Viable Tissue Research Articles

Abstract 5474: Biorepository yields potential for personalized medicine approach in pediatric brain cancer patients

Abstract Childhood brain cancer, with a 12% mortality rate, stands as the most lethal cancer among children. Out of the annual 4,000 diagnoses, 30% involve low-grade gliomas, with a troubling 10% progressing to lethal high-grade gliomas within 2 years. While surgery can often cure the remaining 90%, 50% necessitate adjuvant therapies like chemotherapy and molecular targeting, with radiation sparingly used. Alarmingly, 66% of survivors grapple with long-term health, mental, and cognitive issues due to treatment toxicity, emphasizing the urgent need for innovative biological therapies in pediatric glioma treatment.Our proposed strategy involves the acute analysis of freshly discarded tissue from brain cancer craniotomies, departing from the use of FFPE-preserved tissue, which may exhibit suboptimal quality. All patients provided informed consent, and by prospectively preserving viable tissue, we enable its utilization in ten or more subsequent analyses. These encompass cell-culture-based assays, viability assays, bulk, single cell, and single nucleus RNA sequencing, DNA sequencing, single-cell proteome analyses, flow cytometry, and immunofluorescence.Our findings highlight a direct correlation between surgical sample size and analysis feasibility. Bulk RNA sequencing proves valuable in acutely isolated surgical tissue and cryostored or flash-frozen tissue for identifying novel therapeutic targets in individual patients. Viable cells retrieved from cryopreserved tissue of the same patient allow ex vivo functional validation of therapeutic targets, eliminating the need for establishing patient-derived cell lines and xenografts, particularly challenging for low-grade gliomas. These findings establish the feasibility of a personalized bedside-to-bench approach, contingent on an adequate supply of surgical material.Expanding this approach to encompass all childhood brain cancer types, we plan to investigate using our comprehensive biorepository, housing viable tissue and single-cell suspensions from 122 childhood brain cancer patients. Among these, 51 cases yielded tissue for single-cell RNA-seq and viable cryoprotection. The most common tumor histology identified was low-grade glioma (32), primarily pilocytic astrocytoma (19), followed by embryonal (12) and ependymal tumors (4).In summary, we developed a standard operating procedure for analyzing fresh and cryopreserved surgical discards, identifying novel targets in childhood brain cancer tissue, and functionally validating them in patient-matched samples. This personalized medicine approach holds promise in circumventing the tedious development of patient-derived models. Citation Format: Anna Pronina, Emon Nasajpour, Caitlynn Tran, Troy Schouten, Eden Gibson, Jeffrey Nirschl, Laura Marie Prolo, Kelly Mahaney, Hannes Vogel, Claudia Katharina Petritsch. Biorepository yields potential for personalized medicine approach in pediatric brain cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5474.

Abstract 7550: KB-0742, an oral highly selective CDK9 inhibitor, demonstrates preclinical activity in transcription factor fusion driven adenoid cystic carcinoma patient-derived models

Abstract Adenoid Cystic Carcinoma (ACC) is an aggressive rare type of cancer of the secretory glands with no approved targeted therapy and high unmet clinical need. Deregulated transcription driven by MYB-NFIB or MYBL1-NFIB transcription factor (TF) fusions are a hallmark of ACC pathogenesis. More aggressive disease and resistance to therapy has been associated with co-mutation in NOTCH. Although direct targeting of oncogenic TF fusions has remained challenging, targeting of their activity via transcriptional cofactors has emerged as an attractive and clinically actionable strategy. Cyclin-dependent kinase 9 (CDK9) is a key potentiator of TF activity via its ability to act both as an upstream regulator of TF expression and a downstream cofactor. KB-0742 is a potent, selective, and orally bioavailable inhibitor of CDK9 with a long plasma half-life. KB-0742 is being studied in an ongoing Phase 1/2 study (NCT04718675) in advanced solid tumors including ACC. KB-0742 has demonstrated on-mechanism, single agent anti-tumor activity and a manageable safety profile in heavily pre-treated patients (0-11 prior lines of therapy) with transcriptionally addicted solid tumors. Here we present the preclinical rationale for targeting of oncogenic fusion TF activity in ACC. XenoSTART Patient-Derived Xenograft (XPDX) models were established from viable human tumor tissue for in vivo and ex vivo testing. Cell viability was assessed by CellTiter GLO reagent and phase imaging. Changes in MYB-fusion transcripts were assessed by RNA-Seq. Tumor Fragments (~70 mg) from ACCx5M1, ACCx6, ACCx9, ACCx11 models were implanted into athymic nude mice. Animals were treated with either vehicle (normal saline) or 60 mg/kg KB-0742 on a Q3D/week until tumors reached 2500 mm3 or up to 60 days post start of treatment. In a retrospective analysis using real world data, ACC has a low incidence of genomic instability and high rates of MYB fusions detectable by RNA-seq, making immunotherapy challenging but creating an opportunity for targeting MYB transcription. In primary MYB-fusion positive and NOTCH co-mutated patient-derived spheroid models, KB-0742 treatment induced strong antiproliferative activity and cytotoxicity. Importantly, KB-0742 demonstrated stronger cytotoxic effects when compared to historical control agents. In XPDXs, CDK9 inhibition with KB-0742 resulted in antiproliferative activity and stronger tumor growth inhibition in MYB-fusion positive and NOTCH co-mutated tumor models compared to MYB-fusion positive only models.These data demonstrate KB-0742 is effective in preclinical models of ACC suggesting KB-0742 may be a promising therapeutic option for ACC patients. KB-0742 is currently being evaluated in a phase 1/2 dose-escalation and cohort expansion trial in patients with ACC and other transcriptionally addicted tumors (NCT04718675). Citation Format: Michael R. McKeown, Luis A. Carvajal, Tressa R. Hood, Nicole S. Burr, Jeffrey Kaufman, Adam Boynton, Kameron Mori, Tessa DesRochers, Michael Wick, Charles Y. Lin, Jorge F. DiMartino. KB-0742, an oral highly selective CDK9 inhibitor, demonstrates preclinical activity in transcription factor fusion driven adenoid cystic carcinoma patient-derived models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7550.

