Sustained delivery of celecoxib from nanoparticles embedded in hydrogel injected into the biopsy cavity to prevent biopsy-induced breast cancer metastasis.

PURPOSE: We have previously reported that protracted Cyclooxygenase-2 (COX-2) activity in bone marrow-derived cells (BMDCs) infiltrating into biopsy wounds adjacent to the biopsy cavity of breast tumors in mice promotes M2-shift of macrophages and pro-metastatic changes in cancer cells, effects which were suppressed by oral administration of COX-2 inhibitors. Thus, local control of COX-2 activity in the biopsy wound may mitigate biopsy-induced pro-metastatic changes. METHODS: A combinatorial delivery system-thermosensitive biodegradable poly(lactic acid) hydrogel (PLA-gel) incorporating celecoxib-encapsulated poly(lactic-co-glycolic acid) nanoparticles (Cx-NP/PLA-gel)-was injected into the biopsy cavity of Py230 murine breast tumors to achieve local control of COX-2 activity in the wound stroma. RESULTS: A single intra-biopsy cavity injection of PLA-gel loaded with rhodamine-encapsulated nanoparticles (NPs) showed sustained local delivery of rhodamine preferentially to infiltrating BMDCs with minimal to no rhodamine uptake by the reticuloendothelial organs in mice. Moreover, significant reductions in M2-like macrophage density, cancer cell epithelial-to-mesenchymal transition, and blood vessel density were observed in response to a single intra-biopsy cavity injection of Cx-NP/PLA-gel compared to PLA-gel loaded with NPs containing no payload. Accordingly, intra-biopsy cavity injection of Cx-NP/PLA-gel led to significantly fewer metastatic cells in the lungs than control-treated mice. CONCLUSION: This study provides evidence for the feasibility of sustained, local delivery of payload preferential to BMDCs in the wound stroma adjacent to the biopsy cavity using a combinatorial delivery system to reduce localized inflammation and effectively mitigate breast cancer cell dissemination.

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.

Involvement of ribonuclease Y in pilus production by M49 Streptococcus pyogenes strain via modulation of messenger RNA level of transcriptional regulator.

Streptococcus pyogenes displays a wide variety of pili, which is largely dependent on serotype. A distinct subset of S. pyogenes strains that possess the Nra transcriptional regulator demonstrates thermoregulated pilus production. Findings obtained in the present study of an Nra-positive serotype M49 strain revealed involvement of conserved virulence factor A (CvfA), also referred to as ribonuclease Y (RNase Y), in virulence factor expression and pilus production, while a cvfA deletion strain showed reduced pilus production and adherence to human keratinocytes as compared with wild-type and revertant strains. Furthermore, transcript levels of pilus subunits and srtC2 genes were decreased by cvfA deletion, which was remarkable at 25°C. Likewise, both messenger RNA (mRNA) and protein levels of Nra were remarkably decreased by cvfA deletion. Whether the expression of other pilus-related regulators, including fasX and CovR, was subject to thermoregulation was also examined. While the mRNA level of fasX, which inhibits cpa and fctA translation, was decreased by cvfA deletion at both 37°C and 25°C, CovR mRNA and protein levels, as well as its phosphorylation level were not significantly changed, suggesting that neither fasX nor CovR is necessarily involved in thermosensitive pilus production. Phenotypic analysis of the mutant strains revealed that culture temperature and cvfA deletion had varied effects on streptolysin S and SpeB activities. Furthermore, bactericidal assay data showed that cvfA deletion decreased the rate of survival in human blood. Together, the present findings indicate that CvfA is involved in regulation of pilus production and virulence-related phenotypes of the serotype M49 strain of S. pyogenes.

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.

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.

Risk stratification against inferior alveolar nerve injury after lower third molar extraction by scoring on cone-beam computed tomography image.

The study aimed to stratify the risk of inferior alveolar nerve injury (IANI) after lower third molar (LM3) surgery with a scoring system using identified predictive factors based on cone-beam computed tomography (CBCT) images. In a case-control study, the primary outcome was IANI occurrence. The control group included randomly selected patients without IANI. Predictor variables included patient demographics, surgical situations, Pell-Gregory classification, and inferior alveolar canal (IAC)-associated factors on CBCT. Study variables were analyzed using logistic regression models. Risk stratification was assessed by a scoring system that was constructed using independent predictors. The 858 patients who underwent LM3 surgery (1177 teeth) after CBCT scan were divided into case (25 patients, 2.9%, 27 teeth) and control (235 patients, 300 teeth) groups. In the multivariate model, lingual/inter-radicular position of IAC [odds ratio (OR) 7.21; P < 0.001; assigned score, 2], multiple roots closed to the IAC with cortical perforation (OR 3.72; P = 0.015; 1), and age > 30 years (OR 4.99; P = 0.008; 2) were associated with an increased IANI-risk. The IANI-risk scoring system could be stratified into low- and high-risk groups at a cutoff score of 3 (sensitivity, 68.0%; specificity, 90.6%; positive predictive value, 17.8%; positive likelihood ratio, 7.23). In conclusion, the high-risk group of IANI after LM3 surgery corresponded to individuals with multiple factors: lingual/inter-radicular IAC position to LM3, multiple roots with perforated IAC, and increased age (> 30 years). Raising awareness of the higher probability for IANI is needed for patients with multiple aforementioned factors.

