Evaluation of Antibacterial Effect of Hybrid Nano-coating of Stainless Steel Orthodontic Brackets on Streptococcus Mutans - An In vitro Study.

Nano-coating of orthodontic brackets with a combination or hybrid of metals and metal oxides may reduce the streptococcus mutans count and incidence of enamel decalcification seen around brackets in patients undergoing fixed orthodontic treatment. In total, 255 orthodontic brackets (3M Unitek, Monrovia, California, USA) were divided into one control group (group I) of 60 and three experimental groups of 65 each (groups II, III, and IV). The experimental group brackets were coated with a combination of silver-zinc oxide, copper oxide -zinc oxide, and silver-copper oxide nanoparticles using physical vapour deposition method. The two nanoparticles used for each group were mixed in the ratio of 1:1 by weight for providing a uniform hybrid coating. Sixty brackets from each group were used for microbiological evaluation of antibacterial activity against Streptococcus mutans in blood agar medium, and the remaining five brackets from each experimental group were used for SEM analysis to check the uniformity of the coating. Nano-coated brackets demonstrated better antibacterial properties than uncoated brackets. Copper oxide-zinc oxide nanoparticles coated brackets demonstrated better antibacterial properties than the silver-zinc oxide and silver- copper oxide coated brackets.

Effects of fixation and demineralization on histomorphology and DNA amplification of canine bone marrow.

Fixation and demineralization protocols for bone marrow (BM) across diagnostic laboratories are not standardized. How different protocols affect histomorphology and DNA amplification is incompletely understood. In this study, 2 fixatives and 3 demineralization methods were tested on canine BM samples. Twenty replicate sternal samples obtained within 24 hours of death were fixed overnight in either acetic acid-zinc-formalin (AZF) or 10% neutral-buffered formalin (NBF) and demineralized with formic acid for 12 hours. Another 53 samples were fixed in AZF and demineralized with hydrochloric acid for 1-hour, formic acid for 12 hours, or ethylenediamine tetraacetic acid (EDTA) for 24 hours. Histologic sections were scored by 4 raters as of insufficient, marginal, good, or excellent quality. In addition, DNA samples extracted from sections treated with the different fixation and demineralization methods were amplified with 3 sets of primers to conserved regions of T cell receptor gamma and immunoglobulin heavy chain genes. Amplification efficiency was graded based on review of capillary electrophoretograms. There was no significant difference in the histomorphology scores of sections fixed in AZF or NBF. However, EDTA-based demineralization yielded higher histomorphology scores than demineralization with hydrochloric or formic acid, whereas formic acid resulted in higher scores than hydrochloric acid. Demineralization with EDTA yielded DNA amplification in 29 of 36 (81%) samples, whereas demineralization with either acid yielded amplification in only 2 of 72 (3%) samples. Although slightly more time-consuming and labor-intensive, tissue demineralization with EDTA results in superior morphology and is critical for polymerase chain reaction (PCR) amplification with the DNA extraction method described in this article.

Choroidal Osteoma With Choroidal Excavation Due to Decalcification Five Years After Photodynamic Therapy: A Case Report.

A choroidal osteoma (CO) is a relatively rare, benign tumor with ossification that develops in the choroid and undergoes enlargement and decalcification in its natural course. Photodynamic therapy (PDT) is used to induce decalcification, but there are few reports on individual cases treated with PDT. A 47-year-old Japanese man who had reduced decimal visual acuity (VA) of the right eye to 0.7 due to a CO away from the fovea was treated with PDT. The PDT resulted in a partial decalcification of CO, and the visual acuity improved to 1.0. However, the tumor slowly expanded, eventually reaching the central fovea. Decalcification and focal choroidal excavation occurred during this natural course of the disease. Although his metamorphopsia worsened, his VA was maintained at 1.0. This case highlights that a CO partially decalcified after PDT can still enlarge and decalcify over several years. These findings indicate the need for careful and continuous monitoring of eyes with a CO.

Hypertonic saline- and detergent-accelerated EDTA-based decalcification better preserves mRNA of bones.

