2-Azabicyclo[3.2.1]octane scaffold: synthesis and applications.

2-Azabicyclo[3.2.1]octanes are nitrogen containing heterocycles with significant potential in the field of drug discovery. This core has been applied as key synthetic intermediate in several total synthesis, while their unique structure can make them a challenging scaffold to acquire. This Minireview summarizes the synthetic approaches to access this bicyclic architecture and highlights its presence in the total synthesis of several target molecules.

One nodule-one punction-one slide: Optimizing thyroid fine-needle aspiration for a digital workflow.

OBJECTIVE: Interventional pathologists have expanded their expertise by acquiring proficiency in ultrasound-guided thyroid fine-needle aspiration biopsy (FNAB) and are now required to optimize punction procedures due to low resources and digital workflows. The aim of this study is to compare FNAB sample adequacy in two series with one versus two slides available for cytopathological analysis and its influence on diagnosis categorization, time taken to reach a final diagnosis, scanning time and size of the digital files produced. METHODS: Patients were retrospectively selected based on the sampling of thyroid nodules using either two glass slides (two-slide group) or one slide only (one-slide group) and cytological diagnosis was performed using the second edition of the Bethesda system. For each group, the initial 15 cases were sorted to be scanned. RESULTS: From a total of 713 procedures, 328 were sampled into two slides and 385 on one slide only. No significant differences were found regarding nodule size, location or EU-TIRADS classification between the two groups. The one-slide group did not exhibit a higher prevalence of non-diagnostic or atypia of undetermined significance (AUS) categories. As expected, the mean time taken to finalize diagnoses in cases where only one slide was prepared was 1.2 days faster. Scanning time and total file size were also significantly smaller in the one-slide group. CONCLUSIONS: Adopting the 'one nodule-one puncture-one slide' strategy for thyroid FNAB optimization enhances procedural efficiency in digital workflows, leading to cost savings without compromising diagnostic accuracy.

ClotCatcher: a novel natural language model to accurately adjudicate venous thromboembolism from radiology reports.

INTRODUCTION: Accurate identification of venous thromboembolism (VTE) is critical to develop replicable epidemiological studies and rigorous predictions models. Traditionally, VTE studies have relied on international classification of diseases (ICD) codes which are inaccurate - leading to misclassification bias. Here, we developed ClotCatcher, a novel deep learning model that uses natural language processing to detect VTE from radiology reports. METHODS: Radiology reports to detect VTE were obtained from patients admitted to Emory University Hospital (EUH) and Grady Memorial Hospital (GMH). Data augmentation was performed using the Google PEGASUS paraphraser. This data was then used to fine-tune ClotCatcher, a novel deep learning model. ClotCatcher was validated on both the EUH dataset alone and GMH dataset alone. RESULTS: The dataset contained 1358 studies from EUH and 915 studies from GMH (n = 2273). The dataset contained 1506 ultrasound studies with 528 (35.1%) studies positive for VTE, and 767 CT studies with 91 (11.9%) positive for VTE. When validated on the EUH dataset, ClotCatcher performed best (AUC = 0.980) when trained on both EUH and GMH dataset without paraphrasing. When validated on the GMH dataset, ClotCatcher performed best (AUC = 0.995) when trained on both EUH and GMH dataset with paraphrasing. CONCLUSION: ClotCatcher, a novel deep learning model with data augmentation rapidly and accurately adjudicated the presence of VTE from radiology reports. Applying ClotCatcher to large databases would allow for rapid and accurate adjudication of incident VTE. This would reduce misclassification bias and form the foundation for future studies to estimate individual risk for patient to develop incident VTE.

Synovial Fluid Biomarkers for the Diagnosis of Periprosthetic Joint Infection-A Systematic Review and Meta-Analysis of Their Diagnostic Accuracy According to Different Definitions.

