Fresh Specimens Research Articles

Observations on the anatomy of the funiculus spermaticus and the testis in Brachyteles sp. (SPIX, 1823) primates - Atelidae.

Analysing the literature about the reproductive anatomy on New World Primates, one can see the need of standardisation on the description methods and, more importantly, the lack of detailed information. The problem is aggravated by the difficult access to specimens of the endangered species such as Brachyteles sp. This paper's objective was to extend knowledge on the male reproductive anatomy of these primates. The testis and funiculus spermaticus of Brachyteles are described in detail. We utilised one individual of Brachyteles arachnoides, two hybrids (B. arachnoides × Brachyteles hypoxanthus) and photographs of the testis of a third fresh specimen of a hybrid individual. The septum formed by tunica dartos adheres to the testis and separates the scrotum in two testicular cavities. Passed the spermatic cord, the external spermatic fascia continues with the tunica dartos, covering the cranial half of the testis. The fascia cremasterica divides itself in bundles of fibres and forms loops around the testicles creating a sac like structure that seems to be unique among Neotropical Primates. The appendix testis is described for the first time in platyrrhini. It is presented as a sessile structure in the extremitas capitata of the testis. Previous literature on the reproductive anatomy of platyrrhini is limited. Despite that, it can be said that the large size and volume of the testis, and complex structure of the cremaster, could consist of a heat loss reduction strategy in cold and humid forest environments. Except for the larger size of the testicles in hybrid individuals, no other significant morphological differences were found between B. arachnoides and hybrids (B. arachnoides × B. hypoxanthus).

Evaluation of microRNA expression profiles in human sperm frozen using permeable cryoprotectant-free droplet vitrification and conventional methods.

For sperm cryopreservation, the conventional method, which requires glycerol, has been used for a long time. In addition, the permeable cryoprotectant-free vitrification method has been continuously studied. Although the differences of cryopreservation effects between the two methods have being studied, differences in microRNA (miRNA) profiles between them remain unclear. In this study, we investigated the differences in miRNA expression profiles among conventional freezing sperm, droplet vitrification freezing sperm and fresh human sperm. We also analyzed the differences between these methods in terms of differentially expressed miRNAs (DEmiRs) related to early embryonic development and paternal epigenetics. Our results showed no significant differences between the cryopreservation methods in terms of sperm motility ratio, plasma membrane integrity, DNA integrity, mitochondrial membrane potential, acrosome integrity, and ultrastructural damage. However, sperm miRNA-sequencing showed differences between the two methods in terms of the numbers of DEmiRs (28 and 19 with vitrification using a nonpermeable cryoprotectant and the conventional method, respectively) in postthaw and fresh sperm specimens. DEmiRs related to early embryonic development and paternal epigenetics mainly included common DEmiRs between the groups. Our results showed that the differences between conventional freezing and droplet vitrification were minimal in terms of miRNA expression related to embryonic development and epigenetics. Changes in sperm miRNA expression due to freezing are not always detrimental to embryonic development. This study compared differences in miRNA expression profiles before and after cryopreservation between cryopreservation by conventional and vitrification methods. It offers a new perspective to evaluate various methods of sperm cryopreservation.

Evaluation of Elnady preserved tissues as a teaching aid for undergraduate animal science courses

Abstract The use of tissue specimens for undergraduate instruction is a very valuable tool. However, fresh tissue specimens are not always available and many common preservation techniques can result in discoloration, offensive odors, and/or dangerous chemical residues. The Elnady Technique was developed as a means to produce tissue specimens that “are realistic, durable, have no offensive odor, and are dry, soft and flexible” (Elnady, F.A. 2016 The Elnady Technique: An innovative, new method for tissue preservation. Altex. 33:237-242. doi:10.14573/altex.1511091). Briefly for soft tissue, specimens were preserved by fixing in formalin. The tissue specimen was then dehydrated with a series of acetone baths. Once the tissue was fully dehydrated, the specimen was impregnated in glycerin. Excess glycerin was then removed by draining followed by immersion in cornstarch. Cornstarch residue was removed with a soft brush, and the specimen was stored in a plastic bag. Multiple specimens (including female reproductive tract of the cat, goat, horse, and sow; digestive tract of cat, chicken, and dog; one day old lamb stomach; goat rumen, reticulum, omasum, and abomasum; and sheep heart and kidney) have been preserved and used in various animal science course laboratories (126 laboratory sections and 1696 students at Berry College). Some of the specimens have been in use for seven years and are still in usable condition. Anonymously surveyed Berry College Animal Science Faculty strongly agreed or agreed Elnady preserved tissues are a useful teaching aid (n = 5). The Elnady Technique has proven to be a useful means of preserving tissue samples used in undergraduate animal science courses.

