Endoplasmic reticulum stress in human chronic wound healing: Rescue by 4-phenylbutyrate.

During wound healing, cells have a high rate of protein synthesis and many proteins need to be folded post-translationally to function, which occurs in the endoplasmic reticulum (ER). In addition to proliferation, several cellular stress conditions, such as hypoxia, in the wound micro-environment lead to the accumulation of unfolded or misfolded proteins in the ER, causing ER stress. Eukaryotic cells have a signalling system to manage ER stress called the unfolded protein response (UPR). Mild UPR activation has a beneficial homeostatic effect; however, excessive UPR induces cell death. Herein, we examined venous leg ulcer biopsies versus normal acute incisional wounds in age-matched elderly subjects and found a large increase in ER stress markers. To study the underlying mechanism, we established several cell cultures from amputated legs from the elderly that showed inherent ER stress. While both keratinocytes and fibroblasts migration was impaired by ER stress, migration of elderly leg skin keratinocytes was markedly improved after treatment with the chemical chaperone and clinically established drug 4-phenylbutyrate (4-PBA) and demonstrated a reduction in ER stress markers. In a full-thickness human skin wound healing model, 4-PBA improved the reepithelialisation rate, which suggests it as a promising drug repurposing candidate for wound healing.

SMARCAL1, the annealing helicase and the transcriptional co-regulator.

The ATP-dependent chromatin remodeling proteins play an important role in DNA repair. The energy released by ATP hydrolysis is used for myriad functions ranging from nucleosome repositioning and nucleosome eviction to histone variant exchange. In addition, the distant member of the family, SMARCAL1, uses the energy to reanneal stalled replication forks in response to DNA damage. Biophysical studies have shown that this protein has the unique ability to recognize and bind specifically to DNA structures possessing double-strand to single-strand transition regions. Mutations in SMARCAL1 have been linked to Schimke immuno-osseous dysplasia, an autosomal recessive disorder that exhibits variable penetrance and expressivity. It has long been hypothesized that the variable expressivity and pleiotropic phenotypes observed in the patients might be due to the ability of SMARCAL1 to co-regulate the expression of a subset of genes within the genome. Recently, the role of SMARCAL1 in regulating transcription has been delineated. In this review, we discuss the biophysical and functional properties of the protein that help it to transcriptionally co-regulate DNA damage response as well as to bind to the stalled replication fork and stabilize it, thus ensuring genomic stability. We also discuss the role of SMARCAL1 in cancer and the possibility of using this protein as a chemotherapeutic target.

Near Fatal Poisoning by Isoniazid and Rifampicin.

Since six decades, Isoniazid and Rifampicin are used as first line drugs for treatment of tuberculosis. The minimum acute lethal or toxic dose of Rifampicin is not well established. However, non-fatal acute overdoses in adults have been reported with doses ranging from 9 to 12 gm and fatal acute overdoses with doses ranging from 14 to 60 gm. Isoniazid, if acutely ingested, even 1.5 to 2 gram may cause toxicity in adults. We report a case of Pott's spine on ATT, who took massive overdose of Rifampicin (>18 gm) and Isoniazid (>12 gm) and reported late (almost 36 hours) after ingestion. He was treated successfully with pyridoxine, hemodialysis and supportive care.

Role of the mitochondrial protein cyclophilin D in skin wound healing and collagen secretion.

Central for wound healing is the formation of granulation tissue, which largely consists of collagen and whose importance stretches past wound healing, including being implicated in both fibrosis and skin aging. Cyclophilin D (CyD) is a mitochondrial protein that regulates the permeability transition pore, known for its role in apoptosis and ischemia-reperfusion. To date, the role of CyD in human wound healing and collagen generation has been largely unexplored. Here, we show that CyD was upregulated in normal wounds and venous ulcers, likely adaptive as CyD inhibition impaired reepithelialization, granulation tissue formation, and wound closure in both human and pig models. Overexpression of CyD increased keratinocyte migration and fibroblast proliferation, while its inhibition reduced migration. Independent of wound healing, CyD inhibition in fibroblasts reduced collagen secretion and caused endoplasmic reticulum collagen accumulation, while its overexpression increased collagen secretion. This was confirmed in a Ppif-KO mouse model, which showed a reduction in skin collagen. Overall, this study revealed previously unreported roles of CyD in skin, with implications for wound healing and beyond.

Integrative Analysis of Potential Biomarkers Involved in the Progression of Papillary Thyroid Cancer.

