Insights from multi-omic modeling of neurodegeneration in xeroderma pigmentosum using an induced pluripotent stem cell system.

Xeroderma pigmentosum (XP) is caused by defective nucleotide excision repair of DNA damage. This results in hypersensitivity to ultraviolet light and increased skin cancer risk, as sunlight-induced photoproducts remain unrepaired. However, many XP patients also display early-onset neurodegeneration, which leads to premature death. The mechanism of neurodegeneration is unknown. Here, we investigate XP neurodegeneration using pluripotent stem cells derived from XP patients and healthy relatives, performing functional multi-omics on samples during neuronal differentiation. We show substantially increased levels of 5',8-cyclopurine and 8-oxopurine in XP neuronal DNA secondary to marked oxidative stress. Furthermore, we find that the endoplasmic reticulum stress response is upregulated and reversal of the mutant genotype is associated with phenotypic rescue. Critically, XP neurons exhibit inappropriate downregulation of the protein clearance ubiquitin-proteasome system (UPS). Chemical enhancement of UPS activity in XP neuronal models improves phenotypes, albeit inadequately. Although more work is required, this study presents insights with intervention potential.

A glycolytic metabolite bypasses "two-hit" tumor suppression by BRCA2.

Knudson's "two-hit" paradigm posits that carcinogenesis requires inactivation of both copies of an autosomal tumor suppressor gene. Here, we report that the glycolytic metabolite methylglyoxal (MGO) transiently bypasses Knudson's paradigm by inactivating the breast cancer suppressor protein BRCA2 to elicit a cancer-associated, mutational single-base substitution (SBS) signature in nonmalignant mammary cells or patient-derived organoids. Germline monoallelic BRCA2 mutations predispose to these changes. An analogous SBS signature, again without biallelic BRCA2 inactivation, accompanies MGO accumulation and DNA damage in Kras-driven, Brca2-mutant murine pancreatic cancers and human breast cancers. MGO triggers BRCA2 proteolysis, temporarily disabling BRCA2's tumor suppressive functions in DNA repair and replication, causing functional haploinsufficiency. Intermittent MGO exposure incites episodic SBS mutations without permanent BRCA2 inactivation. Thus, a metabolic mechanism wherein MGO-induced BRCA2 haploinsufficiency transiently bypasses Knudson's two-hit requirement could link glycolysis activation by oncogenes, metabolic disorders, or dietary challenges to mutational signatures implicated in cancer evolution.

An Exploration of Organoid Technology: Present Advancements, Applications, and Obstacles.

BACKGROUND: Organoids are in vitro models that exhibit a three-dimensional structure and effectively replicate the structural and physiological features of human organs. OBJECTIVE: This work examines the potential applications of organoid technology, as well as the challenges and future directions associated with its implementation. METHODS: The manuscript was put together by conducting a comprehensive literature review, which involved an in-depth evaluation of globally renowned scientific research databases. RESULTS: The field of organoids has generated significant attention due to its potential applications in tissue development and disease modelling, as well as its implications for personalised medicine, drug screening, and cell-based therapies. The utilisation of organoids has proven to be effective in the examination of various conditions, encompassing genetic disorders, cancer, neurodevelopmental disorders and infectious diseases. CONCLUSION: The exploration of the wider uses of organoids is still in its early phases. Research shall be conducted to integrate 3D organoid systems as alternatives for current models, potentially improving both fundamental and clinical studies in the future.

A transcriptional response to replication stress selectively expands a subset of Brca2-mutant mammary epithelial cells.

Germline BRCA2 mutation carriers frequently develop luminal-like breast cancers, but it remains unclear how BRCA2 mutations affect mammary epithelial subpopulations. Here, we report that monoallelic Brca2mut/WT mammary organoids subjected to replication stress activate a transcriptional response that selectively expands Brca2mut/WT luminal cells lacking hormone receptor expression (HR-). While CyTOF analyses reveal comparable epithelial compositions among wildtype and Brca2mut/WT mammary glands, Brca2mut/WT HR- luminal cells exhibit greater organoid formation and preferentially survive and expand under replication stress. ScRNA-seq analysis corroborates the expansion of HR- luminal cells which express elevated transcript levels of Tetraspanin-8 (Tspan8) and Thrsp, plus pathways implicated in replication stress survival including Type I interferon responses. Notably, CRISPR/Cas9-mediated deletion of Tspan8 or Thrsp prevents Brca2mut/WT HR- luminal cell expansion. Our findings indicate that Brca2mut/WT cells activate a transcriptional response after replication stress that preferentially favours outgrowth of HR- luminal cells through the expression of interferon-responsive and mammary alveolar genes.

