Comparison of immunohistochemical mesothelial biomarkers in paired biopsies and effusion cytology cell blocks from pleural mesothelioma.

OBJECTIVE: Traditionally, the diagnosis of pleural mesothelioma is based on histological material. Minimally invasive effusion cytology specimens are an alternative that, like biopsies, require ancillary analyses. Validation of immunohistochemical (IHC) analyses on cytology, including the surrogate markers for molecular alterations BAP1 and MTAP, is of interest. METHODS: IHC for eight different markers was performed on 59 paired formalin-fixed, paraffin-embedded pleural biopsies and pleural effusion cell blocks with mesothelioma. Immunoreactivity in ≥10% of tumour cells was considered positive/preserved. The concordance between histological and cytological materials was assessed. RESULTS: The overall percentage of agreement between the histological epithelioid component in 58 biopsies and paired cell blocks was 93% for calretinin, 98% for CK5, 97% for podoplanin, 90% for WT1, 86% for EMA, 100% for desmin, 91% for BAP1, and 72% for MTAP. For 11 cases with biphasic or sarcomatoid histology, the concordance between cytology and the histological sarcomatoid component was low for calretinin, CK5, and WT1 (all ≤45%). For the whole cohort, loss of both BAP1 and MTAP was seen in 40% while both markers were preserved in 11% of the biopsies for epithelioid histology. The corresponding numbers were 54% and 8%, respectively, for the paired cell blocks. CONCLUSIONS: Generally, a high concordance for IHC staining was seen between paired biopsies and pleural effusion cell blocks from mesotheliomas, but the somewhat lower agreement for WT1, EMA, and especially MTAP calls for further investigation and local quality assurance. The lower concordance for the sarcomatoid subtype for some markers may indicate biological differences.

Exploring digital technologies used in the design and manufacture of craniofacial implant surgical guides: A scoping review.

STATEMENT OF PROBLEM: Unlike intraoral implants, digitally planned surgical templates used for guiding the ideal position of the craniofacial implants are not well established, and clear methods and guidelines for their design and construction are lacking. PURPOSE: The purpose of this scoping review was to identify the publications that used a full or partial computer-aided design and computer-aided manufacture (CAD-CAM) protocol to create a surgical guide that achieves the correct positioning of craniofacial implants to retain a silicone facial prosthesis. MATERIAL AND METHODS: A systematic search was conducted in MEDLINE/PubMed, Web of Science, Embase, and Scopus for articles published before November 2021 in the English language. Articles needed to satisfy the eligibility criterion of in vivo articles that created a surgical guide with digital technology for inserting titanium craniofacial implants to hold a silicone facial prosthesis. Articles that inserted implants in the oral cavity or upper alveolus only and articles that did not describe the structure and retention of the surgical guide were excluded. RESULTS: Ten articles were included in the review; all were clinical reports. Two of the articles used a CAD-only approach alongside a conventionally constructed surgical guide. Eight articles described applying a complete CAD-CAM protocol for the implant guides. The digital workflow varied considerably depending on the software program, design, and retention of guides. Only 1 report described a follow-up scanning protocol to verify the accuracy of the final implant positions compared with the planned positions. CONCLUSIONS: Digitally designed surgical guides can be an excellent adjunct to accurately place titanium implants in the craniofacial skeleton for support of silicone prostheses. A sound protocol for the design and retention of the surgical guides will enhance the use and accuracy of craniofacial implants in prosthetic facial rehabilitation.

PD-L1 Expression in Non-Small Cell Lung Cancer Specimens: Association with Clinicopathological Factors and Molecular Alterations.

