p85α): the unliganded heterodimer PI3Kα, PI3Kα bound to the p110α-specific inhibitor BYL-719, and PI3Kα exposed to an activating phosphopeptide. The cryo-EM structures of unbound and of BYL-719-bound PI3Kα are in general accord with published crystal structures. Local deviations are presented and discussed. BYL-719 stabilizes the structure of PI3Kα, but three regions of low-resolution extra density remain and are provisionally assigned to the cSH2, BH, and SH3 domains of p85. One of the extra density regions is in contact with the kinase domain blocking access to the catalytic site. This conformational change indicates that the effects of BYL-719 on PI3Kα activity extend beyond competition with adenosine triphosphate (ATP). In unliganded PI3Kα, the DFG motif occurs in the "in" and "out" positions. In BYL-719-bound PI3Kα, only the DFG-in position, corresponding to the active conformation of the kinase, was observed. The phosphopeptide-bound structure of PI3Kα is composed of a stable core resolved at 3.8 Å. It contains all p110α domains except the adaptor-binding domain (ABD). The p85α domains, linked to the core through the ABD, are no longer resolved, implying that the phosphopeptide activates PI3Kα by fully releasing the niSH2 domain from binding to p110α. The structures presented here show the basal form of the full-length PI3Kα dimer and document conformational changes related to the activated and inhibited states.

Lack of Parkinsonian Pathology and Neurodegeneration in Mice After Long-Term Injections of a Proteasome Inhibitor in Olfactory Bulb and Amygdala.

Proteinaceous inclusions, called Lewy bodies (LBs), are used as a pathological hallmark for Parkinson’s disease (PD). Recent studies suggested a prion-like spreading mechanism for α-synucleinopathy where early neuropathological deposits occur, among others, in the olfactory bulb (OB) and amygdala. LBs contain insoluble α-synuclein and many other ubiquitinated proteins, suggesting a role of protein degradation system failure in PD pathogenesis. Therefore, we wanted to study the effects of a proteasomal inhibitor, lactacystin, on the aggregability and transmissibility of α-synuclein in the OB and amygdala. We performed injections of lactacystin in the OB and amygdala of wild-type mice. Motor behavior, markers of neuroinflammation, α-synuclein, and dopaminergic integrity were assessed by immunohistochemistry. Overall, there were no differences in the number of neurons and α-synuclein expression in these regions following injection of lactacystin into either the OB or amygdala. Microglial and astroglial labeling appeared to be correlated with surgery-induced inflammation or local effects of lactacystin. Consistent with the behavior and pathological findings, there was no loss of dopaminergic cell bodies in the substantia nigra and terminals in the striatum. Our data showed that long-term lactacystin injections in extra nigrostriatal regions may not mimic spreading aspects of PD and reinforce the special vulnerability of dopaminergic neurons of the substantia nigra pars compacta (SNc).

A Novel 3-D-CAD for Breast and Lesion Segmentation of Axial Breast DCE-MRI

Abstract The present paper proposes a fully automated three-dimensional (3-D) system for breast and lesion segmentation of Dynamic Contrast Enhanced MRI (DCE-MRI). Such a system as the Computer-Aided Diagnostic system (CAD) can be used to support radiologists by marking suspicious areas. The proposed 3-D-CAD system includes three modules. The first one concerns breast area segmentation based on image content analysis, the Moore-Neighbor tracing algorithm, and the Dijkstra algorithm. The second one concerns the automation of locating and selecting lesions that starts by preprocessing the already segmented breast regions; then, a K-means algorithm allows extraction of regions including suspicious tissues. The third one is the superimposition of all detected lesions from selected slices to create a 3-D view of the lesion. The 3-D reconstruction is based on the Marching Cube algorithm. The validation of breast area segmentation reveals the robustness of the proposed process versus different breast densities, complex forms, and challenging cases. The segmentation of the breast part from 120 slices with the proposed method is achieved in 20.57 ± 5.2 s, which is faster than existing methods. In addition to calculated metrics as Dice similarity coefficient (DSC) and rates of true positives, the module of lesion extraction is validated by experimented radiologists. The proposed workflow of this module shows competitive results compared to the existing methods of automated lesion segmentation and a total ability for eliminating the extra regions to lesions.