Sinonasal Tumors Masquerading as Invasive Fungal Sinusitis (IFS).

Fungal tissue invasion in the setting of sinonasal malignancy has been rarely described in the literature. Only a handful of studies have discussed cases of suspected chronic and acute IFS (CIFS and AIFS, respectively), having an underlying undifferentiated sinonasal carcinoma, sinonasal teratocarcinosarcoma, and NK/T-cell lymphoma. Here, we describe 3 cases of carcinoma mimicking IFS from a single institution. Each of our patients presented with sinonasal complaints as an outpatient in the setting of immunosuppression. Intranasal biopsies consistently were predominated by necrotic debris, with and without fungal elements, ultimately leading to a delay of oncologic care. The final pathologies included NK/T-cell lymphoma and SNEC. All patients were followed by radiation and chemotherapy, with 1 case of mortality. We aim to emphasize the importance of obtaining viable tissue as pathology specimens as the presence of necrosis with fungal elements may limit the diagnosis and ultimately delay the care of an underlying sinonasal carcinoma.

Long-Term Outcomes of Ascending Sliding Arch Aortoplasty.

Background: Coarctation of the aorta can be associated with significant hypoplasia of the aortic arch. In contrast to patch aortoplasty, ascending sliding arch aortoplasty uses viable autologous tissue for potential growth in children. We reviewed the mid- to long-term outcomes of this technique. Methods: Between 2002 and 2023, 28 patients underwent ascending sliding arch aortoplasty for the patients with coarctation of the aorta (n = 22) and interrupted aortic arch (n = 2). Four patients underwent previous surgical coarctation repair at other institutions. The median patient age and body weight were 28.5 months (3 weeks to 15.6 years) and 13.4 kg (3.7-70 kg), respectively. Results: Although one patient had a recurrent nerve injury postoperatively, there were no other major morbidities or mortalities. The last follow-up echocardiography demonstrated that the mean peak velocity improved from 3.9 ± 0.6 to 0.9 ± 0.8 m/s, and the pressure gradient improved from 63.6 ± 21.5 to 7.1 ± 7.7 mm Hg. The postoperative diameters of the ascending aorta, proximal arch, distal arch, and isthmus all increased significantly. The mean postoperative length of stay was 5.9 ± 2.1 days, and the median follow-up time was 7.3 years (10 days to 20.5 years). No reoperation or catheterization-based intervention was performed for residual coarctation. Conclusions: Ascending sliding arch aortoplasty is safe and effective for treating coarctation of the aorta with aortic arch hypoplasia. This technique is applicable for children ranging in size from neonates to older children (or adolescents), recurrent coarctation cases, and provides complete relief of narrowing by utilizing viable native aortic tissue.

Femtosecond Laser Cutting of Human Crystalline Lens Capsule and Decellularization for Corneal Endothelial Bioengineering.

The bioengineering of corneal endothelial grafts consists of seeding in vitro cultured corneal endothelial cells onto a thin, transparent, biocompatible, and sufficiently robust carrier which can withstand surgical manipulations. This is one of the most realistic alternatives to donor corneas, which are in chronic global shortage. The anterior capsule of the crystalline lens has already been identified as one of the best possible carriers, but its challenging manual preparation has limited its use. In this study, we describe a femtosecond laser cutting process of the anterior capsule of whole lenses in order to obtain capsule discs of 8 mm diameter, similar to conventional endothelial grafts. Circular marks made on the periphery of the disc indicate its orientation. Immersion in water for 3 days is sufficient to completely remove the lens epithelial cells and to enable the seeding of corneal endothelial cells, which remain viable after 27 days of culture. Therefore, this method provides a transparent, decellularized disc ready to form viable tissue engineered endothelial grafts.