Immune checkpoint inhibitors (ICI) targeting programmed cell death-1 or its ligand (PD-L1) have improved outcomes in non-small cell lung cancer (NSCLC). High tumor PD-L1 expression, detected by immunohistochemistry (IHC) typically on formalin-fixed paraffin-embedded (FFPE) histological specimens, is linked to better response. Following our previous investigation on PD-L1 in cytological samples, the aim of this study was to further explore the potential impacts of various clinicopathological and molecular factors on PD-L1 expression. Two retrospective NSCLC cohorts of 1131 and 651 specimens, respectively, were investigated for PD-L1 expression (<1%/1−49%/≥50%), sample type, sample site, histological type, and oncogenic driver status. In both cohorts, PD-L1 was positive (≥1%) in 55% of the cases. Adenocarcinomas exhibited lower PD-L1 expression than squamous cell carcinomas (p < 0.0001), while there was no difference between sample types, tumor locations, or between the two cohorts in multivariate analysis (all p ≥ 0.28). Mutational status correlated significantly with PD-L1 expression (p < 0.0001), with the highest expression for KRAS-mutated cases, the lowest for EGFR-mutated, and the KRAS/EGFR wild-type cases in between. There was no difference in PD-L1 levels between different prevalent KRAS mutations (all p ≥ 0.44), while mucinous KRAS-mutated adenocarcinomas exhibited much lower PD-L1 expression than non-mucinous (p < 0.0001). Our data indicate that cytological and histological specimens are comparable for PD-L1 evaluation. Given the impact of KRAS mutations and the mucinous growth pattern on PD-L1 expression, these factors should be further investigated in studies on ICI response.

Synthesis, Anticancer, Antioxidant, Anti-Inflammatory, Antimicrobial Activities, Molecular Docking, and DFT Studies of Sultams Derived from Saccharin.

A series of N-substituted saccharins namely 2-(1,1-dioxido-3-oxobenzo[d]isothiazol-2(3H)-yl) acetonitrile (2) and (alkyl 1,1-dioxido-3-oxobenzo[d]isothiazol-2(3H)-yl) acetate (3a-g) were synthesized, in moderate to excellent yields, from commercially available starting materials by two different approaches and their chemical structures were characterized by spectroscopic techniques (1H-NMR, 13C-NMR, IR, and MS). All the synthesized compounds were evaluated for their anti-inflammatory toward IL-6 and TNF-α, antioxidant, as well as their anticancer activities against hepatic cancer cells. In addition, their anti-fungal and antibacterial activities against both Gram-positive and Gram-negative bacteria were tested. All the tested compounds have exhibited excellent (3a, d, e) to moderate anti-inflammatory activity. Additionally, esters (3b, f) and nitrile (2) showed excellent antioxidant activity. Furthermore, ester 3f, with isopropyl ester, exhibited the highest cytotoxic activity compared to the other esters. Moreover, all compounds were evaluated as selective inhibitors of the human COX-1 enzyme using molecular docking by calculating the free energy of binding, inhibition constant, and other parameters to find out the binding affinity. The molecular study showed that esters (3d, f) and nitrile (2) revealed the highest binding affinities, hence enhancing the inhibition activity with the active site of the COX-1 enzyme. All the tested compounds have more negative Gibbs free, electrostatic, and total intermolecular energies than the standard inhibitor ASA. These results indicate that, all the tested sultams are potent anti-inflammatory drugs as compared to standard inhibitors. Finally, the chemical properties and the quantum factors of synthesized sultams were calculated based on density functional theory (DFT) to predict reactivity, and then correlated with the experimental data. Ester 3f showed the lowest ionization potential and lowest energy gap (Egap = 7.5691 eV), which was correlated with its cytotoxic activity. Furthermore, the spatial electron distribution of HOMO, LUMO were computed and it clearly indicates the electron donation ability of all the tested compounds.

SP3 is associated with migration, invasion, and Akt/PKB signalling in MDA-MB-231 breast cancer cells.

Specificity proteins (SPs) have pro-oncogenic functions in cancer cells, ranging from cancer cell proliferation, migration, invasion, and angiogenesis. There is strong evidence that several antineoplastic drugs target depletion of SP proteins via different pathways. However, the mode of action of SP3 and the underlying consequences of its depletion are not well understood. Here, we demonstrate that SP3 is overexpressed in invasive breast cancer cells vs normal counterparts. The gene expression analysis from The Cancer Genome Atlas datasets indicated that SP3 is strongly correlated with Akt signalling-related proteins, G protein subunit alpha 13, and RAB33B (RAB33B, member RAS oncogene family). RNA interference of SP3 decreased active phosphorylation of Akt at serine and threonine sites. These findings indicate that SP3 exhibits a pro-oncogenic function, which clearly fits the description of an nononcogene addiction gene. Future analyses are prompted to uncover the SP3 gene regulation function and to reveal downstream targets of SP3 in breast cancer.

