Thiol reductive stress activates the hypoxia response pathway.

Owing to their capability to disrupt the oxidative protein folding environment in the endoplasmic reticulum (ER), thiol antioxidants, such as dithiothreitol (DTT), are used as ER-specific stressors. We recently showed that thiol antioxidants modulate the methionine-homocysteine cycle by upregulating an S-adenosylmethionine-dependent methyltransferase, rips-1, in Caenorhabditis elegans. However, the changes in cellular physiology induced by thiol stress that modulate the methionine-homocysteine cycle remain uncharacterized. Here, using forward genetic screens in C. elegans, we discover that thiol stress enhances rips-1 expression via the hypoxia response pathway. We demonstrate that thiol stress activates the hypoxia response pathway. The activation of the hypoxia response pathway by thiol stress is conserved in human cells. The hypoxia response pathway enhances thiol toxicity via rips-1 expression and confers protection against thiol toxicity via rips-1-independent mechanisms. Finally, we show that DTT might activate the hypoxia response pathway by producing hydrogen sulfide. Our studies reveal an intriguing interaction between thiol-mediated reductive stress and the hypoxia response pathway and challenge the current model that thiol antioxidant DTT disrupts only the ER milieu in the cell.

Iron regulates the quiescence of naive CD4 T cells by controlling mitochondria and cellular metabolism.

In response to an immune challenge, naive T cells undergo a transition from a quiescent to an activated state acquiring the effector function. Concurrently, these T cells reprogram cellular metabolism, which is regulated by iron. We and others have shown that iron homeostasis controls proliferation and mitochondrial function, but the underlying mechanisms are poorly understood. Given that iron derived from heme makes up a large portion of the cellular iron pool, we investigated iron homeostasis in T cells using mice with a T cell-specific deletion of the heme exporter, FLVCR1 [referred to as knockout (KO)]. Our finding revealed that maintaining heme and iron homeostasis is essential to keep naive T cells in a quiescent state. KO naive CD4 T cells exhibited an iron-overloaded phenotype, with increased spontaneous proliferation and hyperactive mitochondria. This was evidenced by reduced IL-7R and IL-15R levels but increased CD5 and Nur77 expression. Upon activation, however, KO CD4 T cells have defects in proliferation, IL-2 production, and mitochondrial functions. Iron-overloaded CD4 T cells failed to induce mitochondrial iron and exhibited more fragmented mitochondria after activation, making them susceptible to ferroptosis. Iron overload also led to inefficient glycolysis and glutaminolysis but heightened activity in the hexosamine biosynthetic pathway. Overall, these findings highlight the essential role of iron in controlling mitochondrial function and cellular metabolism in naive CD4 T cells, critical for maintaining their quiescent state.

Nanoconfinement Effect in Water Processable Discrete Molecular Complex-Based Hybrid Piezo- and Thermo-Electric Nanogenerator.

Water-processable hybrid piezo- and thermo-electric materials have an increasing range of applications. We use the nanoconfinement effect of ferroelectric discrete molecular complex [Cu(l-phe)(bpy)(H2O)]PF6·H2O (1) in a nonpolar polymer 1D-nanofiber to envision the high-performance flexible hybrid piezo- and thermo-electric nanogenerator (TEG). The 1D-nanoconfined crystallization of 1 enhances piezoelectric throughput with a high degree of mechano-sensitivity, i.e., 710 mV/N up to 3 N of applied force with 10,000 cycles of unaffected mechanical endurance. Thermoelectric properties analysis shows a noticeable improvement in Seebeck coefficient (∼4 fold) and power factor (∼6 fold) as compared to its film counterpart, which is attributed to the enhanced density of states near the Fermi edges as evidenced by ultraviolet photoelectric spectroscopy and density functional based theoretical calculations. We report an aqueous processable hybrid TEG that provides an impressive magnitude of Seebeck coefficient (∼793 µV/K) and power factor (∼35 mWm-1K-2) in comparison to a similar class of materials.

Calponin 1 inhibits agonist-induced ERK activation and decreases calcium sensitization in vascular smooth muscle.