Ethylenediaminetetraacetic acid (EDTA), a classically used chelating agent of decalcification, maintains good morphological details, but its slow decalcification limits its wider applications. Many procedures have been reported to accelerate EDTA-based decalcification, involving temperature, concentration, sonication, agitation, vacuum, microwave, or combination. However, these procedures, concentrating on purely tissue-outside physical factors to increase the chemical diffusion, do not enable EDTA to exert its full capacity due to tissue intrinsic chemical resistances around the diffusion passage. The resistances, such as tissue inner lipids and electric charges, impede the penetration of EDTA. We hypothesized that delipidation and shielding electric charges would accelerate EDTA-based penetration and the subsequent decalcification. The hypothesis was verified by the observation of speedy penetration of EDTA with additives of detergents and hypertonic saline, testing on tissue-mimicking gels of collagen and adult mouse bones. Using a 26% EDTA mixture with the additives at 45°C, a conventional 7-day decalcification of adult mouse ankle joints could be completed within 24 h while the tissue morphological structure, antigenicity, enzymes, and DNA were well preserved, and mRNA better retained compared to using 15% EDTA at room temperature. The addition of hypertonic saline and detergents to EDTA decalcification is a simple, rapid, and inexpensive method that doesn't disrupt the current histological workflow. This method is equally or even more effective than the currently most used decalcification methods in preserving the morphological details of tissues. It can be highly beneficial for the related community.

Resin infiltration for white spot lesions: An in vitro experimental trial.

OBJECTIVE: To evaluate the aesthetic outcome by varying the duration allowed for infiltrant penetration when treating white spot lesions with resin infiltration. DESIGN: An in vitro, experimental randomised study. METHODS: Artificially created white spot lesions (WSLs) were induced on 100 extracted anterior teeth (T1). Teeth were divided into enamel and dentine groups depending on the extent of the lesion and then randomly assigned into different treatment protocol groups: penetration times of 3, 6 and 9 min. Resin infiltration treatment was applied according to the treatment protocol assigned (T2). Samples were thermocycled for 10,000 cycles (1 clinical year) (T3). The samples from the 3-min enamel and dentine groups were then randomly assigned into either a repeat treatment or no additional treatment group (T4). Samples were then thermocycled for an additional 10,000 cycles (T5). Spectrophotometric analysis was measured colour change (ΔE) for all groups. RESULTS: Mean ΔE values equal to or greater than the critical value (3.7) indicate a detectable clinical difference in colour of the treated WSL when compared to before WSL formation. Mean ΔE values, for the enamel groups, were slightly above or significantly below the critical value, and for the dentine groups, were significantly above the critical value. Mean ΔE values within the enamel and dentine groups both demonstrated a downward trend with increasing time allowed for resin infiltrant penetration (P < 0.05). No significant mean ΔE difference (P = 0.53) was found between groups that received a single or repeat treatment. After the first thermocycling event, no significant difference in colour change was observed in all groups except for the deep dentine lesion treated for 3 min. There was a significant difference in colour change for all groups except the enamel group that received a single treatment following thermocycling after a single or repeat treatment. CONCLUSION: Increasing the resin infiltrant penetration time to at least 9 min is advised as the most optimised treatment protocol. Resin infiltration treatment should be done only once to treat a particular white spot lesion as subsequent treatment for the same lesion results in marginal colour improvement. The colour improvement of WSLs resulting from the resin infiltration treatment can be expected to last for at least 1 year. Resin infiltration treatment of shallow lesions with a single and optimised infiltration technique can be expected to last an additional year.

Quantitative evaluation of disc degeneration using dual-energy CT: advantages of R-VH, D-VH values and the IVNCa + CT model.

OBJECTIVE: To evaluate the correlation between dual-energy CT (DECT) virtual calcium free (VNCA), CT attenuation, the ratio and difference of VNCA to CT attenuation, and Pfirrmann grading of lumbar disc degeneration. METHODS: A retrospective analysis on 135 intervertebral discs from 30 patients who underwent DECT and MR. Discs was graded using the Pfirrmann system. ROIs on the sagittal plane assessed HU value, VNCA value, Rho value, Z value, R-VH value, and D-VH value. Correlation, grade differences, and multivariate regression models were assessed. Diagnostic performance and cut-off values were determined using AUC. RESULTS: VNCA (r = 0.589, P < 0.001), R-VH (r = 0.622, P < 0.001), and D-VH (r = 0.613, P < 0.001) moderately correlated with Pfirrmann grading. HU (r = 0.388, P < 0.001), Rho (r = 0.142, P = 0.102), and Z (r = -0.125, P = 0.153) showed a weak correlation. R-VH, D-VH, and VNCA had significantly higher correlation than HU. Statistically significant differences were observed in P values of VNCA, HU, R-VH, and D-VH in relative groups (P < 0.05), but not in Rho and Z values (P > 0.05). R-VH and D-VH had significant differences between Pfirrmann grades 1 and 2, and grades 2 and 3 (early stage) (P < 0.05). AUC readings of R-VH and D-VH (≥2, ≥3, ≥4) were higher. The multivariate model IVNCa + CT had the highest AUC. CONCLUSION: The new quantitative indices R-VH value and D-VH value of DECT have advantages over VNCA value and HU value in evaluating early-stage disc degeneration (≥2 grades, ≥3 grades). The multivariate model IVNCa + CT has the best AUC values for evaluating disc degeneration at all stages.

