Effect of Processing Methods of Human Saliva on the Proteomic Profile and Protein-Mediated Biological Processes.

The use of saliva as a protein source prior to microbiological and biological assays requires previous processing. However, the effect of these processing methods on the proteomic profile of saliva has not been tested. Stimulated human saliva was collected from eight healthy volunteers. Non-processed saliva was compared with 0.22 µm filtered, 0.45 µm filtered, and pasteurized saliva, by liquid chromatography-mass spectrometry. Data are available via ProteomeXchange with identifier PXD039248. The effect of processed saliva on microbial adhesion was tested using bacterial and fungus species and in biological cell behavior using HaCaT immortalized human keratinocytes. Two hundred and seventy-eight proteins were identified in non-processed saliva, of which 54 proteins (≈19%) were exclusive. Saliva processing reduced identified proteins to 222 (≈80%) for the 0.22 µm group, 219 (≈79%) for the 0.45 µm group, and 201 (≈72%) for the pasteurized saliva, compared to non-processed saliva. The proteomic profile showed similar molecular functions and biological processes. The different saliva processing methods did not alter microbial adhesion (ANOVA, p > 0.05). Interestingly, pasteurized saliva reduced keratinocyte cell viability. Saliva processing methods tested reduced the proteomic profile diversity of saliva but maintained similar molecular functions and biological processes, not interfering with microbial adhesion and cell viability, except for pasteurization, which reduced cell viability.

A Compilation of Study Models for Dental Pulp Regeneration.

Efforts to heal damaged pulp tissue through tissue engineering have produced positive results in pilot trials. However, the differentiation between real regeneration and mere repair is not possible through clinical measures. Therefore, preclinical study models are still of great importance, both to gain insights into treatment outcomes on tissue and cell levels and to develop further concepts for dental pulp regeneration. This review aims at compiling information about different in vitro and in vivo ectopic, semiorthotopic, and orthotopic models. In this context, the differences between monolayer and three-dimensional cell cultures are discussed, a semiorthotopic transplantation model is introduced as an in vivo model for dental pulp regeneration, and finally, different animal models used for in vivo orthotopic investigations are presented.

A Comparative Analysis of Dental Measurements in Physical and Digital Orthodontic Case Study Models.

Background and Objectives: Study models are essential tools used in the dental teaching process. The aim of the present study was to compare the values obtained by manual and digital orthodontic measurements on physical and digital case study models. Materials and Methods: The physical experimental models were obtained by traditional pouring (improved stone-type IV gypsum products) and by additive manufacturing (resins). The digital experimental models were created by scanning the physical ones, using a white light-emitting diode (LED) source and an L-shaped dental scanner­Swing DOF (DOF, Seoul, Korea). The physical study models were first measured using a digital caliper, and then, they were scanned and evaluated using the DentalCad 3.0 Galway software (exocad GmbH, Darmstadt, Germany). The Pont, Linder−Harth, and Bolton indices, which are used in orthodontics for training students, were derived using the available data. Results: When comparing the linear measurement mean ranks taken on physical study models to those of digital models, no statistically significant differences (p > 0.05) were found. A similar result was also shown when the dentoalveolar growth indicators were analyzed. Conclusions: It can be concluded that dental study models made by direct light processing (DLP) and pouring type IV class gypsum are both acceptable for orthodontic teaching purposes.

Comparison of accuracy of mesiodistal tooth measurements made in conventional study models and digital models obtained from intraoral scan and desktop scan of study models.

OBJECTIVE: To compare the measured values obtained from the plaster model, digital models created by scanning the plaster models and direct intraoral scanning with the values obtained from direct intraoral measurements. DESIGN: This was a prospective clinical study. SETTING: The study was conducted in Department of Orthodontics, Saveetha Dental College and Hospital, Tamil Nadu, India. PARTICIPANTS: Ten patients before the start of orthodontic treatment were selected for the study. METHODS: A computer-aided design and manufacturing (CAD-CAM) system is an advanced technology that is being adopted in the field of orthodontics for diagnosis, treatment planning and documentation of patient records. Mesiodistal tooth width measurements of first premolars, canines, lateral incisors and central incisors, and transverse width measurement from mesial pit of right first premolar to mesial pit of left first premolar in both maxilla and mandible were obtained from direct intraoral measurement (gold standard), study model obtained from alginate impression, intraoral scanned image, and desktop scanned image of the study model. Descriptive statistics and ANOVA was performed to find the difference in mean among the groups. RESULTS: A P value > 0.05 was obtained in ANOVA indicating that there is no statistically significant difference in the measurements obtained by either of the methods. CONCLUSION: Conventional stone models and digital models obtained from intraoral scan and desktop scanning of plaster models are clinically reliable as the variations in measurements obtained from these methods were clinically negligible.