Dual inhibition of glycolysis and autophagy as a therapeutic strategy in the treatment of Ehrlich ascites carcinoma.

Cancer cells have extra biosynthetic demands to sustain cell growth and redox homeostasis. Glycolysis and autophagy are crucial to fuel and recycle these biosynthetic demands. This plasticity of cancer cell metabolism participates in therapy resistances. The current study was designed to assess the therapeutic efficacy of dual targeting of glycolysis and autophagy in cancer. Using 3-bromopyruvate (3-BP; antiglycolytic inhibitor) and hydroxychloroquine (HCQ; autophagy inhibitor), we demonstrate their antitumor activity in Ehrlich ascites carcinoma (EAC)-bearing mice. A combination of 3-BP and HCQ significantly decreases tumor ascitic volume and cell count as compared with the EAC group and individual treatment groups. The enhanced antitumor activity is accompanied by hexokinase inactivation, inhibition of cellular protective autophagy, elevated antioxidant activity, and reduced oxidative stress levels. Together, these results suggest targeting both pathways in cancer as an effective therapeutic strategy. Further studies are required to validate this strategy in different cancer models and preclinical trials.

Combination of arsenic trioxide and cisplatin synergistically inhibits both hexokinase activity and viability of Ehrlich ascites carcinoma cells.

Hexokinase-2 is overexpressed in several carcinomas including breast cancer to sustain energy for rapidly dividing cells and associates with chemoresistance. However, the impact of chemo drugs (alone or in combination) on hexokinase activity and autophagic cell death is unclear. In this report, we used an in vivo murine adenocarcinoma model to validate the effects of As2 O3 and cisplatin on hexokinase activity and autophagic cancer cell death. We found that the two drugs inhibit hexokinase activity and induce autophagic marker, beclin 1 expression. Interestingly, combining As2 O3 with cisplatin synergistically enhanced these effects and alleviated oxidative stress often encountered in As2 O3 treatment. Altogether, our data provide direct evidence that inhibition of hexokinase activity and induction of autophagic cell death are mediating the antineoplastic effects of As2 O3 and cisplatin. Our findings raise the potential of combining As2 O3 with cisplatin as an approach to augment cisplatin-induced cell death and combat cisplatin chemoresistance in cancer.

FAM98A is a novel substrate of PRMT1 required for tumor cell migration, invasion, and colony formation.

Protein arginine methylation, which is mediated by a family of protein arginine methyltransferases (PRMTs), is associated with numerous fundamental cellular processes. Accumulating studies have revealed that the expression of multiple PRMTs promotes cancer progression. In this study, we examined the role of PRMT1 in ovarian cancer cells. PRMT1 is expressed in multiple ovarian cancer cells, and the depletion of its expression suppressed colony formation, in vivo proliferation, migration, and invasion. To gain insight into PRMT1-mediated cancer progression, we searched for novel substrates of PRMT1. We found that FAM98A, whose physiological function is unknown, was arginine-methylated by PRMT1. FAM98A is expressed in numerous ovarian cancer cell lines and is important for the malignant characteristics of ovarian cancer cells. Our results indicate the possible role of the PRMT1-FAM98A pathway in cancer progression.

UBE2S is associated with malignant characteristics of breast cancer cells.