BACKGROUND: Different synovial fluid biomarkers have emerged to improve periprosthetic joint infection (PJI) diagnosis. The goals of this paper were (i) to assess their diagnostic accuracy and (ii) to evaluate their performance according to different PJI definitions. METHODS: A systematic review and meta-analysis was performed using studies that reported diagnostic accuracy of synovial fluid biomarkers using validated PJI definitions published from 2010 to March 2022. A database search was performed through PubMed, Ovid MEDLINE, Central, and Embase. The search identified 43 different biomarkers with four being the more commonly studied, with 75 papers overall: alpha-defensin; leukocyte esterase; synovial fluid C-reactive protein; and calprotectin. RESULTS: Overall accuracy was higher for calprotectin, followed by alpha-defensin, leukocyte esterase, and synovial fluid C-reactive protein with sensitivities of 78 to 92% and specificities of 90 to 95%. Their diagnostic performance was different according to which definition was adopted as the reference. Specificity was consistently high across definitions for all four biomarkers. Sensitivity varied the most with lower values for the more sensitive European Bone and Joint Infection Society or Infectious Diseases Society of America definitions with higher values for the Musculoskeletal Infection Society definition. The International Consensus Meeting 2018 definition showed intermediate values. CONCLUSION: All evaluated biomarkers had good specificity and sensitivity, making their use acceptable in the diagnosis of PJI. Biomarkers perform differently according to the selected PJI definitions.

Functionalized Cyclopentenes via the Formal [4+1] Cycloaddition of Photogenerated Siloxycarbenes from Acyl Silanes.

This work describes the first formal cycloaddition reaction of photogenerated nucleophilic carbenes derived from acylsilanes with electrophilic dienes. The resulting transient donor-acceptor cyclopropane rearranges to its stable and highly functionalized cyclopentene isomer in an unprecedented metal-free process. The cyclopropanation-vinyl cyclopropane rearrangement sequence was corroborated by computational calculations. The cyclopropane formation corresponds to a higher energetic barrier, and the vinylcyclopropane-cyclopentene rearrangement proceeds through different mechanisms, although of comparable energies, depending on the stereochemistry of the cyclopropane.

Comparison of short-stem versus conventional stem for hip arthroplasty in patients younger than 60 years: 7-14 years follow-up.

PURPOSE: Short-stem prostheses in hip arthroplasty have emerged as an alternative to conventional stems, especially in younger patients. The purpose of this study was to compare functional and radiological results of a short metaphyseal fitting cementless stem versus a conventional stem implant, in patients younger than 60 years. METHODS: All patients operated from January 2006 to April 2013 were included, obtaining a minimum follow-up of 7 years. Harris Hip Score (HHS) and SF-36 (quality of life) questionnaires were applied and the presence of "thigh pain" was specifically assessed. We also compared complication rate, revision rate and average prosthesis survival. Femoral stress shielding (Gruen scale), stem subsidence, varus-valgus tilt and implant stability (Engh scale) were also compared. RESULTS: A total of 101 short-stem and 74 conventional arthroplasties were included, with an average follow-up of 9.82 (7-14) years. HHS functional score and SF-36 were excellent in both implants and no significant difference between them (p > 0.05) was found. However, "thigh pain" was present in 7 patients with conventional stems and none with short-stems (p < 0.001). The survival rate at 13 years was 99%, for both implants, and no significant differences were found between them (χ2(2) = 0.178; p = 0.673). Conventional stems had stress shielding at the greater trochanter in 72% of the cases and 43% at the calcar, being statistically superior (p < 0.001) to the stress shielding observed in the short stems. CONCLUSION: According to our results, this short-stem seems to allow preservation of bone stock, with decreased stress shielding and also a lower incidence of thigh pain compared to conventional stems. LEVEL OF EVIDENCE: Level III retrospective comparative study.

Nanoaggregate-forming lipid-conjugated AS1411 aptamer as a promising tumor-targeted delivery system of anticancer agents in vitro.

Nanoparticles offer targeted delivery of drugs with minimal toxicity to surrounding healthy tissue and have great potential in the management of human papillomavirus (HPV)-related diseases. We synthesized lipid-modified AS1411 aptamers capable of forming nanoaggregates in solution containing Mg2+. The nanoaggregates presented suitable properties for pharmaceutical applications such as small size (100 nm), negative charge, and drug release. The nanoaggregates were loaded with acridine orange derivative C8 for its specific delivery into cervical cancer cell lines and HPV-positive tissue biopsies. This improved inhibition of HeLa proliferation and cell uptake without significantly affecting healthy cells. Finally, the nanoaggregates were incorporated in a gel formulation with promising tissue retention properties aiming at developing a local delivery strategy of the nanoaggregates in the female genital tract. Collectively, these findings suggest that the nanoformulation protocol has great potential for the delivery of both anticancer and antiviral agents, becoming a novel modality for cervical cancer management.