Melanocortin Derivatives Induced Vascularization and Neuroglial Proliferation in the Rat Brain under Conditions of Cerebral Ischemia.

Stroke remains the second leading cause of death worldwide. The development of new therapeutic agents focused on restoring vascular function and neuroprotection of viable tissues is required. In this study the neuroprotective activity of melanocortin-like ACTH(4-7)PGP and ACTH(6-9)PGP peptides was investigated in rat brain at 24 h after transient middle cerebral artery occlusion (tMCAO). The severity of ischemic damage, changes in the proliferative activity of neuroglial cells and vascularization of rat brain tissue were analyzed. The administration of peptides resulted in a significant increase in the volume density of neurons in the perifocal zone of infarction compared to rats subjected to ischemia and receiving saline. Immunohistochemical analysis of the proliferative activity of neuroglia cells using PCNA antibodies showed a significant increase in the number of proliferating cells in the penumbra and in the intact cerebral cortex of rats receiving peptide treatment. The effect of peptides on vascularization was examined using CD31 antibodies under tMCAO conditions, revealing a significant increase in the volume density of vessels and their sizes in the penumbra after administration of ACTH(4-7)PGP and ACTH(6-9)PGP. These findings confirm the neuroprotective effect of peptides due to the activation of neuroglia proliferation and the enhancement of collateral blood flow.

(159) Factors and Management Strategies for Addressing Reduced Glans Engorgement Following Inflatable Penile Prosthesis: A Scoping Review

Abstract Introduction Inflatable penile prosthesis (IPP) provides a reliable stiffness along the length of the penile shaft, yet patients may express dissatisfaction as it may still differ from their recollection of their previously normal erections. A common complaint is reduced engorgement of the glans penis, which can manifest as subjective reduction in penile length or the feeling of a cold insensitive glans. Patient factors often ensure a sound justification for this complaint, namely vasculogenic erectile dysfunction (ED) or their corporo-glandular morphology despite a well-sized and well-positioned implant. Objective In this study, we sought to identify factors that may contribute to reduced glans engorgement and possible management strategies. Methods Intervention: Any method of treating or avoiding reduced glans engorgement post-IPP. Population: Men undergoing IPP insertion or men with glans complaints post-operatively Inclusion Criteria: Articles published in English with full text available and objective outcome measures Search Strategy: Literature search was performed in June 2023 using Medline, PubMed, Embase and Scopus. PubMed search included both “Mesh” protocols and “free text” to capture the correct study population and intervention utilized. Review Type & Analysis: Although an initial systematic model was considered, a scoping model was ultimately selected due to the limited literature and inconsistent outcome measures. This approach aimed to identify gaps in knowledge and provide an overview of the existing literature, thus informing future studies in this area. The variability in outcome measures across studies prevented higher-powered analyses from being conducted. Results Initial search strategy retrieved 98 non-duplicate studies with 40 being selected for initial screening after applying inclusion criteria. Ultimately, a total of 7 articles were included and categorized by theme: Cavernous-sparing technique (1–3); Pharmacotherapy for post-IPP glans complaints (4–7) (Table 1 & 2). The cavernous-sparing technique described aims to limit disruption of cavernous tissue around the implanted cylinder. This was achieved by omitting dilatation and using Furlow directly for measurements, or by intraoperative intracavernosal (IC) prostaglandin followed by dilatation only with size 7 or 8 Hegar alone. In all studies that compared penile dimensions, girth was significantly greater in the cavernous-sparing group. A higher proportion of men in this group also had detectable cavernous arteries, thicker residual corporal tissue, better residual tumescence post-IPP and demonstrated significant increases in both PSV and EDV post-PDE5i, compared to no PDE5i. Men who had standard dilatation also responded to post-IPP PDE5i but fewer had identifiable cavernous artery and had significant increase in only PSV, compared to no PDE5i. Other articles relating to post-IPP pharmacotherapy unanimously demonstrated that post-IPP glans engorgement was improved. This equated to reported statistically improved satisfaction when compared to the IPP alone groups, as measured by IIEF and EDITS questionnaires. Post-IPP PDE5i were better tolerated than intraurethral alprostadil, which was associated with discomfort. Conclusions Post-IPP pharmacotherapy appears to enhance residual erectile tissue engorgement. It can be used as an adjunctive after-care measure. Cavernous-sparing techniques show the importance of viable residual tissue for improved prosthetic erections. More comprehensive studies are needed to enhance patient experience and explore the microanatomical and hemodynamic role of residual tissue post-IPP. Disclosure No.