Twelve more bulky genomes in the Polyneoptera: characterizing the Order Embioptera

Abstract Polyneopteran insects have relatively large genomes compared to holometabolous insects, which appear constrained by a threshold of 2 pg/1C DNA. This threshold may be due to costly complex development and higher energy demands. Genome sizes (GSs) are particularly bulky in various species of Orthoptera, reaching sizes up to 18.64 pg/1C DNA, however, recent analyses comparing insect GSs neglected the order Embioptera (webspinners). Our access to fresh specimens of 12 species from 6 of the 13 named taxonomic families provided a chance to determine if embiopterans also support bulky genomes. Flow cytometry results revealed that embiopteran GSs ranged from 2.41 to 7.56 pg/1C, similar to other polyneopterans. Based on previous studies reporting correlations with GS, we tested for correlations between GS and body length using independent contrasts. The ancestral state of the root was estimated at 4.57 pg/1C. A positive relationship was detected whereby larger-bodied webspinners displayed larger genomes, with female Antipaluria urichi (Saussure) (Clothodidae) having the largest genome at 7.56 pg/1C. This GS is approximately 3 times larger than the previously reported embiopteran GS of pg = 2.66 for a male Oligotoma saundersii (Westwood) (Oligotomidae). GS showed no consistent patterns of phylogenetic signal for Embioptera. The underlying causes for large genomes are briefly reviewed.

Sensitivity of magnetic resonance elastography to extracellular matrix and cell motility in human prostate cancer cell line-derived xenograft models

Prostate cancer (PCa) is a significant health problem in the male population of the Western world. Magnetic resonance elastography (MRE), an emerging medical imaging technique sensitive to mechanical properties of biological tissues, detects PCa based on abnormally high stiffness and viscosity values. Yet, the origin of these changes in tissue properties and how they correlate with histopathological markers and tumor aggressiveness are largely unknown, hindering the use of tumor biomechanical properties for establishing a noninvasive PCa staging system.To infer the contributions of extracellular matrix (ECM) components and cell motility, we investigated fresh tissue specimens from two PCa xenograft mouse models, PC3 and LNCaP, using magnetic resonance elastography (MRE), diffusion-weighted imaging (DWI), quantitative histology, and nuclear shape analysis. Increased tumor stiffness and impaired water diffusion were observed to be associated with collagen and elastin accumulation and decreased cell motility. Overall, LNCaP, while more representative of clinical PCa than PC3, accumulated fewer ECM components, induced less restriction of water diffusion, and exhibited increased cell motility, resulting in overall softer and less viscous properties. Taken together, our results suggest that prostate tumor stiffness increases with ECM accumulation and cell adhesion - characteristics that influence critical biological processes of cancer development.MRE paired with DWI provides a powerful set of imaging markers that can potentially predict prostate tumor development from benign masses to aggressive malignancies in patients. Statement of significanceXenograft models of human prostate tumor cell lines, allowing correlation of microstructure-sensitive biophysical imaging parameters with quantitative histological methods, can be investigated to identify hallmarks of cancer.

Direct Implantation of Patient Brain Tumor Cells into Matching Locations in Mouse Brains for Patient-Derived Orthotopic Xenograft Model Development.

Background: Despite multimodality therapies, the prognosis of patients with malignant brain tumors remains extremely poor. One of the major obstacles that hinders development of effective therapies is the limited availability of clinically relevant and biologically accurate (CRBA) mouse models. Methods: We have developed a freehand surgical technique that allows for rapid and safe injection of fresh human brain tumor specimens directly into the matching locations (cerebrum, cerebellum, or brainstem) in the brains of SCID mice. Results: Using this technique, we successfully developed 188 PDOX models from 408 brain tumor patient samples (both high-and low-grade) with a success rate of 72.3% in high-grade glioma, 64.2% in medulloblastoma, 50% in ATRT, 33.8% in ependymoma, and 11.6% in low-grade gliomas. Detailed characterization confirmed their replication of the histopathological and genetic abnormalities of the original patient tumors. Conclusions: The protocol is easy to follow, without a sterotactic frame, in order to generate large cohorts of tumor-bearing mice to meet the needs of biological studies and preclinical drug testing.