Anatomical targets and expected outcomes of catheter-based ablation of atrial fibrillation in 2020.

Anatomical-based approaches, targeting either pulmonary vein isolation (PVI) or additional extra PV regions, represent the most commonly used ablation treatments in symptomatic patients with atrial fibrillation (AF) recurrences despite antiarrhythmic drug therapy. PVI remains the main anatomical target during catheter-based AF ablation, with the aid of new technological advances as contact force monitoring to increase safety and effective radiofrequency (RF) lesions. Nowadays, cryoballoon ablation has also achieved the same level of scientific evidence in patients with paroxysmal AF undergoing PVI. In parallel, electrical isolation of extra PV targets has progressively increased, which is associated with a steady increase in complex cases undergoing ablation. Several atrial regions as the left atrial posterior wall, the vein of Marshall, the left atrial appendage, or the coronary sinus have been described in different series as locations potentially involved in AF initiation and maintenance. Targeting these regions may be challenging using conventional point-by-point RF delivery, which has opened new opportunities for coadjuvant alternatives as balloon ablation or selective ethanol injection. Although more extensive ablation may increase intraprocedural AF termination and freedom from arrhythmias during the follow-up, some of the targets to achieve such outcomes are not exempt of potential severe complications. Here, we review and discuss current anatomical approaches and the main ablation technologies to target atrial regions associated with AF initiation and maintenance.

Non-Rotating Black Holes, Dark Matter and Dark Energy in a Unifying Theory

The theory here developed, makes use of the decomposition of matter (mass) in different spatial frequencies k’s using spatial Fourier transforms, and the posterior use of modified inverse Fourier transforms to construct an accurate description of the classical Newtonian gravitational field. Introducing the concept of quantization of the spatial frequency k, which means allowing only discrete values, such as km, 2km, 3km, leads to the appearance of extra gravitational force regions that occur at distances equally spaced apart in 2π/km. These areas of extra gravitational force decrease inscribed in an inverse of the distance envelope (1/r). The value of 2π/km can be adjusted to be of the order of kiloparsec (kpc), being this way a plausible explanation for the effect of the dark matter since this causes practically flat rotation curves for most of the galaxies. As these regions of extra gravitational force also have adjacent areas of negative values (repulsive gravitational force), it is possible to show that any mass placed in the gravitational field far from the galaxy center will acquire, on average, a null acceleration, thereby remains the “light push,” or in other words, the “mean luminosity density” between galaxies as an explanation for the accelerating expansion of the universe, today being considered mainly due to dark energy. Along with the article, it is showed that the effect of light push is sufficient to explain the expansion of the universe. The present work also explains the nonlinear behavior of gravitational fields near massive objects such as blackholes, not contradicting the theory of general relativity, instead giving a complementary description of how black holes work, even describing the gravitational field internally to it, which is not available in the GR theory.

Structural behavior of a two length scale core-softened fluid in two dimensions

2D competitive systems shows a large variety of solid and aggregate patterns, re-entrant fluid phase, a continuous melting as predicted by the KTHNY theory and, in some cases, waterlike anomalies. In this paper, we employ Langevin Dynamics simulations of a ramp-like core-softened fluid that have two characteristic length scales and all the features mentioned above. Analyzing the fluid phase of this system, Bordin and Barbosa (2018) reported the existence of two waterlike anomalous region. Now, we revisit this problem focusing in the low temperature regime and a larger range of densities looking for a relation between the origin of the anomalous behavior, the solid phases and the re-entrant melting. Now, not two, but three anomalous regions were observed. The extra anomalous regions are related to the re-entrant melting of a amorphous to a cluster fluid. They are ruled not only by the competition between the characteristic length scales in the potential, but also by extra competitions induced by the variety of particles conformations. These competitions extends from the solid to the fluid phase, reflecting in the structural waterlike anomaly. Our results shade some light in the complex behavior of two length scales competitive potential, and helps to elucidate the relation between the large number of solid phases and the existence of more than one waterlike anomalies region in these systems.