TOXICITY TESTING OF CHITOSAN OF PAPUYU FISH SCALE (Anabas testudineus) TOWARD BHK-21 FIBROBLAST CELLS

       Background: Pulp capping is a treatment that aims to maintain the viability of the pulp tissue. Calcium hydroxide (Ca(OH)2 is a material that is often used for pulp capping procedures. However, this material does not provide a good adaptation to dentin. Therefore, an alternative material that is more biocompatible is needed, one of which is by using papuyu fish scale chitosan. papuyu fish as an alternative material must be proven to have no toxic effects. Purpose: to analyze whether the chitosan of papuyu fish scales (Anabas testudineus) is toxic to BHK-21 fibroblast cells using the MTT assay method. Methods: This study was a pure experimental study with a post-test-only design with a control group design consisting of 12 groups. The group given the chitosan treatment of papuyu fish scales consisted of 10 groups with concentrations of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% and 100% with 2 control groups namely control cells. and media control. Results: The results showed that the chitosan of papuyu fish scales did not have a toxic effect on BHK-21 fibroblast cells because the percentage value of cell viability of the entire treatment group was > 60% and the IC50 value was > 0.1%, which was 5.405 %. Conclusion: There was no toxic effect after administration of papuyu fish scales chitosan (Anabas testudineus) with concentrations of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% on cells BHK-21 fibroblasts. Keywords: BHK-21 fibroblast cells, Papuyu fish scale chitosan, Toxicity test

Traumatic scrotal sac hematoma after a fall injury

Introduction and Importance: Traumatic scrotal sac hematoma is an uncommon medical condition following a fall injury. Symptoms include swelling, pain, and discoloration. Accurate and immediate diagnosis is crucial to avoid complications and prevent testis loss, abscess formation, erectile dysfunction, and infection. This case highlights the importance of prompt evaluation and treatment to prevent complications and ensure optimal patient outcomes. Case Presentation: A 56-year-old man without prior medical history presented to the emergency department with scrotal pain and swelling 10 h after a traumatic event. The patient underwent a series of examinations, including radiological studies, before undergoing scrotal exploration and hematoma evacuation under subarachnoid block anesthesia. As of now, the patient has not experienced any complications that have impacted his quality of life. Clinical Discussion: Scrotal sac hematoma is formed following a blunt trauma when a significant force is transferred during contact with a solid object, impact from sports equipment, road traffic accidents, or falls from a height. If early exploration is delayed, then it can lead to complications like testicular atrophy, abscess formation, and infection. Hence, prompt diagnosis with clinical and radiological evaluation followed by immediate surgical intervention can result in better outcomes. Conclusion: Scrotal sac hematoma from blunt trauma is rare but can cause significant morbidity and psychological distress. Early surgical intervention is often necessary, especially if a testicle has ruptured, to salvage viable testicular tissue and control bleeding, resolve pain faster, shorten hospital stays, and improve the quality of life for patients.

Unmasking the unexpected: Testicular tumour uncovered as a result of scrotal trauma

AbstractWe present a case of a 17‐year‐old male with a testicular tumour diagnosed after trauma. A 17‐year‐old male patient came to our clinic with swelling in the right hemiscrotum. The patient had a history of right scrotal blunt trauma 3 weeks before the admission and had no history of urological surgery or chronic disease. The patient doesn't use tobacco, alcohol, or illicit drugs. Genitourinary system examination revealed swelling and tenderness on palpation in the right hemiscrotum. The right testicle was not palpable. The left hemiscrotum was normal. The left testicle had a normal size and shape. Testicular tumour markers were high (AFP:196 μg/L (N: 0–8 μg/L), b‐HCG: 916 mIU/mL (N: <10 mIU/mL), LDH: 364 U/L (N: 0–265 U/L). Scrotal ultrasonography and magnetic resonance imaging revealed that there were areas of 9 × 9 × 11 cm in size of the right scrotal region without viable testis tissue, hematoma area, and areas compatible with necrosis. These findings were related to intratesticular rupture due to trauma. Abdominal tomography revealed lymphadenopathies of approximately 6 × 3 cm in the anterior paracaval area and approximately 4 × 3 cm in the anterior aspect of the psoas muscle. We performed right radical inguinal orchiectomy on the patient. Testicular tumour markers were still high on the 8th postoperative day (AFP:120 μg/L (N: 0–8 μg/L), bHCG: 680 mIU/mL (N: <10 mIU/mL), LDH: 200 U/L (N: 0–265 U/L)). The testicular tumour was diagnosed as mixed germ cell tumour (50% Yolk Sac, 30% Embryonal Carcinoma, 20% Teratoma). The tumour diameter was 9 cm, surgical margins were intact, there was rete testis invasion, and no tumour was detected in the tunica vaginalis. No tumour invasion was seen in the soft tissue around the spermatic cord (pT2). The patient received 3 cycles of chemotherapy with bleomycin, etoposide, and cisplatin. We suggest that patients presenting with scrotal trauma should be carefully evaluated for testicular malignancy, and if malignancy is suspected, radical inguinal orchiectomy should be preferred for the operation.