The tibial capsular reflection and septum in posterior compartment are safe and reliable soft-tissue landmark for tibial tunnel drilling in posterior cruciate ligament reconstruction.

To investigate the validity of using tibial capsular reflection and septum in the posterior compartment as landmark during posterior cruciate ligament (PCL) reconstruction (PCLR). Anatomic measurements were obtained for 12 fresh human cadaveric knee specimens to observe the spatial position of the tibial insertion of the PCL in relation to the posterior septum and the capsular reflection in the posterior compartment. Sixty patients who underwent reconstruction of the PCL between 2020 and 2023 were also retrospectively investigated. The tibial tunnel was replaced in all patients using the same method (with reference to the tibial capsular reflection and the posterior septum). The placement of the tibial tunnel was assessed using X-ray fluoroscopy intraoperatively and computed tomography and three-dimensional reconstruction postoperatively. All fibres in the tibial insertion of the PCL in the 12 cadaveric specimens were located in the posteromedial compartment, adjacent to the posterior septum. The inferior border of the PCL insertion is adjacent to the tibial capsular reflection, which is attached at the champagne glass drop-off of the posterior tibia. In our previous cases, none of the patients experienced postoperative or intraoperative complications such as neurovascular injury, and the angle between the pin and the PCL facet was 93.1 ± 3.9° as measured on intraoperative radiographs. The mean distance from the centre of the tibial tunnel outlet to the inferior border of the PCL insertion was 5.6 ± 1.1 mm, and the distance from the centre of the tibial tunnel outlet to the outer border of the PCL insertion as a percentage of the length of the inferior border of PCL insertion was 42.2 ± 6.3%. The tibial capsular reflection and septum in the posterior compartment are safe and reliable soft-tissue landmark for tibial tunnel drilling in PCLR. Level Ⅳ.

Severe Venous Calcifications in Phlebosclerotic Colitis and Significantly Decreased Expression in Betaine.

Phlebosclerotic colitis (PC) is a rare type of chronic ischemic colitis. Its etiology is still unknown, and PC is also known as idiopathic mesenteric phlebosclerosis colitis. Currently, many studies have reported that long-term use of Chinese herbal medicine and drinking history are related to its pathogenesis. In the early stage of the lesion, due to insufficient understanding of PC, it is difficult to distinguish it from inflammatory bowel disease and other nonneoplastic intestinal diseases. We reported a case of severe diffuse total colon calcification with multiple misdiagnosis, summarizing and analyzing the clinical pathological characteristics to increase clinical and pathological physicians' understanding of the disease and reduce misdiagnosis. Moreover, for the first time, we conducted metabolomics sequencing on fresh intestinal specimens of PC, in order to explore the possible mechanism of severe calcification in the patient. We found that betaine was significantly decreased in the intestinal specimens of the patient, which is an amino acid that has been shown to improve vascular risk factors, and may be one of the mechanisms underlying severe calcification in the patient.

Experimental Study on Erosion Modeling of Architectural Red Sandstone under the Action of the Natural Environment

When buildings are exposed to erosion from the natural environment, erosion behaviors such as surface damage and structural instability occur, which greatly affect the aesthetic value and service life of the buildings. The study of erosion behaviors and the establishment of a suitable erosion model are constructive references for the protection and restoration of buildings. In order to establish a suitable erosion model for architectural red sandstone, two types of red sandstone specimens were selected in this paper to carry out dry and wet cycle tests. Combining the theoretical analysis and the actual erosion situation, a unidirectional corrosion model is proposed to describe the erosion of buildings by the natural environment. In this model, it is assumed that only the outer surface of the building is in contact with external erosion factors for a long period of time, so this situation can be considered a unidirectional erosion process. The paper uses XRD, SEM, and ultrasonic methods to record changes in the properties of the red sandstone samples. Finally, the rationality of the unidirectional erosion model was verified numerically. The test results show that the red sandstone specimens subjected to erosion by the natural environment will be accompanied by the development of defects, such as cracks, fissures, and holes, as well as the generation of fresh material. The demarcation point of different erosion stages exists in both the in-service red sandstone specimens and the fresh red sandstone specimens, which is consistent with the results of the unidirectional erosion model. In this paper, a calculation model for the demarcation point of different erosion stages is established, and the model estimation shows that the demarcation point of different erosion stages of the in-service red sandstone sample is 1.1528 cm from the erosion surface, and the demarcation point of different erosion stages of the fresh red sandstone sample is 1.67 cm.