This study aims to explore key prognostic and diagnostic biomarkers involved in the pathogenesis of papillary thyroid cancer (PTC) which is one of the most common endocrine cancers and whose occurrence is rapidly increasing. Papillary thyroid cancer datasets containing normal and tumor samples were collected from Gene Expression Omnibus. Protein-protein interaction (PPI) network for common upregulated differentially expressed genes (DEGs) was constructed, and hub genes were studied. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed to identify the vital biological behaviors and pathways involved in PTC. PPI network analysis demonstrated the interaction between 134 common upregulated DEGs, and top 15 pivotal genes with highest degree of connectivity were retrieved. Three of the hub genes (DPP4, ITGA2, FN1) were linked to the prognosis of PTC patients and considered clinically relevant core genes via survival analysis. We suggest that the identification of key genes associated with PTC development help us in understanding molecular mechanisms related to disease. These genes could also be considered the diagnostic biomarkers or as therapeutic targets in the future treatment for PTC.

Metabolic Reprogramming and Reliance in Human Skin Wound Healing.

Impaired skin wound healing is a significant global health issue, especially among the elderly. Wound healing is a well-orchestrated process involving the sequential phases of inflammation, proliferation, and tissue remodeling. Although wound healing is a highly dynamic and energy-requiring process, the role of metabolism remains largely unexplored. By combining transcriptomics and metabolomics of human skin biopsy samples, we mapped the core bioenergetic and metabolic changes in normal acute as well as chronic wounds in elderly subjects. We found upregulation of glycolysis, the tricarboxylic acid cycle, glutaminolysis, and ß-oxidation in the later stages of acute wound healing and in chronic wounds. To ascertain the role of these metabolic pathways on wound healing, we targeted each pathway in a wound healing assay as well as in a human skin explant model using metabolic inhibitors and stimulants. Enhancement or inhibition of glycolysis and, to a lesser extent, glutaminolysis had a far greater impact on wound healing than similar manipulations of oxidative phosphorylation and fatty acid ß-oxidation. These findings increase the understanding of wound metabolism and identify glycolysis and glutaminolysis as potential targets for therapeutic intervention.

Knowledge, attitude and use of fluorides among dentists in Texas.

AIM: The centers for disease control and prevention (CDC) recommendations on fluoride use were published in 2001. This study examines how this information has diffused to practicing dentists and the level of fluoride knowledge and use among Texas dentists. MATERIALS AND METHODS: A questionnaire was sent to dentists who self-identified as being in pediatric (343), dental public health (72), and general practices (980); a 12% sample of registered dentists in Texas. RESULTS: Response rate was 42.9%. About 90% of surveyed dentists reported using fluorides routinely. Only 18.8% reported fluoride varnish as the topical fluoride most often used. About 57% incorrectly identified primary effect of fluoride. 'Makes enamel stronger while tooth is developing prior to eruption' was the most commonly cited wrong answer (44%). Only 5% identified that posteruptive effect exceeds any preeruptive effect. CONCLUSION: Despite the evidence for fluoride varnish preventing and controlling dental caries being Grade I, its use is still uncommon. Dentists are expected to be knowledgeable about products they use, but this study reflects lack of understanding about fluoride's predominant mode of action. More accurate understanding enables dentists to make informed and appropriate judgment on treatment options and effective use of fluoride based on risk assessment of dental caries. CLINICAL SIGNIFICANCE: Lack of knowledge of, or failure of adherence to evidence based guidelines in caries prevention by use of appropriate fluoride regimens may adversely affect caries incidence in the population.

Evaluation of two methods in controlling dental treatment water contamination.

UNLABELLED: Dental unit water systems are contaminated with biofilms that amplify bacterial counts in dental treatment water in excess of a million colony forming units per milliliter (cfu/ml). The Centers for Disease Control and Prevention and the American Dental Association have agreed that the maximum allowable contamination of dental treatment water not exceed 500 cfu/ml. This study was conducted to evaluate two protocols in controlling contamination of dental unit water systems and dental treatment water. Both methods used an antimicrobial self-dissolving chlorine dioxide (ClO2) tablet at a high concentration (50 ppm) to shock the dental unit water system biofilms initially followed by periodic exposure. To treat dental treatment source water for patient care, 3 parts per million (ppm) ClO2 in municipal/tap water was compared to use of a citrus botanical extract dissolved in municipal water. Heterotrophic microbial counts of effluent water and laser scanning confocal microscopy were performed to evaluate effects of the two treatments. Results from this study indicated that both treatments were effective in controlling biofilm contamination and reducing heterotrophic plate counts <500 cfu/ml. A comprehensive study addressing compatibility of 50 ppm ClO2 on the metals and nonmetal components of the dental water system and effects of low-grade chemicals used on composite bonding to dentin and enamel is warranted before translation from efficacy studies to common clinical use. CLINICAL SIGNIFICANCE: This study provides evidence-based information of using two methods of controlling dental treatment water contamination. The study was conducted in a clinical practice setting in an active dental clinic and the results are meaningful to a clinician who is interested in providing safe dental treatment water for patient care. KEYWORDS: Dental waterline biofilms, Dental treatment water contamination control, Chlorine dioxide, Emulsifiers, Heterotrophic plate counts, Laser scanning confocal microscopy. How to cite this article: Bansal R, Puttaiah R, Harris R, Reddy A. Evaluation of Two Methods in Controlling Dental Treatment Water Contamination. J Contemp Dent Pract 2011;12(2):73-83. Source of support: Nil Conflict of interest: None declared.