Point-of-care devices based on fluorescence imaging and spectroscopy for tumor margin detection during breast cancer surgery: Towards breast conservation treatment.

OBJECTIVE: Fluorescence-based methods are highly specific and sensitive and have potential in breast cancer detection. Simultaneous fluorescence imaging and spectroscopy during intraoperative procedures of breast cancer have great advantages in detection of tumor margin as well as in classification of tumor to healthy tissues. Intra-operative real-time confirmation of breast cancer tumor margin is the aim of surgeons, and therefore, there is an urgent need for such techniques and devices which fulfill the surgeon's priorities. METHODS: In this article, we propose the development of fluorescence-based smartphone imaging and spectroscopic point-of-care multi-modal devices for detection of invasive ductal carcinoma in tumor margin during removal of tumor. These multimodal devices are portable, cost-effective, noninvasive, and user-friendly. Molecular level sensitivity of fluorescence process shows different behavior in normal, cancerous and marginal tissues. We observed significant spectral changes, such as, red-shift, full-width half maximum (FWHM), and increased intensity as we go towards tumor center from normal tissue. High contrast in fluorescence images and spectra are also recorded for cancer tissues compared to healthy tissues. Preliminary results for the initial trial of the devices are reported in this article. RESULTS: A total 44 spectra from 11 patients of invasive ductal carcinoma (11 spectra for invasive ductal carcinoma and rest are normal and negative margins) are used. Principle component analysis is used for the classification of invasive ductal carcinoma with an accuracy of 93%, specificity of 75% and sensitivity of 92.8%. We obtained an average 6.17 ± 1.66 nm red shift for IDC with respect to normal tissue. The red shift and maximum fluorescence intensity indicates p < 0.01. These results described here are supported by histopathological examination of the same sample. CONCLUSION: In the present manuscript, simultaneous fluorescence-based imaging and spectroscopy is accomplished for the classification of IDC tissues and breast cancer margin detection.

Posterior Mediastinal Goiter Removal Through Cervical Incision.

Intrathoracic goiter when encountered can be treated by thyroidectomy using cervical incision, only occasionally requiring extra cervical approach. We are reporting one such case in a patient with pituitary macroadenoma with extension of the adenomatous goiter into the posterior mediastinum. It was removed through the cervical collar incision using a vessel sealing device. There were no intraoperative and postoperative complications during the procedure. The need for extra cervical incision should be decided on a case-to-case basis to avoid the increased morbidity associated with sternotomy and lateral thoracotomy incision.

Complex chest wall reconstruction after excision of malignant phyllodes tumour.

Phyllodes tumour is a rare breast tumour which is locally invasive, and full thickness chest wall involvement by phyllodes is rarely reported. A woman presented with recurrent phyllodes tumour and multilobulated exophytic mass of size 18.5×5.3 cm in anterior chest wall which was abutting the right 2nd-7th rib, left 3rd-5th rib and costal cartilages. She underwent wide excision of the left chest wall tumour with excision of left 3rd-5th rib, with polypropylene mesh and pedicled lattismus dorsi myocutaneous flap reconstruction. After recovery, 3 weeks later, she underwent wide excision of the phyllodes tumour on the right chest wall with excision of 2nd-7th rib, lateral border of sternum, right crus of the diaphragm and the lower lobe of the right lung. Anterior chest wall was reconstructed using scaffold made from Steinmann pins and polypropylene mesh with greater omentum pedicled flap and split thickness skin graft.

An In Vitro Study to Compare the Release of Fluoride from Glass Ionomer Cement (Fuji IX) and Zirconomer.

