CRISPR-Cas13d screens identify KILR, a breast cancer risk-associated lncRNA that regulates DNA replication and repair.

BACKGROUND: Long noncoding RNAs (lncRNAs) have surpassed the number of protein-coding genes, yet the majority have no known function. We previously discovered 844 lncRNAs that were genetically linked to breast cancer through genome-wide association studies (GWAS). Here, we show that a subset of these lncRNAs alter breast cancer risk by modulating cell proliferation, and provide evidence that a reduced expression on one lncRNA increases breast cancer risk through aberrant DNA replication and repair. METHODS: We performed pooled CRISPR-Cas13d-based knockdown screens in breast cells to identify which of the 844 breast cancer-associated lncRNAs alter cell proliferation. We selected one of the lncRNAs that increased cell proliferation, KILR, for follow-up functional studies. KILR pull-down followed by mass spectrometry was used to identify binding proteins. Knockdown and overexpression studies were performed to assess the mechanism by which KILR regulates proliferation. RESULTS: We show that KILR functions as a tumor suppressor, safeguarding breast cells against uncontrolled proliferation. The half-life of KILR is significantly reduced by the risk haplotype, revealing an alternative mechanism by which variants alter cancer risk. Mechanistically, KILR sequesters RPA1, a subunit of the RPA complex required for DNA replication and repair. Reduced KILR expression promotes breast cancer cell proliferation by increasing the available pool of RPA1 and speed of DNA replication. Conversely, KILR overexpression promotes apoptosis in breast cancer cells, but not normal breast cells. CONCLUSIONS: Our results confirm lncRNAs as mediators of breast cancer risk, emphasize the need to annotate noncoding transcripts in relevant cell types when investigating GWAS variants and provide a scalable platform for mapping phenotypes associated with lncRNAs.

Enhanced lung cancer detection: Integrating improved random walker segmentation with artificial neural network and random forest classifier.

Background: Medical image segmentation is a vital yet difficult job because of the multimodality of the acquired images. It is difficult to locate the polluted area before it spreads. Methods: This research makes use of several machine learning tools, including an artificial neural network as well as a random forest classifier, to increase the system's reliability of pulmonary nodule classification. Anisotropic diffusion filtering is initially used to remove noise from a picture. After that, a modified random walk method is used to get the region of interest inside the lung parenchyma. Finally, the features corresponding to the consistency of the picture segments are extracted using texture-based feature extraction for pulmonary nodules. The final stage is to identify and classify the pulmonary nodules using a classifier algorithm. Results: The studies employ cross-validation to demonstrate the validity of the diagnosis framework. In this instance, the proposed method is tested using CT scan information provided by the Lung Image Database Consortium. A random forest classifier showed 99.6 percent accuracy rate for detecting lung cancer, compared to a artificial neural network's 94.8 percent accuracy rate. Conclusions: Due to this, current research is now primarily concerned with identifying lung nodules and classifying them as benign or malignant. The diagnostic potential of machine learning as well as image processing approaches are enormous for the categorization of lung cancer.

Effectiveness of picture-based interventions of toothbrush training on improvement of oral hygiene in children with autism spectrum disorders: A systematic review and meta-analysis.

BACKGROUND: Children with autism exhibit a higher risk of poor oral health due to difficulty in the performance of simple tasks such as toothbrushing. AIM: This current systematic review aims to evaluate the effectiveness of Picture based intervention of toothbrush training on improvement of oral hygiene in children with autism spectrum disorders (ASD) MATERIALS AND METHODS: Prospero registered (CRD42023450156). PubMed, Cochrane, Scopus databases are searched from years January 1, 1980 to August 1, 2023 using broad search terms (brush) AND (autism). RESULTS: The search queries have identified 853 titles, from three databases (PubMed, Scopus, Cochrane), after application of filters for exclusion of systematic reviews and meta-analysis, duplicate exclusion and removal of irrelevant titles led to the final inclusion of 24 articles for full text screening. From the 24 included studies, 10 studies (four RCTs and six non-randomized clinical studies) sustained the final rigorous PICO search. Quantitative pooling of data were performed for limited articles. CONCLUSION: Low quality evidence suggest that picture-based intervention of toothbrush training has significant improvement (p ≤ .05) in improving toothbrushing habit as well as performance as indicated by the Plaque Index score (PI), Gingival index (GI) and Oral hygiene index score (OHI-S).