Development of a hydrogen peroxide fluorescent probe for the rapid discrimination of IDH1 mutant gliomas from the wild-type

Isocitrate dehydrogenase (IDH) mutations are common in gliomas, and recent studies have suggested that these mutants have distinct microenvironments characterized by higher levels of reactive oxygen species (ROS) and a higher pH. The identification of IDH mutants is important for the diagnosis, individualized treatment, and clinical prognosis of gliomas. Sanger sequencing and immunohistochemistry are the standard methods for IDH mutation detection. As the mutations have several types, the detection of multiple gene sites and proteins are required, which means high cost and tedious operations. The high levels of ROS observed in mutIDH gliomas have not been used for mutIDH detection. Fluorescent probe-based detection techniques have emerged as a prevalent method to detect diverse analytes in living systems due to their distinctive benefits of simple operation, high sensitivity and selectivity, fast response, and real-time applicability. In the present study, we report the rapid discrimination of mutIDH1 from wild-type gliomas using the novel fluorescent probe BOD-G that varies in its fluorescence response to different levels of H2O2. BOD-G exhibits high sensitivity and selectivity in its turn-on response to H2O2, and its performance improves as the pH increases. The probe exhibits a much stronger fluorescence in IDH1 mutant U87 cells than in wild-type U87MG cells after 5 min. Importantly, BOD-G displays a bright fluorescence when exposed to fresh mutIDH1 human glioma specimens after 15 min but not with wild-type samples. Thus, BOD-G can be used to discriminate mutIDH1 from wild-type human glioma specimens rapidly by simple fluorescent imaging.

The taxonomy of Heteropanope indica De Man, 1887 from the Indian Ocean, with a note on Pilumnopeus pearsei (Rathbun, 1932) (Crustacea: Decapoda: Brachyura: Pilumnidae)

Benthopanope indica (De Man, 1887) was originally described from the Mergui Archipelago and has subsequently been reported from many Indo-West Pacific localities. The identity of this species is herein clarified. The re-examination of the type female, fresh specimens collected from Chilika Lake, Odisha State, India, as well as material from other locations, revealed that B. indica is not a Benthopanope species but should be assigned to Pilumnopeus instead. Pilumnopeus indicus comb. nov. is now restricted to the Indian Ocean and is reported from the western coast of Peninsular Malaysia for the first time. Heteropanope pearsei Rathbun, 1932, described from East Asia, previously synonymised under P. indicus by Sakai (1965), is here shown to be valid species of Pilumnopeus, and all previous records of “B. indica” from Japan, China, Taiwan, and Korea must be referred to this species. The two species can be differentiated by characters of the carapace, epistome, chelipeds and gonopods. Lectotypes are also designated for both species.

Assessment of myofascial medialization following intraoperative fascial traction (IFT) in a cadaveric model

PurposeIntraoperative fascial traction (IFT) for the treatment of large ventral hernias and loss of domain (LOD) hernias is a promising tool in abdominal wall surgery. However, little is known about the extent of gain in myofascial advancement especially for the anterior rectus sheath. We, therefore, used a cadaveric model to determine the medialization during IFT.Methods4 fresh frozen specimens were used. Retromuscular preparation was carried out followed by IFT with diagonal vertical traction for 30 min. Medial advancement of the anterior rectus sheath was measured after 15 and 30 min as well as traction forces.ResultsTotal medialization for anterior rectus sheath after 30 min of IFT was 10.5 cm (mean). The mean traction force was 16.28 kg. Total medialization was significantly higher during the first 15 min of vertical fascial traction (p < 0.05).ConclusionsIFT provides significant medialization for the anterior rectus sheath in the cadaveric model. The findings align with results from a retrospective case study. Therefore, we see IFT as a beneficial tool in abdominal wall surgery.

