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Introduction Testis microenvironment which is comprised of cells like Sertoli and peritubular cells releases paracrine factors which could potentially modulate testicular functions. In our previous studies, we identified Leptin as a key paracrine factor in the testicular microenvironment, released by Sertoli and peritubular cells and goes to bind its receptor in Leydig cells. Once this association happens it further induces the expression of the desert hedgehog signaling pathway, which is known to play an important role in the Leydig Stem cell’s differentiation to Adult Lydig Cells, which are capable of producing testosterone. However, one of the limitations in our study was that the model we used were from human cells that were extracted from testes’ biopsies which lacked an intact immune microenvironment. In the present study we explored the influence of Leptin on an intact testicular immune microenvironment. Objective To explore the influence of Leptin on an intact testicular immune microenvironment. Methods We used 6-weeks old C57BL6 mice which were exposed to two different concentrations of Leptin, 10 and 100 mg/ml for seven days. After seven days of treatment, these animals were humanly sacrificed and we collected their blood and testes. Blood was subjected to the CBC differential profile performed using EDTA anti-coagulated blood samples with a Hemavet Mascot Multispecies Hematology System Counter 1500R (CDC Technologies, Inc., Oxford, CT). Samples were counted no longer than five minutes after blood was drawn. In addition to that, the testes were used and subjected to comprehensive immunophenotyping that allowed us to evaluate in detail the impact of Leptin treatment at different doses on different immune cell types which are found within the testes. Results Results showed that Leptin at low dose (10mg/ml) showed increased levels of serum Testosterone (13.20 + 0.37 ng/dl) but were not significant, LH and FSH (LH 0.016 + 0.02 ng/ml, FSH in 51.71 + 12.93 ng/ml) showed significant (p Conclusions The observation made in this study highlights unexplored aspects of influence of paracrine factors released by Sertoli and peritubular myoid cells on modulating immune microenvironment within the testis to potentially influence testosterone production. In future studies we will be exploring how the changes in the different immune cell types are correlated directly with the increase in T cells and how this correlation can influence the Leydig Stem Cell differentiation into Leydig Adult Cells. Disclosure No

Introduction Congenital Bilateral Absence of the Vas Deferens (CBAVD) is a condition leading to obstructive azoospermia and male infertility. CBAVD is associated with mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Congenital Unilateral Absence of the Vas Deferens (CUAVD), where only one vas deferens is missing, is also associated with mutations in the CFTR gene. However, the genetic causes of both conditions are not fully understood and warrant further investigation. We present two brothers, one with CBAVD and one with CUAVD, who were subjected to whole exome sequencing. Objective To assess genetic similarities using whole exome sequencing between two brothers with differing reproductive capacities. Methods Whole exome sequencing was performed in two brothers. Subject A was 39-year-old-male who had previously fathered a child and was diagnosed with CUAVD during his vasectomy. Subject B was a 37-year-old male diagnosed with CBAVD, and genetic testing significant for one copy of a mutated cystic fibrosis gene (unaffected CF carrier). Whole blood was collected. DNA was extracted and quality assessed. Finally, whole exome sequencing was completed. Results Whole exome sequencing results were subjected to annotation and filtering. Both subjects were noted to share a rare mutation in the CFTR gene (p.R347H). CFTR mutations are the most common cause of CBAVD and have also been linked to CUAVD. However, this is a unique case of two brothers with the same genetic mutation but different phenotypic expressions of the mutation. Conclusions We have identified a rare CFTR variant (p.R347H) in two brothers with different reproductive capacities. We present evidence that absence of the vas deferens (unilateral or bilateral) may be associated with CFTR mutations, even in the absence of diagnosed cystic fibrosis. Understanding the phenotypic spectrum of CFTR gene mutations is pivotal to treating associated male infertility. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus Pharmaceuticals, Boston Scientific, Coloplast, Endo Pharmaceuticals, Empower Pharmacy, Nestle Health, Olympus, Hims Inc.

Introduction Testosterone (T) plays an important role in properly developing and maintaining male reproductive tissues. The normal serum testosterone levels can vary between 300 to 1000 ng/dL or 10.4 to 34.7 nmol/L. However, it is unclear whether androgen receptor signaling can vary based on serum T levels or the half-life of testosterone therapy. What remains unknown is whether there is a uniform serum T level above which androgen receptor signaling within the tissue remains similar (saturation hypothesis). Objective We first evaluated whether varying serum levels of T alter androgen receptor (AR) signaling in human penile tissue. Then, we evaluated and compared AR signaling in corpus cavernosum tissue of men treated with short-acting vs. long-acting testosterone. We hypothesized that we would find similar AR signaling in the penile tissue of both eugonadal and hypogonadal men as well as those men receiving short and long-acting testosterone. Methods We obtained human corpus cavernosum (hCC) tissue biopsies during penile surgery (for erectile dysfunction) from 14 men. We recorded their serum T levels on week prior to surgery before 10AM. We used western blot to evaluate adrenergic receptor (AR) and VEGF protein levels. Men without erectile dysfunction undergoing penile surgery for Peyronie's disease were used as controls. Results The median age of participants was 61 (IQR 12) and the median T level was 325ng/dL (IQR 201.45). The results of western blot showed expression of similar levels of VEGF in hCC samples from men with T >200ng/dL. However, men with serum T levels lower than 200 ng/dL had decreased expression of VEGF and AR. Conclusions This finding is significant because it demonstrates that serum testosterone beyond a certain threshold may have a minimal impact on erectile function. If a serum testosterone threshold is identified, this threshold could be a valuable target for future studies on the effects of testosterone on ED. It may alter the treatment threshold for men who have ED as their primary symptom of hypogonadism. This data also suggests the saturation value for penile androgen receptors. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus Pharmaceuticals, Boston Scientific, Coloplast, Endo Pharmaceuticals, Empower Pharmacy, Nestle Health, Olympus, Hims, Inc