Gingival Neurofibroma With Teardrop-Shaped Defects of the Interdental Alveolar Bone: An Unusual Oral Manifestation of Neurofibromatosis Type 1.

Gingival enlargement, although frequently encountered in clinical settings, is rarely associated with systemic diseases or syndromes. Among the diverse pathological conditions of neurofibromatosis type 1 (NF-1), minor manifestations in the orofacial region are occasionally overlooked. Herein, the authors present an unusual case of gingival neurofibroma in a patient with NF-1 associated with characteristic osseous defects in the alveolus in the long-term course of 17 years from the first examination.A 5-year-old boy with NF-1 was referred for the evaluation of gingival enlargement in the posterior left maxilla. An incisal biopsy led to the diagnosis of neurofibroma. At 22 years of age, the patient was referred again with a complaint of bleeding and pain in the same region refractory to periodontal therapy. The gingiva and tuberosity were swollen, and the second molar was affected by the tumor. Radiography revealed a low level of the interdental septum beneath the tumor with a relatively intact periodontal cortical bone, exhibiting a teardrop-shaped bone defect. The lesion was completely resected with the periosteum.Gingival neurofibroma in NF-1 may be associated with osseous and dental abnormalities and can be mistaken for periodontitis. Raising awareness of this clinical entity can lead to proper management of the esthetic and functional problems in the oral and maxillofacial region.

Exfoliative toxin A staphylococcal scalded skin syndrome in preterm infants.

UNLABELLED: Staphylococcal scalded skin syndrome (SSSS) demonstrates dermal symptoms due to exfoliative toxin (ET) A or ETB produced by Staphylococcus aureus. We examined the association between anti-ETA antibodies and SSSS onset in neonates. Three preterm infants carried an ETA-producing strain of S. aureus, manifesting as either SSSS or bullous impetigo; a full-term infant carrying the same strain was asymptomatic. The infants (n=106) were categorized into three groups according to their gestational age (GA) as follows: <30 weeks, 30-37 weeks, and >37 weeks. The measured levels of anti-ETA antibody in the three infants displaying SSSS were low before the onset of dermal symptoms; only the asymptomatic full-term infant displayed a high antibody level. Anti-ETA antibody levels in the preterm group with a GA of <30 weeks were statistically lower than those in the term infant group; the prevalences of anti-ETA antibodies above a cutoff value in the three groups of neonates were 55 % (18/33) among preterm infants with a GA <30 weeks, 73 % (25/34) among those with a GA of 30-37 weeks, and 90 % (35/39) among infants with a GA >37 weeks. CONCLUSION: The presence of anti-ETA antibodies below a particular cutoff level might be associated with SSSS onset in preterm infants.

Helicobacter heilmannii sensu stricto-related gastric ulcers: a case report.

A spiral bacterium (SH9), morphologically different from Helicobacter pylori (H. pylori), was found in a 62-year-old woman's gastric mucosa. Gastroscopic examination revealed multiple gastric ulcers near the pyloric ring; mapping gastric biopsy showed mild mononuclear infiltration with large lymphoid follicles in the antrum, without corpus atrophy. Urea breath test and H. pylori culture were negative, but Giemsa staining of biopsies revealed tightly coiled bacteria that immunostained with anti-H. pylori antibody. Sequencing of SH9 16S rRNA and the partial urease A and B subunit genes showed that the former sequence had highest similarity (99%; 1302/1315 bp) to Helicobacter heilmannii (H. heilmannii) sensu stricto (H. heilmannii s.s.) BC1 obtained from a bobcat, while the latter sequence confirmed highest similarity (98.3%; 1467/1493 bp) to H. heilmannii s.s. HU2 obtained from a human. The patient was diagnosed with multiple gastric ulcers associated with H. heilmannii s.s. infection. After triple therapy (amoxicillin, clarithromycin, and lansoprazole) with regimen for eradicating H. pylori, gastroscopy showed ulcer improvement and no H. heilmannii s.s. upon biopsy.

Factor H gene variants in Japanese: its relation to atypical hemolytic uremic syndrome.

Mutations and polymorphisms of factor H gene (FH1) are known to be closely involved in the development of atypical hemolytic uremic syndrome (aHUS). Several groups have identified disease risk mutations and polymorphisms of FH1 for the development of aHUS, and have investigated frequencies of aHUS in a number of ethnic groups. However, such studies on Japanese populations are limited. In the present study, we analyzed FH1 in Japanese aHUS patients and healthy volunteers, and examined whether those variants impacted on a tendency for the development of aHUS in Japanese populations. Similar to previous studies, we found that a high frequency of FH1 mutations, located in exon 23 of FH1, encodes short consensus repeat 20 in C-terminal end of factor H molecule in patients with aHUS (40%), but not in healthy volunteers. Interestingly, no significant differences in frequency of well-known disease risk polymorphisms for aHUS were observed between healthy volunteers and aHUS patients. Our results suggested that although FH1 mutations relates to the development of Japanese aHUS in accordance with other ethnic studies, other factor may be required for factor H polymorphism to be a risk factor of Japanese aHUS.