Molecular phylogeny and biogeography of the aquatic dance fly subfamily Clinocerinae (Diptera: Empididae)

AbstractThis study presents the first molecular phylogenetic analysis of the Clinocerinae, challenging the traditionally accepted monophyly of this subfamily. DNA was extracted from fresh and museum specimens representing all biogeographical regions. Maximum likelihood (ML) and Bayesian inference (BI) phylogenetic analyses were performed based on sequences from two mitochondrial genes, cytochrome c oxidase subunit I (COI) and cytochrome β, and three nuclear genes, carbomoylphosphate synthase domain of rudimentary, elongation factor‐1α and isocitrate dehydrogenase. Through molecular data and morphological examination, our results reveal a division within Clinocerinae, distinguishing ‘typical’ or Clinocerinae (s.s.) from several genera, specifically Afroclinocera Sinclair, Asymphyloptera Collin and Proagomyia Collin, possibly lending support for a reclassification of these genera outside Clinocerinae. Bergenstammia Mik is proposed as a junior synonym of Phaeobalia Mik, syn. n., and the following new combinations are recognized: Phaeobalia albanica (Wagner) comb. n., Phaeobalia aurinae (Pusch &amp; Wagner) comb. n., Phaeobalia carniolica (Horvat) comb. n., Phaeobalia frigida (Vaillant) comb. n., Phaeobalia glacialis (Palaczyk &amp; Słowińska) comb. n., Phaeobalia multiseta (Strobl) comb. n., Phaeobalia nudimana (Vaillant) comb. n., Phaeobalia nudipes (Loew) comb. n., Phaeobalia pulla (Vaillant &amp; Wagner) comb. n., Phaeobalia pyrenaica (Vaillant &amp; Vinçon) comb. n., Phaeobalia slovaca (Wagner) comb. n. and Phaeobalia thomasi (Vaillant &amp; Vinçon) comb. n. Re‐evaluation of the genus Roederiodes resulted in the following new combinations: Clinocerella macedonicus (Wagner &amp; Horvat) comb. n. and Clinocerella montenegrinus (Wagner &amp; Horvat) comb. n. The origins of Clinocerinae (s.s.) are traced back to the Holarctic region, Laurasian origin, with a likely complex history of dispersal events into the Southern Hemisphere. Based on current knowledge, the greatest generic and species richness is confined to the Palaearctic Region. These findings provide valuable insights into the evolutionary relationships and distribution patterns of Clinocerinae (s.s.), challenging existing taxonomic classifications and shedding light on their historical biogeography.

Biomechanics Explains Variability of Response of Small Hypertropia to Graded Vertical Rectus Tenotomy

PurposeSmall angle hypertropia in sagging eye syndrome is conveniently treated by graded vertical rectus tenotomy, yet adjustable technique under topical anesthesia has been recommended due to variability of effect. We performed graded tenotomy in an experimental model to elucidate the effect reason for variability of response to this surgical procedure. DesignExperimental study: Methods: 32 fresh bovine rectus musculotendon specimens were prepared including continuity with insertional sclera, and extending for a total 40mm length to the proximal muscle bellies, and trimmed to 16mm width. Specimens were anchored by the clamps at the scleral insertion and muscle belly ends within a physiological chamber. After preconditioning and elongation to 10% strain was imposed by a linear motor, tensile force was allowed to stabilize at a plateau state. Then 25, 50, 75, 90, 100% marginal tenotomies were perormed progressively as remnant forces were measured. ResultsTendon thickness averaged 0.29±0.05mm and width 19.71±2.25mm. On average, remnant force decreased linearly (R2=0.985) from 4.23±1.34, 2.76±0.88, 1.70±0.73, 1.01±0.49, 0.39±0.10, and 0 N, at 0, 25, 50, 75, 90, 100% tenotomy. However, there was marked individual variability in effect among specimens, with coefficient of variation 32, 32, 43, 49, and 27%, respectively ConclusionOn average there is a linear relationship between graded rectus tenotomy and percentage force reduction, but the effect among individual tendons is large, paralleling the reported variation in surgical effect. This explains and implies continued advisability of adjustable technique in this procedure.

Piston-based specimen holder for rapid surgical and biopsy specimen imaging.