Study on the effect of different durations of decalcification and depigmentation on programmed death ligand 1 immunohistochemical staining.

Background: Programmed death ligand 1 (PD-L1) score is an important companion diagnosis to predict the response to immunotherapy. Immunohistochemistry can accurately assess the expression of PD-L1 in routine paraffin-embedded tissue. However, whether decalcified or depigmented tissue is still accurate and can be used as a companion diagnosis is controversial. This study attempts to resolve this controversy by analyzing the effects of decalcification and depigmentation at different times on PD-L1 expression. Methods: Placental tissues were selected for tissue microarray, decalcification was performed according to time gradients of 6, 12, 24, 36, and 48 h, and depigmentation was performed according to time gradients of 1, 5, 15, 30, and 60 min. The intensity of PD-L1 expression at different time points was observed and quantified. Ten PD-L1-positive esophageal squamous carcinoma samples were selected for decalcification treatment, and the PD-L1. Combined Positive Score (CPS), Tumor Proportion Score (TPS) and Immunocyte Proportion Score (IPS) and the positivity rates were compared before and after decalcification. Results: After the placenta was decalcified, the intensity of PD-L1 positivity diminished, and the average optical density (AOD) value decreased with the prolongation of decalcification time and decreased significantly (P<0.05) at 24 h compared with the control group, and significantly (P<0.01) at 36 and 48 h compared with the control group. The intensity of PD-L1 positivity was weakened considerably after the treatment with potassium permanganate depigmentation. In addition, the AOD value decreased significantly (P<0.01) after the depigmentation time reached 5 min compared with the control group. Ten cases of PD-L1 positive esophageal squamous carcinoma were treated with 24 h decalcification, although the PD-L1 score decreased to a certain degree (P>0.05), and the positivity rate could reach 90%. After 36 h treatment, PD-L1 scores decreased, the CPS and IPS scores decreased significantly (P<0.05), and the positive rate was only 50%. Conclusions: Potassium permanganate depigmentation significantly reduces PD-L1 expression, even for a shorter time, affecting the accuracy of the results. The accuracy of PD-L1 remained high within 24 h decalcification. The above results have certain reference value for clinical selection of immunotherapy.

Clinicopathological analysis of thyroid carcinomas with the RET and NTRK fusion genes: characterization for genetic analysis.

Thyroid carcinomas exhibit various genetic alterations, including the RET and NTRK fusion genes that are targets for molecular therapies. Thus, detecting fusion genes is crucial for devising effective treatment plans. This study characterized the pathological findings associated with these genes to identify the specimens suitable for genetic analysis. Thyroid carcinoma cases positive for the fusion genes were analyzed using the Oncomine Dx Target Test. Clinicopathological data were collected and assessed. Among the 74 patients tested, 8 had RET and 1 had NTRK3 fusion gene. Specifically, of the RET fusion gene cases, 6 exhibited "BRAF-like" atypia and 2 showed "RAS-like" atypia, while the single case with an NTRK3 fusion gene presented "RAS-like" atypia. Apart from one poorly differentiated thyroid carcinoma, most cases involved papillary thyroid carcinomas (PTCs). Primary tumors showed varied structural patterns and exhibited a high proportion of non-papillary structures. Dysmorphic clear cells were frequently observed. BRAF V600E immunoreactivity was negative in all cases. Interestingly, some cases exhibited similarities to diffuse sclerosing variant of PTC characteristics. While calcification in lymph node metastases was mild, primary tumors typically required hydrochloric acid-based decalcification for tissue preparation. This study highlights the benefits of combining morphological and immunohistochemical analyses for gene detection and posits that lymph node metastases are more suitable for genetic analysis owing to their mild calcification. Our results emphasize the importance of accurate sample processing in diagnosing and treating thyroid carcinomas.