Introduction The primary source of testosterone (T) in men are Leydig cells (LCs) and their dysfunction can lead to T deficiency. Testicular microenvironment (TME) which is comparised of cells such as Sertoli and Peritubular myoid cells releases the paracrine factors to influence the growth and differentiation of LCs. Leptin is a 16-kDa protein produced primarily by adipose tissue and exerts a significant influence on hormonal regulation. In our recent study, we demonstrated that Leptin, a paracrine factor secreted by TME, is critical for Leydig stem cell (LSCs) differentiation and subsequent T production under the regulation of the desert hedgehog signalling pathway. We also showed that this regulation is inverse concentration dependent such as Leptin increases LSC differentiation at low doses and decrease it at higher doses. However, an in-depth understanding of mechanisms that derive the Leptin secretion and decide the fate of LCs are still unclear and is the focus of the present study. Objective To study the mechanisms that derive the Leptin secretion and decide the fate of LCs. Methods A total of 18 men with testicular failure underwent testis biopsies. The testicular cells were characterized using Flow cytometry, Immunostaining and RNA sequencing then the cells were also treated with Leptin in a dose dependent (0, 1, 10 ng/mL respectively) and time dependent (24, 48, 72, and 96 hours) manner. For the functional studies, UCSC genome browser was used to study the transcription factor genes (TFs) binding the leptin receptor (LEPR) promoter. The list of TFs was subjected to enrichment analysis. To evaluate the effects of shortlisted TFs on leptin-LEPR-DHH to induce EMT in LSCs, qPCR and western blotting were performed. For statistical analysis GraphPad Prism was used. For statistical analysis GraphPad Prism was used. All data were presented as the means ± SEM. All data were presented as the means ± SEM Results Over 60 TF’s were identified by UCSC Genome browser, which were subjected to enrichment analysis using ENRICHR and Ingenuity pathway analysis. Output revealed TFs; SOX2, SOX8 and SOX9 that were enriched for mechanisms influencing hormonal regulation via hypothalamic pituitary gonadal axis. The role of these TFs has been well established in epithelial to mesenchymal transition in different cancer types, however their impact on LSC differentiation remained underexplored. RNA sequencing data from cells exposed to different concentrations of Leptin showed an inducing effect of Leptin on SOX2, SOX8 Sox9 respectively. This was confirmed at RNA as well as at protein levels. Consequently, upon inhibition of LEPR, the expression of SOX2, SOX8 and SOX9 was reduced as well. Furthermore we used SOX8, SOX9 and SOX2 antagonists in the presence or absence of Leptin to validate the effects on EMT of LCs. Conclusions Our results demonstrate that Leptin-LEPR-DHH induced Leydig stem cells differentiation in testis is modulated by EMT inducing transcription factors which binds to LEPR. Further studies are ongoing to explore the potential therapeutic effects of leptin treatment in improving fertility. Disclosure No

Introduction Platelet Rich Plasma (PRP) is a novel restorative therapy rich in growth factors and cytokines used to target the underlying causes of erectile dysfunction (ED). It is not known, however, if the composition of growth factors in PRP varies between men. Objective This study aimed to evaluate PRP growth factor variability among men with ED. Methods Whole blood was collected from sixteen men, eight participants with at least a six-month history of ED. The others were the control group, including seven men with Peyronie’s Disease (PD) and one healthy man without sexual dysfunction. PRP was extracted from whole blood using the Arthrex Angel system. A Human Growth Factor Antibody Array for 41 proteins was performed. Using all 16 samples, quantitative detection of GM-CSF, VEGF, and TGF-β were validated by western blot. Results From the growth factor array, two growth factors - Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) and Transforming Growth Factor-beta (TGF-β) - were identified as having a 1.5-fold decrease between the participants and the control. In addition to the shortlisted growth factors, VEGF was selected because androgens can upregulate VEGF production. Other than a weak negative correlation between VEGF expression and age, we found no correlation between growth factor expression for GM-CSF or TGF-β and age, BMI, or comorbidities. Conclusions PRP growth factor concentration appears to vary among men with ED. Depending on individual growth factor concentration, PRP treatment for ED may need to be personalized for patients. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus Pharmaceuticals, Boston Scientific, Coloplast, Endo Pharmaceuticals, Empower Pharmacy, Nestle Health, Olympus, Hims, Inc.