Ubiquitination is essential for various biological processes, such as signal transduction, intracellular trafficking, and protein degradation. Accumulating evidence has demonstrated that ubiquitination plays a crucial role in cancer development. In this report, we examine the expression and function of ubiquitin-conjugating enzyme E2S (UBE2S) in breast cancer. Immunohistochemical analysis revealed that UBE2S is highly expressed in breast cancer. The depletion of UBE2S by siRNA induced disruption of the actin cytoskeleton and focal adhesions. Interestingly, phosphorylation of FAK at Tyr397, which is important for the transduction of integrin-mediated signaling, was significantly reduced by UBE2S knockdown. We also show that UBE2S knockdown suppressed the malignant characteristics of breast cancer cells, such as migration, invasion, and anchorage-independent growth. Our results indicate that UBE2S could be a potential target for breast cancer treatment.

Special AT-rich sequence-binding protein 2 suppresses invadopodia formation in HCT116 cells via palladin inhibition.

Invadopodia are specialized actin-based microdomains of the plasma membrane that combine adhesive properties with matrix degrading activities. Proper functioning of the bone, immune, and vascular systems depend on these organelles, and their relevance in cancer cells is linked to tumor metastasis. The elucidation of the mechanisms driving invadopodia formation is a prerequisite to understanding their role and ultimately to controlling their functions. Special AT-rich sequence-binding protein 2 (SATB2) was reported to suppress tumor cell migration and metastasis. However, the mechanism of action of SATB2 is unknown. Here, we show that SATB2 inhibits invadopodia formation in HCT116 cells and that the molecular scaffold palladin is inhibited by exogenous expression of SATB2. To confirm this association, we elucidated the function of palladin in HCT116 using a knock down strategy. Palladin knock down reduced cell migration and invasion and inhibited invadopodia formation. This phenotype was confirmed by a rescue experiment. We then demonstrated that palladin expression in SATB2-expressing cells restored invasion and invadopodia formation. Our results showed that SATB2 action is mediated by palladin inhibition and the SATB2/palladin pathway is associated with invadopodia formation in colorectal cancer cells.

Corrigendum to:"Deep learning based suture training system" [Surgery Open Science 15 (2023) 1-11/1].

[This corrects the article DOI: 10.1016/j.sopen.2023.07.023.].

The impact of different fixatives on immunostaining of lung adenocarcinomas in pleural effusion cell blocks.

BACKGROUND: Cell blocks (CBs) are widely used for biomarker analyses such as immunostaining. Although immunohistochemistry on formalin-fixed paraffin-embedded tissues is standardized, there are multiple preparation methods and fixatives for cytology. Our objective was to investigate the effect of different common fixatives on the immunoreactivity of pleural effusion CBs with metastatic lung adenocarcinomas. METHODS: This prospective study included 24 malignant pleural effusions from different patients with lung adenocarcinoma. From each case, four identical CBs were fixed in 10% neutral buffered formalin, PreservCyt, CytoLyt, and CytoRich Red (only 17 of the cases), respectively. Samples containing <100 malignant cells were excluded. All CBs were stained with thyroid transcription factor 1 (TTF-1; clones 8G7G3/1 and SPT24), napsin A, claudin 4, CEA, CK7, and epithelial cell adhesion molecule (EpCAM; clones BS14, Ber-Ep4, and MOC-31). The fraction and intensity of stained cells were evaluated. RESULTS: Of the investigated markers, a significant difference in staining proportion was seen for TTF-1 clone 8G7G3/1 and EpCAM clone MOC-31, especially with cases being negative in CytoLyt (33.3% and 83.3% positive, respectively) and PreservCyt (62.5% and 83.3%) whereas being positive in CytoRich Red (76.5% and 94.1%) and formalin (both 95.8%). A significantly weaker intensity of staining was seen for all alcohol-based fixatives compared to formalin for TTF-1 clone 8G7G3/1, napsin A, and EpCAM clone MOC-31, whereas EpCAM clone Ber-Ep4 was significantly weaker only in PreservCyt compared with formalin. CONCLUSIONS: Immunocytochemical expression and concordance with formalin-fixed CBs differ depending on the used fixative as well as the antibody and clone, warranting investigation of the reliability of each biomarker for non-formalin-fixed cytology.

Sarcoidosis and Sjögren's syndrome: clinical and salivary evaluation.