Lung Cancer Detection from CT Images: Modified Adaptive Threshold Segmentation with Support Vector Machines and Artificial Neural Network Classifier.

OBJECTIVE: The objective of the research is to implement an advanced modified threshold segmentation and classification model for early and accurate detection of lung cancer from CT images. METHODS: Using the Support Vector Machines (SVM) classifier as well as the Artificial Neural Network (ANN) classifier, the authors propose using Modified adaptive threshold segmentation as a segmentation approach for cancer detection. Here, Lung Image Database Consortium (LIDC) datasets, a collection of CT scans, are used as the video frames in an investigation to authorize the recitation of the suggested technique. RESULTS: Both quantitative as well as qualitative analyses are used to analyze the segmentation function of the anticipated algorithm. Both the ANN and SVM classifiers used in the suggested technique for lung cancer diagnosis achieve world-record levels of accuracy, with the former achieving a 96.3% detection rate and the latter a 97% rate of accuracy. CONCLUSION: This innovation may have a major impact on the worldwide rate of lung cancer rate due to its ability to detect lung tumors in their earliest stages when they are most amenable to being avoided and treated. This method is useful because it provides more information and facilitates quick, precise decision-making for doctors diagnosing lung cancer in their patients.

Preventing recurrence in Sonic Hedgehog Subgroup Medulloblastoma using the OLIG2 inhibitor CT-179.

Recurrence is the primary life-threatening complication for medulloblastoma (MB). In Sonic Hedgehog (SHH)-subgroup MB, OLIG2-expressing tumor stem cells drive recurrence. We investigated the anti-tumor potential of the small-molecule OLIG2 inhibitor CT-179, using SHH-MB patient-derived organoids, patient-derived xenograft (PDX) tumors and mice genetically-engineered to develop SHH-MB. CT-179 disrupted OLIG2 dimerization, DNA binding and phosphorylation and altered tumor cell cycle kinetics in vitro and in vivo, increasing differentiation and apoptosis. CT-179 increased survival time in GEMM and PDX models of SHH-MB, and potentiated radiotherapy in both organoid and mouse models, delaying post-radiation recurrence. Single cell transcriptomic studies (scRNA-seq) confirmed that CT-179 increased differentiation and showed that tumors up-regulated Cdk4 post-treatment. Consistent with increased CDK4 mediating CT-179 resistance, CT-179 combined with CDK4/6 inhibitor palbociclib delayed recurrence compared to either single-agent. These data show that targeting treatment-resistant MB stem cell populations by adding the OLIG2 inhibitor CT-179 to initial MB treatment can reduce recurrence.

Influence of Birth Characteristics and Feeding Practices on Eruption of Primary Teeth in Children: A Preliminary Cross-sectional Observational Study.

