Evaluating the Expression of Candidate Homeobox Genes and Their Role in Local-Site Inflammation in Mucosal Tissue Obtained from Children with Non-Syndromic Cleft Lip and Palate

Funding Information: Funding: The present study was funded by Riga Stradinš University (RSU). Funding Information: The present study was funded by Riga Stradin? University (RSU).We would like to extend our gratitude to Ilze Akota, Department of Oral & Maxillofacial Surgery, Faculty of Dentistry, RSU for providing the patient material. We would also like to acknowledge the parents of the patients for consenting to participate in the present study. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Craniofacial development including palatogenesis is a complex process which requires an orchestrated and spatiotemporal expression of various genes and factors for proper embryogenesis and organogenesis. One such group of genes essential for craniofacial development is the homeobox genes, transcriptional factors that are commonly associated with congenital abnormalities. Amongst these genes, DLX4, HOXB3, and MSX2 have been recently shown to be involved in the etiology of non-syndromic cleft lip and palate. Hence, we investigated the gene and protein expression of these genes in normal and cleft affected mucosal tissue obtained from 22 children, along with analyzing their role in promoting local-site inflammation using NF-κB. Additionally, we investigated the role of PTX3, which plays a critical role in tissue remodeling and wound repair. We found a residual gene and protein expression of DLX4 in cleft mucosa, although no differences in gene expression levels of HOXB3 and MSX2 were noted. However, a significant increase in protein expression for these genes was noted in the cleft mucosa (p < 0.05), indicating increased cellular proliferation. This was coupled with a significant increase in NF-κB protein expression in cleft mucosa (p < 0.05), highlighting the role of these genes in promotion of pro-inflammatory environment. Finally, no differences in gene expression of PTX3 were noted.publishersversionPeer reviewe

Debunking the myth of using “quiet” in clinical departments : an integrative overview of available literature

Funding Information: We would like to acknowledge the support and contributions of The ECOMSIR Collaborative (European Collaboration of Medical Students in Research), a non-profit, non-governmental student collaboration. The support of Riga Stradins University (RSU) is also greatly acknowledged.publishersversionPeer reviewe

Anatomical variations in position of mandibular foramen : An East European morphometric study in dry adult human mandibles for achieving a successful inferior alveolar nerve block

Publisher Copyright: © 2019 Firenze University Press Copyright: Copyright 2020 Elsevier B.V., All rights reserved.In today’s era of modern dentistry, the race towards developing painless and trauma-less procedures remain an important goal of all major companies and dental researchers. One such technique of focus is the Inferior Alveolar Nerve Block (IANB) which remains by far the most common anesthetic technique followed worldwide prior to any submaxillary treatment procedure. Despite of it being so popular, the failure rates remain significantly high with estimates at 15-60% among dental practitioners and dental students alike. Major reasons for failure include inaccurate determination of Mandibular foramen, variations in location of foramen, presence of accessory foramen, etc. to name a few. The present article aims to present the morpho-anatomical variations in the position of Mandibular Foramen in East European population. For this reason, the distance of the foramen was measured from 5 different bony landmarks using digital Vernier caliper on dry human adult mandibles. Analyzing the results, the foramen was found to be positioned at a mean distance of 16.88 ± 2.43 mm on the right side and 17.33 ± 2.24 mm on the left side from the anterior border of the ramus. Similarly, it was found to be 12.31 ± 2.49 mm and 11.75 ± 2.47 mm on right and left sides respectively from posterior border of ramus. It was found to be 17.41 ± 3.22 mm and 18.01 ± 3.44 mm and 19.80 ± 3.86 mm and 20.11 ± 4.08 mm on right and left sides from mandibular notch and angle of mandible respectively.publishersversionPeer reviewe

Group A streptococcal (GAS) infections amongst children in Europe : Taming the rising tide

A rising tide of Group A streptococcal infections is being reported in several European countries including the UK, Spain, Sweden, France, and Ireland. In this editorial, we shed light on the current scenario and present the discussion on the latest available interim clinical guidelines. We further explore the cause of this sudden rise and the interventional measures needed to tame the rising tide.publishersversio