BACKGROUND: Sarcoidosis and Sjögren's syndrome are two different diseases; however, when affecting the salivary glands, both diseases exhibit similar clinical signs and symptoms, which often complicates the diagnosis. The purpose of this study was to investigate the possibility of using salivary electrophoresis to differentiate between the two diseases. METHODS: Saliva was collected from patients with sarcoidosis and patients with Sjögren's syndrome. Salivary flow rate, total protein, and electrophoretic profiles were examined. RESULTS: Mean salivary flow rate was 0.41 ± 0.07 ml/min/gland vs. 0.43 ± 0.07 ml/min/gland; total salivary protein was 130.0 ± 29.2 mg% vs. 104.0 ± 8.8 mg% for sarcoidosis vs. Sjögren's syndrome, respectively. No differences were observed in salivary flow rate, total salivary protein, or electrophoretic profile between patients with sarcoidosis and patients with Sjögren's syndrome (P = 0.768, 0.718, and 1.000, respectively). CONCLUSIONS: Salivary protein electrophoresis does not appear to be useful to differentiate between sarcoidosis and Sjögren's syndrome.

Deep learning based suture training system.

Background and objectives: Surgical suturing is a fundamental skill that all medical and dental students learn during their education. Currently, the grading of students' suture skills in the medical faculty during general surgery training is relative, and students do not have the opportunity to learn specific techniques. Recent technological advances, however, have made it possible to classify and measure suture skills using artificial intelligence methods, such as Deep Learning (DL). This work aims to evaluate the success of surgical suture using DL techniques. Methods: Six Convolutional Neural Network (CNN) models: VGG16, VGG19, Xception, Inception, MobileNet, and DensNet. We used a dataset of suture images containing two classes: successful and unsuccessful, and applied statistical metrics to compare the precision, recall, and F1 scores of the models. Results: The results showed that Xception had the highest accuracy at 95 %, followed by MobileNet at 91 %, DensNet at 90 %, Inception at 84 %, VGG16 at 73 %, and VGG19 at 61 %. We also developed a graphical user interface that allows users to evaluate suture images by uploading them or using the camera. The images are then interpreted by the DL models, and the results are displayed on the screen. Conclusions: The initial findings suggest that the use of DL techniques can minimize errors due to inexperience and allow physicians to use their time more efficiently by digitizing the process.

Estimation of lower limb joint moments based on the inverse dynamics approach: a comparison of machine learning algorithms for rapid estimation.

The aim of this study is to estimate the joint moments of the ankle, knee, and hip joints during walking. A sit-to-stand (STS) movement analysis was first performed on 20 participants with different anthropometric characteristics. Then, analysis of the dynamics of the STS motion was used to develop a biomechanical model. Decision tree (DT), linear regression (LR), support vector machine (SVM), random forest (RF), and three deep learning (DL) algorithms and deep neural network (DNN), long-short-term memory (LSTM), and convolutional neural network (CNN) are examined in this work to estimate three joint moments: ankle, knee, and hip. The results of the seven algorithms were evaluated using four statistical benchmarks: MSR, RMSE, correlation coefficient (R), and MAE to find the most accurate one. The results show that the most successful algorithms were LSTM in estimating knee, hip, and ankle joint moments using 19 and 7 inputs. The R value was 0.9990 using 19 inputs and 0.9972 using 7 inputs. The other algorithms have a correlation coefficient (R) success of 0.9902, 0.9770, 0.9884, 0.9577, 0.9786, and 0.9022 for RF, CNN, DT, DNN, SVM, and LR, respectively. The prediction of joint moments plays a crucial role in the design of the biomechanical system with the desired mechanical properties. Especially, the need has arisen to predict joint moments in a shorter time to utilize in real-time active prosthesis/orthosis controllers.

Non-invasive detection of anemia using lip mucosa images transfer learning convolutional neural networks.