Smooth muscle cell (SMC) contraction and vascular tone are modulated by phosphorylation and multiple modifications of the thick filament, and thin filament regulation of SMC contraction has been reported to involve extracellular regulated kinase (ERK). Previous studies in ferrets suggest that the actin-binding protein, calponin 1 (CNN1), acts as a scaffold linking protein kinase C (PKC), Raf, MEK and ERK, promoting PKC-dependent ERK activation. To gain further insight into this function of CNN1 in ERK activation and the regulation of SMC contractility in mice, we generated a novel Calponin 1 knockout mouse (Cnn1 KO) by a single base substitution in an intronic CArG box that preferentially abolishes expression of CNN1 in vascular SMCs. Using this new Cnn1 KO mouse, we show that ablation of CNN1 has two effects, depending on the cytosolic free calcium level: (1) in the presence of elevated intracellular calcium caused by agonist stimulation, Cnn1 KO mice display a reduced amplitude of stress and stiffness but an increase in agonist-induced ERK activation; and (2) during intracellular calcium depletion, in the presence of an agonist, Cnn1 KO mice exhibit increased duration of SM tone maintenance. Together, these results suggest that CNN1 plays an important and complex modulatory role in SMC contractile tone amplitude and maintenance.

Barriers to engagement in the care cascade for tuberculosis disease in India: A systematic review of quantitative studies.

BACKGROUND: India accounts for about one-quarter of people contracting tuberculosis (TB) disease annually and nearly one-third of TB deaths globally. Many Indians do not navigate all care cascade stages to receive TB treatment and achieve recurrence-free survival. Guided by a population/exposure/comparison/outcomes (PECO) framework, we report findings of a systematic review to identify factors contributing to unfavorable outcomes across each care cascade gap for TB disease in India. METHODS AND FINDINGS: We defined care cascade gaps as comprising people with confirmed or presumptive TB who did not: start the TB diagnostic workup (Gap 1), complete the workup (Gap 2), start treatment (Gap 3), achieve treatment success (Gap 4), or achieve TB recurrence-free survival (Gap 5). Three systematic searches of PubMed, Embase, and Web of Science from January 1, 2000 to August 14, 2023 were conducted. We identified articles evaluating factors associated with unfavorable outcomes for each gap (reported as adjusted odds, relative risk, or hazard ratios) and, among people experiencing unfavorable outcomes, reasons for these outcomes (reported as proportions), with specific quality or risk of bias criteria for each gap. Findings were organized into person-, family-, and society-, or health system-related factors, using a social-ecological framework. Factors associated with unfavorable outcomes across multiple cascade stages included: male sex, older age, poverty-related factors, lower symptom severity or duration, undernutrition, alcohol use, smoking, and distrust of (or dissatisfaction with) health services. People previously treated for TB were more likely to seek care and engage in the diagnostic workup (Gaps 1 and 2) but more likely to suffer pretreatment loss to follow-up (Gap 3) and unfavorable treatment outcomes (Gap 4), especially those who were lost to follow-up during their prior treatment. For individual care cascade gaps, multiple studies highlighted lack of TB knowledge and structural barriers (e.g., transportation challenges) as contributing to lack of care-seeking for TB symptoms (Gap 1, 14 studies); lack of access to diagnostics (e.g., X-ray), non-identification of eligible people for testing, and failure of providers to communicate concern for TB as contributing to non-completion of the diagnostic workup (Gap 2, 17 studies); stigma, poor recording of patient contact information by providers, and early death from diagnostic delays as contributing to pretreatment loss to follow-up (Gap 3, 15 studies); and lack of TB knowledge, stigma, depression, and medication adverse effects as contributing to unfavorable treatment outcomes (Gap 4, 86 studies). Medication nonadherence contributed to unfavorable treatment outcomes (Gap 4) and TB recurrence (Gap 5, 14 studies). Limitations include lack of meta-analyses due to the heterogeneity of findings and limited generalizability to some Indian regions, given the country's diverse population. CONCLUSIONS: This systematic review illuminates common patterns of risk that shape outcomes for Indians with TB, while highlighting knowledge gaps-particularly regarding TB care for children or in the private sector-to guide future research. Findings may inform targeting of support services to people with TB who have higher risk of poor outcomes and inform multicomponent interventions to close gaps in the care cascade.

varAmpliCNV: analyzing variance of amplicons to detect CNVs in targeted NGS data.