Synthesis of 6,12-Disubstituted Methanodibenzo[b,f][1,5]dioxocins: Pyrrolidine Catalyzed Self-Condensation of 2'-Hydroxyacetophenones.

The preparation of unprecedented 6,12-disubstituted methanodibenzo[b,f][1,5]dioxocins from pyrrolidine catalyzed self-condensation of 2'-hydroxyacetophenones is herein described. This method provides easy access to this highly bridged complex core, resulting in construction of two C-O and two C-C bonds, a methylene bridge and two quaternary centers in a single step. The intricate methanodibenzo[b,f][1,5]dioxocin compounds were obtained in up to moderate yields after optimization of the reaction conditions concerning solvent, reaction times and the use of additives. Several halide substituted methanodibenzo[b,f][1,5]dioxocins could be prepared from correspondent 2'-hydroxyacetophenones.

Pot-Economy Autooxidative Condensation of 2-Aryl-2-lithio-1,3-dithianes.

The autoxidative condensation of 2-aryl-2-lithio-1,3-dithianes is here reported. Treatment of 2-aryl-1,3-dithianes with n-BuLi in the absence of any electrophile leads to condensation of three molecules of 1,3-dithianes and formation of highly functionalized α-thioether ketones orthothioesters in 51-89% yields upon air exposure. The method was further expanded to benzaldehyde dithioacetals, affording corresponding orthothioesters and α-thioether ketones in 48-97% yields. The experimental results combined with density functional theory studies support a mechanism triggered by the autoxidation of 2-aryl-2-lithio-1,3-dithianes to yield a highly reactive thioester that undergoes condensation with two other molecules of 2-aryl-2-lithio-1,3-dithiane.

Optimizing the management of thyroid specimens to efficiently generate whole slide images for diagnosis.

Transition from optical to digital observation requires an additional procedure in the pathology laboratory, the scanning of glass slides, leading to increased time and digital archive consumption. Thyroid surgical samples often carry the need to collect several tissue fragments that generate many slides to be scanned. This study evaluated the impact of using different inking colours for the surgical margin, section thickness, and glass slide type, in the consumption of time and archive. The series comprehended 40 nodules from 30 patients, including 34 benign nodules in follicular nodular disease, 1 NIFTP, and 5 papillary carcinomas. In 12 nodules, the dominant pattern was microfollicular/solid and in 28 it was macrofollicular. Scanning times/mm2 were longer in red-inked fragments in comparison to green (p = 0.04) and black ones (p = 0.024), and in blue-inked in comparison to green ones (p = 0.043). File sizes/mm2 were larger in red-inked fragments in comparison to green (p = 0.008) and black ones (p = 0.002). The dominant pattern microfollicular/solid was associated with bigger file size/mm2 in comparison with the macrofollicular one (p < 0.001). All scanner outputs increase significantly with the thickness of the section. All scanning outputs increase with the usage of adhesive glass slides in comparison to non-adhesive ones. Small interventions in thyroid sample management that can help optimizing the digital workflow include to prefer black and green inking colours for the surgical margins and 2 µm section in non-adhesive glass slides for increased efficiency.

Analysis of Greek prehistoric combat in full body armour based on physiological principles: A series of studies using thematic analysis, human experiments, and numerical simulations.

One of the oldest complete suits of European armour was discovered in 1960 near the village of Dendra, in Southern Greece, but it remained unknown whether this armour was suitable for extended use in battle or was purely ceremonial. This had limited our understanding of the ancient Greek-Late Bronze Age-warfare and its consequences that have underpinned the social transformations of prehistoric Europe and Eastern Mediterranean. In a series of archeo-physiological studies, merging knowledge in archaeology, history, human physiology, and numerical simulation, we provide supporting evidence that the Mycenaean armour found at Dendra was entirely compatible with use in extended combat, and we provide a free software enabling simulation of Late Bronze Age warfare. A group of special armed-forces personnel wearing a replica of the Dendra armour were able to complete an 11-hour simulated Late Bronze Age combat protocol that we developed from a series of studies based on the available evidence. Numerical simulation of the thermal exchanges in Late Bronze Age warfare extended this conclusion across different environmental conditions and fighting intensities. Our results support the notion that the Mycenaeans had such a powerful impact in Eastern Mediterranean at least partly as a result of their armour technology.