Advanced fluorescence imaging modalities such as confocal microscopy and two photon fluorescence microscopy can provide rapid, real-time histology images, but the mounting of fresh tissue specimens in standard orientations required for diagnosis without embedding and sectioning remains an unsolved problem. Here, we introduce a piston-based specimen holder designed for consistent, even pressure distribution. We improve upon previous designs by incorporating an air piston system with a flexible membrane and wick that extracts fluid during compression. We combine this with support fixtures to aid in the distribution of pressure, enabling imaging of specimens with small surface areas relative to their thickness, such as bisected shave skin biopsies in standard orientation without embedding or sectioning. We image both fresh biopsy specimens and diagnostic Mohs first stage specimens during clinical procedures, demonstrating improved visualization of the tissue surface in real time. Finally, we show that conventional cryosectioning can exaggerate the extent of margin positivity, which can be avoided using the piston-based holder.

A review of clinical use of surface-enhanced Raman scattering-based biosensing for glioma.

Glioma is the most common malignant tumor of the nervous system in recent centuries, and the incidence rate of glioma is increasing year by year. Its invasive growth and malignant biological behaviors make it one of the most challenging malignant tumors. Maximizing the resection range (EOR) while minimizing the impact on normal brain tissue is crucial for patient prognosis. Changes in metabolites produced by tumor cells and their microenvironments might be important indicators. As a powerful spectroscopic technique, surface-enhanced Raman scattering (SERS) has many advantages, including ultra-high sensitivity, high specificity, and non-invasive features, which allow SERS technology to be widely applied in biomedicine, especially in the differential diagnosis of malignant tumor tissues. This review first introduced the clinical use of responsive SERS probes. Next, the sensing mechanisms of microenvironment-responsive SERS probes were summarized. Finally, the biomedical applications of these responsive SERS probes were listed in four sections, detecting tumor boundaries due to the changes of pH-responsive SERS probes, SERS probes to guide tumor resection, SERS for liquid biopsy to achieve early diagnosis of tumors, and the application of free-label SERS technology to detect fresh glioma specimens. Finally, the challenges and prospects of responsive SERS detections were summarized for clinical use.

Effects of standardized sampling of fresh specimens on lymph nodes detection in radical pancreatic surgery specimens

Effects of standardized sampling of fresh specimens on lymph nodes detection in radical pancreatic surgery specimens

Abstract CT137: Efficacy and immunological outcomes of non-fucosylated anti-CTLA-4 (BMS-986218) + degarelix acetate vs. degarelix acetate alone in men with high-risk localized prostate cancer (Neo-Red-P)