MOTIVATION: Computational identification of copy number variants (CNVs) in sequencing data is a challenging task. Existing CNV-detection methods account for various sources of variation and perform different normalization strategies. However, their applicability and predictions are restricted to specific enrichment protocols. Here, we introduce a novel tool named varAmpliCNV, specifically designed for CNV-detection in amplicon-based targeted resequencing data (Haloplex™ enrichment protocol) in the absence of matched controls. VarAmpliCNV utilizes principal component analysis (PCA) and/or metric dimensional scaling (MDS) to control variances of amplicon associated read counts enabling effective detection of CNV signals. RESULTS: Performance of VarAmpliCNV was compared against three existing methods (ConVaDING, ONCOCNV and DECoN) on data of 167 samples run with an aortic aneurysm gene panel (n = 30), including 9 positive control samples. Additionally, we validated the performance on a large deafness gene panel (n = 145) run on 138 samples, containing 4 positive controls. VarAmpliCNV achieved higher sensitivity (100%) and specificity (99.78%) in comparison to competing methods. In addition, unsupervised clustering of CNV segments and visualization plots of amplicons spanning these regions are included as a downstream strategy to filter out false positives. AVAILABILITY AND IMPLEMENTATION: The tool is freely available through galaxy toolshed and at: https://hub.docker.com/r/cmgantwerpen/varamplicnv. Supplementary Data File S1: https://tinyurl.com/2yzswyhh; Supplementary Data File S2: https://tinyurl.com/ycyf2fb4. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Unveiling the potential of native arbuscular mycorrhizal fungi for growth promotion and phytochemical enrichment in Valeriana jatamansi Jones.

Medicinal plants are rich sources of pharmaceutically important compounds and have been utilized for the treatment of various diseases since ancient times. Valeriana jatamansi Jones, also known as Indian valerian, holds a special place among temperate Himalayan medicinal plants and is renowned for its therapeutic properties in addressing a variety of ailments. The therapeutic potential of V. jatamansi is attributed to the presence of valuable compounds such as valepotriates, sesquiterpenoids, valeriananoids, jatamanins, lignans, cryptomeridiol, maaliol, xanthorrhizzol, and patchouli alcohol found in its rhizome and roots. This study employed various treatments, including the cultivation of V. jatamansi with the inoculation of Funneliformis mosseae, F. constrictus, and a consortium of arbuscular mycorrhizal fungi (AMF), to investigate their influence on biomass production, chlorophyll content, and the accumulation of bioactive compounds in V. jatamansi. The results revealed significant improvement in these parameters in the inoculated plants. The parameters of plants inoculated with F. mosseae were the highest, followed by those of plants inoculated with F. constrictus and a mixture of AMFs. This study not only underscores the potential of native AMF for promoting the growth of V. jatamansi but also elucidates their role in influencing the synthesis of bioactive compounds. The cultivation of V. jatamansi with native AMF has emerged as a sustainable and eco-friendly approach, providing the dual benefit of enhancing both the medicinal and economic value of this valuable plant. This research contributes valuable insights into the practical application of mycorrhizal associations for the cultivation of medicinal plants, bridging the realms of agriculture and pharmaceuticals.

Decoding the molecular mechanism underlying salicylic acid (SA)-mediated plant immunity: an integrated overview from its biosynthesis to the mode of action.