Anemia is defined as a drop in the number of erythrocytes or hemoglobin concentration below normal levels in healthy people. The increase in paleness of the skin might vary based on the color of the skin, although there is currently no quantifiable measurement. The pallor of the skin is best visible in locations where the cuticle is thin, such as the interior of the mouth, lips, or conjunctiva. This work focuses on anemia-related pallors and their relationship to blood count values and artificial intelligence. In this study, a deep learning approach using transfer learning and Convolutional Neural Networks (CNN) was implemented in which VGG16, Xception, MobileNet, and ResNet50 architectures, were pre-trained to predict anemia using lip mucous images. A total of 138 volunteers (100 women and 38 men) participated in the work to develop the dataset that contains two image classes: healthy and anemic. Image processing was first performed on a single frame with only the mouth area visible, data argumentation was preformed, and then CNN models were applied to classify the dataset lip images. Statistical metrics were employed to discriminate the performance of the models in terms of Accuracy, Precision, Recal, and F1 Score. Among the CNN algorithms used, Xception was found to categorize the lip images with 99.28% accuracy, providing the best results. The other CNN architectures had accuracies of 96.38% for MobileNet, 95.65% for ResNet %, and 92.39% for VGG16. Our findings show that anemia may be diagnosed using deep learning approaches from a single lip image. This data set will be enhanced in the future to allow for real-time classification.

Profile of orthopaedic day-case procedures at a district general hospital (retrospective study).

BACKGROUND: The outcomes of orthopaedic day-case procedures have been reported widely, but there is a lack of reports from secondary health facilities such as district hospitals. AIM: We aimed to perform a retrospective analysis of patient records to capture the profile of day-case procedures performed. MATERIALS AND METHODS: We conducted a retrospective analysis of day-case procedures at the dedicated Day Surgery Unit of a moderate-sized district hospital in Saudi Arabia between January 2021 and December 2022. The medical records of all the patients who had day-case procedures by the hospital's orthopaedic unit were analyzed. RESULTS: Within the study period, 71 out of 914 elective orthopaedic procedures were carried out as day-cases, giving a day-case surgery rate of 7.8%. The mean age was 25.3 ± 12.2 (range, 4-55 years), and the male-to-female ratio was 6:1. The spectrum of the procedures was dominated by implant removal in 59 cases (83.1%). Whilst the anaesthetic technique varied, all the patients were ASA class I or II. There were minor complications in 10 patients (14.0%), with 7 of them (9.8%) needing inpatient admission. There was no cancellation of cases in our study. CONCLUSION: We found day-case procedures to be safe and effective but with low utilisation of the Day Surgery Unit, which can be improved through the development of a detailed protocol for day surgery in the hospital.

Anemia detection through non-invasive analysis of lip mucosa images.

This paper aims to detect anemia using images of the lip mucosa, where the skin tissue is thin, and to confirm the feasibility of detecting anemia noninvasively and in the home environment using machine learning (ML). Data were collected from 138 patients, including 100 women and 38 men. Six ML algorithms: artificial neural network (ANN), decision tree (DT), k-nearest neighbors (KNN), logistic regression (LR), naive bayes (NB), and support vector machine (SVM) which are widely used in medical applications, were used to classify the collected data. Two different data types were obtained from participants' images (RGB red color values and HSV saturation values) as features, with age, sex, and hemoglobin levels utilized to perform classification. The ML algorithm was used to analyze and classify images of the lip mucosa quickly and accurately, potentially increasing the efficiency of anemia screening programs. The accuracy, precision, recall, and F-measure were evaluated to assess how well ML models performed in predicting anemia. The results showed that NB reported the highest accuracy (96%) among the other ML models used. DT, KNN and ANN reported an accuracies of (93%), while LR and SVM had an accuracy of (79%) and (75%) receptively. This research suggests that employing ML approaches to identify anemia will help classify the diagnosis, which will then help to create efficient preventive measures. Compared to blood tests, this noninvasive procedure is more practical and accessible to patients. Furthermore, ML algorithms may be created and trained to assess lip mucosa photos at a minimal cost, making it an affordable screening method in regions with a shortage of healthcare resources.

Corrigendum: Anemia detection through non-invasive analysis of lip mucosa images.

[This corrects the article DOI: 10.3389/fdata.2023.1241899.].