Abstract Background: Men with high-risk localized prostate cancer are at significant risk for disease recurrence and there is currently no role for neoadjuvant therapy. Although androgen deprivation therapy (ADT) is immunomodulatory and can increase CD8 T cell infiltration, these effects are counterbalanced by the expansion of suppressive regulatory T cells (Tregs). We therefore designed a correlative-rich clinical trial for men with high-risk localized prostate cancer to test whether BMS-986218 - a novel afucosylated anti-CTLA-4 antibody optimized for antibody-dependent cell-mediated cytotoxicity (ADCC) - depletes Tregs and augments response to ADT. Methods: Neo-Red-P is a single-center, two-arm, open-label pilot study assessing degarelix (ADT) versus ADT + anti-CTLA-4-NF (BMS-986218) in patients with treatment naïve high-risk localized prostate cancer. Key eligibility criteria included histologically confirmed adenocarcinoma with Gleason sum of ≥4+3 and clinical stage T1c-T3bN0M0. All patients received degarelix (ADT; 240mg SQ) 2 weeks before radical prostatectomy (RP). Patients were randomized to Arm A (ADT only), or Arm B (ADT + BMS-986218 [20mg IV] 2 doses Q2w starting 3 weeks before RP). The primary endpoint was the safety and feasibility of neoadjuvant BMS-986218 in prostate cancer. Secondary endpoints included the pathologic complete response rate, rate of undetectable PSA at 12 months, PSA response rate (&amp;gt;50% decrease pre-RP), and time-to-PSA recurrence. Fresh tumor specimens were collected at RP for single-cell RNA-sequencing, cytometry by time-of-flight, next-generation sequencing, and multiplex immunofluorescence. A treatment naïve cohort of matching demographic and pathological characteristics was collected for comparative analysis (n=14). Results: Between February 26, 2020, and November 15, 2022, 24 patients were enrolled. Severe treatment-related adverse events (TRAE; Grade ≥ 3) were rare in both arms; 0% in the ADT alone arm and 4% in the combination arm, suggesting a tolerable safety profile with the addition of BMS-986218. In the combination arm, 54% of subjects achieved a 50% reduction in PSA versus 33% in the ADT alone arm. In the ADT alone arm, 80% of patients had an undetectable PSA at 12 months, with a median recurrence-free survival of 1.61 years (IQR: 1.50-1.99 years). In the combination group, 75% had an undetectable PSA at 12 months, with a median recurrence-free survival of 1.82 years (IQR: 0.89-2.07 years). The overall recurrence rate in patients treated with combination therapy was 25% (3 of 12 patients) versus 20% (2 of 10 patients) in the ADT alone arm. Correlative studies suggest that aCTLA4-NF mitigates ADT-induced Treg accumulation through enhanced Fc receptor activity and augmented CTL activation, indicating an enhanced anti-tumor response with combination therapy. Conclusions: The neoadjuvant combination of ADT + aCTLA-4-NF (BMS-986218) is feasible, well tolerated, and induces significant remodeling of the immune TME through enhanced Fc receptor and CTL activity. Rates of disease recurrence following surgery were low in this high-risk population, warranting further investigation of combination immuno-hormonal therapy in the neoadjuvant setting for high-risk prostate cancer. Clinical Trial Information: NCT04301414 Citation Format: Patrick J. McCann, Casey R. Ager, Aleksandar Z. Obradovic, Matthew G. Chaimowitz, Samuel Pan, Caroline J. Laplaca, Parin Shah, Renu K. Virk, Christopher B. Anderson, Catherine S. Spina, Karie D. Runcie, Mark N. Stein, Charles G. Drake, Matthew C. Dallos. Efficacy and immunological outcomes of non-fucosylated anti-CTLA-4 (BMS-986218) + degarelix acetate vs. degarelix acetate alone in men with high-risk localized prostate cancer (Neo-Red-P) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT137.

Expression of Concern: Enterobacter timonensis sp. nov., a new bacterium isolated from a fresh human stool specimen

Expression of Concern: Enterobacter timonensis sp. nov., a new bacterium isolated from a fresh human stool specimen

293 The Impact of Tumor Mutational Status on Protoporphyrin IX Accumulation in Tumor-Associated Macrophages in Glioblastoma Patients

INTRODUCTION: Immunotherapies provide promising new treatment options for lung cancer and melanoma, however success in glioblastoma (GBM) patients has been much rarer and uneven. Identifying candidates with GBM that are most likely to benefit from immunotherapy remains a challenge. The abundance of tumor-associated macrophages (TAMs) correlates response to immunotherapy in GBM patients. Here we leverage a new imaging technique for quantifying TAMs via tandem stimulated Raman histology (SRH) and two-photon excitation fluorescence microscopy (TPEF) to assess the impact of genetic classification on the abundance of TAMs. METHODS: We conducted tandem SRH and TPEF on specimens from 79 patients undergoing high-grade glioma resection with 5-ALA. We trained a MaskR-CNN deep neural network to identify TAMs and compute their density in each specimen. Corresponding clinical, demographic, genomic and methylomic data were collected for each patient. Independent sample t-tests were used to identify significant correlations between genetic markers, treatment, and TAM density. RESULTS: We analyzed 3,121,524 300 x 300 micron fields of view, from 151 fresh specimens, enabling segmentation of 611,789 TAMs. IDH-mutant tumors had significantly lower TAM density than IDH-wildtype tumors (p &lt; 0.01). The absence of MGMT promoter hypermethylation was associated with significantly higher TAM density (p = 0.03). Tumors with EGFR amplification averaged significantly lower TAM density than EGFR wildtype specimens (p &lt; 0.01). Recurrent tumors possessed 31.5% lower TAM density than primary resections. CONCLUSIONS: Tandem SRH and TPEF can estimate variations in TAM density in human GBMs. We found TAM density to be associated with IDH status, MGMT promoter methylation, and EGFR amplification. This imaging technique creates a new avenue for forecasting response to immunotherapy in GBM patients.