The new face of monkeypox virus : an emerging global emergency

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Fostering Public Health Ethics Awareness among Medical Students through Interactive Web-Based Values Exchange Learning : A Cross-Sectional Study

Background Medical ethics have traditionally encountered resistance in medical education, with doubts regarding their necessity during preclinical years. Concerns include the practical application of theoretical ethics and favoring the learning of ethics through observation and imitation. Furthermore, ethics courses often focus on codes and regulations and neglect the promotion of moral reasoning and decision-making skills. Therefore, enhancing ethical researcher-training modules is essential for advancing instructional methods and promoting a unified and interactive learning atmosphere. Methods A three-week course was conducted using the Values Exchange (VX) online platform to assess the effectiveness of the VX system in involving students and its impact on their perceptions of the importance of research ethics. Accordingly, a blended course was provided as an optional course to the students, and a qualitative course evaluation survey was administered at the end of the course. Results Sixty-one medical students participated in a pilot-blended research ethics program and the majority (93%) reported a positive learning experience. The course was preferred over traditional classroom-based courses by 80% of the respondents, with 77% reporting increased interest and motivation to study Research Ethics. Over half of the students (57%) reported that the course stimulated their thinking and changed their perspectives on handling medical research issues. Many students (20%) reported improvements in their social and information technology skills. The VX platform was able to meet the expectations of both faculty and students, and fulfilled all dimensions of the Health Professions Learning Environment Conceptual Framework. Conclusions The VX platform provides an interactive and effective simulated learning environment that promotes ethical research Aligned with the Health Professions Learning Environment Framework, VX fosters core competencies, such as ethical reasoning, decision-making, and cultural respect. Medical schools are encouraged to implement VX to enhance the teaching of research ethics.Peer reviewe

Expression Analysis of FGF/FGFR and FOX Family Proteins in Mucosal Tissue Obtained from Orofacial Cleft-Affected Children

Funding Information: Funding: The present study was funded by Riga Stradinš University (RSU), Project Nb. 5-1/384/2020 (dated 10 September 2020). Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Orofacial clefts affect hundreds of thousands of children worldwide annually and are usually corrected by a series of surgeries extending to childhood. The underlying mechanisms that lead to clefts are still unknown, mainly because of the multifactorial etiology and the myriad of interactions between genes and environmental factors. In the present study, we investigated the role and expression of candidate genes belonging to the FGF/FGFR signaling pathway and FOX family in tissue material obtained from 12 pediatric patients undergoing cleft correction surgery. The expression was investigated using immunohistochemistry (IHC) and chromogenic in-situ hybridization (CISH) in three cell/tissue types—epithelial cells, connective tissue, and endothelial cells. We found elevated expression of FGFR1 in epithelial cells while no expression was observed in endothelial cells. Further, our results elucidate the potential pathogenetic role of FGFR1 in cellular proliferation, local site inflammation, and fibrosis in cleft patients. Along with bFGF (also called FGF2), FGFR1 could play a pro-inflammatory role in clefts. Over-amplification of FGFR2 in some patients, along with bFGF, could potentially suggest roles for these genes in angiogenesis. Additionally, increased expression of FOXE1 (also called TTF2) contributes to local site inflammation. Finally, zero to low amplification of FOXO1 could suggest its potential role in inducing oxidative stress in the endothelium along with reduced epithelial apoptosis.publishersversionPeer reviewe

Singer’s Nodules : Investigating the Etiopathogenetic Markers Progressing Their Pathogenesis and Clinical Manifestations