Background: The eruption process is under strong genetic control, external environmental factors also significantly contribute to this process. Therefore the present study was undertaken to identify the influence of birth characteristics and feeding practices on the eruption of the first primary tooth among healthy children in Pune, Maharashtra, India. Materials and methods: The study was undertaken among 200 children aged 3-24 months. Children with any underlying conditions, those not accompanied by their mother and with neonatal teeth/natal teeth were excluded. The birth weight, gestational age, and type of delivery were obtained from medical records of the child. Data on feeding practices and the age of eruption of the first primary tooth was recorded after a structured interview of the mother followed by oral examination of the child. Gestational age was categorized into >37 weeks and <37 weeks, birth weight was divided into three groups-<1500, 1500-2500, and >2500 gm and four groups according to feeding practice during the first 6 months of life (exclusively breastfed, exclusively bottle fed, combination of breastfeeding and bottle feeding, and spoon-feeding). Results: The mean age of eruption in the study population was 9.17 months. A9.05 months in males and 9.44 in females. A statistically significant correlation was noted with gestational age, birth weight, and feeding practices. Infants with a gestational age of >37 weeks, higher birth weight and those who were exclusively breastfed for the first 6 months had an earlier eruption of first primary teeth. Conclusion: External factors seem to influence the eruption of the primary tooth. More longitudinal studies with larger sample sizes and matching confounders are needed to clearly understand the factors affecting the primary tooth eruption. How to cite this article: Shetty SS, Mathur A, Gopalakrishnan D, et al. Influence of Birth Characteristics and Feeding Practices on Eruption of Primary Teeth in Children: A Preliminary Cross-sectional Observational Study. Int J Clin Pediatr Dent 2023;16(5):724-727.

Immunogenicity, reactogenicity and safety of an inactivated quadrivalent influenza vaccine in children and adolescents 6 months through 17 years of age in India.

Efficacy and safety data on quadrivalent influenza vaccines (QIVs) for immunization of Indian children are scarce. This phase 3, registration study evaluated the immunogenicity, safety, and tolerability of a QIV in Indian children aged 6-35 months (Group 1) and 3-17 y (Group 2). Subjects received one or two doses (0.5 mL each) of the study vaccine based on their priming status. Immunogenicity (post-vaccination geometric mean fold increase in hemagglutination inhibition [HI] titers and proportion of patients with seroprotection and seroconversion against the four influenza strains), unsolicited adverse events (AEs), and tolerability were analyzed. Among 118 subjects enrolled in each group, the geometric mean(standard deviation) fold increase in HI titers against A(H3N2), A(H1N1), B(Victoria), and B(Yamagata) strains were 31.7(5.33), 10.5(6.06), 4.1(5.70), and 8.6(5.34) in Group 1 and 14.0(4.37), 9.2(4.26), 14.3(6.73), and 14.4(5.41) in Group 2, respectively. Seroprotection was achieved by 91.2%, 83.3%, 41.2%, and 68.4% subjects in Group 1 and 100%, 95.8%, 73.7%, and 89.8% subjects in Group 2, respectively. Seroconversion was achieved by 87.7%, 66.7%, 41.2%, and 64.9% subjects in Group 1 and 89.0%, 78.8%, 69.5%, and 75.4% subjects in Group 2, respectively. Vaccination site pain and fever were the most common local and systemic reactions, respectively. Systemic reactions were more frequent in Group 1 (16.9% vs 7.6%). Most subjects (>90%) did not experience inconvenience within 7 d of vaccination; <10% in both groups reported unsolicited AEs. Thus, the QIV had a positive benefit/risk profile in Indian children/adolescents aged 6 months to 17 y.CTRI Registry No: CTRI/2018/05/014191Registry Name: Clinical Trials Registry - IndiaDate of Trial Registration: May 29, 2018Study Dates: August 03, 2018 (first subject first visit) to January 31, 2019 (last subject last visit)Drugs Controller General of India [DCGI] permission letter number: CT-03/2018.

Monte Carlo calculated detector-specific correction factors for Elekta radiosurgery cones.