Cardiac and perivascular myofibroblasts, matrifibrocytes, and immune fibrocytes in hypertension; commonalities and differences with myocardial infarction and other cardiovascular diseases.

Hypertension is a major cause of cardiovascular diseases such as myocardial infarction and stroke. Cardiovascular fibrosis occurs with hypertension and contributes to vascular resistance, aortic stiffness, and cardiac hypertrophy. However, the molecular mechanisms leading to the fibroblast activation in hypertension remain largely unknown. There are two types of fibrosis: replacement fibrosis and reactive fibrosis. Replacement fibrosis occurs in response to the loss of viable tissue to form a scar. Reactive fibrosis occurs in response to increase in mechanical and neurohormonal stress. Although both types of fibrosis are considered as adaptive processes, they become mal-adaptive when the tissue loss is too large, or the stress persists. Myofibroblasts represent a subpopulation of activated fibroblasts that have gained contractile function to promote wound healing. Therefore, myofibroblasts are a critical cell type that promotes replacement fibrosis. Although myofibroblasts were recognized as the fibroblasts participating in reactive fibrosis, recent experimental evidence indicated there are distinct fibroblast populations in cardiovascular reactive fibrosis. Accordingly, we will discuss the updated definition of fibroblast subpopulations, the regulatory mechanisms, and their potential roles in cardiovascular pathophysiology utilizing new knowledge from various lineage tracing and single-cell RNA sequencing studies. Among the fibroblast subpopulations, we will highlight the novel roles of matrifibrocytes and immune fibrocytes in cardiovascular fibrosis including experimental models of hypertension, pressure overload, myocardial infarction, atherosclerosis, aortic aneurysm, and nephrosclerosis. Exploration into the molecular mechanisms involved in the differentiation and activation of those fibroblast subpopulations may lead to novel treatments for end-organ damage associated with hypertension and other cardiovascular diseases.

Detection of lymphoproliferative disease virus in Iowa Wild Turkeys (Meleagris gallopavo): Comparison of two sections of the proviral genome.

An accurate diagnostic test is an essential aspect of successfully monitoring and managing wildlife diseases. Lymphoproliferative Disease Virus (LPDV) is an avian retrovirus that was first identified in domestic turkeys in Europe and was first reported in a Wild Turkey (Meleagris gallopavo) in the United States in 2009. It has since been found to be widely distributed throughout North America. The majority of studies have utilized bone marrow and PCR primers targeting a 413-nucleotide sequence of the gag gene of the provirus to detect infection. While prior studies have evaluated the viability of other tissues for LPDV detection (whole blood, spleen, liver, cloacal swabs) none to date have studied differences in detection rates when utilizing different genomic regions of the provirus. This study examined the effectiveness of another section of the provirus, a 335-nucleotide sequence starting in the U3 region of the LTR (Long Terminal Repeat) and extending into the Matrix of the gag region (henceforth LTR), for detecting LPDV. Bone marrow samples from hunter-harvested Wild Turkeys (n = 925) were tested for LPDV with the gag gene and a subset (n = 417) including both those testing positive and those where LPDV was not detected was re-tested with LTR. The positive percent agreement (PPA) was 97.1% (68 of 70 gag positive samples tested positive with LTR) while the negative percent agreement (NPA) was only 68.0% (236 of 347 gag negative samples tested negative with LTR). Cohen's Kappa (κ = 0.402, Z = 10.26, p<0.0001) and the McNemar test (OR = 55.5, p<0.0001) indicated weak agreement between the two gene regions. We found that in Iowa Wild Turkeys use of the LTR region identified LPDV in many samples in which we failed to detect LPDV using the gag region and that LTR may be more appropriate for LPDV surveillance and monitoring. However, neither region of the provirus resulted in perfect detection and additional work is necessary to determine if LTR is more reliable in other geographic regions where LPDV occurs.

(311) Reduced Glans Engorgement After Inflatable Penile Prosthesis – A Scoping Review of Factors Involved and Management Strategies