Background The epicardial-mesothelial cells (EMCs) are essential regulators of cardiac growth and repairment process. The epicardial-to-mesenchymal transition (EMT) of epicardial fate determination are controlled by both cell autonomous and cardiomyocyte-originated mechanisms. Here, by using an in vitro and in vivo model of epicardial EMT, we investigated the role of miRNAs as regulators of these process and their potential targets. Methods EMT was induced in mice embryonic tcf21+EMCs through an angiogenic cytokine, prokineticin treatments. Cre-dependent tracing of TCF21-tm-iCRE-EMCs was utilized to abrogate prokineticin receptor-1 (PKR1) in mice epicardium. Human tcf21+ cardiac fibroblast (mainly originated from epicardial origin also utilized to study for their repairment signaling. Results Upon EMC-specific abrogation of PKR1 in early stages of cardiac development the TCF21-tm-iCREPKR1−/− mice exhibited 13±4% embryonic lethality due to a disconnection of epicardial cells from compact layer, failed expansion of the sub-epicardial space, and disruption of heterotypic cell interaction between epicardium and myocardium. EMT-RT profiler and enrichment analysis revealed an impaired EMT in TCF21-tm-iCREPKR1−/− hearts. Ingenuity Pathway Analysis on the EMT molecular network (e.g., Snai1, Snai2, B-catenin, N-cadherin, vimentin), specific to cardiac development and defects revealed MiR-124 as a part of this network that was inversely associated and down-stream of PKR1 in cultured Tcf21+ cells. Furthermore, protein expression of SNAIL2 was significantly downregulated in TCF21-tm-iCREPKR1−/− hearts and cultured Tcf21+ cells upon treatment with mir124 mimic. The luciferase reporter assay showed that miR124 directly targeted the 3'-untranslated region of SNAIL2 in EMCs. In a counter experiment, PKR1 gene replacement or mir124 inhibitor was able to rescue the impaired EMT-genes, and an increase in apoptosis and impaired proliferation in epicardium of embryonic TCF21-tm-iCREPKR1−/− hearts. Importantly, miRNA mimic in cultured tcf21 cells maintained epithelial features in EMCs, and increased EMT-associated transcripts and traits, such as apoptosis. In contrast, MiR124 is involved in wound healing process in tcf21+human CFs and EMCs. Conclusion This study shows the importance of a crosstalk between PKR1 and miR-124 with respect to EMT regulation during cardiac development and repair. Controlling fibroblast-committed EMT and pathological fibrotic remodeling is of increasing interest within the field of regenerative tissue engineering and development of interventional strategies after cardiac injury. Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): Fondation de France

Iron deficiency anaemia in orthopaedic surgery is common and there is increased risk of blood transfusion and associated adverse reactions. The management involves administration of iron (oral or intravenous) and erythropoietin stimulating agents.We searched for PubMed, Embase, Google Scholar and Cochrane database to identify the studies from inception to April 2021. Randomized controlled trials with adult patients undergoing orthopedic surgery were included. The metanalysis compared patients who were administered combination of erythropoietin stimulating agents and iron in one group and iron alone. The primary outcome was the rate of blood transfusion and the secondary outcome studied were postoperative hemoglobin concentration, after treatment hemoglobin levels, and complications like mortality, stroke, myocardial infarction, deep vein thrombosis, pulmonary embolism and renal dysfunction.Eleven studies were included. The combination of ESA and iron decreased number of patients who required blood transfusion in comparison to patients treated with iron therapy alone (RR, 0.73; 95% CI, 0.59 to 0.91, I.ref id="2"2/ref= 65%; p = 0.005). In subgroup analysis with oral and intravenous iron, the difference was not statistically significant (p = 0.24). Administration of erythropoietin either in high ( 80,000 IU) or low dose ( 80,000 IU) resulted in lower blood transfusion rates (p = 0.0007) with no significant difference between groups. The risk of mortality, myocardial infarction, stroke, deep vein thrombosis or pulmonary embolism did not significantly increase. Combined administration of ESA and iron versus iron only reduces the number of red blood cell transfusions in the postoperative period in orthopedic procedures with minimal risk of complications.La anemia por deficiencia de hierro en la cirugía ortopédica es común y existe un mayor riesgo de transfusión de sangre y reacciones adversas asociadas. El tratamiento implica la administración de hierro (oral o intravenoso) y agentes estimulantes de la eritropoyetina.Se realizaron búsquedas en PubMed, Embase, Google Académico y la base de datos Cochrane para identificar los estudios desde su inicio hasta Abril de 2021. Se incluyeron ensayos controlados aleatorios con pacientes adultos sometidos a cirugía ortopédica. El metaanálisis comparó pacientes a los que se les administró una combinación de agentes estimulantes de la eritropoyetina y hierro en un grupo y hierro solo. El resultado primario fue la tasa de transfusión de sangre y el resultado secundario estudiado fue la concentración de hemoglobina postoperatoria, los niveles de hemoglobina después del tratamiento y complicaciones como mortalidad, accidente cerebrovascular, infarto de miocardio, trombosis venosa profunda, embolia pulmonar y disfunción renal.Se incluyeron 11 estudios. La combinación de AEE y hierro disminuyó el número de pacientes que requirieron transfusión de sangre en comparación con los pacientes tratados con tratamiento con hierro solo (RR, 0.73; IC del 95%, 0.59 a 0.91, I.ref id="2"2/ref= 65%; p = 0.005). En el análisis de subgrupos con hierro oral e intravenoso, la diferencia no fue estadísticamente significativa (p = 0.24). La administración de eritropoyetina en dosis altas ( 80,000 UI) o bajas ( 80,000 UI) dio lugar a tasas de transfusión de sangre más bajas (p = 0.0007) sin diferencias significativas entre los grupos. El riesgo de mortalidad, infarto de miocardio, accidente cerebrovascular, trombosis venosa profunda o embolia pulmonar no aumentó significativamente. La administración combinada de AEE y hierro frente al hierro solo reduce el número de transfusiones de glóbulos rojos en el período postoperatorio en procedimientos ortopédicos con un riesgo mínimo de complicaciones.