Funding Information: The support of Riga Stradins University (RSU) is greatly acknowledged. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Vocal nodules (or Singer’s nodules) are benign vocal cord structures which are commonly encountered by clinicians. Though phonetic trauma/abuse is thought to be the main cause of the development of vocal nodules, the exact etiopathogenesis remains unknown. Hence, we compared the immunohistochemical markers for proliferation (Ki-67), apoptosis (TUNEL), growth (EGFR), ischemia (VEGF), inflammation (IL-1α and 10), and immunoreactive innervation (PGP 9.5), in vocal nodule tissue samples obtained from 10 females (17–56 years) and vocal cord tissue from seven controls. A statistically significant increase in Ki-67, TUNEL, EGFR, VEGF and IL-1α expression was noted (p 0.05). All markers demonstrated moderate to strong positive correlations, except for IL-10. These findings suggest increased cellular growth and proliferation in vocal nodules coupled with a persistent presence of inflammatory and ischemic environment. Furthermore, global prevalence of apoptotic cells and decreased anti-inflammatory cytokines highlight the presence of underlying complex mechanisms in the etiopathogenesis of vocal nodules, with age having a negligible impact on the marker levels. Our results could potentially further our knowledge in understanding the effects of different treatment modalities available at the cellular level.publishersversionPeer reviewe

Alcohol-Induced Alterations in the Vascular Basement Membrane in the Substantia Nigra of the Adult Human Brain

Funding Information: Funding: This research was funded by the Latvian Council of Science, “The Role of Human Herpesvirus-6t Infection and Alcohol Abuse in the Development of Neuroinflammation”, project no. lzp-2020/2-0069. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.The blood–brain barrier (BBB) represents a highly specialized interface that acts as the first line of defense against toxins. Herein, we investigated the structural and ultrastructural changes in the basement membrane (BM), which is responsible for maintaining the integrity of the BBB, in the context of chronic alcoholism. Human post-mortem tissues from the Substantia Nigra (SN) region were obtained from 44 individuals, then grouped into controls, age-matched alcoholics, and non-age-matched alcoholics and assessed using light and electron microscopy. We found significantly less CD31+ vessels in alcoholic groups compared to controls in both gray and white matter samples. Alcoholics showed increased expression levels of collagen-IV, laminin-111, and fibronectin, which were coupled with a loss of BM integrity in comparison with controls. The BM of the gray matter was found to be more disintegrated than the white matter in alcoholics, as demonstrated by the expression of both collagen-IV and laminin-111, thereby indicating a breakdown in the BM’s structural composition. Furthermore, we observed that the expression of fibronectin was upregulated in the BM of the white matter vasculature in both alcoholic groups compared to controls. Taken together, our findings highlight some sort of aggregation or clumping of BM proteins that occurs in response to chronic alcohol consumption.publishersversionPeer reviewe

Selecting suitable reference genes for qPCR normalization : A comprehensive analysis in MCF-7 breast cancer cell line

Publisher Copyright: © 2020 The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Background: MCF-7 breast cancer cell line is undoubtedly amongst the most extensively studied patient-derived research models, providing pivotal results that have over the decades translated to constantly improving patient care. Many research groups, have previously identified suitable reference genes for qPCR normalization in MCF-7 cell line. However, over the course of identification of suitable reference genes, a comparative analysis comprising these genes together in a single study has not been reported. Furthermore, the expression dynamics of these reference genes within sub-clones cultured over multiple passages (p) has attracted limited attention from research groups. Therefore, we investigated the expression dynamics of 12 previously suggested reference genes within two sub-clones (culture A1 and A2) cultured identically over multiple passages. Additionally, the effect of nutrient stress on reference gene expression was examined to postulate an evidence-based recommendation of the least variable reference genes that could be employed in future gene expression studies. Results: The analysis revealed the presence of differential reference gene expression within the sub-clones of MCF-7. In culture A1, GAPDH-CCSER2 were identified as the least variable reference genes while for culture A2, GAPDH-RNA28S were identified. However, upon validation using genes of interest, both these pairs were found to be unsuitable control pairs. Normalization of AURKA and KRT19 with triplet pair GAPDH-CCSER2-PCBP1 yielded successful results. The triplet also proved its capability to handle variations arising from nutrient stress. Conclusions: The variance in expression behavior amongst sub-clones highlights the potential need for exercising caution while selecting reference genes for MCF-7. GAPDH-CCSER2-PCBP1 triplet offers a reliable alternative to otherwise traditionally used internal controls for optimizing intra- and inter-assay gene expression differences. Furthermore, we suggest avoiding the use of ACTB, GAPDH and PGK1 as single internal controls.publishersversionPeer reviewe