Salicylic acid (SA) is an important phytohormone, well-known for its regulatory role in shaping plant immune responses. In recent years, significant progress has been made in unravelling the molecular mechanisms underlying SA biosynthesis, perception, and downstream signalling cascades. Through the concerted efforts employing genetic, biochemical, and omics approaches, our understanding of SA-mediated defence responses has undergone remarkable expansion. In general, following SA biosynthesis through Avr effectors of the pathogens, newly synthesized SA undergoes various biochemical changes to achieve its active/inactive forms (e.g. methyl salicylate). The activated SA subsequently triggers signalling pathways associated with the perception of pathogen-derived signals, expression of defence genes, and induction of systemic acquired resistance (SAR) to tailor the intricate regulatory networks that coordinate plant immune responses. Nonetheless, the mechanistic understanding of SA-mediated plant immune regulation is currently limited because of its crosstalk with other signalling networks, which makes understanding this hormone signalling more challenging. This comprehensive review aims to provide an integrated overview of SA-mediated plant immunity, deriving current knowledge from diverse research outcomes. Through the integration of case studies, experimental evidence, and emerging trends, this review offers insights into the regulatory mechanisms governing SA-mediated immunity and signalling. Additionally, this review discusses the potential applications of SA-mediated defence strategies in crop improvement, disease management, and sustainable agricultural practices.

Mutagenic primer-based novel multiplex PCR-RFLP technique to genotype BECN1 SNPs rs10512488 and rs11552192.

BACKGROUND: Single Nucleotide Polymorphisms (SNPs) in candidate autophagy gene BECN1 could influence its functions thereby autophagy process. BECN1 noncoding SNPs were found to be significantly associated with neurodegenerative disease and type 2 diabetes mellitus. This study aimed to develop a simultaneous genotyping technique for two BECN1 SNPs (rs10512488 and rs11552192). METHODS: A mutagenic primer-based approach was used to introduce a NdeI restriction site to genotype rs10512488 by Artificial-Restriction Fragment Length Polymorphism (A-RFLP) along with rs11552192 by Polymerase Chain Reaction (PCR)-RFLP. Multiplexing PCR and restriction digestion reactions were set up for simultaneous genotyping of both SNPs in 100 healthy individuals. Genotypic and allele frequencies were manually calculated, and the Hardy-Weinberg Equilibrium was assessed using the chi-square test. RESULTS: We successfully developed PCR and RFLP conditions for the amplification and restriction digestion of both SNPs within the same tube for genotyping. The results of genotyping by newly developed multiplexing PCR-RFLP technique were concordant with the genotypes obtained by Sanger sequencing of samples. Allelic frequencies of rs10512488 obtained were 0.15 (A) and 0.85 (G), whereas allelic frequencies of rs11552192 were 0.16 (T) and 0.84 (A). CONCLUSION: The newly developed technique is rapid, cost-effective and time-saving for large-scale applications compared to sequencing methods and would play an important role in low-income settings. For the first time, allelic frequencies of rs10512488 and rs11552192 were reported among the North Indian population.

Angiopoietin-like 4 is upregulated by amphiregulin and activates cell proliferation and migration through p38 kinase in head and neck squamous cell carcinoma.

BACKGROUND: Angiopoietin-like 4 is a molecular hallmark that correlates with the growth and metastasis of head and neck squamous cell carcinoma, one of the most prevalent cancers worldwide. However, the molecular mechanisms by which angiopoietin-like 4 promotes head and neck squamous cell carcinoma tumorigenesis are unclear. METHODS: Using well-characterized cell lines of head and neck squamous cell carcinoma development, including human normal oral keratinocytes, dysplastic oral keratinocytes, oral leukoplakia-derived oral keratinocytes, and head and neck squamous cell carcinoma cell lines, HN13, HN6, HN4, HN12, and CAL27, we investigated the signaling pathways upstream and downstream of angiopoietin-like 4-induced head and neck squamous cell carcinoma tumorigenesis. RESULTS: We found that both epidermal growth factor receptor ligands, epithelial growth factor, and amphiregulin led to angiopoietin-like 4 upregulation in normal oral keratinocytes and dysplastic oral keratinocytes and cooperated with the activation of hypoxia-inducible factor-1 in this effect. Interestingly, amphiregulin and angiopoietin-like 4 were increased in dysplastic oral keratinocytes and head and neck squamous cell carcinoma cell lines, and amphiregulin-induced activation of cell proliferation was dependent on angiopoietin-like 4. Although both p38 mitogen-activated protein kinases (p38 MAPK) and protein kinase B (AKT) were activated by angiopoietin-like 4, only pharmacological inhibition of p38 MAPK was sufficient to prevent head and neck squamous cell carcinoma cell proliferation and migration. We further observed that angiopoietin-like 4 promoted the secretion of interleukin 11 (IL-11), interleukin 12 (IL-12), interleukin-1 alpha (IL-1α), vascular endothelial growth factor, platelet-derived growth factor (PDGF), and tumour necrosis factor alpha (TNF-α), cytokines and chemokines previously implicated in head and neck squamous cell carcinoma pathogenesis. CONCLUSION: Our results demonstrate that angiopoietin-like 4 is a downstream effector of amphiregulin and promotes head and neck squamous cell carcinoma development both through direct activation of p38 kinase as well as paracrine mechanisms.