A promising route for the development of opto-elelctronic technology is to use terahertz radiation to modulate the optical properties of semiconductors. Here we demonstrate the dynamical control of photoluminescence (PL) emission in few-layer InSe using picosecond terahertz pulses. We observe a strong PL quenching (up to 50%) after the arrival of the terahertz pulse followed by a reversible recovery of the emission on the time scale of 50ps at T =10K. Microscopic calculations reveal that the origin of the photoluminescence quenching is the terahertz absorption by photo-excited carriers: this leads to a heating of the carriers and a broadening of their distribution, which reduces the probability of bimolecular electron-hole recombination and, therefore, the luminescence. By numerically evaluating the Boltzmann equation, we are able to clarify the individual roles of optical and acoustic phonons in the subsequent cooling process. The same PL quenchingmechanismis expected in other van derWaals semiconductors and the effectwill be particularly strong for materials with low carrier masses and long carrier relaxation time, which is the case for InSe. This work gives a solid background for the development of opto-electronic applications based on InSe, such as THz detectors and optical modulators., Comment: The following article has been accepted by Applied Physics Letters. After it is published, it will be found at https://publishing.aip.org/resources/librarians/products/journals/

Introduction Adult Leydig cells in the testes are essential for testosterone production. The growth and differentiation of Leydig cells could be affected by paracrine factors released by testicular microenvironment (TME) (Sertoli and peritubular myoid cells). In our recent study, we demonstrated that Leptin, a paracrine factor secreted by TME, is critical for Leydig stem cells (LSCs) differentiation and subsequent testosterone production via its interaction with Leptin receptor on LSCs, followed by downstream regulation of desert hedgehog (DHH) signaling. Although it is well-known that obesity adversely affects male fertility and testosterone production, the endogenous effects of Leptin on Leydig stem cell differentiation and transcription factors genes (TFs) binding to leptin receptor (LEPR) and the mechanism that are specific to patient's BMI (normal, obese and lean men) are understudied. Additionally, the underlying mechanism behind the differential effects of Leptin (if any) is unknown. Therefore, In the present study, we evaluated the impacts and mechanism of Leptin as a paracrine factor on LSC differentiation in patients with different BMI's. Objective To uncover the mechanism of Leptin as a paracrine factor on LSC differentiation in patients with different BMI's. Methods A total of 13 men with testicular failure that were subcategorized as obese (BMI >35), normal (BMI 25-30), and lean (BMI Results Results showed that a strong relationship exists between BMI and leptin levels. Immunostaining, flow cytometry and RNA sequencing results showed that in the men with normal BMI, in the presence of low doses of leptin, there was a shift of the testicular cell population towards adult Leydig cells (increased population of cells staining positive for 3BHSD, LHR) and increased DHH signaling (GLI and SMO). On the contrary, at higher concentrations, the positive effects of leptin on cells from men with normal BMI were neutralized. Furthermore, in cells from lean patients, leptin treatment had a consistent, linear and positive impact on LSC differentiation markers and DHH signaling. Moreover, in cells from obese patients, leptin had a consistent, linear and negative impact on LSC differentiation and markers of DHH signaling. Enrichment analysis identified 50 TFs, of which SOX2, SOX8 and SOX9 were enriched for abnormal pituitary function and therefore for hypothalamic pituitary gonadal axis. The role of these TFs on LSC differentiation remained under-explored. RNA sequencing data from cells exposed to different concentrations of Leptin showed an inducing effect of Leptin on SOX2, SOX8 Sox9 respectively. Consequently, with increasing leptin doses SOX2, SOX8 Sox9 protein and expression has been upregulated. Mechanistically, Leptin-LEPR-TFs induced DHH signaling regulates LSCs differentiation in a different manner which is specific to patient's BMI. Conclusions Our results demonstrate the influence, BMI has on Leptin induced LSC differentiation. These findings suggest the future potential use of Leptin as a personalized therapy for inducing LSC differentiation and overcoming low Testosterone levels. Further studies are necessary to identify potential therapeutic effects of leptin treatment in improving fertility in the setting of leptin resistance and obesity. Financial Support: Supported by the American Urological Association Research Scholar Award to H.A. J.M.H. is supported by NIH grants 1R01 HL137355, 1R01 HL107110, 1R01 HL134558, 5R01 CA136387, 5UM1 HL113460 and Soffer Family Foundation. Disclosure No