Mercury toxicity from amalgam dental fillings and their potential for creating problems in the environment and for human health have prompted the development of new restorative materials. The leading alternatives among these are glass ionomer cements. According to current understanding, restorative materials that slowly release fluoride exert a local cariostatic effect. For this purpose, glass ionomer cements have desirable properties in that they help prevent recurrence of caries by releasing fluoride over a long period. Thus, they function in accord with the major cariostatic mechanism of fluoride, which is believed to be its action to promote remineralization and to influence the morphology of teeth by reducing enamel solubility and by suppressing oral cariogenic bacteria. Although the minimum local concentration of fluoride release required to inhibit demineralization has not been determined, it is reported that the cariostatic ability of fluoride releasing restorative materials is significant. Zirconomer defines a new class of restorative that promises the strength and durability of amalgam with the protective benefits of glass ionomer while completely eliminating the hazards of mercury. The inclusion of specially micronized zirconia fillers in the glass component of zirconomer reinforces the structural integrity of the restoration and imparts superior mechanical properties for the restoration of load-bearing permanent teeth. Combination of outstanding strength, durability, and sustained fluoride protection deems it ideal for multiple applications. The aim of the present study was to determine the fluoride release from glass ionomer cements and compare it with new material zirconomer. Materials and methods: Sample preparation: Tablets of glass-ionomer cements and zirconomer were prepared. A dental floss was incorporated into the tablets during fabrication to allow suspension into the test medium. Each disk specimen was immersed in airtight polyethylene bottle containing 20 mL of deionized water and incubated at 37°C and stored for 24 hours. Determination of fluoride ion release: Fluoride ion measurement was performed after 6 hours, 24 hours, 48 hours, 7 days, and 14 days under normal atmospheric conditions by fluoride ion selective electrode connected to an ion selective electrode meter. Result and conclusion: Both the material tested in the study had the ability to release fluoride but higher fluoride release was observed by zirconomer as compared to GIC at all time intervals. Clinical significance: From a clinical point of view, both the restorative materials release fluoride at all time intervals; however, addition of zirconia particles in zirconomer increases its strength and provides superior mechanical properties. Therefore, due to the combination of both good structural integrity and fluoride releasing properties, zirconomer can be used for restoration of load bearing teeth. How to cite this article: Kukreja R, Singla S, Bhadoria N, et al. An In Vitro Study to Compare the Release of Fluoride from Glass Ionomer Cement (Fuji IX) and Zirconomer. Int J Clin Pediatr Dent 2022;15(1):35-37.

Interdisciplinary Management of Teeth With a Complicated Crown Fracture: A Case Report With Follow-Up Checklist.

Traumatic injuries resulting in damage to the teeth and associated structures have a grave psychological impact on the patient and hence, these require timely intervention. The fracture extent determines the appropriate treatment plan for the patient. Crown fractures can be treated aesthetically without the invasion of the biological width with meticulous interdisciplinary management. The objective of this article is to report a case of a 27-year-old Indian male patient who presented to our hospital the following day, post-trauma to the anterior teeth. On clinical examination, two teeth were found to have undergone fracture that involved the pulp chamber. Considering the overall oral health, pulpal and periodontal health, availability of the fragments, and invasion of the biological width, a tailor-made treatment plan was devised. The fractured crown fragments were reattached surgically, using a glass fiber post that resulted in an aesthetic biological restoration. This treatment has been successfully managed by an interdisciplinary approach. At the 18th-month follow-up, clinical and radiological examinations suggest a successful outcome.

Protocolized approach saves the limb in peripheral arterial injury: A decade experience.

PURPOSE: Outcomes of peripheral arterial injury (PAI) depend on various factors, such as warm ischemia time and concomitant injuries. Suboptimal prehospital care may lead to delayed presentation, and a lack of dedicated trauma system may lead to poorer outcome. Also, there are few reports of these outcomes. The aim of this study was to review our experience of PAI management for more than a decade, and identify the predictors of limb loss in these patients. METHODS: This is a retrospective analysis of prospectively maintained database of trauma admissions at a level I trauma center from January 2008 to December 2019. Patients with acute upper limb arterial injuries or lower limb arterial injuries at or above the level of popliteal artery were included. Association of limb loss with ischemia time, mechanism of injury and concomitant injuries was studied using multiple logistic regressions. Statistical analysis was performed using STATA version 15.0 (Stata Corp LLC, Texas). RESULTS: Out of 716 patients with PAI, the majority (92%) were young males. Blunt trauma was the most common mechanism of injury. Median ischemia time was 4 h (interquartile range 2-7 h). Brachial artery (28%) was the most common injured vessel followed by popliteal artery (18%) and femoral artery (17%). Limb salvage rate was 78%. Out of them, 158 (22%) patients needed amputation, and 53 (7%) had undergone primary amputation. The majority (86%) of patients who required primary or secondary amputations had blunt trauma. On multivariate analysis, blunt trauma, ischemia time more than 6 h and concomitant venous, skeletal, and soft tissue injuries were associated with higher odds of amputation. CONCLUSION: Over all limb salvage rates was 78% in our series. Blunt mechanism of injury and associated skeletal and soft tissue injury, ischemia time more than 6 h portend a poor prognosis. Injury prevention, robust prehospital care, and rapid referral to specialized trauma center are few efficient measures, which can decrease the morbidity associated with vascular injury.