Potential Application of Non-Invasive Optical Imaging Methods in Orthodontic Diagnosis.

During orthodontic treatment, the early diagnosis of microscopic changes in soft and hard tissues, including periodontal tissue, is very important to prevent iatrogenic side effects like root resorption and periodontal diseases. Cervical periodontal tissue is the most critical area that reacts first to mal-habits or orthodontic forces, and it is also the place where bacteria deposits in the early stage of periodontal diseases. The early diagnosis of hard tissue changes, such as demineralization, is also very important in maintaining a patient's health during orthodontic treatment. Many diagnostic devices, including radiographic equipment and intra-oral scanners, are helpful in diagnosing these problems, but have certain limitations in invasiveness and precision. The purpose of this study is to verify the possible utilities of non-invasive diagnostic devices in the orthodontic field that can compensate for these limitations. For this, non-invasive optical diagnostic devices, including optical coherence tomography and optical Doppler tomography, were used in vivo with animal and human examination for hard and soft tissues. These devices can provide real-time three-dimensional images at the histological scale. The results of this study verified these devices can be used in clinical practice during orthodontic treatment and introduced a new diagnostic paradigm differentiating microstructural changes in tissues in orthodontic diagnosis.

Comparative Histomorphometric Analysis of Bone Regeneration According to Bone Graft Type.

PURPOSE: To use histomorphometric analysis to evaluate bone reconstruction in rabbit calvaria with autogenous bone, anorganic bovine bone, undecalcified human tooth bone (UdTB), and decalcified human tooth bone (dTB) grafts. MATERIALS AND METHODS: Extracted human teeth were crushed, and tooth bone with and without decalcification was prepared. Bony defects were made in 10 rabbit calvaria and allocated to one of the following four groups: group 1, in which UdTB was grafted; group 2, in which dTB was grafted; group 3, in which anorganic bovine bone was grafted; group 4, in which autogenous bone was grafted. The rabbits were sacrificed at 2 or 8 weeks postoperatively, and histomorphometric comparison was performed. RESULTS: Histologically, new bone formation was observed at the defect margin and around all graft materials. The dTB group revealed significantly greater new bone areas at 2 and 8 weeks compared to the UdTB group and the anorganic bovine bone group (P < .05). The dTB group revealed no significant difference in the new bone area at 2 weeks but revealed significantly less new bone area at 8 weeks compared to the autogenous bone group (P < .05). The dTB group also revealed significantly less graft material area compared to the anorganic bovine bone group at 8 weeks (P < .05). The autogenous bone group revealed significantly less graft material area and significantly greater bone marrow area compared to other groups at 8 weeks (P < .05). CONCLUSIONS: Grafting with dTB resulted in better bone regeneration than UdTB and anorganic bovine bone grafting at 8 weeks and addresses the potential disadvantages of autogenous bone grafting.

Direct specimen collection during routine operation improves nucleic acid quality in genetic analysis samples for head and neck tumors: A retrospective study.

The nucleic acid integrity of head and neck squamous cell carcinoma (HNSCC) samples is poor, and the material available for genetic analysis is limited. Therefore, to expand the effectiveness of personalized medicine in patients with HNSCC, a new sampling method is needed. In total, 128 samples from 44 patients with HNSCC were studied: 32 genetic analysis samples (GASs) collected as 5 × 5 × 5 mm tissue fragments from resected large tumors and immediately embedded in a small formalin bottle within 10 min (i.e., the ischemic time), 43 primary tumor components (primary), 14 decalcified tumor (DC) samples, 32 metastatic tumors in lymph nodes (LNs), and 7 parakeratinized components (PKCs). The nucleic acid quality in the GAS, primary, DC, LN, and PKC groups was compared and next-generation sequencing (NGS) was performed. DNA integrity number and percentage of RNA fragments with > 200 nucleotides were significantly higher in the GAS group than those in the other groups. RNA integrity number decreased first in LN, followed by GAS, primary, and DC. No significant differences were observed in DIN, RIN and DV200 among the PKC, primary and LN. Following methyl green-pyronin staining, preserved DNA and RNA were not visualized in DC samples. Most NGS metrics did not differ significantly among primary, LN, and PKC samples. In conclusion, GASs should be collected during routine hospital activities. When the volume of viable materials is limited, PKCs should be considered for genetic analysis.