Introduction Although testosterone deficiency (TD) may be present in 1 out of 5 men 40 years or older, the factors responsible for TD remain largely unknown. Leydig stem cells (LSCs) differentiate into adult Leydig cells (ALC) and produce testosterone in the testes under the pulsatile control of luteinizing hormone (LH) from the pituitary gland. However, our recent studies have suggested that the testicular microenvironment (TME), which is comprised of Sertoli and peritubular myoid cells (PMC), plays an instrumental role in LSC differentiation and testosterone production under the regulation of the desert hedgehog signaling pathway (DHH). In the present study, we hypothesized that testicular TME releases paracrine factors that modulate LSC differentiation. Objective To further examine the TME its see if its release of paracrine factors modulates LSC differentiation. Methods A total of 13 men with testicular failure underwent testis biopsies for sperm retrieval. Using an IRB approved protocol, about 10mg of testicular tissue from each of these men were processed for Leydig stem cell isolation, culture and characterized. Cytokine antibody array was performed to identify the paracrine factors released by TME. The cells were treated with hedgehog signaling agonist and antagonist in the presence or absence of identified paracrine factors from the conditioned media. Two independent biopsies were treated with siRNA against LEPR (25pmol) for 48hrs. This was followed by evaluating the extent of knockdown on LEPR and B3HSD. Further, to evaluate the functional aspects (cell proliferation) modulated by Leptin-DHH signalling by first sorting the Sertoli cells, Peritubular myoid cells, Leydig stem cells and Adult Leydig cells using surface markers. These cells were then treated with Hedgehog agonist (SAG) and antagonist (Vismodigib), in the presence or absence of increasing doses of Leptin (0, 1, 10 ng/ml respectively) 48 hours post treatment, these cells were used for checking the impacts of Leptin-DHH treatments on cell proliferation by MTT assay. GraphPad Prism (GraphPad Software) was used for statistical analysis. All data were presented as the means ± SEM. The statistical significance between two groups was estimated by unpaired two-tailed t-test. Results We successfully isolated and cultured up to 5 × 106million LSC's / biopsy from all 13 human testis biopsies. These cells were characterized for the presence or absence of different cell types in testis such as adult Leydig cells, Leydig stem cells, Sertoli cells, peritublar myoid cells and germ cells. Results showed that, of the cells cultured, up to 70% of the cells were Leydig stem cells and 10% of them were Sertoli-cell in origin on day 14. IF and qPCR data showed as the majority of cell population was undifferentiated (PDGFR-α). Upon stimulation by LH, the expression of 3βHSD (mature Leydig cells) was increased and that of PDGFR-α was decreased. Results from siRNA against LEPR has shown the down-regulated LEPR and B3HSD expression levels. The results from cell proliferation showed that there is minimal to no impact of leptin treatment on upon DHH agonist (SAG) or antagonist (Vismodigib) treatment in either cell types therefore suggesting that Leptin is not involving in the proliferation mechanism. Furthermore, the results demonstrated that TME secretes leptin which induces LSC differentiation and increases T production. Leptin's effects on LSC differentiation and T production, however, are inversely concentration-dependent: positive at low doses and negative at higher doses. Mechanistically, leptin acts on LSCs upstream of DHH; leptin-DHH regulation functions unidirectionally insofar as DHH gain or loss of function has no effects on leptin levels. Conclusions Taken together, results from this study demonstrate the instrumental role of leptin as a paracrine factor secreted by TME on human LSC function and differentiation. Moreover, this study established the molecular events (DHH signaling) that underlie the leptin-modulated effects on LSCs. Further studies are ongoing to validate the implications of these Leptin in terms of its role in LSCs function, differentiation and survival Disclosure No

Microwave ablation (MWA) therapy is a hyperthermic treatment for cancerous tumors whereby microwave energy is dispersed into a target tissue region. Absorption of these waves leads to temperature rise through microwave heating with the aim of exposing cancerous cells to a thermal dose sufficient to kill them whilst minimizing the damage to the surrounding healthy tissue. Being able to predict the progression of ablation during a procedure is of high value when designing equipment and planning patient-specific care, as current dosimetry is given in guidelines by the equipment provider. From a modelling standpoint this poses a complex multi-physics problem coupling electro-magnetics and heat conduction. Added to this there are existence of multiple temperature sensitive properties of tissue, as well as physiological processes such as blood perfusion which are heavily influential in the overall temperature profile. Here, we create a two dimensional axisymmetric geometry of a probe embedded within a tissue material, solving the coupled electromagnetic and bioheat equation using the finite element method, utilizing hp discretisation and the NGSolve library. Temperature, electric field strength and energy absorption can be calculated and investigations into the effects of the highly temperature-sensitive properties of tissue on the distribution of temperature throughout ablation.

Introduction Leydig cells (LCs) are the primary source of testosterone (T) in men, and their dysfunction can lead to T deficiency. The growth and differentiation of LCs is influenced by the paracrine factors released by the testicular microenvironment (TME). Leptin is a 16-kDa protein produced primarily by adipose tissue and exerts a significant influence on hormonal regulation. In our recent study, we demonstrated that Leptin, a paracrine factor secreted by TME, is critical for Leydig stem cell (LSCs) differentiation and subsequent T production under the regulation of the desert hedgehog signaling pathway. Objective To gain an in-depth understanding of mechanisms that derive the Leptin secretion and decide the fate of LCs are still unclear and is the focus of the present study. Methods A total of 13 men with testicular failure underwent testis biopsies. The testicular cells were characterized using Flow cytometry, Immunostaining and RNA sequencing then the cells were also treated with Leptin in a dose dependent (0, 1, 10 ng/mL respectively) and time dependent (24, 48, 72, and 96 hours) manner. For the functional studies, UCSC genome browser was used to study the transcription factor genes (TFs) binding the leptin receptor (LEPR) promoter. The list of TFs was subjected to enrichment analysis. To evaluate the effects of shortlisted TFs on leptin-LEPR-DHH to induce EMT in LSCs, qPCR and western blotting were performed. For statistical analysis GraphPad Prism was used. For statistical analysis GraphPad Prism was used. All data were presented as the means ± SEM. All data were presented as the means ± SEM. Results UCSC Genome browser revealed over 50 TFs, which were subjected to enrichment analysis using ENRICHR. Output revealed TFs; SOX2, SOX8 and SOX9 that were enriched for mechanisms influencing hormonal regulation via hypothalamic pituitary gonadal axis. The role of these TFs has been well established in epithelial to mesenchymal transition in different cancer types, however their impact on LSC differentiation remained underexplored. RNA sequencing data from cells exposed to different concentrations of Leptin showed an inducing effect of Leptin on SOX2, SOX8, Sox9 respectively. This was confirmed at RNA as well as at protein levels. Consequently, upon inhibition of LEPR, the expression of SOX2, SOX8 and SOX9 was reduced as well. Moreover, leptin treatment in conjunction with SOX2, SOX8 and SOX9 was found to modulate epithelial to mesenchymal transition in LCs. Conclusions Our results demonstrate that Leptin-LEPR-DHH induced Leydig stem cells differentiation in testis is modulated by EMT inducing transcription factors which binds to LEPR. Further studies are ongoing to explore the potential therapeutic effects of leptin treatment in improving fertility. Financial Support: Supported by the Clinician Scientist Development award from American Cancer Society to RR and Research Scholar Award from American Urological Association to HA. J.M.H. is supported by NIH grants 1R01 HL137355, 1R01 HL107110, 1R01 HL134558, 5R01 CA136387, 5UM1 HL113460 and Soffer Family Foundation Disclosure No