Iron-arsenide monolayers as an anode material for lithium-ion batteries: a first-principles study.

This theoretical investigation delves into the structural, electronic, and electrochemical properties of two hexagonal iron-arsenide monolayers, 1T-FeAs and 1H-FeAs, focusing on their potential as anode materials for lithium-ion batteries. Previous studies have highlighted the ferromagnetic nature of 1T-FeAs at room temperature. Our calculations reveal that both phases exhibit metallic behaviour with spin-polarized electronic band structures. Electrochemical studies show that the 1T-FeAs monolayer has better ionic conductivity for Li ions than the 1H-FeAs phase, attributed to a lower activation barrier of 0.38 eV. This characteristic suggests a faster charge/discharge rate. Both FeAs phases exhibit comparable theoretical capacities (374 mA h g-1), outperforming commercial graphite anodes. The average open-circuit voltage for maximum Li atom adsorption is 0.61 V for 1H-FeAs and 0.44 V for 1T-FeAs. The volume expansion over the maximum adsorption of Li atoms on both phases is also remarkably less than the commercially used anode material such as graphite. Furthermore, the adsorption of Li atoms onto 1H-FeAs induces a remarkable transition from ferromagnetism to anti-ferromagnetism, with minimal impact on the electronic band structure. In contrast, the original state of 1T-FeAs remains unaffected by Li adsorption. To summarize, both 1T-FeAs and 1H-FeAs monolayers have potential as promising anode materials for lithium-ion batteries, offering valuable insights into their electrochemical performance and phase transition behaviour upon Li adsorption.

Theoretical insights into the structural, electronic and thermoelectric properties of the inorganic biphenylene monolayer.

Being motivated by a recently synthesized biphenylene carbon monolayer (BPN), using first principles methods, we have studied its inorganic analogue (B-N analogue) named I-BPN. A comparative study of structural, electronic and mechanical properties between BPN and I-BPN was carried out. Like BPN, the stability of I-BPN was verified in terms of formation energy, phonon dispersion calculations, and mechanical parameters (Young's modulus and Poisson's ratio). The chemical inertness of I-BPN was also investigated by adsorbing an oxygen molecule in an oxygen-rich environment. It has been found that the B-B bond favours the oxygen molecule to be adsorbed through chemisorption. The lattice transport properties reveal that the phonon thermal conductivity of I-BPN is ten times lower than that of BPN. The electronic band structure reveals that I-BPN is a semiconductor with an indirect bandgap of 1.88 eV, while BPN shows metallic behaviour. In addition, we investigated various thermoelectric properties of I-BPN for possible thermoelectric applications. The thermoelectric parameters, such as the Seebeck coefficient, show the highest peak value of 0.00289 V K-1 at 300 K. Electronic transport properties reveal that I-BPN is highly anisotropic along the x and y-axes. Furthermore, the thermoelectric power factor as a function of chemical potential shows a peak value of 0.057 W m-1 K-2 along the x-axis in the p-type doping region. The electronic figure of merit shows a peak value of approximately unity. However, considering lattice thermal conductivity, the peak value of the total figure of merit (ZT) reduces to 0.68(0.46) for p-type and 0.56(0.48) for n-type doping regions along the x(y) direction at 900 K. It is worth noting that our calculated ZT value of I-BPN is higher than that of many other reported B-N composite materials.

Blockade of phosphodiesterase 5 by sildenafil reduces tumor growth and potentiates tumor-killing ability of cisplatin in vivo against T cell lymphoma: Implication of modulated apoptosis, reactive oxygen species homeostasis, glucose metabolism, and pH regulation.

