CDDP-induced desmoplasia-like changes in oral cancer tissues are related to SASP-related factors induced by the senescence of cancer cells.

The tumor microenvironment (TME) concept has been proposed and is currently being actively studied. The development of extracellular matrix (ECM) in the TME is known as desmoplasia and is observed in many solid tumors. It has also been strongly associated with poor prognosis and resistance to drug therapy. Recently, cellular senescence has gained attention as an effect of drug therapy on cancer cells. Cellular senescence is a phenomenon wherein proliferating cells become resistant to growth-promoting stimuli, secrete the SASP (senescence-associated phenotypic) factors, and stably arrest the cell cycle. These proteins are rich in pro-inflammatory factors, such as interleukin (IL)-6, IL-8, C-X-C motif chemokine ligand 1, C-C motif chemokine ligand (CCL)2, CCL5, and matrix metalloproteinase 3. This study aimed to investigate the desmoplasia-like changes in the TME before and after cancer drug therapy in oral squamous cell carcinomas, evaluate the effect of anticancer drugs on the TME, and the potential involvement of cancer cell senescence. Using a syngeneic oral cancer transplant mouse model, we confirmed that cis-diamminedichloroplatinum (II) (CDDP) administration caused desmoplasia-like changes in cancer tissues. Furthermore, CDDP treatment-induced senescence in tumor-bearing mouse tumor tissues and cultured cancer cells. These results suggest CDDP administration-induced desmoplasia-like structural changes in the TME are related to cellular senescence. Our findings suggest that the administration of anticancer drugs alters the TME of oral cancer cells. Additionally, oral cancer cells undergo senescence, which may influence the TME through the production of SASP factors.

Exploring animal protein sources against prevent age-related short-term memory decline in mice: Dietary fish (Alaska Pollock) protein exhibits superior protective effects.

Dietary factors, particularly proteins, have been extensively explored to combat cognitive impairment. We have previously reported that dietary fish (Alaska Pollock) protein (APP) is more effective than casein (CAS) or fish oil in maintaining short-term memory in senescence-accelerated mice prone 10 (SAMP10). To examine the specificity of the protective effect of APP intakes against short-term memory decline, we assessed the impact of various dietary animal proteins, including APP, CAS, chicken breast protein (CP), and whey protein (WP), against age-related cognitive function in SAMP10 mice. After feeding the experimental diets for 5 months, memory was assessed using the Y-maze. The APP group exhibited a significant increase in spontaneous alternation behavior as an indicator of working memory when group compared with groups fed with other protein source. Additionally, the APP group displayed significantly higher neurofilament heavy chain positivity than the CAS and CP groups, as evidenced immunohistochemical analysis. Gut microbiota analysis indicated that dietary APP significantly enhanced the relative abundance of Lactobacillus, which positively correlated with spontaneous alternation behavior. Collectively, these findings suggest that dietary APP is more effective than CAS, CP, or WP in preventing age-related short-term memory decline and morphological abnormalities in the hippocampal axons of SAMP10 mice. Moreover, APP-mediated improvements in cognitive deficits may be associated with changes in microbiota diversity. PRACTICAL APPLICATION: This research suggests that dietary fish protein from Alaska Pollock may be more efficient in prevention short-term memory decline in mice, compared to other animal proteins. This finding has practical implications for nutritional optimization, developing the new health food products, and elucidating the relationship between the impact of specific proteins on gut microbiota and prevention of age-related cognitive decline.

Risk factors for impacted lower third molar root perforation through the undercut-shaped lingual plate: preoperative insights from panoramic radiography.

This study aimed to identify risk factors associated with perforation of the undercut (U)-shaped lingual plate (LP) by the lower third molar (LM3) root using panoramic radiography (PAN). We retrospectively examined 468 impacted LM3s from 468 individuals, categorizing LM3-LP associations and LP morphology in the coronal section of cone-beam computed tomography as perforation or nonperforation and U-type or non-U-type, respectively. The outcome was the combination of perforation and U-type, and study variables included patient demographics (age and sex) and PAN-associated features (Winter's classification, Pell-Gregory classification, and two major Rood signs). Multivariate logistic regression methods were used for analysis. Perforated and U-type LPs were observed in 205 (43.8%) and 212 (45.3%) cases, respectively. The double-positive outcome was observed in 126 LM3s (26.9%). In the multivariate model, age ≥ 26 years [odds ratio (OR), 2.66; p = 0.002], men (OR, 2.01; p = 0.002), mesioangular (OR, 2.74; p = 0.038) and horizontal impaction (OR, 3.05; p = 0.019), and root darkening (OR, 1.73; p = 0.039) were independently associated with the risk. Class III impaction (OR, 0.35; p = 0.021) and interruption of the white line (OR, 0.55; p = 0.017) were negatively correlated with the risk. In conclusion, this study highlights the importance of identifying the higher probability of U-type LP perforation by the LM3 root in men aged over their midtwenties with Class I/II impaction and mesioangularly or horizontally impacted LM3s, along with root darkening and no interruption of the white line on PAN.