Introduction Erectile Dysfunction (ED) affects 40% of men over 40 and every decade after increases the likelihood by 10% (1). Platelet Rich Plasma (PRP) is a restorative therapy rich in cytokines and growth factors that may reverse the underlying pathology of ED (2). Before PRP can be used effectively in clinic, it is necessary to understand if its components are similar among men with ED. Objective To determine whether growth factors vary among PRP of men with ED Methods Blood was obtained from three men with ED and one healthy man as a control. The Arthrex Angel system was used to extract their PRP. The sample was immediately stored in ice and kept in -80 C. Bradford assay was ran to ensure equal protein concentrations were used. The Abcam Human Growth Factor Antibody Array kit (ab134002) was used to simultaneously detect 41 human growth factors. Results Subject 1 was 43 years old with ED only. Subject 2 was 43 years old with obstructive sleep apnea, hyperlipidemia, and delayed sleep phase syndrome, and BMI 27.1. Subject 3 was 49 years old with hypertension, diabetes mellitus (not on insulin), asthma, high cholesterol, and BMI 32.4.The results of evaluating the 41 growth factor array showed that several growth factors had a fold change greater than 1.5 compared to the control. In PRP of adults with ED, PDGF-AA, PDGF- AB, and PDGF-AB had a greater than 1.5-fold change in at least 2 out of the 3 samples when compared to control. In FGF-6, GM-CSF, NT-3, TGF-α, TGF-β, and TGF-β2, there was a negative fold change of greater than 1.5 in at least 2 out of the 3 samples when compared to control. Conclusions Based on these results, the use of PRP as a therapeutic agent may need to be personalized based on individuals' age, comorbidities, or growth factor content. Individuals with underlying comorbidities may need different volumes of PRP. Our future ongoing research is focusing on particular growth factors within PRP that are responsible for improvement in erectile function, thus being able to personalize PRP therapy for me with ED. Citations: 1. Feldman HA, Goldstein I, Hatzichristou DG, Krane RJ, McKinlay JB. Impotence and its medical and psychosocial correlates: results of the Massachusetts Male Aging Study. J Urol. 1994;151(1):54-61. 2. Liu MC, Chang ML, Wang YC, Chen WH, Wu CC, Yeh SD. Revisiting the Regenerative Therapeutic Advances Towards Erectile Dysfunction. Cells. 2020;9(5). 3. Fang J, Huang X, Han X, Zheng Z, Hu C, Chen T, et al. Endothelial progenitor cells promote viability and nerve regenerative ability of mesenchymal stem cells through PDGF-BB/PDGFR-beta signaling. Aging (Albany NY). 2020;12(1):106-21. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus Pharmaceutical

Self supporting targets of 64Zn having thickness range (448 ± 27 – 1294 ± 40) μ g/cm 2 were fabricated from ∼ 50 mg of the target material. Vacuum evaporation by resistive heating technique was used to achieve the high evaporation yield. Use of a specially designed pinhole type crucible source minimized the solid angle of evaporation during the fabrication process. Thickness and purity of the fabricated targets were determined by techniques viz., α -transmission method, Rutherford Backscattering Spectrometry (RBS) and Energy Dispersive X-ray Analysis (EDXA). The fabricated targets contained negligible impurities. A target of thickness (513 ± 20) μ g/cm 2 , utilized for a fusion-evaporation study was found to be durable.

With increasing awareness about the highly communicable bacteria and viruses, medical professionals and associations are making it mandatory to use protective surgical gowns to barrier the spreading of various diseases like SARS, HIV, hepatitis, etc. These diseases, which may be caused by a variety of microorganisms, can pose significant risks to human life and health unless sufficient measures are taken. Are medical professionals really feeling safe with existing surgical gowns prevalent in the market? Do these gowns provide absolute barrier to viruses along with providing comfort? Can we rely on the performance or is there something else we need to understand and modify? This article reveals some facts pertaining to these questions and precisely highlights various issues related to the development, application, and quality evaluation of surgical gowns. It also deals with the types of gowns available, their classification, technology behind them, technical aspects, testing means, their limitations, and future course of action to satisfy the growing needs of our health care professionals.

Template assisted direct Growth of Bit Patterned Media (TG BPM) is a novel alternative approach of producing Bit Patterned Media (BPM). Unlike conventional BPM, TG BPM directly grows the magnetic bit patterns with oxide bit boundaries by co-depositing magnetic alloy and oxide materials onto an assisting template[1]. This assisting growth template contains pre-designed topographic and/or chemical features, which act as a growth guidance for the sequentially deposited multi under layers and magnetic/oxide layer.