In the past years, PDE5 has emerged as a promising therapeutic target for many cancers due to its highly upregulated expression. Interestingly, a recent in vitro study by our group has shown the antitumor and chemopotentiating action of sildenafil against T cell lymphoma. Our study showed that lower doses of sildenafil (50 µM) and cisplatin (0.5 µg/mL) exhibited 4% and 23% cytotoxicity against HuT78 cells, respectively, which was dramatically increased up to 50% when treated with both. Hence, the present study was designed to evaluate the antitumor and chemo-potentiating action of sildenafil in a murine model of T cell lymphoma (popularly called as Dalton's lymphoma [DL]). In the present study, DL-bearing mice were administered with vehicle (PBS), sildenafil (5 mg/kg bw), cisplatin (5 mg/kg bw), and sildenafil and cisplatin followed by evaluation of their impact on tumor growth by analyzing various parameters. The apoptosis was assessed by Wright-Giemsa, annexin-V, and DAPI staining. Reactive oxygen species (ROS) level was examined through DCFDA staining. The expression of genes and proteins were estimated by RT-PCR and Western blotting, respectively. The experimental findings of the study demonstrate for the first time that sildenafil inhibits tumor growth and potentiates tumor inhibitory ability of cisplatin by altering apoptosis, glycolysis, ROS homeostasis, and pH regulation in T cell lymphoma-carrying host. In addition, our investigation also showed amelioration of tumor-induced liver and kidney damage by sildenafil. Overall, the experimental data of our study strongly advocate the use and repurposing of SDF in designing promising chemotherapeutic regimens against malignancies of T cells.

Clinical Profile and Outcome of Young Infants With Hypernatremic Dehydration Presenting to the Emergency Department.

OBJECTIVES: The aim of this study was to evaluate the clinical profile and outcome of young infants presenting to the pediatric emergency department with hypernatremic dehydration. METHODS: A prospective observational study was conducted at a tertiary care teaching hospital over a period of 18 months. All outborn sick young infants aged 2 months or younger who presented to the emergency department with symptoms and signs of possible sepsis and/or dehydration were screened, and those with hypernatremia were enrolled in the study. Those infants born at less than 37 weeks of gestation and gross congenital anomaly were excluded. Hypernatremic dehydration was defined as serum sodium levels (Se Na+)higher than 145 mEq/L. Variables used in the study were defined as per standard definitions. Acute kidney injury was defined and staged using serum creatinine as per modified neonatal Kidney Disease Improving Global Outcome guidelines. Clinical presentation, laboratory parameters, and comorbidities were compared among outcome groups (survived and died). RESULTS: Of 1124 outborn young infants who met the eligibility criteria for screening, 63 were diagnosed to have hypernatremic dehydration and 55 were enrolled. The hospital-based period prevalence of hypernatremic dehydration in young infants was 4.89%. The median age of presentation was 17 days (10-30). Male-to-female ratio was 1.1:1. Seventy-three percent were first in birth order. Feeding pattern showed 61.8%, 30.9%, and 7.3% of infants were exclusively breastfed, top fed, and mixed fed, respectively. The median serum sodium at the time of admission was 160 (153.5-167) mg/dL. Three (5.5%) infants had mild, 39 (70.9%) had moderate, and 13 (23.6%) had severe hypernatremic dehydration. There was statistically significant correlation between median platelet count with severity of hypernatremic dehydration. The mean time taken to correct serum sodium level was 3.30 ± 1.60 days. The case fatality rate was 41.8%. Those who died had statistically more severe hypernatremic dehydration, acute kidney injury, sepsis, and need for ventilation. CONCLUSIONS: Acute kidney injury stage 3, shock, and need for ventilation are associated with poor outcome in infants with hypernatremic dehydration.

Ectomycorrhizas of Rhizopogon himalayensis on Cedrus deodara.