Deep learning model for the automated evaluation of contact between the lower third molar and inferior alveolar nerve on panoramic radiography.

Background/purpose: In lower third molar (LM3) surgery, panoramic radiography (PAN) is important for the initial assessment of the anatomical association between LM3 and the inferior alveolar nerve (IAN). This study aimed to develop a deep learning model for the automated evaluation of the LM3-IAN association on PAN. Further, its performance was compared with that of oral surgeons using original and external datasets. Materials and methods: In total, 579 panoramic images of LM3 from 384 patients in the original dataset were utilized. The images were divided into 483 images for the training dataset and 96 for the testing dataset at a ratio of 83:17. The external dataset comprising 58 images from an independent institution was used for testing only. The LM3-IAN associations on PAN were categorized into direct or indirect contact based on cone-beam computed tomography (CBCT). The You Only Look Once (YOLO) version 3 algorithm, a fast object detection system, was applied. To increase the amount of training data for deep learning, PAN images were augmented using the rotation and flip techniques. Results: The final YOLO model had high accuracy (0.894 in the original dataset and 0.927 in the external dataset), recall (0.925, 0.919), precision (0.891, 0.971), and f1-score (0.908, 0.944). Meanwhile, oral surgeons had lower accuracy (0.628, 0.615), recall (0.821, 0.497), precision (0.607, 0.876), and f1-score (0.698, 0.634). Conclusion: The YOLO-driven deep learning model can help oral surgeons in the decision-making process of applying additional CBCT to confirm the LM3-IAN association based on PAN images.

External validation of cone-beam computed tomography- and panoramic radiography-featured prediction models for inferior alveolar nerve injury after lower third molar removal: proposal of a risk calculator.

We previously developed basic and extended models to predict inferior alveolar nerve injuries (IANI) after lower third molar (LM3) removal based on cone-beam computed tomography (CBCT) images. Although these models comprised predictors, including increased age and inferior alveolar canal-related CBCT factors, external validations were lacking. Therefore, this study externally validated these models and compared them with other related models based on their performance. Original and newly validated samples included patients who underwent LM3 removal following CBCT. Subsequently, 39 and 25 patients with IANI, then 457 and 295 randomly selected patients without IANI were chosen of the observed 1573 and 1052 patients, respectively. CBCT- and panoramic radiograph (PAN)-featured models were validated. Then, models' discrimination and calibration abilities were assessed using C-statistics and calibration plots, respectively. Brier scores were also quantified, after which logistic recalibration was achieved to optimize calibration, and a risk calculator was developed. During the external validation, the extended model exhibited the best C-statistic (0.822) and Brier score (0.064), whereas two CBCT- and two PAN-featured models showed lower performances with C-statistics (0.764, 0.706, 0.584, and 0.627) and Brier scores (0.069, 0.074, 0.075, and 0.072). Besides, all models showed a tendency to overpredict its high-risk range. However, recalibration of the extended model resulted in excellent calibration performance. CBCT-featured models, especially the extended model, conclusively showed a superior predictive performance to PAN models. Therefore, the risk calculator on the extended CBCT model is proposed to be a clinical decision-aid tool that preoperatively predicts IANI risk.

Protective Effects of Fish (Alaska Pollock) Protein Intake against Short-Term Memory Decline in Senescence-Accelerated Mice.

Dietary fish intake has proven to have health benefits in humans. n-3 polyunsaturated fatty acids (PUFAs) in fish oil (FO), especially, may provide protection against age-related cognitive disorders. Owing to the unique benefits of n-3 PUFAs, other nutrients, such as fish protein (FP), have not been well studied. To clarify the effects of FO and FP on brain function, we investigated whether FO or FP feeding can prevent age-related cognitive dysfunction in senescence-accelerated mouse-prone 10 (SAMP10) mice. The FP group maintained a better working memory compared to the control and FO groups in the Y-maze test, but not episodic memory in the novel object recognition test. To evaluate demyelination levels, we measured neurofilament H (NfH) and myelin basic protein (MBP) immunoreactivity in the hippocampus (Hipp). Axon morphology was maintained in the FP group, but not in the control and FO groups. Additionally, the percentage of positive area for double-staining with NfH/MPB was significantly higher in the Hipp of FP-fed mice than in the control (p < 0.05). These results suggest that FP intake prevents age-related cognitive dysfunction by maintaining axonal morphology in the Hipp of SAMP10 mice.