Revisiting the utility of identifying nuclear grooves as unique nuclear changes by an object detector model.

BACKGROUND: Among other structures, nuclear grooves are vastly found in papillary thyroid carcinoma (PTC). Considering that the application of artificial intelligence in thyroid cytology has potential for diagnostic routine, our goal was to develop a new supervised convolutional neural network capable of identifying nuclear grooves in Diff-Quik stained whole-slide images (WSI) obtained from thyroid fineneedle aspiration. METHODS: We selected 22 Diff-Quik stained cytological slides with cytological diagnosis of PTC and concordant histological diagnosis. Each of the slides was scanned, forming a WSI. Images that contained the region of interest were obtained, followed by pre-formatting, annotation of the nuclear grooves and data augmentation techniques. The final dataset was divided into training and validation groups in a 7:3 ratio. RESULTS: This is the first artificial intelligence model based on object detection applied to nuclear structures in thyroid cytopathology. A total of 7,255 images were obtained from 22 WSI, totaling 7,242 annotated nuclear grooves. The best model was obtained after it was submitted 15 times with the train dataset (14th epoch), with 67% true positives, 49.8% for sensitivity and 43.1% for predictive positive value. CONCLUSIONS: The model was able to develop a structure predictor rule, indicating that the application of an artificial intelligence model based on object detection in the identification of nuclear grooves is feasible. Associated with a reduction in interobserver variability and in time per slide, this demonstrates that nuclear evaluation constitutes one of the possibilities for refining the diagnosis through computational models.

Encasement Of Popliteal Vascular Bundle Due Local Osteosarcoma Recurrence Treated With Wide Resection And Vascular Reconstruction.

Osteosarcoma is the most common primary bone malignancy. The prognosis of patients after local recurrence is generally poor and management of the locally recurrent disease is not well defined, especially in patients who have undergone limb-sparing surgery. A 20-year-old male presented a local recurrence of conventional osteosarcoma at the popliteal fossa with encasement of popliteal vascular bundle after previous tumor-wide resection and reconstruction with proximal tibia endoprosthesis. A wide resection "en bloc" of the lesion included part of the popliteal vessel. A bypass of both popliteal vessels, the vein with polytetrafluoroethylene (PTFE) prosthesis, and the artery with contralateral saphenous vein were performed to allow a limb salvage surgery. Local management of recurrent osteosarcoma in a previously reconstructed limb is highly individualized. This case confirms that preservation of lower limb function is possible using reconstruction techniques of bone and vessels in the sarcoma of the musculoskeletal system.

Laparoscopic Versus Robotic Gastric Cancer Surgery: Short-Term Outcomes-Systematic Review and Meta-Analysis of 25,521 Patients.

Background: Gastric cancer has the third highest cancer-related mortality worldwide. There is no consensus regarding the optimal surgical technique to perform curative resection surgery. Objective: Compare laparoscopic gastrectomy (LG) and robotic gastrectomy (RG) regarding short-term outcomes in patients with gastric cancer. Materials and Methods: This systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We searched the following topics: "Gastrectomy," "Laparoscopic," and "Robotic Surgical Procedures." The included studies compared short-term outcomes between LG and RG. Individual risk of bias was assessed with the Methodological Index for Non-Randomized Studies (MINORS) scale. Results: There was no significant difference between RG and LG regarding conversion rate, reoperation rate, mortality, overall complications, anastomotic leakage, distal and proximal resection margin distances, and recurrence rate. However, mean blood loss (mean difference [MD] -19.43 mL, P < .00001), length of hospital stay (MD -0.50 days, P = .0007), time to first flatus (MD -0.52 days, P < .00001), time to oral intake (MD -0.17 days, P = .0001), surgical complications with a Clavien-Dindo grade ≥III (risk ratio [RR] 0.68, P < .0001), and pancreatic complications (RR 0.51, P = .007) were significantly lower in the RG group. Furthermore, the number of retrieved lymph nodes was significantly higher in the RG group. Nevertheless, the RG group showed a significantly higher operation time (MD 41.19 minutes, P < .00001) and cost (MD 3684.27 U.S. Dollars, P < .00001). Conclusion: This meta-analysis supports the choice of robotic surgery over laparoscopy concerning relevant surgical complications. However, longer operation time and higher cost remain crucial limitations. Randomized clinical trials are required to clarify the advantages and disadvantages of RG.