The explosion of knowledge management systems (KMS) and the need for their wide accessibility and availability has created an urgent need for reassessing the security practices and policies in organizations. Security of these assets is a day-to-day job placing a tremendous cognitive load on information-technology (IT) professionals, which can make it almost impossible to manage the security aspects of KMS. Autonomic-computing systems are well suited to manage KMS, as they use high-level system objectives provided by administrators as the basis for managing the security of KMS. The authors model the self-protection and self-healing configuration attributes in autonomic systems through game-theoretic models. The proposed modeling approach progressively moves from a manual intervention-oriented security setup to an autonomic security setup. This allows the authors to compare and contrast the different approaches and provide insights on their applicability to different security environments. The authors find that moving to a partial autonomic system with self-healing mechanisms can provide a stable environment for securing enterprise knowledge assets and can reduce hacking. It is beneficial to implement an autonomic system when manual investigation costs are higher and/or when the volume of malicious traffic is very low. An autonomic approach is especially attractive when it is difficult to impose penalties on malicious users. Autonomic systems can be effective in securing organizational knowledge assets and in reducing the potential damage from malicious users

Mathematical models for the behavior of fractional-N phase-locked-loop frequency synthesizers (Frac-N) are presented. The models are intended for calculating rms phase error and determining spurs in the output of Frac-N. The models describe noise contributions due to the charge pump (CP), the phase frequency detector (PFD), the loop filter, the voltage control oscillator, and the delta-sigma modulator. Models are presented for the effects of static CP gain mismatch, CP dynamic mismatch and PFD reset delay mismatch. A simple analytic expression shows the level of /spl Delta//spl Sigma/ sequence noise caused by static CP current mismatch. We further show that un-equal rise time and fall time constants of the CP result in dynamic mismatch noise. Reset delay mismatch in PFD is shown to also contribute significantly to close-in phase noise. The model takes into account the reduction in CP thermal and flicker noise due to the changing duty cycle of Frac-N CP. Our model is therefore useful in characterizing the noise performance of Frac-N at the system-level, simplifying the design of fractional-N synthesizers and transmitters. Analytical and simulated results are compared and show good agreement with prior published data on Frac-N realizations.

Thirty-one cases of untreated `mild hypertension' and equal number of age and sex matched controls with `normal' blood pressure were evaluated by echocardiography. Patients with mild hypertension had significantly increased left ventricular mass index, concentric remodeling, and diastolic dysfunction. Thickness of right ventricular anterior wall, flow velocities across tricuspid and pulmonary valves were also significantly higher in hypertensives. Pulmonary flow acceleration time was significantly less in hypertensives. On multiple regression analysis, mitral valve `A' wave velocity alone correlated with systolic blood pressure. Other echocardiographic variables did not have any relation with blood pressure readings. Height, weight, body surface area and body mass index could also explain only around 50% of variability in echocardiographic parameters. Cardiac structure and functions in hypertensives are affected by factors other than blood pressure reading, body surface area or body mass index. Routine echocardiography can be useful in identifying those patients of mild hypertension who have disproportionate increase in left ventricular mass or disproportionate impairment of diastolic functions.

The Itanium processor is the first implementation of the IA-64 instruction set architecture (ISA). The design team optimized the processor to meet a wide range of requirements: high performance on Internet servers and workstations, support for 64-bit addressing, reliability for mission-critical applications, full IA-32 instruction set compatibility in hardware, and scalability across a range of operating systems and platforms. The processor employs EPIC (explicitly parallel instruction computing) design concepts for a tighter coupling between hardware and software. In this design style the hardware-software interface lets the software exploit all available compilation time information and efficiently deliver this information to the hardware. It addresses several fundamental performance bottlenecks in modern computers, such as memory latency, memory address disambiguation, and control flow dependencies.

Congenital infiltrating lipomatosis of the face (CIL-F) is a rare lipomatous lesion with diffuse fatty infiltration of tissues and hyperplasia of underlying bone. We report clinical and CT findings in an unusual case of CIL-F presenting with progressive hemifacial asymmetry, manifesting as severely restricted mouth opening owing to exophytic temporomandibular joint ankylosis. The role of imaging in diagnosis is presented with a review of the literature. Differential diagnosis of CIL-F and its exclusion as a cause of hemifacial hyperplasia are also discussed.

Constructing a high-resolution (HR) image from low-resolution (LR) image(s) has been a very active research topic recently with focus shifting from multi-frames to learning based single-frame super-resolution (SR). Multi-frame SR algorithms attempt the exact reconstruction of reality, but are limited to small magnification factors. Learning based SR algorithms learn the correspondences between LR and HR patches. Accurate replacements or revealing the exact underlying information is not guaranteed in many scenarios. In this paper we propose an alternate solution. We propose to capture images at right zoom such that it has just sufficient amount of information so that further resolution enhancements can be easily achieved using any off the shelf single-frame SR algorithm. This is true under the assumption that such a zoom factor is not very high, which is true for most man-made structures. The low-resolution image is divided into small patches and ideal resolution is predicted for every patch. The contextual information is incorporated using a Markov Random Field based prior. Training data is generated from high-quality images and can use any single-frame SR algorithm. Several constraints are proposed to minimize the extent of zoom-in. We validate the proposed approach on synthetic data and real world images to show the robustness.

Projector-camera systems are extensively used for various applications in computer vision, immersive environments, visual servoing, etc. Due to gaps between neighboring pixels on the projectorpsilas image plane and variations in scene depth, the image projected onto a scene shows pixelation and blurring artifacts. In certain vision and graphics applications, it is desirable that a high quality composition of the scene and the projected image, excluding the artifacts, is captured, while retaining the scene characteristics. Localization of projected pixels can also help in dense estimation of scene shape. In this paper, we address the problem of localizing each of the projected pixels from a captured scene and restoring the captured image so that the pixelation and blurring artifacts of the projector are removed.We improve the quality of the captured image further by virtualizing a high resolution projector. i.e., we modify the captured image as if the scene were illuminated with a high-resolution projector. We propose robust solutions to these problems and demonstrate their effectiveness on scenes of different complexities.