Lovastatin corrects ERK pathway hyperactivation in fragile X syndrome: potential of platelet's signaling cascades as new outcome measures in clinical trials.

AIM: To establish whether platelets from fragile X syndrome (FXS) individuals recapitulate FXS mouse neurons' defects in ERK and Akt pathways, and to evaluate the effect of lovastatin on these pathways. METHODS: ERK and Akt phosphorylation (pERK, pAkt) statuses were assessed with quantitative Western blotting before and after a 12-week lovastatin trial. RESULTS: Levels of pERK and pAkt were increased in FXS platelets, and lovastatin specifically normalized ERK activity. Changes in ERK phosphorylation were correlated with clinical response to lovastatin. CONCLUSIONS: Platelets' signaling pathways provide biomarkers that can be used as treatment outcome measures in FXS clinical trials.

Genetic and mechanistic evaluation for the weak A phenotype in Ael blood type with IVS6 + 5G>A ABO gene mutation.

BACKGROUND AND OBJECTIVES: Ael is a rare blood type that is characterized by weak agglutination of RBCs when reacts with anti-A antibody in adsorption-elution test. Although IVS6 + 5G→A mutation is known to associate with the Ael blood type, genetic and mechanistic evaluation for the weak agglutination of Ael with IVS6 + 5G→A mutation has not yet been completely addressed. MATERIALS AND METHODS: In this study, five cases of confirmed Ael individuals were analysed. The cDNAs for the A(el) alleles were obtained by cloning method for sequence analyses. The erythroleukemia K562 cells were used as the cell study model and were transfected with the A(el) expression construct. Flow cytometry analysis was then performed to determine the levels of surface antigen expression. RESULTS: The results indicated that IVS6 + 5G→A attributes to all cases of Ael . RT-PCR analyses revealed the presence of at least 10 types of aberrant A(el) splicing transcripts. Most of the transcripts caused early termination and produced non-functional protein during translation. Nevertheless, the transcript without exons 5-6 was predicted to generate functional Ael glycosyltransferase lacking 57 amino acids at the N-terminal segment. When the exons 5-6 deletion transcript was stably expressed in the K562 cells, weak agglutination of the cells can be induced by adding anti-A antibody followed by adsorption-elution test. CONCLUSION: This study demonstrates that aberrant splicing of A transcripts contributes to weak A expression and the weak agglutination of Ael -RBCs, adding to the complexity for the regulatory mechanisms of ABO gene expression.

Human EWS-FLI protein levels and neomorphic functions show a complex, function-specific dose-response relationship in Drosophila.

Ewing sarcoma (EwS) is a cancer that arises in the bones and soft tissues, typically driven by the Ewing's sarcoma breakpoint region 1-Friend leukemia virus integration 1 (EWS-FLI) oncogene. Implementation of genetically modified animal models of EwS has proved difficult largely owing to EWS-FLI's high toxicity. The EWS-FLI1FS frameshift variant that circumvents toxicity but is still able to perform key oncogenic functions provided the first study model in Drosophila. However, the quest for Drosophila lines expressing full-length, unmodified EWS-FLI remained open. Here, we show that EWS-FLI1FS's lower toxicity is owed to reduced protein levels caused by its frameshifted C-terminal peptide, and report new strategies through which we have generated Drosophila lines that express full-length, unmodified EWS-FLI. Using these lines, we have found that the upregulation of transcription from GGAA-microsatellites (GGAAµSats) presents a positive linear correlation within a wide range of EWS-FLI protein concentrations. In contrast, rather counterintuitively, GGAAµSats-independent transcriptomic dysregulation presents relatively minor differences across the same range, suggesting that GGAAµSat-dependent and -independent transcriptional upregulation present different kinetics of response with regards to changing EWS-FLI protein concentration. Our results underpin the functional relevance of varying EWS-FLI expression levels and provide experimental tools to investigate, in Drosophila, the effect of the EWS-FLI 'high' and 'low' states that have been reported and are suspected to be important for EwS in humans.