Influence of Concomitant Injuries on Outcome of Traumatic Brain Injury

Traumatic brain injury (TBI) is a leading cause of death and disability in trauma patients. Patients with TBI frequently sustain concomitant injuries in extra cranial regions. The effect of severe extracranial injury (SEI) on the outcome of TBI is controversial. We retrospectively enrolled 129 patients with head injury Total patients were dichotomized into isolated TBI (n = 90) and TBI associated with SEI (n = 39). The differences in severity and outcome between these two groups were analyzed. Mortality was 8.89 % in the isolated TBI group and 20.28 % in TBI with SEI group (P = 0.0719), but the Glasgow Outcome Scale (GOS) in terms of Good Recovery (GR) was more in the isolated TBI group (P = 0.0004). Adjusting for age, GCS, and length of hospital stay, SEI was a strong prognostic factor for mortality. It is important to manage not only the brain but the whole body in the treatment of TBI patients with SEI.&#x0D; Bangladesh Crit Care J March 2020; 8(1): 24-28

Experimental Investigation of Phase Equilibria in the Cr-Mo-Hf Ternary System

In this study, the phase equilibria of the Cr-Mo-Hf ternary system at 1100 and 1300 °C were experimentally investigated by means of back-scattered electron, electron probe microanalysis and x-ray diffraction. The two C15 phases (α(HfMo2) and α(HfCr2)) link up with each other from Mo-Hf side to Cr-Hf side across the isothermal sections both at 1100 and 1300 °C. The isothermal section at 1300 °C consists of one three-phase region and three two-phase regions, while no ternary compound is found. One single-phase region of bcc(βHf) not connected with Mo-Hf binary boundary is found unexpectedly at 1100 °C, which results in the appearance of two extra three-phase regions.

LAMPA, LArge Multidomain Protein Annotator, and its application to RNA virus polyproteins.

MotivationTo facilitate accurate estimation of statistical significance of sequence similarity in profile–profile searches, queries should ideally correspond to protein domains. For multidomain proteins, using domains as queries depends on delineation of domain borders, which may be unknown. Thus, proteins are commonly used as queries that complicate establishing homology for similarities close to cutoff levels of statistical significance.ResultsIn this article, we describe an iterative approach, called LAMPA, LArge Multidomain Protein Annotator, that resolves the above conundrum by gradual expansion of hit coverage of multidomain proteins through re-evaluating statistical significance of hit similarity using ever smaller queries defined at each iteration. LAMPA employs TMHMM and HHsearch for recognition of transmembrane regions and homology, respectively. We used Pfam database for annotating 2985 multidomain proteins (polyproteins) composed of >1000 amino acid residues, which dominate proteomes of RNA viruses. Under strict cutoffs, LAMPA outperformed HHsearch-mediated runs using intact polyproteins as queries by three measures: number of and coverage by identified homologous regions, and number of hit Pfam profiles. Compared to HHsearch, LAMPA identified 507 extra homologous regions in 14.4% of polyproteins. This Pfam-based annotation of RNA virus polyproteins by LAMPA was also superior to RefSeq expert annotation by two measures, region number and annotated length, for 69.3% of RNA virus polyprotein entries. We rationalized the obtained results based on dependencies of HHsearch hit statistical significance for local alignment similarity score from lengths and diversities of query-target pairs in computational experiments.Availability and implementationLAMPA 1.0.0 R package is placed at github (https://github.com/Gorbalenya-Lab/LAMPA).Supplementary information Supplementary data are available at Bioinformatics online.

High-Efficiency Ultra-Broadband Multi-Tip Edge Couplers for Integration of Distributed Feedback Laser With Silicon-on-Insulator Waveguide

A high-efficiency ultra-broadband multi-tip edge coupler based on a silicon-on-insulator platform for direct coupling with the elliptic beam of a distributed feedback laser was developed. The device is composed of a multi-tip section and a combiner section with extra offset regions to reduce the mode mismatch caused by the structural discontinuity which results from a limitation of the fabrication process that creates an inevitable gap width at the junction between the two sections. The widths and the spacing of the tips for the multi-tip section and the extra offset region for the combiner section are fine-tuned by using the particle swarm optimization method to reduce the mode mismatch. A high overall coupling efficiency up to 90.68% (0.4249 dB) at 1550 nm can be achieved for the optimized 90-μm-long four-tip edge coupler. The device can be operated over a broad spectral range of 1260–1675 nm with less than 1 dB extra loss. With its high-efficiency ultra-broadband operation and small device footprint, the proposed device is promising for laser-to-chip edge coupling in silicon photonics.