In-silico study of peptide-protein interaction of antimicrobial peptides potentially targeting SARS and SARS-CoV-2 nucleocapsid protein.

This study is an attempt to find a suitable therapy using antimicrobial peptides (AMPs) by identifying peptide-protein interaction of AMPs and nucleocapsid protein of SARS and SARS-CoV- 2. The AMPs were shortlisted from the APD3 database (Antimicrobial peptide database) based on various physicochemical parameters. The binding efficacy of AMPs was measured using the lowest energy score of the docked complexes with 10 selected AMPs. For SARS-CoV, AP00180 showed the best pose with a binding affinity value of - 6.4 kcal/mol. Prominent hydrogen bonding interactions were observed between Lys85 (nucleocapsid receptor) and Arg13 (antimicrobial peptide ligand) having the least intermolecular distance of 1.759 Å. For SARS-CoV-2, AP00549 was docked with a binding affinity value of - 3.4 kcal/mol and Arg119 and Glu14 of receptor nucleocapsid protein and ligand AMP having the least intermolecular distance of 2.104 The dynamic simulation was performed at 50 ns to check the stability of the final docked complexes, one with each protein. The two best AMPs were AP00180 (Human Defensin-5) for SARS and AP00549 (Plectasin) for SARS-CoV-2. From positive results of dynamic simulation and previously known knowledge that some AMPs interact with the nucleocapsid of coronaviruses, these AMPs might be used as a potential therapeutic agent for the treatment regime of SARS-CoV-2 and SARS infection. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40203-021-00103-z.

RecA-like domain 2 of DNA-dependent ATPase A domain, a SWI2/SNF2 protein, mediates conformational integrity and ATP hydrolysis.

ATP-dependent chromatin remodeling proteins use the energy released from ATP hydrolysis to reposition nucleosomes in DNA-dependent processes. These proteins are classified as SF2 helicases. SMARCAL1, a member of this protein family, is known to modulate both DNA repair and transcription by specifically recognizing DNA molecules possessing double-strand to single-strand transition regions. Mutations in this gene cause a rare autosomal recessive disorder known as Schimke Immuno-Osseous Dysplasia (SIOD).Structural studies have shown that the ATP-dependent chromatin remodeling proteins possess two RecA-like domains termed as RecA-like domain 1 and RecA-like domain 2. Using Active DNA-dependent ATPase A domain (ADAAD), the bovine homolog of SMARCAL1, as a model system we had previously shown that the RecA-like domain 1 containing helicase motifs Q, I, Ia, II, and III are sufficient for ligand binding; however, the Rec A-like domain 2 containing motifs IV, V, and VI are needed for ATP hydrolysis. In the present study, we have focused on the motifs present in the RecA-like domain 2. Our studies demonstrate that the presence of an aromatic residue in motif IV is needed for interaction with DNA in the presence of ATP. We also show that the motif V is required for the catalytic efficiency of the protein and motif VI is needed for interaction with DNA in the presence of ATP. Finally, we show that the SIOD-associated mutation, R820H, present in motif VI results in loss of ATPase activity, and therefore, reduced response to DNA damage.

SMARCAL1 Negatively Regulates C-Myc Transcription By Altering The Conformation Of The Promoter Region.

SMARCAL1, a member of the SWI2/SNF2 protein family, stabilizes replication forks during DNA damage. In this manuscript, we provide the first evidence that SMARCAL1 is also a transcriptional co-regulator modulating the expression of c-Myc, a transcription factor that regulates 10-15% genes in the human genome. BRG1, SMARCAL1 and RNAPII were found localized onto the c-myc promoter. When HeLa cells were serum starved, the occupancy of SMARCAL1 on the c-myc promoter increased while that of BRG1 and RNAPII decreased correlating with repression of c-myc transcription. Using Active DNA-dependent ATPase A Domain (ADAAD), the bovine homolog of SMARCAL1, we show that the protein can hydrolyze ATP using a specific region upstream of the CT element of the c-myc promoter as a DNA effector. The energy, thereby, released is harnessed to alter the conformation of the promoter DNA. We propose that SMARCAL1 negatively regulates c-myc transcription by altering the conformation of its promoter region during differentiation.

A Review on Mechanisms of Anti Tumor Activity of Chalcones.

Chalcones comprise a characteristic framework of 1, 3-diaryl-2-propen-1-one. They have been heralded as promising anti cancer drugs and have received much attention in the field of cancer research and drug development. The cytotoxicity of these potent pharmacophores is attributable to a wide spectrum of biological activities like anti inflammatory, anti proliferative, anti fungal, etc. These anti tumor activities are effectuated through apoptosis, cell cycle arrest, anti tubulin and so forth. This review summarizes the recent developments on anti tumor activity of synthetic and natural chalcones and their detailed underlying mechanisms as reported in the past.