Silver Nanoprism for Selective and Sensitive Detection of Hg+2 Ions.

In this article, we propose high-performance colorimetric detection of Hg+2 using silver nanoprisms. The spherical and triangular AgNPs were synthesized using varied concentration of NaBH4. Pristine AgNPs without any further modification were used for the detection of various metal ions including Hg2+, Pb2+, Cl-, Cd2+, Co2+, Cu2+, Ba2+, Pb2+, Cr3+, Cr2O2-7 , Fe2+, Fe3+ etc. AgNPs were not only selective in detecting the ions of Cl- and Hg+2 ions but also highly sensitive. Minimum detection limit was observed to be as low as 10-7 ppm for both Hg+2 and Cl-. Water samples collected from various locations detected for the presence of various heavy metals. Silver nanoprisms owing to their surface plasmon resonance exhibit highly selective tendency towards detection against Hg+2.

Synthetic Strategies Towards Safer NSAIDs Through Prodrug Approach: A Review.

Non-steroidal anti-inflammatory drugs (NSAIDs) are agents that are used for various properties such as analgesic activity, anti-inflammatory activity, antipyretic activity, etc. But this class of drugs is associated with different side effects such as dyspepsia, gastroduodenal ulcers, gastrointestinal (GI) bleeding and perforation. The prodrug approach is quite beneficial to curb these side effects. Prodrugs are the inactive compounds that, on metabolism get converted into an active metabolite exhibiting desired activities. Commonly used approaches for synthesizing prodrugs are amide, esters, and mutual prodrugs by suppressing the free carboxylic groups responsible for these side effects. In this review, different schemes reported for the synthesis of NSAIDs that are devoid of undesired side effects such as irritation to gastric mucosa, gastrotoxicity, and ulcerogenicity have been compiled. Docking studies and the structure-activity relationship of some compounds are also discussed. The paper shall help the researchers to understand the methods to expedite the synthesis by carrying out substitutions of various groups of the parent compound and establish the mechanism of action of these derivatives of masking the unwanted side effects.

DETECTing Merkel Cell Polyomavirus in Merkel Tumors.

Merkel cell carcinoma (MCC) is a rare, aggressive skin cancer caused either by Merkel cell polyomavirus (MCV) T antigen expression, post-integration (~80% cases), or by UV-mediated DNA damage. Interestingly, overall survival of MCV-positive Merkel cell carcinoma patients is better, making this differential information of significant diagnostic and prognostic value. Also, MCV provides a direct target for therapy in MCC patients. Currently, the methods used for diagnosis of MCV in tumors are often discordant and unreliable. Here we used a guided molecular scissors based-DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) technique to develop an in vitro molecular diagnostic tool for MCV-positive MCC. DETECTR couples recombinase polymerase based amplification of target MCV DNA with Cas12a mediated detection. CRISPR diagnostics couple specific detection followed by cutting of the pathogenic DNA by the Cas enzyme-gRNA complex, with non-specific cutting of ssDNA that provides a measurable visual cue. To detect MCV DNA in MCC, we designed Cas12a gRNAs targeting the MCV DNA and tested their targeting efficiency, and sensitivity using a fluorophore quencher labeled reporter assay. We show that MCV DETECTR system can detect MCV integrated in Merkel tumor rapidly, specifically and with femto-molar sensitivity. Our study is a preliminary, proof-of-principle analysis showing the use of CRISPR for MCV diagnosis. Further validation in human tumor samples is needed for its clinical use in the near future. This new system is promising and we hope it can be coupled with immunohistochemical studies to diagnose the viral status of MCC in clinics soon.

Control of Anxiety in Pediatric Patients using "Tell Show Do" Method and Audiovisual Distraction.