Tectonostratigraphic evolution and timing deformation in the Miocene Paso del Sapo Basin: Implications for the Patagonian broken foreland

Several segments of the Central and Southern Andes show deformation evidences located far away from the orogenic front. As a result, a landscape constituted by fault-blocks mountains and intermontane basins occurs. The spatial-temporal distribution of the deformation in these Broken Foreland Systems is still uncertain. Some works suggest that change in space and time of rock uplift along the structures is controlled mainly by older crustal discontinuities. However, other authors propose, a particular deformation pattern between the Andean and extra Andean regions, such as eastward-westward migration of the orogenic front related to subduction zone dynamics. In this work we analyse new structural, geomorphological and tectono-stratigraphical data together with previous chronological scheme (based on U-Pb and magnetostratigraphic ages) of one of the Patagonian Broken Foreland depocenters: the Paso del Sapo Basin. The results allow to perform the tectonostratigraphic evolution of the basin and constrain the timing of deformation in Paso del Sapo Basin at 15–11.5 Ma. Different tectonic structures were activated and de-activated in this time interval, while an older crustal discontinuity – the Río Chubut Medio fault, located at the eastern border of the basin – recorded a continuous tectonic activity throughout the entire evolution of the basin. These evidences show a synchronous tectonic activity for the Andean and extra-Andean Patagonian regions at those times. In addition, the presence of a climax deformation constrained to 15–14.6 Ma and the deceleration of the Río Chubut Medio Fault at 12.2 Ma could be related to eastward expansion and westward retraction of the orogenic front respectively. Therefore, our data indicate that both the presence and absence of a deformation pattern in extra-Andean regions should be considered to suggest alternative lithospheric-scale models.

11P - Expression of erythropoietin receptor variant forms is associated with lymphatic invasion and metastasis formation in breast cancer

Background: Anemia not only impairs the quality of life of patients, but it leads to tumor hypoxia, resistance to therapy and reduces survival. Although recombinant Epo has revolutionized the treatment of anemia, clinical trials suggested a potential adverse effect of Epo on tumor recurrence and patient survival. Previously we identified altered EpoR mRNA transcripts in breast cancer cells lacking ligand binding domain exon 1-3 sequences in vitro. Our hypothesis is that altered expression of these proteins may be associated with unique clinicopathologic features. Methods: The expression level of C-and N-terminal portion of EpoR were measured by quantitative RT-PCR in 157 retrospective cohort of archived breast cancers. In order to identify the variant EpoR mRNA forms we have established a SMART RACE PCR assay using EpoR specific primer sets producing large overlapping sequences. Total RNA was frozen sections of 20 primary breast cancers after the H&E diagnoses.The products of the SMART RACE and nested PCR reactions was subjected to sequence analysis. Results: Out of the 157 samples 95 (61%) showed more than 2-fold change between the expression levels of the C and N-terminal regions of EpoR by quantitative RT-PCR. Significant correlation was found between the expression levels of EpoR mRNA variants and lymphatic invasion of the tumors. Logistic regression model demonstrated the association of metastatic capacity and expression differences of extra and intracellular regions of EpoR in primary breast cancers (P < 0.05, greater than 2-fold change). Using a SMART RACE PCR assay a single EpoR mRNA species at ∼1500 kb, corresponding to the full length wild type EpoR mRNA, and several distinct bands at ∼1300 kb, ∼1000 kb and ∼800 kb, lacking exon 1-4 N-terminal ligand binding domain, were seen in the samples. Conclusions: We showed that different variant forms of EPOR present not only in vitro but also in the individual breast tumor samples. These altered forms of EpoR may be responsible for the observed differences in the Epo responsiveness of EpoR bearing breast cancers. Legal entity responsible for the study: Zsuzsa Rakosy. Funding: OTKA PD 116996, 2015-2018. Disclosure: The author has declared no conflicts of interest.