Abstract Introduction Inflatable penile prosthesis (IPP) provides a reliable stiffness along the length of the penile shaft, yet patients may express dissatisfaction as it may still differ from their recollection of their previously normal erections. A common complaint is reduced engorgement of the glans penis, which can manifest as subjective reduction in penile length or the feeling of a cold insensitive glans. Patient factors often ensure a sound justification for this complaint, namely vasculogenic erectile dysfunction (ED) or their corporo-glandular morphology despite a well-sized and well-positioned implant. Objective To identify factors that may contribute to reduced glans engorgement and possible management strategies. Methods Intervention: Any method of treating or avoiding reduced glans engorgement post-IPP. Population: Men undergoing IPP insertion or men with glans complaints post-operatively Inclusion Criteria: Articles published in English with full text available and objective outcome measures Search Strategy: Literature search was performed in June 2023 using Medline, PubMed, Embase and Scopus. PubMed search included both “Mesh” protocols and “free text” to capture the correct study population and intervention utilized. Review Type &amp; Analysis: Although an initial systematic model was considered, a scoping model was ultimately selected due to the limited literature and inconsistent outcome measures. This approach aimed to identify gaps in knowledge and provide an overview of the existing literature, thus informing future studies in this area. The variability in outcome measures across studies prevented higher-powered analyses from being conducted. Results Initial search strategy retrieved 98 non-duplicate studies with 40 being selected for initial screening after applying inclusion criteria. Ultimately, a total of 7 articles were included and categorized by theme: Cavernous-sparing technique (1-3); Pharmacotherapy for post-IPP glans complaints (4-7) (Table 1 &amp; 2). The cavernous-sparing technique described aims to limit disruption of cavernous tissue around the implanted cylinder. This was achieved by omitting dilatation and using Furlow directly for measurements, or by intraoperative intracavernosal (IC) prostaglandin followed by dilatation only with size 7 or 8 Hegar alone. In all studies that compared penile dimensions, girth was significantly greater in the cavernous-sparing group. A higher proportion of men in this group also had detectable cavernous arteries, thicker residual corporal tissue, better residual tumescence post-IPP and demonstrated significant increases in both PSV and EDV post-PDE5i, compared to no PDE5i. Men who had standard dilatation also responded to post-IPP PDE5i but fewer had identifiable cavernous artery and had significant increase in only PSV, compared to no PDE5i. Other articles relating to post-IPP pharmacotherapy unanimously demonstrated that post-IPP glans engorgement was improved. This equated to reported statistically improved satisfaction when compared to the IPP alone groups, as measured by IIEF and EDITS questionnaires. Post-IPP PDE5i were better tolerated than intraurethral alprostadil, which was associated with discomfort. Conclusions Post-IPP pharmacotherapy appears to enhance residual erectile tissue engorgement. It can be used as an adjunctive after-care measure. Cavernous-sparing techniques show the importance of viable residual tissue for improved prosthetic erections. More comprehensive studies are needed to enhance patient experience and explore the microanatomical and hemodynamic role of residual tissue post-IPP. Disclosure No.

Brush Sign on pre-treatment imaging is associated with good functional outcome in stroke patients treated with mechanical thrombectomy: a prospective monocentric study.

BackgroundThe Brush Sign (BrS) is a radiological biomarker (MRI) showing signal decrease of subependymal and deep medullary veins on paramagnetic-sensitive magnetic resonance sequences. Previous studies have shown controversial results regarding the prognostic value of BrS. We aimed to assess whether BrS on T2*-weighted sequences could predict functional prognosis in patients treated with mechanical thrombectomy (MT). MethodsWe included all consecutive patients with large artery occlusion related stroke in anterior circulation treated with MT between February 2020 and August 2022 at Reims University Hospital. Multivariable logistic regression models were used to investigate factors associated with BrS and its impact on outcomes. ResultsOf the 327 included patients, 124 (37,9 %) had a BrS on baseline MRI. Mean age was 72 ± 16 years and 184 (56,2%) were female. In univariate analysis, BrS was associated with a younger age (67 vs 74; p<0.001), a higher NIHSS score (16(10–20) vs 13(8–19); p=0.047) history of diabetes (15.3% vs 26.1%; p=0.022) and a shorter onset to MRI time (145.5 (111.3-188.5) vs 162 (126-220) p=0.008). In multivariate analyses, patients with a BrS were younger (OR:0.970 (0.951 – 0.989)), tend to have a higher NIHSS score at baseline (OR:1.046 (1.000 – 1.094) and were less likely to have diabetes (OR: 0.433; 0.214 – 0.879). The presence of BrS was independently associated with functional independence (OR: 2.234(1.158 – 4,505) at 3 months but not with mortality nor with symptomatic intracerebral hemorrhage. ConclusionBrS on pre-treatment imaging could be considered as a biomarker of physiological adaptation to cerebral ischemia, allowing prolonged viability of brain tissue and might participate in the therapeutic decision.

Wound Debridement in Pyoderma Gangrenosum.

Wound debridement improves healing in a variety of acute and chronic ulcers. However, there is concern that debridement may trigger pathergy and worsen pyoderma gangrenosum (PG). To determine whether published evidence supports conservative wound debridement for PG. The authors reviewed the literature published in MEDLINE through January 2023 using the search germs "pyoderma gangrenosum" and "debridement." Articles reporting sharp surgical debridement or maggot debridement for PG were included in the review. The authors also searched the reference sections of the reviewed articles for additional reports on debridement for PG. Clinical data regarding patient status, procedures performed, and patient outcomes were extracted from the selected articles. There are multiple reports of uncontrolled, active-phase PG wounds worsening after aggressive excisional debridement of viable inflamed tissues. In contrast, there is no evidence indicating that conservative debridement of nonviable necrotic tissue worsens PG wounds, regardless of the disease activity. There are multiple reports of successful debridement and surgical grafting for PG in remission. There is no evidence in favor of or against using conservative debridement of nonviable necrotic tissue for a PG wound. Therefore, it should not be considered contraindicated, even in the active phase of the disease.