A radiation field is considered small if its dimension is lower than the range of secondary electrons and the collimating devices partially occlude the source. Different detector types, such as unshielded diodes, diamond detectors, and small-volume ion chambers, are used for small-field measurements. Although the active volumes of these detectors are small, their non-water equivalent materials cause response variations. Herein, we aim to calculate the correction factors for our clinical detectors, EDGE detector (Sun Nuclear), 60017 diode (PTW), and CC01 ion chamber (IBA), for stereotactic radiosurgery cones of diameters of 5-15 mm in an Elekta Synergy linear accelerator using a Monte Carlo simulation. An Elekta Synergy linear accelerator treatment head was simulated using BEAMnrc Monte Carlo code as per the manufacturer specification. All three detectors were simulated as per the manufacturer specification. Three EGSnrc user codes were used for the detector simulation based on the detector geometry. The Monte Carlo model of the treatment head was validated against the measured data for a standard field size of 10 × 10 cm2. The off-axis profile, percentage depth dose, and tissue phantom ratioTPR1020were verified in the validation procedure. The measured and Monte Carlo calculated relative output factors (ROFs) were not consistent. In a 5 mm field size, EDGE diode overestimated the ROF by 7.06%, and 60017 diode to 4.611%. In a 7.5 mm field size, the variations were 4.295% and 3.691% for EDGE and 60017 diodes, respectively. CC01 ion chamber under-responded up to 10% because of its low-density active volume. The maximum corrections were obtained in the smallest field size, which were 0.939(0.007), 0.962(0.006), and 1.117(0.008) for EDGE, PTW T60017, and CC01 detectors, respectively. After applying the Monte Carlo calculated correction factor to the measured ROF, it became consistent with the Monte Carlo calculated ROF.

Development of new drug-regimens against multidrug-resistant tuberculosis.

Tuberculosis (TB) being the leading infectious killer in the domain wherein globally, almost 20% of all TB strains are resistant to at least 1 major TB drug and there's a growing incidence of multi-drug resistance tuberculosis (MDR-TB). Looking at the current scenario and challenges the existing strategies fall back in terms of treatment of TB. So, to overcome this new, stronger, improved TB drug pipeline and a new standard for the development of novel anti-TB drugs are required in order to make more drug-resistant and efficient drug which also lower the duration period of the treatment of the TB. This review article aims to highlight the recent developments in the anti-tuberculosis agents, those are currently in the clinical development stage.

Ultra-performance liquid chromatography-tandem mass spectrometry assay for determination of plasma nomegestrol acetate and estradiol in healthy postmenopausal women.

A highly sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry method is described for the simultaneous determination of nomegestrol acetate (NOMAC), a highly selective progestogen, and estradiol (E2), a natural estrogen in human plasma. NOMAC was obtained from plasma by solid-phase extraction, while E2 was first separated by liquid-liquid extraction with methyl tert-butyl ether followed by derivatization with dansyl chloride. Deuterated internal standards, NOMAC-d5 and E2-d4 were used for better control of extraction conditions and ionization efficiency. The assay recovery of the analytes was within 90-99%. The analytes were separated on UPLC BEH C18 (50 × 2.1 mm, 1.7 µm) column using a mobile phase comprising of acetonitrile and 3.0 mm ammonium trifluoroacetate in water (80:20, v/v) with a resolution factor (Rs ) of 3.21. The calibration curves were linear from 0.01 to 10.0 ng/mL for NOMAC and from 1.00 to 1000 pg/mL for E2, respectively. The intra- and inter-batch precision was ≤5.8% and the accuracy of quality control samples ranged from 96.7 to 103.4% for both analytes. The practical applicability of the method is demonstrated by analyzing samples from 18 healthy postmenopausal women after oral administration of 2.5 mg nomegestrol acetate and 1.5 mg estradiol film-coated tablets under fasting.

Simultaneous determination of etonogestrel and ethinyl estradiol in human plasma by UPLC-MS/MS and its pharmacokinetic study.