Effectiveness of Flapless Cortico-Alveolar Perforations Using Mechanical Drills Versus Traditional Corticotomy on the Retraction of Maxillary Canines in Class II Division 1 Malocclusion: A Three-Arm Randomized Controlled Clinical Trial.

BACKGROUND AND OBJECTIVES: Both invasive and minimally invasive surgical methods have recently gained popularity in accelerating orthodontic tooth movement. Traditional corticotomy (TC) was one of the first effective invasive surgical techniques in shortening orthodontic treatment time, whereas the flapless cortico-alveolar perforations (FCAPs) technique is a modern minimally invasive method that has recently shown good results in different types of orthodontic tooth movement. Therefore, this study aimed to compare the effectiveness of TC versus FCAPs in maxillary canine retraction when treating Class II division 1 malocclusion patients. MATERIALS AND METHODS: This was a single-blinded, single-center, three-arm randomized controlled trial. A total of 51 patients (22 males, 29 females, mean age 20.98 ± 1.95) whose treatment planning included the extraction of maxillary first premolars were enrolled and randomly divided into three groups: the TC group, the FCAPs group, and the control group. The assessed outcomes were the amount of canine retraction, anchorage loss, and canines' rotation, which was evaluated at five-time points till the completion of canine retraction. RESULTS: There were statistically significant differences in the amount of canine retraction between the three groups in the first two months (p < 0.001), with greater mean values in the TC group (p < 0.001) in the first month. However, the amount of canine retraction in the FCAPs group was significantly greater in the second month compared to the TC group (p = 0.003) and the control group (p < 0.001). In the first month of canine retraction, anchorage loss, and canine rotation were significantly lesser in the TC and FCAPs groups than in the control group (p < 0.001). On the contrary, the canines' rotation amount after the completion of retraction was greater in the TC group than in the other two groups (p < 0.001). CONCLUSION: TC and FCAPs are efficient adjunctive surgical methods for accelerating canine retraction. At the end of the first month, the TC accelerated canine retraction by 59.85% and FCAPs by 44% compared to the conventional retraction. At the end of the second month, the acceleration was less than recorded in the first month (35.44% and 50.20%, respectively). The acceleration effect of the surgical interventions appeared transient and did not last in the following observation period.

Melanocytes derived from mouse hair follicles: A novel study model to assess pigmentation disorders.

Melanocytes are the major cells responsible for skin and fair pigmentation in vertebrates. They localize to hair follicles(HFs) and the epidermis during embryonic development. A reduced number or dysfunction of melanocytes results in pigmentation disorders.Thus, methods for isolation, culture, and identification of melanocytes in mouse hair follicles provide an experimental basis for thestudy of of pigmentation disorders. In the current work, we harvested the melanocytes from the anagen phase dorsal skin of C57BL/6 mice.After its separation from the skin, the dermis was digested, and the HFs were released. HFs were then also digested, and the cells released from HFs were cultured in melanocyte growth medium. Immunofluorescence and immunohistochemistry staining were used to localize the distribution of melanocytes in HFs . Reverse transcription polymerase chain reaction was performed to detect the expression of specific melanocyte marker genes. Immunofluorescence, immunohistochemistry, flow cytometry, and western blot were carried out to detect the expression of marker proteins in cells. 3,4-Dihydroxy-L-phenylalanine (L-DOPA) staining was used to detect the pigmentation functionality of melaonocytes. Based on our results, we conclude that mature and functional melanocytes can be successfully obtained from theHFs, providing a cell model to study pigmentation disorders. The current findings provide novel insights for the treatment of pigmentation disorders by autologous cell transplantation. Further, we believe that issues related to skin damage, insufficient numbers of autologous cells, and autoimmune problems can be resolved in future though the use of functional melanocytes.

Morphological analysis of various rugae patterns among Dravidian population.