Novel Noninvasive Hybrid Flap Preconditioning (HFP) Surpasses Surgical Delay in the Murine Model.

Ischemic necrosis in the distal portion of the flap is a challenging complication in plastic surgery. We hypothesize a novel hybrid flap preconditioning (HFP) device combining foam-mediated external suction and non-surgical delay can promote skin flap survival better than surgical delay. Twenty-eight mouse were divided into four groups. Control group: a 4*1.5 dorsal flap was made with no preconditioning. Surgical delay (SD) group: surgical delay was made 7 days before flap elevation. Foam-mediated external suction (FMES) group: foam-mediated external suction at -100mmHg was employed 5 hours per day for 6 days, and the flap was elevated on the seventh day. Hybrid Flap Preconditioning (HFP) group: silicone strips was applied along the contour of the foam interface. Same negative pressure protocol was used as the FMES group. Seven days after flap elevation, macroscopic, histologic, and Western blot analyses were performed. The flap survival rate was 46.25% (8.12%) in the control group, 68.72% (7.00%) in the SD group, 57.03% (8.17%) in the FMES group and 80.66% (3.27%) in the HFP group. Immunohistologic analysis of CD 31+ cell in distal end of viable tissue procured seven days after flap elevation showed a significantly higher angiogenesis in SD group and HFP group. Western Blot results showed an increased expression of VEGF in SD group and HFP group. We have developed and fabricated a novel hybrid flap precondition (HFP) device combining foam-mediated external suction and non-surgical delay. The concept of HFP is proved to promote flap survival better than surgical delay.

Success of Endoscopic Laser Cyclophotocoagulation vs Repeat Transscleral Treatment after Prior Transscleral Cycloablation.

To compare the efficacy of endoscopic cyclophotocoagulation (ECP) vs repeat transscleral cyclophotocoagulation (TCP) in eyes with persistent glaucoma despite prior treatment with TCP. This was a retrospective chart review of glaucoma patients at the American University of Beirut Medical Center over 10 years who underwent ECP or repeat TCP. We reported qualified and complete success; success was defined as postoperative intraocular pressure (IOP) ≤21 mm Hg, with (qualified) or without medications (complete) and without procedure-related complications. This study included 23 eyes of 21 patients with various forms of uncontrolled glaucoma who had failed TCP. A total of 13 eyes of 12 patients underwent ECP with a mean age of 39.9 ± 23.2 years, and 10 eyes of nine patients underwent repeat TCP with a mean age of 27.2 ± 22.6 years. A significant decrease in IOP was observed from 38.5 ± 7.9 mm Hg preoperatively to 25.2 ± 8.8 mm Hg postrepeat TCP (p = 0.006) and from 33.0 ± 9.5 to 12.8 ± 3.9 mm Hg post-ECP (p < 0.001), noted at a mean follow-up time of 39.2 ± 44.4 and 41.5 ± 37.4 months, respectively. The mean number of antiglaucoma medications decreased in the two groups (from 3.8 ± 1.0 preoperatively to 1.8 ± 0.9 postoperatively for ECP and from 3.5 ± 1.3 to 3.1 ± 0.9 postoperatively for TCP); however, the drop was only statistically significant post-ECP. Qualified success was significantly higher after ECP vs repeat TCP (91.7 vs 40%, respectively). Complete success was achieved only in 1/12 (8.3%) eyes in the ECP group. Endoscopic cyclophotocoagulation (ECP) performed in glaucomatous eyes previously treated with transscleral cycloablation provided more IOP control as compared to repeat TCP by directly treating viable tissue in previously skipped ciliary processes and in between processes. In glaucomatous eyes previously treated with transscleral cycloablation, ECP attained better IOP control than repeat transscleral cycloablation. Al-Haddad C, Barikian A, Moussawi ZE, et al. Success of Endoscopic Laser Cyclophotocoagulation vs Repeat Transscleral Treatment after Prior Transscleral Cycloablation. J Curr Glaucoma Pract 2023;17(4):191-196.