Pathophysiological investigation of the gastric surface mucous gel layer of patients with Helicobacter pylori infection by using immunoassays for trefoil factor family 2 and gastric gland mucous cell-type mucin in gastric juice.

BACKGROUND: The trefoil factor family (TFF) 2 protein is produced by gastric gland mucous cells (GMCs), and the secreted TFF2 shares a mucosal barrier function with GMC-type mucin. Recently, we presented an enzyme-linked immunosorbent assay (ELISA) method for measurement of GMC-type mucin in the gastric juice. AIMS: We aimed to develop an ELISA for TFF2 and to assess pathophysiological changes in the gastric surface mucous gel layer (SMGL) of patients with Helicobacter pylori infection. METHODS: The distribution of TFF2 and GMC-type mucin in the SMGL was immunohistochemically determined. The ELISA for TFF2 was based on a polyclonal goat antibody. Recombinant TFF2 was employed to prepare the calibrators. TFF2 and GMC-type mucin in the gastric juice in healthy individuals (n = 33) and patients with gastritis (n = 37), gastric ulcer (n = 16), and duodenal ulcer (n = 10) were assayed using ELISA. RESULTS: TFF2 and GMC-type mucin were immunohistochemically co-localized in the gastric SMGL and GMCs. The TFF2 levels in the patients were significantly higher than those in the healthy individuals. Further, the TFF2 levels in the H. pylori-positive patients were significantly higher than those in the H. pylori-negative patients, and decreased after the eradication of the infection. GMC-type mucin levels showed a tendency similar to that of TFF2 levels. CONCLUSIONS: The upregulation of TFF2 and GMC-type mucin secretion may reflect the response of the gastric mucosa to H. pylori-induced injuries. TFF2 and GMC-type mucin secreted into the SMGL may protect the gastric mucosa against H. pylori.

Quantitative determination of gland mucous cells-type mucin using a monoclonal antibody, HIK1083: its pathophysiological changes in human gastric juice.

BACKGROUND: Pathological alteration in gastric mucosa is caused by Helicobacter pylori infection and is detectable by histological analysis. In particular, the alteration of gland mucous cells (GMCs)-type mucin, which plays a protective role against H. pylori infection, is critical in the pathogenesis of H. pylori-related gastritis. We established an assay for GMCs-type mucin and quantitatively assessed the pathophysiological changes in its content in human gastric juice samples. METHODS: The assay method for GMCs-type mucin was based on ELISA using a monoclonal antibody (HIK1083), and was used it to measure GMCs-type mucin in gastric juice obtained from patients with or without H. pylori infection. RESULTS: All the basic characteristics of the current method were satisfactory to quantify the GMCs-type mucin content in gastric juice. The GMCs-type mucin content, but not total mucin content, was significantly higher in patients with H. pylori infection (n=17; 437+/-476 U, mean+/-SD) than in those without H. pylori infection (n=55; 168+/-322 U, p<0.05). CONCLUSIONS: The current method is suitable for the quantitative analysis of GMCs-type mucin in gastric juice. The change in GMCs-type mucin content in gastric juice may be possibly implicated in the pathophysiology of the gastric mucosa and in the patient's gastric mucosal lesions.

Discrimination of normal gastric mucosa from Helicobacter pylori gastritis using standard endoscopes and a single observation site: studies in children and young adults.

BACKGROUND: In the Helicobacter pylori-negative normal stomach, collecting venules are visible in the gastric corpus as numerous minute points. This finding has been termed 'regular arrangement of collecting venules' (RAC). The aim of the present study was to investigate the reliability of the presence of the RAC pattern for discrimination of normal gastric mucosa from H. pylori gastritis in pediatric patients. METHODS: Fifty-two consecutive children, adolescents and young adults (male:female 24 : 28; median age 15 years, range 8-29 years) referred for endoscopy and assessed for H. pylori infection were prospectively studied. The lower lesser curvature of the corpus near the incisura was evaluated for the RAC pattern using a standard endoscope with the tip close to, but not in contact with, the gastric surface. Gastric biopsies were taken after the endoscopic observation. RESULTS: In all the 29 RAC-positive patients, active H. pylori gastritis was absent, whereas H. pylori gastritis was found in 20 of 23 RAC-negative patients (86.9%). CONCLUSIONS: Identification of the RAC pattern at the lower lesser curvature of the corpus using close observation with a standard endoscope proved to be an effective and practical marker to discriminate normal histology from H. pylori gastritis among both children and young adults. Absence of the RAC pattern should prompt gastric mucosal biopsies despite otherwise normal-appearing gastric mucosa.