Inking cell blocks improves scanner detection for diagnosis in pathology.

Cell blocks may be hard to be totally automatically detected by the scanner (ADS), generating incomplete whole slide images (WSIs), with areas that are not scanned, leading to possible false negative diagnosis. The aim of this study is to test if inking the cell blocks helps increasing ADS. Test 1: 15 cell blocks were sectioned, one half inked black (1HB) and the other inked green (1HG). Each of the halves was individually processed to generate a WSI stained by the H&E. 1HBs and 1HGs had similar scanning time (median 59 s vs. 65 s, p = .126) and file sizes (median 382 Mb vs. 381 Mb, p = .567). The black ink interfered less in the observation (2.2% vs. 44.4%; p < .001) than in the green one. Test 2: 15 cell blocks were sectioned, one half inked black (2HB) and the other left unstained/null (2HN). Each of the halves was individually processed to generate three WSIs-one HE, one periodic-acid Schiff (PAS), and one immunostained by cytokeratin AE1&AE3 (CKAE1AE3). HE and PAS WSIs from both 2HN and 2HB groups were all totally ADS and had similar scanning times and file sizes. Concerning immunostaining with CKAE1AE3: ADS (46.7% vs. 93.3%; p = .014), median time for scanning (57 s vs. 83 s; p < .001) and file size (178 Mb vs. 338 Mb; p < .001) were reduced significantly in the 2HN group in comparison with the 2HB. Although increasing scanning time and file size, inking the cell blocks helps increasing ADS after immunostaining, improving the safety and efficiency of the digital pathology workflow.

Artificial intelligence-assisted cancer diagnosis improves the efficiency of pathologists in prostatic biopsies.

Paige Prostate is a clinical-grade artificial intelligence tool designed to assist the pathologist in detecting, grading, and quantifying prostate cancer. In this work, a cohort of 105 prostate core needle biopsies (CNBs) was evaluated through digital pathology. Then, we compared the diagnostic performance of four pathologists diagnosing prostatic CNB unaided and, in a second phase, assisted by Paige Prostate. In phase 1, pathologists had a diagnostic accuracy for prostate cancer of 95.00%, maintaining their performance in phase 2 (93.81%), with an intraobserver concordance rate between phases of 98.81%. In phase 2, pathologists reported atypical small acinar proliferation (ASAP) less often (about 30% less). Additionally, they requested significantly fewer immunohistochemistry (IHC) studies (about 20% less) and second opinions (about 40% less). The median time required for reading and reporting each slide was about 20% lower in phase 2, in both negative and cancer cases. Lastly, the average total agreement with the software performance was observed in about 70% of the cases, being significantly higher in negative cases (about 90%) than in cancer cases (about 30%). Most of the diagnostic discordances occurred in distinguishing negative cases with ASAP from small foci of well-differentiated (less than 1.5 mm) acinar adenocarcinoma. In conclusion, the synergic usage of Paige Prostate contributes to a significant decrease in IHC studies, second opinion requests, and time for reporting while maintaining highly accurate diagnostic standards.

The Role of Systemic Steroids in Sciatica Due to Herniated Lumbar Disc: A Systematic Review and Meta-analysis.

STUDY DESIGN: Systematic review and meta-analysis. OBJECTIVE: Evaluate the role of systemic steroids in treating patients with sciatica due to lumbar disk herniation (LDH). SUMMARY OF BACKGROUND DATA: The association between LDH and sciatica has been well described. The use of steroids seems logical in this context; however, their efficacy is not well described, and their use remains controversial. METHODS: A comprehensive search on PubMed, EMBASE, and Scopus databases (up to February 15, 2022) was performed to identify randomized clinical trials that included patients with symptoms of sciatica due to LDH that were treated with systemic steroids. The risk of bias was judged using the Cochrane risk-of-Bias2 tool. Meta-analysis was conducted using a random-effects model to estimate the between-group effect size for pain and functional outcomes. The risk of developing adverse events (AE) was computed using relative risks. All pooled results are reported with their 95% confidence interval (CI) and certainty of evidence analyzed using the Grading of Recommendations Assessment, Development, and Evaluation framework. RESULTS: Ten studies met inclusion criteria, comprising a total of 1017 participants: 540 in the treatment group and 477 in the control group. Steroid treatment was associated with a significant superior reduction of pain (SMD = -0.42, 95% CI -0.76 to -0.08, weak effect, very-low certainty) and reduction in disability (SMD = -0.30, 95% CI -0.51 to -0.10, weak effect, very-low certainty). Corticosteroid administration was associated with a significant increased risk of developing an AE (relative risks = 2.00, 95% CI 1.40 to 2.85, low certainty). CONCLUSION: The use of systemic steroids in the treatment of sciatica due to LDH seems reasonable despite a 2-fold higher risk of developing mild AEs. However, the effect size is small for reducing pain in the short term and improving functional outcomes at long-term follow-up.