GATA6 regulates expression of annexin A10 (ANXA10) associated with epithelial-mesenchymal transition of oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) can disturb oral function and quality of life and is associated with poor survival, likely due to the development of cervical lymph node metastases. Epithelial-mesenchymal transition (EMT) is a process in which cells acquire molecular alterations that facilitate cell motility and invasion, and has been associated with tumor metastasis. EMT changes also play important roles in the induction of lymph node metastasis in OSCC. GATA6 is known as the earliest marker of the primitive endoderm lineages. GATA6 inhibits de-differentiation and EMT in human pancreatic ductal adenocarcinoma cells and promotes EMT. However, in OSCC, the expression and function of GATA6 in EMT and lymph node metastasis remains unclear. Therefore, this study aimed to clarify the targets of GATA6 in OSCC cells and whether the change in GATA6 expression affects EMT in OSCC cells, as well as the association between GATA6 and lymph node metastasis. The results showed that GATA6 knockdown OSCC cells promoted EMT and increased lymph node metastasis compared with control cells, whereas the overexpression of GATA6 inhibited the induction of EMT and reduced lymph node metastasis. In addition, annexin A10 (ANXA10) which is the largest type of Ca2+-regulated phospholipid-binding protein in eukaryotic cells was detected as a target gene for GATA6 and ANXA10 suppressed Vimentin expression in EMT in OSCC. Therefore, the GATA6/ANXA10 cascade may be a potential therapeutic approach for the treatment of lymph node metastases in OSCC patients.

Predictive scoring model for inferior alveolar nerve injury after lower third molar removal based on features of cone-beam computed tomography image.

INTRODUCTION: This study aimed to construct a predictive scoring system for inferior alveolar nerve injury (IANI) following lower third molar (LM3) surgery based on cone-beam computed tomography (CBCT) images. MATERIAL AND METHODS: Of the 1573 patients who underwent LM3 removal following the CBCT, 39 with IANI and 457 randomly selected patients without IANI were enrolled. We collected information regarding the demographic characteristics of the patients, surgical situations, and inferior alveolar canal (IAC)-related CBCT factors. The association with IANI-risk was evaluated with a backward stepwise logistic regression model as per the Akaike information criterion. Scoring models' abilities of discrimination (area under the curve) and calibration (Hosmer-Lemeshow test and calibration plots) were assessed, followed by evaluation of the clinical usefulness using decision curve analysis. RESULTS: As per the multivariate analysis, the coronal positioned IAC on the enlarged root (odds ratio [OR], 3.78; P = 0.001), the length of perforated IAC (>3.4 mm) (OR, 3.05; P = 0.012), lingual/inter-radicular position of the IAC (OR, 3.96; P = 0.001), multiple roots closed to the perforated IAC (OR, 2.78; P = 0.025), and age >30 y (OR, 2.31; P = 0.076) were identified in the extended scoring model ranging from 0 to 12. This model was compared with our previously constructed baseline model that involved the latter three variables mentioned above, resulting in superior performance than that of the baseline model. CONCLUSION: The extended model would be a useful tool for reliable determination of the preoperative probability of IANI.

Dietary Alaska pollock protein alters insulin sensitivity and gut microbiota composition in rats.

Fish protein is not only nutritional but also promotes health by improving insulin sensitivity and hypercholesterolemia. Few studies have examined the relationship between gut microbiota and the enhanced insulin sensitivity due to the intake of Alaska pollock protein (APP). Hence, we assessed the glycolytic enzyme inhibitory activity of APP in in vitro study and the alteration of blood glucose level in insulin tolerance test (ITT) and glucose tolerance test (GTT) and gut microbiota following APP intake in the in vivo study. In initial experiments, the glycolytic enzyme (α-amylase, α-glucosidase, and sucrase) inhibitory activities of APP and its digest were not drastically altered compared with that of casein and its digests. In further experiments, rats fed an AIN-93G diet containing 20% (w/w) casein or APP for 8 weeks, and the composition of fecal microbiota analyzed by 16S rRNA amplicon sequence analysis. In addition, at 6 and 7 weeks of administration of experimental diet, insulin and glucose tolerance tests were evaluated, respectively. Compared with dietary casein, dietary APP has blood glucose-lowering activity as evident in the ITT and GTT. Moreover, APP group altered the structure of fecal microbiota, and area under the curves of the ITT and GTT and the relative abundance of Blautia, which is associated with glucose metabolism, tended to be positively correlated (P = 0.08 and 0.10, respectively). This study illustrates a novel finding that APP intake could alter the composition of gut microbiota and improve insulin sensitivity. PRACTICAL APPLICATION: Studies in animals and humans have shown that Alaska pollock protein (APP) intake improves insulin sensitivity, allowing the body to utilize blood glucose more effectively, thereby keeping blood sugar levels under control. Microorganisms residing in the human gut are associated with glucose metabolism. This study shows that the relative APP intake alters the composition of these gut microorganisms, more than casein intake and therefore might prevent hyperglycemia and type 2 diabetes.