A selective, sensitive and rapid ultra-performance liquid chromatography tandem mass spectrometry method was developed and validated for the simultaneous determination of etonogestrel (ENG) and ethinyl estradiol (EE) in human plasma. The analytes and their deuterated internal standards, ENG-d7 and EE-d4, were extracted from plasma samples by solid-phase extraction on HyperSep™ Retain PEP cartridges. The chromatographic analysis was performed on an Acquity UPLC HSS Cyano column, 100 Å (50 × 2.1 mm, 1.8 µm), column using gradient mobile phase, acetonitrile and 2.0 mm ammonium trifluoroacetate at 0-1.7 min (65:35, v/v) and 1.8-2.7 min (95:5, v/v) with 0.250 mL/min flow rate. Analytes and IS protonated precursor → product ion transitions (ENG, m/z 325.2 → 257.2; EE, m/z 530.2 → 171.2; ENG-d7, m/z 332.2 → 263.2; EE-d4, m/z 534.2 → 171.2) were monitored on a Triple Quadrupole Mass spectrometer (TQMS), operating in multiple reaction monitoring and positive ionization mode. The calibration curves were established at 10.00-2500 pg/mL for ENG and 1.500-150.0 pg/mL for EE with a correlation coefficient (r2 ) ≥0.9996 for both. The validated method was successfully applied to support a bioequivalence study of 0.15 mg ENG and EE 0.03 mg tablet formulation, administered in 24 healthy Indian females. Method reliability was assessed by reanalysis of 94 incurred study samples.

Drug development against tuberculosis: Past, present and future.

Infection of Mycobacterium tuberculosis (MTB) was observed as early as 5000 years ago with evidence, which is a primeval enemy of the humanoid race. MTB is the pathogen which is responsible for causing the infectious disease tuberculosis; it remains a major cause of morbidity and mortality in poor low-income countries as well as in developing countries because of non-availability of reliable laboratory facilities. The current treatment for drug-resistant tuberculosis (TB) is lengthy, complex, and connected with severe harmful side effects and poor outcomes. The present cure against tuberculosis has substantial restrictions, in terms of their efficiency, side-effect outline, and complication of handling. Furthermore, the emergence of multi-drug resistant tuberculosis (MDR-TB) outbreaks during the 1990s and additionally in recent times the vast deadly strains of extensively drug-resistant tuberculosis (XDR-TB) and totally drug resistance tuberculosis (TDR-TB) is hampering efforts to control and manage tuberculosis (TB). As a result, novel methodologies for the treatment of multi-drug-resistant and extensive drug-resistant tuberculosis (TB) are severely desired. A number of new potential anti-tuberculosis drug candidates with novel modes of action have been entered in clinical trials in recent years. These agents are most likely to be effective against resistant strains. The treatment landscape is beginning to shift, with the recent approvals by Food and Drug Administration to the new TB drugs bedaquiline and delamanid. Also, the pipeline of potential new treatments has been fulfilled with several compounds in clinical trials or preclinical development with promising activities against sensitive and resistant MTB bacteria. An additional new chemical entity is also under development. The already existing drugs with their suggested mode of treatment as well as new probable anti-tuberculosis drug moieties which are at present in the pipeline has been summarized in this review.

A dosimetric analysis of cardiac dose with or without active breath coordinator moderate deep inspiratory breath hold in left sided breast cancer radiotherapy.

BACKGROUND AND AIM: Cardiac toxicity is a major concern for left breast tangential field irradiation. The left ventricle and left anterior descending (LAD) artery are suggested to be radiosensitive and radiation to these structures leads to late lethal cardiotoxicity. Moderate deep inspiration breath hold (mDIBH) during radiation treatment delivery helps in reducing the cardiac dose. This study compares dosimetric parameters of heart with and without active breath coordinator (ABC) mDIBH during tangential field breast cancer radiation. STUDY TYPE: This is a dosimetric comparative study. MATERIALS AND METHODS: Forty-five consecutive patients with left-sided breast cancer who underwent breast-conserving surgery and adjuvant tangential field and radiotherapy with ABC mDIBH between November 2013 and September 2015 in our center were analyzed in this study. The ABC device was used for respiratory control and patients who could hold their breath for 20-30 s were considered for radiation with ABC mDIBH. Simulation scans of both free breathing (FB) and ABC mDIBH were done. Tangent field treatment plans with a dose prescription of 40 Gy/15 Fr were generated for each patient, in both scans. Target coverage, dose to the heart, LAD, and the left lung were documented with dose-volume histograms. RESULTS: Statistical Package for the Social Sciences, version 20 software, was used for analysis and the level of significance was set at P < 0.05. Mean heart dose was 308.5cGy with FB and 159cGy with ABC (P < 0.0001). Mean dose to the LAD was reduced by 53.81% (1320.64 cGy vs. 606.56 cGy, P < 0.001). Target coverage was equal in both the plans. CONCLUSION: We report that the use of ABC mDIBH technique resulted in a significant reduction in cardiac dose and hence can be considered as a promising tool for cardiac sparing.