AIM: The aim of this study was to analyze and identify differences in the palatal rugae patterns and genderwise distributions in three different populations (Kerala, Mahe and Puducherry). MATERIALS AND METHODS: Ninety maxillary study models (30 from each group including males and females) were examined in the age group ranging from 18 to 25 years. The palatal rugae pattern was analyzed for shape. RESULTS: After analyzing the rugae patterns among the groups, the most common pattern was the wavy pattern (53.57%) followed by curved (18.22%) and straight (13.66%). The least was circular (1.3%). When compared between sex, the most common pattern was found to be wavy (male - 54.3% and female - 53.09%), while the curved pattern was more common among the females (21.09%) than males (13.97%). The straight pattern was more common among the males (18.8%) than females (10.18%). The least common pattern was found to be the circular in both sexes which accounted for around 1%. CONCLUSION: This study shows no two palates are identical in terms of their rugae pattern. Palatal rugae possess unique characteristics as they are absolutely individualistic. Study also confirms that the "wavy" type of palatal rugae pattern was the most predominant among these three populations.

Cell death pathways of particulate matter toxicity.

Humans are exposed to various complex mixtures of particulate matter (PM) from different sources. Long-term exposure to high levels of these particulates has been linked to a diverse range of respiratory and cardiovascular diseases that have resulted in hospital admission. The evaluation of the effects of PM exposure on the mechanisms related to cell death has been a challenge for many researchers. Therefore, in this review, we have discussed the effects of airborne PM exposure on mechanisms related to cell death. For this purpose, we have compiled literature data on PM sources, the effects of exposure, and the assays and models used for evaluation, in order to establish comparisons between various studies. The analysis of this collected data suggested divergent responses to PM exposure that resulted in different cell death types (apoptosis, autophagy, and necrosis). In addition, PM induced oxidative stress within cells, which appeared to be an important factor in the determination of cell fate. When the levels of reactive oxygen species were overpowering, the cellular fate was directed toward cell death. This may be the underlying mechanism of the development or exacerbation of respiratory diseases, such as emphysema and chronic obstructive pulmonary diseases. In addition, PM was shown to cause DNA damage and the resulting mutations increased the risk of cancer. Furthermore, several conditions should be considered in the assessment of cell death in PM-exposed models, including the cell culture line, PM composition, and the interaction of the different cells types in in vivo models.

Evaluating the Correlation between Anteroposterior and Mediolateral Compensatory Curves and their Influence on Dentoskeletal Morphology-An In vitro CBCT Study.

INTRODUCTION: To date, many orthodontist corrects malocclusion based on patients aesthetic concern and fails to correct the compensatory curves. This scenario is due to less insight on understanding relationship of compensatory curves and its correlation in treatment prognosis. AIM: The purpose of this study was to evaluate the correlation between the curve of Spee, curve of Monson and curve of Wilson, their influence on dentoskeletal morphology and their contribution to occlusal stability. MATERIALS AND METHODS: This study included 104 non-orthodontic models. The study casts were subdivided into two groups, Group-I consist 52 non- orthodontic models with Class-I molar relationship and Group-II consist of 52 non- orthodontic models with Class-II molar relationship. Curve of Spee was measured with digital vernier caliper, curve of Monson estimated using specially made sphere (7″inch, 8″ inch and 9″inch) and curve of Wilson was evaluated using Cone Beam Computed Technology (CBCT). RESULTS: Mean value for curve of Spee obtained for Group I and Group II is 1.844 mm and 3.188 mm. For curve of Monson, the mean value obtained for Group I and Group-II is 7.65 inches and 7.40 inches. The mean degree obtained for the curve of Wilson for Group I and Group-II is 12.05 and 16.49. The result showed positive correlation between curve of Spee and curve of Wilson and no correlation between curve of Monson and curve of Wilson and no correlation between curve of Spee and curve of Monson. The Pearson correlation coefficient analysis from the study confirmed these results. CONCLUSION: The results showed positive correlation between curve of spee and curve of Wilson. The data found in this study can be applied clinically for Class I and Class II malocclusion patients on diagnosis and treatment planning.

DISR: Dental Image Segmentation and Retrieval.

In this paper, we propose novel algorithms for retrieving dental images from databases by their contents. Based on special information of dental images, for better content-based dental image retrieval and representation, the image attributes are used. We propose Dental Image Segmentation and Retrieval (DISR), a content-based image retrieval method that is robust to translation and scaling of the objects in the images. A novel model is used to calculate the features of the image. We implemented the dentition plaster casts and proposed a special technique for segmenting teeth in our dental study models. For testing the efficiency of the presented algorithm, a software system is developed and 60 dental study models are used. The models are covering different kinds of malocclusions. Our experiments show that 95% of the extracted results are accurate and the presented algorithm is efficient.