Experimental Study on Effects of CO2 Curing Conditions on Mechanical Properties of Cement Paste Containing CO2 Reactive Hardening Calcium Silicate Cement.

Human survival is threatened by the rapid climate change due to global warming caused by the increase in CO2 emissions since the Second Industrial Revolution. This study developed a secondary cement product production technology by replacing cement, a conventional binder, with calcium silicate cement (CSC), i.e., CO2 reactive hardening cement, to reduce CO2 emissions and utilize CO2 from the cement industry, which emits CO2 in large quantities. Results showed that the carbonation depth, compressive strength increase rate, and CO2 sequestration rate increased as the CSC content increased, suggesting that CSC can be applied as a secondary cement product.

Mortui vivos docent: a modern revival of temporal bone plug harvests.

Human temporal bones (HTBs) are invaluable resources for the study of otologic disorders and for evaluating novel treatment approaches. Given the high costs and technical expertise required to collect and process HTBs, there has been a decline in the number of otopathology laboratories. Our objective is to encourage ongoing study of HTBs by outlining the necessary steps to establish a pipeline for collection and processing of HTBs. In this methods manuscript, we: (1) provide the design of a temporal bone plug sawblade that can be used to collect specimens from autopsy donors; (2) establish that decalcification time can be dramatically reduced from 9 to 3 months if ethylenediaminetetraacetic acid is combined with microwave tissue processing and periodic bone trimming; (3) show that serial sections of relatively-rapidly decalcified HTBs can be successfully immunostained for key inner ear proteins; (4) demonstrate how to drill down a HTB to the otic capsule within a few hours so that subsequent decalcification time can be further reduced to only weeks. We include photographs and videos to facilitate rapid dissemination of the developed methods. Collected HTBs can be used for many purposes, including, but not limited to device testing, imaging studies, education, histopathology, and molecular studies. As new technology develops, it is imperative to continue studying HTBs to further our understanding of the cellular and molecular underpinnings of otologic disorders.

Evaluation of Different Decalcification Agents on Extracted Tooth: A Comparative Study.

Introduction: The pulp is the most negatively impacted tissue during decalcification since it comprises the soft tissue components. The most effective decalcifying agent would be safest for cells and tissues while yet removing all traces of calcium. It has to get the job done quickly and have good staining properties. Aim and Objective: The goal of this research was to identify the most effective decalcifying agent for diagnostic purposes via a qualitative investigation of tissue preservation and a comparison of the efficiency of several decalcifying agents on human permanent teeth, covering both hard and soft tissue components. Materials and Methods: Fifty premolars from people aged 14 to 30 who needed them pulled for orthodontics were included in the research. Participants in the research were divided into five groups of ten. Group A, Group B, Group C, Group D, and Group E make up the total of five groups. In this investigation, we compared the efficiency of five decalcifying chemicals and analyzed their staining patterns and effects on tooth tissue. Fifty premolar teeth from participants aged 14-30 years old were removed for orthodontic therapy. For the research, they were split up into five groups of ten. Group A contains 5% ethylenediaminetetraacetic acid (EDTA), Group B contains 10% formic acid, Group C contains 5% Trichoraticectic acid, Group D contains 5% nitric acid, and Group E contains 5% formalin-nitric acid. Result: Regardless of the specifics of the chosen decalcification solution, all procedures benefit from the inclusion of external stimuli. None of the variables were used in the current investigation; it was conducted only to compare various decalcifying chemicals. Conclusion: When time is not a concern, neutral EDTA may be recommended for preservation and presentation because of its ability to maintain soft-tissue integrity and provide high-quality staining. The formalin-nitric acid solution was one agent that appeared to strike a good compromise between speed and tissue preservation.

Changes in Pulp and Roots of Deciduous Teeth during Different Stages of Physiologic Resorption: A Histologic Study.