Simultaneous analysis of glucocorticosteroid fluticasone propionate and its metabolite fluticasone propionate 17ß-carboxylic acid in human plasma by UPLC-MS/MS at sub pg/mL level.

A highly sensitive and rapid ultra performance liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous determination of fluticasone propionate (FP) and its major metabolite, fluticasone propionate-17beta-carboxylic acid (FP 17ß-CA) in human plasma. The analytes and their deuterated internal standards, FP-d3 and FP 17ß-CA-d3 were extracted from 500µL plasma samples by solid phase extraction on Oasis MAX cartridges. The chromatographic analysis was performed on ACQUITY UPLC BEH C18 (50mm×2.1mm, 1.7µm) column using methanol-acetonitrile (50:50, v/v) and 2.0mM ammonium trifluroacetate (ATFA) (85:15, v/v) as the mobile phase. Following separation of the analytes, protonated precursor→product ion transitions (FP: m/z 501.1→293.2, FP17ß-CA: m/z 453.3→293.2, FP-d3: m/z 504.2→293.2, FP 17ß-CA-d3: m/z 456.3→293.2) were monitored on FP 17ß-CA a triple quadrupole mass spectrometer, operating in multiple reaction monitoring (MRM) and positive ionization mode. The calibration curves were established in the range of 0.5-100pg/mL with a correlation coefficient (r2)≥0.9992 for both the analytes. The intra-batch and inter-batch accuracy and precision varied from 95.5-103.4% and 0.74-5.06% across quality controls for both the analytes. The mean assay recoveries for FP and FP 17ß-CA were 84.2% and 93.5% respectively. The validated method was successfully applied to support a bioequivalence study of 200µg FP, administered using nasal spray formulation in 18 healthy Indian subjects. Reproducibility of the method was assessed by reanalysis of 98 incurred study samples.

A novel, sensitive and selective method of UPLC/MS-MS for rapid simultaneous determination of midodrine and its active metabolite desglymidodrine in human plasma: Application to support bioequivalence study in healthy human volunteers.

A specific, rapid, sensitive and selective ultra-performance liquid chromatography - tandem mass spectrometry has been developed for the simultaneous determination of midodrine and desglymidodrine in human plasma. The analytes and its deuterated analogs were quantitatively extracted from 100µL of human plasma by solid phase extraction technique. Separation of analytes was achieved on the Waters Acquity UPLC BEH C18 (50×2.1mm, 1.7µm) column using acetonitrile-4.0mM ammonium formate, pH 2.5(90:10, v/v) as mobile phase. The protonated analytes were quantified by selected reaction monitoring in the positive ionization mode by triple quadrupole mass spectrometer. The calibration plots were linear over the concentration range of 0.050-50.0ng/mL. The intra-batch and inter-batch precision (%CV) across quality control levels was <4.0 and the% mean relative recovery was ≥96%. Various other parameters like stability in different conditions; matrix effect and reproducibility of the method were performed in accordance with the guidelines specified by the USFDA for bioanalytical method development and validation. The developed method was successfully administered to the pharmacokinetics study of 5 mg midodrine tablet in 12 healthy subjects. Reproducibility of assay was proved by reanalysis of 48 incurred samples.