The ectomycorrhizal (EcM) roots of Cedrus deodara associated with a unique hypogeous EcM fungus-Rhizopogon himalayensis is meticulously characterized and comprehensively described based on well-established standard morphological and anatomical features. The mycobiont-R. himalayensis was found organically associated with the roots of C. deodara. The EcM morphotypes are distinguished by differences in the shape and color of the roots, type of ramification, surface texture, type of mantle, as well as different chemical reactions. All the examined morphotypes were having similar mycorrhizal system and anatomically (Mantle and Hartig net) no disparities were seen, that is, nonsignificant (p > 0.05) variations were observed. The majority of mycorrhizal systems were irregularly pinnate, dichotomous type with 0-1 order of ramification and occasional coralloid type. Mantle surface was densely cottony to loosely wooly. The outer and inner mantles were H & Q type. Hartig net was a complex net-like structure with uniseriate to mutiseriate type of hyphal cell arrangement. Rhizomorph were smooth and round, consistently growing along roots. Moreover, extraradical hyphae were hyaline, septate, and without clamp connections. Sclerotia and cystidia were absent. Our findings will contribute to the biology of ectomycorrhizae associated with primitive and economically valuable conifers, thriving in the face of shifting environmental conditions in the northwestern Himalayas.

Stereomicroscopic evaluation of sealing ability of four different root canal sealers: an in-vitro study.

AIM: To compare and evaluate the sealing ability of four different commercially available sealers to provide seal against the dye penetration test using a stereomicroscope-an in-vitro study. MATERIAL/METHOD: 80 extracted single rooted mandibular premolar with single canal were used in this study. The samples were divided in 4 groups (20 in each) based on sealer. Group I (Diaproseal), Group II (apexit Plus), Group III (MTA Fillapex) and Group IV (Bio-C). The samples were analyzed using a stereomicroscope and data analysis was done with one-way Anova And post hoc Tukey's test. RESULT: The mean dye penetration score was 1.2400 ± 0.778 mm for Group I. 2.6000 ± 0.897 mm for Group II, 4.2000 ± 0.923 mm for Group III and 4.225 ± 2.055 mm for Group IV. One-way Anova analysis shows that intergroup comparison was statistically significant between the four groups. The post hoc Tukey's test reveals that the difference was statistically non-significant between group III and group IV. CONCLUSION: It was concluded that between the four groups the Group I (Diaproseal) showed the least dye penetration followed by Group II (Apexit Pus), Group III (MTA Fillapex) and then Group IV (Bio-C), where there was no significant difference between the Group III (MTA Fillapex) and Group IV (Bio-C).

Functionalized-DNA nanostructures as potential targeted drug delivery systems for cancer therapy.

Seeman's pioneer idea has led to the foundation of DNA nanostructures, resulting in a remarkable advancement in DNA nanotechnology. Over the last few decades, remarkable advances in drug delivery techniques have resulted in the self-assembly of DNA for encapsulating candidate drug molecules. The nuclear targeting capability of DNA nanostructures is lies within their high spatial addressability and tremendous potential for active targeting. However, effective programming and assembling those DNA molecules remains a challenge, making the path to DNA nanostructures for real-world applications difficult. Because of their small size, most nanostructures are self-capable of infiltrating into the tumor cellular environment. Furthermore, to enable controlled and site-specific delivery of encapsulated drug molecules, DNA nanostructures are functionalized with special moieties that allow them to bind specific targets and release cargo only at targeted sites rather than non-specific sites, resulting in the prevention/limitation of cellular toxicity. In light of this, the current review seeks to shed light on the versatility of the DNA molecule as a targeting and encapsulating moiety for active drugs in order to achieve controlled and specific drug release with spatial and temporal precision. Furthermore, this review focused on the challenges associated with the construction of DNA nanostructures as well as the most recent advances in the functionalization of DNA nanostructures using various materials for controlled and targeted delivery of medications for cancer therapy.

Understanding the Proteomes of Plant Development and Stress Responses in Brassica Crops.