Background: Resorption of primary teeth and eruption of permanent teeth involves a complex series of changes. The cellular and histological changes occurring during the process of resorption vary stagewise. The knowledge of the changes occurring in the pulp of deciduous teeth would provide information about the resorptive process. Aim: To evaluate the histologic changes of the pulp of deciduous teeth related to different stages of physiologic root resorption. Study setting and design: To establish the cause and effect relationship, a contrived histologic study design was planned. Materials and methods: A total of 60 extracted deciduous incisors, canines, and molars were included in the study. The remaining root length (RRL) was determined based on the standardized photographs. The teeth were then grouped into three based on the percentage of RRL. The teeth were subjected to decalcification with 5% nitric acid, following which histological processing was performed. Statistical analysis: The present study being a qualitative study design, descriptively explains the histologic findings, and no statistical tests have been applied. Results: During the initial stages of resorption, there were no histological alterations noted in the pulp, particularly in the cervical 3rd, with the cellular structure relatively maintained. As the resorption progresses, reversal lines were evident, indicating a process of repair occurring simultaneously during the process of resorption. With further advancement, the repair is overtaken by the resorption indicated by the presence of resorptive cells. Neovascularization and an increase in immune cells are also evident in advanced stages. Conclusion: The pulp exhibits progressive changes as the resorption continues from stage I to stage III. The changes vary from a smaller number of immune cells and odontoclasts in stage I to increasing number of the same as resorption progresses. How to cite this article: Murthy P, Bhojraj N, Hegde U. Changes in Pulp and Roots of Deciduous Teeth during Different Stages of Physiologic Resorption: A Histologic Study. Int J Clin Pediatr Dent 2023;16(3):437-443.

Studying the use of a proposed three-level grossing protocol in the histopathological diagnosis of COVID-19-associated rhinomaxillary mucormycosis.

Introduction: Mucormycosis is an acute and rapidly progressing opportunistic fungal infection. COVID-19-associated mucormycosis (CAM) had re-emerged as a complication of COVID-19 infection during the second wave of the pandemic in 2021. The rhinomaxillary form is a variant of the rhino-cerebral mucormycosis that presents a diagnostic challenge to the dentist and the oral and maxillofacial pathologist. Gross examination of pathological specimens is the most undermined step even though it plays a vital role in the final diagnosis. No studies have described this post-clinical step for the maxillofacial soft and hard tissue submitted for examination. Material and Methods: A prospective comparative study was carried out on 52 COVID-19-associated rhinomaxillary mucormycosis (CARM) cases to achieve complete, representative, and informative sampling of the submitted tissue and establish a three-level gross macroscopic examination protocol. Complete clinical and radiological histories were recorded after informed, written consent from every patient was received. Details of the number and type of samples received were recorded, grossing procedure was done as per the proposed three-level grossing protocol and were then compared to the presence of fungal hyphae in the soft tissue or decalcified hard tissue. Result: All 100% of the samples consisted of soft tissue (maxillary sinus lining), while 90.4% of the samples contained different hard tissue specimens. Seventy percent of the grossing workload was carried out by first-year oral pathology residents. Sixty-seven point three percent of the total soft tissue samples submitted showed no presence of fungal hyphae, while 69.2% of total decalcified sections of hard tissue were positive for fungal hyphae with a positive correlation. Out of the 29 cases grossed via the three-level grossing protocol, 89.6% of the cases were histopathologically positive for fungal hyphae. Thus a positive association (P < 0.05) between histopathological diagnosis and the proposed three-level grossing protocol was found. Conclusion: It is imperative to recognise that no mucormycosis report is to be signed out without multi-site (three-level grossed) bone decalcified reports. There is an immediate need to realise how vital documentation, correct laboratory practices, and grossing are for accurate histopathological diagnosis.

Impact of Decalcification, Cold Ischemia, and Deglycosylation on Performance of Programmed Cell Death Ligand-1 Antibodies With Different Binding Epitopes: Comparison of 7 Clones.

Programmed cell death ligand-1 (PD-L1) expression levels in patients' tumors have demonstrated clinical utility across many cancer types and are used to determine treatment eligibility. Several independently developed PD-L1 immunohistochemical (IHC) predictive assays are commercially available and have demonstrated different levels of staining between assays, generating interest in understanding the similarities and differences between assays. Previously, we identified epitopes in the internal and external domains of PD-L1, bound by antibodies in routine clinical use (SP263, SP142, 22C3, and 28-8). Variance in performance of assays utilizing these antibodies, observed following exposure to preanalytical factors such as decalcification, cold ischemia, and duration of fixation, encouraged additional investigation of antibody-binding sites, to understand whether binding site structures/conformations contribute to differential PD-L1 IHC assay staining. We proceeded to further investigate the epitopes on PD-L1 bound by these antibodies, alongside the major clones utilized in laboratory-developed tests (E1L3N, QR1, and 73-10). Characterization of QR1 and 73-10 clones demonstrated that both bind the PD-L1 C-terminal internal domain, similar to SP263/SP142. Our results also demonstrate that under suboptimal decalcification or fixation conditions, the performance of internal domain antibodies is less detrimentally affected than that of external domain antibodies 22C3/28-8. Furthermore, we show that the binding sites of external domain antibodies are susceptible to deglycosylation and conformational structural changes, which directly result in IHC staining reduction or loss. The binding sites of internal domain antibodies were unaffected by deglycosylation or conformational structural change. This study demonstrates that the location and conformation of binding sites, recognized by antibodies employed in PD-L1 diagnostic assays, differ significantly and exhibit differing degrees of robustness. These findings should reinforce the need for vigilance when performing clinical testing with different PD-L1 IHC assays, particularly in the control of cold ischemia and the selection of fixation and decalcification conditions.