Orthogonally Protected Diaminocyclopentenones as Synthons: Total Synthesis of (±)-Agelastatin A.

Natural products containing aminocyclopentanes are common secondary metabolites, often biologically active. This work aims at the preparation of a useful synthon for total synthesis containing orthogonally protected amines. To this end, furfural and two amines were employed to form mixed trans-4,5-diaminocyclopentenones promoted by Cu(OTf)2. The selected amines can be orthogonally deprotected, allowing selective modification of the amines on the cyclopentane core. Their utility was showcased for the total synthesis of highly complex (±)-Agelastatin A.

Total knee arthroplasty coronal alignment and tibial base stress-a new numerical evaluation.

Background: Total knee arthroplasty (TKA) is one of the most frequently performed orthopedic procedures. The correct positioning and alignment of the components significantly affects prosthesis survival. Considering the current controversy regarding the target of postoperative alignment of TKA, this study evaluated the tension at tibial component interface using two numerical methods. Methods: The stress of the prosthesis/bone interface of the proximal tibial component was evaluated using two numerical methods: the finite element method (FEM) and the new meshless method: natural neighbor radial point interpolation method (NNRPIM). The construction of the model was based on Zimmers NexGen LPS-Flex Mobile® prosthesis and simulated the forces by using a free-body diagram. Results: Tibiofemoral mechanical axis (TFMA) for which a higher number of nodes are under optimal mechanical tension is between 1° valgus 2° varus. For values outside the interval, there are regions under the tibial plate at risk of bone absorption. At the extremities of the tibial plate of the prosthesis, both medial and lateral, independent of the alignment, are under a low stress. In all nodes evaluated for all TFMA, the values of the effective stresses were higher in the NNRPIM when compared with the FEM. Conclusion: Through this study, we can corroborate that the optimal postoperative alignment is within the values that are currently considered of 0 ± 3° varus. It was verified that the meshless methods obtain smoother and more conservative results, which may make them safer when transposed to the clinical practice.

Demonstrating the interference of tissue processing in the evaluation of tissue biomarkers: The case of PD-L1.

The immunohistochemical (IHC) expression of PD-L1 in cancer models is used as a predictive biomarker of response to immunotherapy. We aimed to evaluate the impact of the usage of 3 different tissue processors in the IHC expression of PD-L1 antibody clones: 22C3 and SP142. Three different topographies of samples (n = 73) were selected at the macroscopy room: 39 uterine leiomyomas, 17 placentas and 17 palatine tonsils. Three fragments were collected from each sample and were inked with a specific color that represented their separate processing in a different tissue processor (A, B or C). During embedding, the 3 fragments with distinct processing were ensemble in the same cassette for sectioning of 3 slides/each: hematoxylin-eosin, 22C3 PDL1 IHC staining and SP142 PD-L1 IHC staining, that were blindly observed by 2 pathologists under digital environment. All but one set of 3 fragments were considered adequate for observation even in the presence of artifacts associated with processing issues that were recorded as high as 50.7 % for processor C. The occurrence of background non-specific staining and the presence of false positive results appear to be unrelated with the PD-L1 clone or the type of tissue processing. 22C3 PD-L1 was more frequently considered adequate for evaluation than SP142 PD-L1 that, in 29.2 % of WSIs (after tissue processor C) were considered not adequate for observation due to lack of the typical pattern of expression. Similarly, the intensity of PD-L1 staining was significantly decreased in fragments processed by C (both PD-L1 clones) in tonsil and placenta specimens, and by A (both clones) in comparison with those processed by B. This study demonstrates the need to standardize the tissue processing in pathology to cope with the growing needs of precision medicine quantifications and the production of high-quality material necessary for computational pathology usage.