Abstract C050: The unique metabolic signatures of keratin-17 expressing pancreatic ductal adenocarcinomas

Abstract Background: Overexpression of Keratin 17 (K17) is associated with upregulation of de novo pyrimidine synthesis, which is linked to resistance to gemcitabine (dC analog) and 5-fluorouracil (dT analog) in pancreatic ductal adenocarcinomas (PDAC). The metabolic rewiring associated with K17 in upstream pathways has not been defined, and the spatial relationships between K17 overexpression and the metabolome are unknown. Understanding these relationships may elucidate the roles of K17 in drug resistance and metabolic phenotypes of PDACs. We test the hypothesis that K17 is spatially linked to alterations of various metabolites, and that there exists an association with the upregulation of upstream pathways to de novo pyrimidine synthesis. Methods: KPC cells transduced to express wtK17 versus an empty vector were orthotopically implanted into the head of the pancreas of mice. Mice were administered 5-FU at 50 mg/kg twice a week. To evaluate the metabolic profile in situ, MALDI-MSI (Matrix-assisted laser desorption/ionization mass spectrometry imaging) was performed on snap-frozen tumor specimens. Following removal of the MALDI matrix, immunohistochemistry (IHC) was performed on the same sections for K17 to define elevated or low expressing regions. ROIs were selected within the MALDI-MSI for High K17, Low K17, and empty vector viable tissue. Principal component analysis was then performed on the mass spectra of these regions to provide unbiased pattern recognition. Results: The principal component analysis segregated K17-positive ROIs from empty vector (EV) ROIs, revealing that K17-positive tissue has a unique metabolic signature compared to EV tissue. Metabolites with loadings outside the 95th percentiles in the 5-component PCA included UMP, UDP, orotate, sedoheptulose 7-phosphate, glucose 6-phosphate, glutamate, malate, phosphoglycerate, and lactate. Conclusion: These metabolites, therefore, distinguish between the EV and K17-positive clusters. Further hierarchical clustering will be performed to provide a more comprehensive evaluation of the rewired pathways in pancreatic cancer. Citation Format: Mahmoud M. Salem, Michael Horowitz, Lyanne Oblein, Chun-Hao Pan, Natalia Marchenko, Luisa Escobar-Hoyos, Bo Chen, Kenneth Shroyer. The unique metabolic signatures of keratin-17 expressing pancreatic ductal adenocarcinomas [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr C050.

Danger Zone for Paramedian Forehead Flap Elevation: Maximizing Flap Length and Viability

The supratrochlear artery (STA) demonstrates anatomical variability that impacts facial reconstruction with a paramedian forehead flap. STA branching patterns and the distance to the midline have been reported, but the STA pedicle has not been characterized. Our aim was to triangulate the STA pedicle relative to known anatomical landmarks and identify a danger zone to aid surgeons in creating viable tissue flaps. The upper facial region was dissected bilaterally on 38 cadaveric donors. Measurements from the supraorbital neurovascular bundle, orbital rim, and medial canthus to the STA pedicle were collected. Data were tallied and statistically analyzed. Measurement means, range, and standard deviations were calculated; no significant differences were found in the laterality of the measurements (p &gt; 0.05). Statistically significant, sex-based differences were identified for all measurements collected among male and female donors. This study characterizes a surgical danger zone for the STA pedicle specific to a paramedian forehead flap and identifies important differences within this danger zone among male versus female donors that surgeons should consider to prevent pedicle violation and enhance surgical success while maximizing flap length and mobility.

Osteogenic-angiogenic coupled response of cobalt-containing mesoporous bioactive glasses in vivo

The incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of interest for bone regeneration purposes. In the present work, we have designed a set of bioactive mesoporous glasses SiO2-CaO-P2O5-CoO (Co-MBGs) with different amounts of cobalt. The physicochemical changes introduced by the Co2+ ion, the in vitro effects of Co-MBGs on preosteoblasts and endothelial cells and their in vivo behaviour using them as bone grafts in a sheep model were studied. The results show that Co2+ ions neither destroy mesoporous ordering nor inhibit in vitro bioactive behaviour, exerting a dual role as network former and modifier for CoO concentrations above 3 % mol. On the other hand, the activity of Co-MBGs on MC3T3-E1 preosteoblasts and HUVEC vascular endothelial cells is dependent on the concentration of CoO present in the glass. For low Co-MBGs concentrations (1mg/ml) cell viability is not affected, while the expression of osteogenic (ALP, RUNX2 and OC) and angiogenic (VEGF) genes is stimulated. For Co-MBGs concentration of 5 mg/ml, cell viability decreases as a function of the CoO content. In vivo studies show that the incorporation of Co2+ ions to the MBGs improves the bone regeneration activity of these materials, despite the deleterious effect that this ion has on bone-forming cells for any of the Co-MBG compositions studied. This contradictory effect is explained by the marked increase in angiogenesis that takes place inside the bone defect, leading to an angiogenesis-osteogenesis coupling that compensates for the partial decrease in osteoblast cells. Statement of significanceThe development of new bone grafts implies to address the need for osteogenesis-angiogenesis coupling that allows bone regeneration with viable tissue in the long term. In this sense the incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of great interest in this field. Due to the potential cytotoxic effect of cobalt ions, there is an important controversy regarding the suitability of their incorporation in bone grafts. In this work, we address this controversy after the implantation of cobalt-doped mesoporous bioactive glasses in a sheep model. The incorporation of cobalt ions in bioactive glasses improves the bone regeneration ability of these bone grafts, due to enhancement of the angiogenesis-osteogenesis coupling.