Shockwaves delivery for aortic valve therapy-Realistic perspective for clinical translation?

Calcific aortic valve disease (CAVD) is the most frequent valvular heart disorder, and the one with the highest impact and burden in the elderly population. While the quality and standardization of the current aortic valve replacements has reached unprecedented levels with the commercialization of minimally-invasive implants and the design of procedures for valve repair, the need of supplementary therapies able to block or retard the course of the pathology before patients need the intervention is still awaited. In this contribution, we will discuss the emerging opportunity to set up devices to mechanically rupture the calcium deposits accumulating in the aortic valve and restore, at least in part, the pliability and the mechanical function of the calcified leaflets. Starting from the evidences gained by mechanical decalcification of coronary arteries in interventional cardiology procedures, a practice already in the clinical setting, we will discuss the advantages and the potential drawbacks of valve lithotripsy devices and their potential applicability in the clinical scenario.

Analysis of Dental Enamel Remineralization: A Systematic Review of Technique Comparisons.

The demineralization process conditions the structure of the enamel and begins with a superficial decalcification procedure that makes the enamel surface porous and gives it a chalky appearance. White spot lesions (WSLs) are the first clinical sign that can be appreciated before caries evolves into cavitated lesions. The years of research have led to the testing of several remineralization techniques. This study's objective is to investigate and assess the various methods for remineralizing enamel. The dental enamel remineralization techniques have been evaluated. A literature search on PubMed, Scopus, and Web of Science was performed. After screening, identification, and eligibility processes 17 papers were selected for the qualitative analysis. This systematic review identified several materials that, whether used singly or in combination, can be effective in the process of remineralizing enamel. All methods have a potential for remineralization when they come into contact with tooth enamel surfaces that have early-stage caries (white spot lesions). From the studies conducted in the test, all of the substances used to which fluoride has been added contribute to remineralization. It is believed that by developing and researching new remineralization techniques, this process might develop even more successfully.

Rapid decalcification of articular cartilage and subchondral bone using an ultrasonic cleaner with EDTA.

Articular cartilage and subchondral bones were used to be the samples for studying effects of drugs in the joint degenerative diseases such as osteoarthritis. Because of the deposition of mineral salts, articular cartilage and subchondral bones require decalcification process to soften the tissues. EDTA is a chelating agent that is commonly used to remove mineral salts, but this step is time-consuming and can take as long as 45 days. Commercial ultrasonic cleaner and microwave oven were reported to reduce the decalcification timing. The aim of this study is to determine and compare the decalcification of human articular cartilage and subchondral bone using EDTA together with ultrasonic cleaner or microwave oven. Hundred pieces of articular cartilage and subchondral bones obtained from osteoarthritis patients undergone total-knee-replacement were divided into 10 groups according to decalcification method (ultrasonic cleaner or microwave) and timing (2, 4, 6, 8, and 10 h). In each group, all cartilage and subchondral bone pieces were decalcified and sectioned, and subsequently stained with haematoxylin and eosin, Von Kossa, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, or caspase-3 immunohistochemistry. The optimal timing of decalcification of articular cartilage and subchondral bones using EDTA together with ultrasonic cleaner was at 8 and 10 h, while the timing using EDTA together with microwave oven was more than 10 h. Clear TUNEL and caspase-3 signals were obtained from samples decalcified using EDTA together with ultrasonic cleaner for 8 h. In summary, to our knowledge, this is the first study that compared EDTA decalcification between ultrasonic cleaner and microwave oven. Here, we report a new methodology for decalcification for articular cartilage and subchondral bones that reduces decalcification time from weeks to hours and is suitable for further pathological analyses.