INTRODUCTION: Visiting a dentist can easily evoke strong fear reactions and acute anxiety in children. It is one of the most basic reasons for avoidance and neglect of dental care. It may obstruct delivery of dental care, as the child may be unwilling to accept the treatment being provided by the dentist. AIM: To evaluate and compare reduction in anxiety level in patients undergoing dental treatment at first dental visit. TECHNIQUE: The study was conducted on 400 patients coming to the Department of Pedodontics and Preventive Dentistry, Guru Teg Bahadur Hospital, University College of Medical Sciences, New Delhi, for their first dental visit. Anxiety was recorded using facial image scale (FIS), Venham's picture test (VPT), blood pressure, pulse rate (PR), and oxygen saturation (SpO2) at different stages of the visit. Patients coming for the first dental visit were subjected to restorative treatment under Tell show do (TSD) method and audiovisual distraction (AVD). The data collected were tabulated and subjected to statistical analysis. CONCLUSION: The AVD was found to be more capable in reducing anxiety than TSD. Combination of TSD and AVD had an additive effect in reduction of anxiety level and it proved to be more beneficiary. CLINICAL SIGNIFICANCE: If a child's behavior in the dental office cannot be managed, then it is difficult to hold out any dental treatment that is needed. Bringing positivity in the child's behavior would not only increase efficiency of work but would also make the experience for child undergoing treatment more pleasant.

In vitro Comparative Evaluation of Various Restorative Materials used for restoring Class III Cavities in Deciduous Anterior Teeth: A Clinical Study.

INTRODUCTION: Beauty standards in today's modernized world scenario are formed by well-aligned and well-designed bright white teeth. One of the major reasons behind patients reporting to dental clinics is pain. Caries in the anterior primary teeth forms one of the major concerns from a restorative point of view. Very few studies are quoted in literature which stresses on the follow-up of anterior restorations in primary teeth. Hence, we evaluated and compared the efficacy of composite resin and resin-modified glass ionomer cement (RGIC) for class III restorations in primary anterior teeth. MATERIALS AND METHODS: The present study was conducted in the pediatric dental wing and included a total of 80 patients aged 3 to 5½ years who reported with the chief complaint of carious lesions in the primary anterior teeth. Patients having minimal of a pair of similar appearing small carious lesions on the same proximal surfaces of the deciduous maxillary incisors were included for the study. All the patients were randomly divided into two groups: One in which RGIC restoration was done and other in which composite restoration was done. Cavity preparation was done and filling of the cavity with the restorative materials was carried out. Assessment of the restorations was done at 4, 8, and 12 months time following criteria given by Ryge et al. All the results were analyzed by Statistical Package for the Social Sciences (SPSS) software. Mann-Whitney test and one-way analysis of variance (ANOVA) were used to evaluate the level of significance; p value less than 0.05 was considered as significant. RESULTS: For composite and RGIC restorations, the mean score for anatomic shape was 1.21 and 1.10 respectively. While comparing the clinical parameters, nonsignificant results were obtained between composite and RGIC restorative materials at 4-, 8-, and 12-month interval. On comparing the clinical parameters for individual restorative materials at different time intervals, statistically significant results were obtained only for anatomical shape and form. CONCLUSION: Both RGIC and composite resin restorative materials showed acceptable clinical outcomes after 12 months of follow-up in deciduous anterior teeth. In restoring class III restorations in primary anterior teeth, both the restorative materials showed similar outcome. CLINICAL SIGNIFICANCE: Keywords: Composite, Marginal, Resin-modified glass ionomer cement.

Enhanced production of napthoquinone metabolite (shikonin) from cell suspension culture of Arnebia sp. and its up-scaling through bioreactor.

Cell culture in shake flask and air-lift bioreactor was carried out to exploit the potential of Arnebia sp. for napthoquinone metabolite production. Cell suspension cultures of Arnebia were established from friable callus in liquid MS medium supplemented with 6-benzylaminopurine (BAP) (10 µM) and indole-3-butyric acid (IBA) (5 µM). Growth kinetic studies were done by using settled cell volume and fresh/dry cell weight method. Suspension cultures were maintained by sub-culturing at 10 days interval. A two-stage culture system is employed using growth medium (GM) and modified M9 medium (production medium) for cell biomass and naphthoquinone pigment production, respectively. Results showed that cultivation of cells under dark conditions at room temperature (25 ± 2 °C) enhanced the cell biomass from 100 to 625 g l-1. The pigment production was also found to be increased in dark conditions at room temperature. Alkaline pH found to have positive effect on pigment yield. In case of M9 medium constituents, absence of Na2SO4 does not affect the pigment yield. The current approaches have the cumulative effect to meet an increased level of (25.5 µg/ml) metabolite production in air-lift bioreactor.

Pharyngeal and peritonsillar abscess due to Leclercia adecarboxylata in an immunocompetant patient.

Leclercia adecarboxylata was isolated in pure culture from a peritonsillar and lateral pharyngeal abscess in an immunocompetent host. To our knowledge, this is the first case of infection caused by this microorganism being found in an odontogenic area of the head and neck.