Oxidative DNA Damage Induced by Pirarubicin, an Anthracycline Anticancer Agent, in the Presence of Copper(II).

BACKGROUND/AIM: One mechanism of the anticancer action of anthracyclines is believed to be oxidative DNA damage. Previously, we reported that doxorubicin induced oxidative DNA damage in the presence of Cu(II). However, the mechanism of pirarubicin-induced oxidative DNA damage has not been well clarified. MATERIALS AND METHODS: DNA damage by pirarubicin in the presence of Cu(II) was analyzed using pBR322 plasmid DNA. O2•- derived from pirarubicin in the presence of Cu(II) was detected by cytochrome c reduction. RESULTS: Pirarubicin induced DNA damage in the presence of Cu(II). Scavenger experiments suggest that reactive species are generated from H2O2 and Cu(I). Pirarubicin induced O2•- production in the presence of Cu(II). CONCLUSION: These findings suggest that pirarubicin plus Cu(II) induces oxidative DNA damage in a similar manner to doxorubicin, and Cu(II)-mediated oxidative DNA damage may serve as a common mechanism for antitumor effects of anthracyclines.

Pirarubicin, an Anthracycline Anticancer Agent, Induces Apoptosis Through Generation of Hydrogen Peroxide.

BACKGROUND/AIM: Pirarubicin (THP) has shown equal or superior cytotoxicity compared to doxorubicin. One of the main anticancer actions of doxorubicin is believed to be involved in ROS (reactive oxygen species) generation. Therefore, the anticancer mechanisms of THP may involve ROS generation. The aim of this study was to clarify the mechanisms of THP-induced apoptosis through ROS generation. MATERIALS AND METHODS: We analyzed the apoptotic events induced by THP in HL-60 cells and HP100 cells, hydrogen peroxide (H2O2)-resistant cells derived from HL-60. RESULTS: The apparent cytotoxicity could be detected at above 0.1 µM in HL-60 cells after 24-h incubation, whereas it was suppressed under these conditions in HP100 cells. In HP100 cells, THP-induced apoptosis, evaluated by DNA ladder formation, H2O2 generation, mitochondrial membrane potential decrease and caspase-3/7 activity, was suppressed or delayed compared to those of HL-60 cells. CONCLUSION: These findings can be explained by the involvement of H2O2 generation in the THP apoptotic pathway. This is the first report on THP-induced apoptosis through the H2O2 generation.

Expression of the estrogen receptor alpha gene in the anal fin of Japanese medaka, Oryzias latipes, by environmental concentrations of bisphenol A.

The anal fin in Japanese medaka, Oryzias latipes, is a typical sexual secondary character. In the present study, we focused on this organ and examined the effects of low doses of a natural estrogen, 17beta-estradiol (E(2)), and an environmental xenoestrogen, bisphenol A (BPA), in vivo by monitoring estrogen receptor (ER) alpha gene expression. Groups of adult male and female medaka were immersed in 10(-9) M E(2) or 10(-10) to 10(-8) M BPA and the levels of ERalpha gene transcripts in the anal fins were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). One day of treatment with each concentration of BPA examined and 10(-9) M E(2) increased the levels of ERalpha mRNA in female anal fins by 3-fold as compared with controls. In the male specimens, neither 10(-9) M E(2) nor 10(-10) M BPA showed remarkable effects on the anal fins as compared with the results in females, but 10(-9) and 10(-8) M BPA increased the levels of ERalpha mRNA by 2.3- and 3.3-fold with 1 day of exposure, respectively. The present results showed that medaka anal fins may be a sensitive bio-indicator for screening of environmental estrogenic chemicals.