Super-resolution reconstruction algorithms assume the availability of exact registration and blur parameters. Inaccurate estimation of these parameters adversely affects the quality of the reconstructed image. However, traditional approaches for image registration are either sensitive to image degradations such as variations in blur, illumination and noise, or are limited in the class of image transformations that can be estimated. We propose an accurate registration algorithm that uses the local phase information, which is robust to the above degradations. We derive the theoretical error rate of the estimates in presence of non-ideal band-pass behavior of the filter and show that the error converges to zero over iterations. We also show the invariance of local phase to a class of blur kernels. Experimental results on images taken under varying conditions clearly demonstrates the robustness of our approach.

This paper presents a memory efficient version of set partitioning in hierarchical tree (SPIHT). The proposed coder termed as memory efficient SPIHT (MESH), uses a single re-usable list instead of three continuously growing linked lists as in conventional SPIHT. The list is re-initialized at the beginning of each bit-plane (coding pass) and is exhausted within that bit-plane itself. Another feature of the proposed coder is that it uses a single pass for each bit-plane by merging the sorting and refinement passes of conventional SPIHT together. The reinitialization of the list in each bit-plane makes the proposed coder inherently error resilient. The performance of the proposed coder is measured in terms of coding efficiency and the worst case memory requirements for list entries in each bit-plane. The performance comparison with SPIHT shows that the proposed algorithm results in 50-70% memory saving while retaining a coding efficiency comparable to SPIHT.

This paper illustrates how networking protocols can inadvertently exacerbate obstacles to providing real-time guarantees for distributed problem solving in wireless mobile and sensor networks. We analyze the effects of control packet timing on providing quality of service guarantees. Inappropriate timing of control packets gives rise to synchronizations that result in sharp increases and decreases in throughput with small changes in node speed. Such synchronizations can seriously jeopardize network performance with direct effect on real-time guarantees. This paper introduces these synchronizations, analyzes them and suggests ways to modify the control packet timing to overcome them. These analyses include investigating the role of buffering at the network layer and its impact on network throughput. We analyze these effects and evaluate our protocol enhancements through simulation studies.

The ability of a mobile ad hoc network (MANET) to provide adequate quality of service (QoS) is limited by the ability of the underlying routing protocol to provide consistent behavior despite the inherent dynamics of a mobile computing environment. In this paper we study three MANET routing protocols: OLSR, DSR and AODV, with an emphasis on the effect they have on various QoS metrics. We describe and analyze how the protocols differ in the mechanisms they use to select paths, detect broken links, and buffer messages during periods of link outage. The effects of these differences are quantified in terms of packet delivery ratio, end-to-end hop count, end-to-end latency, and mechanism overhead. We show that the proactive protocol, OLSR, builds paths with consistently lower hop counts than the reactive protocols, AODV and DSR, a fact that leads to a reduction in end-to-end latency that assists a QoS model in meeting timing requirements and improves global network performance. We further show the impact of broken link detection latency on the packet delivery ratio. A routing protocol that can not quickly recover from link breakage caused by mobility renders a QoS model incapable of meeting delivery requirements. Finally, we analyze the effect of mobility on the distribution of end-to-end latencies. Traditionally, reactive protocols are criticized for buffering during the building of routes, however we also study buffering phenomenon caused by the proactive mechanisms of OLSR.

This paper presents the exploration of novel mechanisms toward providing quality of service (QoS) guarantees in mobile ad hoc networks. We study mechanisms that provide differentiated services to packets of varying priority traffic flows. These mechanisms do not require any central coordination and do not depend on any specific protocols at the physical, MAC, or network layers. Nodes independently monitor the rates of the highest priority flows and signal corrective mechanisms when these rates fall outside of specified local bounds. Triggering conditions for network-wide corrective mechanisms are designed to trade-off rapid reactive response to local QoS violations with control packet overhead. A range of corrective mechanisms are explored that attempt to maintain reactive response while improving total network utilization, including resources consumed by lower priority traffic. We provide simulation results that demonstrate the effectiveness of monitoring, reactive triggering, and basic and advanced corrective mechanisms. We discuss the extension of these novel mechanisms to a complete QoS solution for mobile ad hoc networks.

The major determinant of a patient’s safety and outcome is the skill and judgment of the surgeon. While knowledge base and decision processing are evaluated during residency, technical skills—which are at the core of the profession—are not evaluated. Innovative state of the art simulation devices that train both surgical tasks and skills, without risk to patients, should allow for the detection and analysis of errors and “near misses”. Studies have validated the use of a sophisticated endoscopic sinus surgery simulator (ES3) for training residents on a procedural basis. Assessments are proceeding as to whether the integration of a comprehensive ES3 training programme into the residency curriculum will have long term effects on surgical performance and patient outcomes. Using various otolaryngology residencies, subjects are exposed to mentored training on the ES3 as well as to minimally invasive trainers such as the MIST-VR. Technical errors are identified and quantified on the simulator and intraoperatively. Through a web based database, individual performance can be compared against a national standard. An upgraded version of the ES3 will be developed which will support patient specific anatomical models. This advance will allow study of the effects of simulated rehearsal of patient specific procedures (mission rehearsal) on patient outcomes and surgical errors during the actual procedure. The information gained from these studies will help usher in the next generation of surgical simulators that are anticipated to have significant impact on patient safety.