Brassica crops have great economic value due to their rich nutritional content and are therefore grown worldwide as oilseeds, vegetables, and condiments. Deciphering the molecular mechanisms associated with the advantageous phenotype is the major objective of various Brassica improvement programs. As large technological advancements have been achieved in the past decade, the methods to understand molecular mechanisms underlying the traits of interest have also taken a sharp upturn in plant breeding practices. Proteomics has emerged as one of the preferred choices nowadays along with genomics and other molecular approaches, as proteins are the ultimate effector molecules responsible for phenotypic changes in living systems, and allow plants to resist variable environmental stresses. In the last two decades, rapid progress has been made in the field of proteomics research in Brassica crops, but a comprehensive review that collates the different studies is lacking. This review provides an inclusive summary of different proteomic studies undertaken in Brassica crops for cytoplasmic male sterility, oil content, and proteomics of floral organs and seeds, under different biotic and abiotic stresses including post-translational modifications of proteins. This comprehensive review will help in understanding the role of different proteins in controlling plant phenotypes, and provides information for initiating future studies on Brassica breeding and improvement programs.

Integration of miRNA dynamics and drought tolerant QTLs in rice reveals the role of miR2919 in drought stress response.

To combat drought stress in rice, a major threat to global food security, three major quantitative trait loci for 'yield under drought stress' (qDTYs) were successfully exploited in the last decade. However, their molecular basis still remains unknown. To understand the role of secondary regulation by miRNA in drought stress response and their relation, if any, with the three qDTYs, the miRNA dynamics under drought stress was studied at booting stage in two drought tolerant (Sahbaghi Dhan and Vandana) and one drought sensitive (IR 20) cultivars. In total, 53 known and 40 novel differentially expressed (DE) miRNAs were identified. The primary drought responsive miRNAs were Osa-MIR2919, Osa-MIR3979, Osa-MIR159f, Osa-MIR156k, Osa-MIR528, Osa-MIR530, Osa-MIR2091, Osa-MIR531a, Osa-MIR531b as well as three novel ones. Sixty-one target genes that corresponded to 11 known and 4 novel DE miRNAs were found to be co-localized with the three qDTYs, out of the 1746 target genes identified. We could validate miRNA-mRNA expression under drought for nine known and three novel miRNAs in eight different rice genotypes showing varying degree of tolerance. From our study, Osa-MIR2919, Osa-MIR3979, Osa-MIR528, Osa-MIR2091-5p and Chr01_11911S14Astr and their target genes LOC_Os01g72000, LOC_Os01g66890, LOC_Os01g57990, LOC_Os01g56780, LOC_Os01g72834, LOC_Os01g61880 and LOC_Os01g72780 were identified as the most promising candidates for drought tolerance at booting stage. Of these, Osa-MIR2919 with 19 target genes in the qDTYs is being reported for the first time. It acts as a negative regulator of drought stress tolerance by modulating the cytokinin and brassinosteroid signalling pathway.

Deciphering the RNA-binding protein interaction with the mRNAs encoded from human chromosome 15q11.2 BP1-BP2 microdeletion region.

Microdeletion of the 15q11.2 BP1-BP2 region, also known as Burnside-Butler susceptibility region, is associated with phenotypes like delayed developmental language abilities along with motor skill disabilities, combined with behavioral and emotional problems. The 15q11.2 microdeletion region harbors four evolutionarily conserved and non-imprinted protein-coding genes: NIPA1, NIPA2, CYFIP1, and TUBGCP5. This microdeletion is a rare copy number variation frequently associated with several pathogenic conditions in humans. The aim of this study is to investigate the RNA-binding proteins binding with the four genes present in 15q11.2 BP1-BP2 microdeletion region. The results of this study will help to better understand the molecular intricacies of the Burnside-Butler Syndrome and also the possible involvement of these interactions in the disease aetiology. Our results of enhanced crosslinking and immunoprecipitation data analysis indicate that most of the RBPs interacting with the 15q11.2 region are involved in the post-transcriptional regulation of the concerned genes. The RBPs binding to this region are found from the in silico analysis, and the interaction of RBPs like FASTKD2 and EFTUD2 with exon-intron junction sequence of CYFIP1 and TUBGCP5 has also been validated by combined EMSA and western blotting experiment. The exon-intron junction binding nature of these proteins suggests their potential involvement in splicing process. This study may help to understand the intricate relationship of RBPs with mRNAs within this region, along with their functional significance in normal development, and lack thereof, in neurodevelopmental disorders. This understanding will help in the formulation of better therapeutic approaches.