Uracil as a biomarker for spatial pyrimidine metabolism in the development of gingivobuccal oral squamous cell carcinoma.

No biomarker has yet been identified that allows accurate diagnosis and prognosis of oral cancers. In this study, we investigated the presence of key metabolites in oral cancer using proton nuclear magnetic resonance (NMR) spectroscopy to identify metabolic biomarkers of gingivobuccal oral squamous cell carcinoma (GB-OSCC). NMR spectroscopy revealed that uracil was expressed in 83.09% of tumor tissues and pyrimidine metabolism was active in GB-OSCC; these results correlated well with immunohistochemistry (IHC) and RNA sequencing data. Based on further gene and protein analyses, we proposed a pathway for the production of uracil in GB-OSCC tissues. Uridinetriphosphate (UTP) is hydrolyzed to uridine diphosphate (UDP) by CD39 in the tumor microenvironment (TME). We hypothesized that UDP enters the cell with the help of the UDP-specific P2Y6 receptor for further processing by ENTPD4/5 to produce uracil. As the ATP reserves diminish, the weakened immune cells in the TME utilize pyrimidine metabolism as fuel for antitumor activity, and the same mechanism is hijacked by the tumor cells to promote their survival. Correspondingly, the differential expression of ENTPD4 and ENTPD5 in immune and tumor cells, respectively, indicatedtheir involvement in disease progression. Furthermore, higher uracil levels were detected in patients with lymph node metastasis, indicating that metastatic potential is increased in the presence of uracil. The presence of uracil and/or expression patterns of intermediate molecules in purine and pyrimidine pathways, such asCD39, CD73, and P2Y6 receptors together with ENTPD4 and ENTPD5, hold promise as biomarker(s) for oral cancer diagnosis and prognosis.

Regulation of chloroplast biogenesis, development, and signaling by endogenous and exogenous cues.

Chloroplasts are one of the defining features in most plants, primarily known for their unique property to carry out photosynthesis. Besides this, chloroplasts are also associated with hormone and metabolite productions. For this, biogenesis and development of chloroplast are required to be synchronized with the seedling growth to corroborate the maximum rate of photosynthesis following the emergence of seedlings. Chloroplast biogenesis and development are dependent on the signaling to and from the chloroplast, which are in turn regulated by several endogenous and exogenous cues. Light and hormones play a crucial role in chloroplast maturation and development. Chloroplast signaling involves a coordinated two-way connection between the chloroplast and nucleus, termed retrograde and anterograde signaling, respectively. Anterograde and retrograde signaling are involved in regulation at the transcriptional level and downstream modifications and are modulated by several metabolic and external cues. The communication between chloroplast and nucleus is essential for plants to develop strategies to cope with various stresses including high light or high heat. In this review, we have summarized several aspects of chloroplast development and its regulation through the interplay of various external and internal factors. We have also discussed the involvement of chloroplasts as sensors of various external environment stress factors including high light and temperature, and communicate via a series of retrograde signals to the nucleus, thus playing an essential role in plants' abiotic stress response.

Interplay of Light and ABA signaling to modulate plant development.

The exogenous light cues and the phytohormone Abscisic acid (ABA) regulate several aspects of plant growth and development. In recent years, the role of the crosstalk between the light and ABA signaling pathways in regulating different physiological processes has become increasingly evident. This includes the regulation of germination and early seedling development, control of stomatal development and conductance, growth and development of roots, buds, branches, and regulation of flowering. Light and ABA signaling cascades have various convergence points at both DNA and protein levels. The molecular crosstalk involves several light signaling factors like HY5, COP1, PIFs and BBXs that integrate with ABA signaling components like the PYL receptors and ABI5. Especially, ABI5 and PIF4 promoters serve as key "hotspots" for the integration of these two pathways. Plants acquired both light and ABA signaling pathways before they colonized land almost 500 million years ago. In this review, we discuss the recent advances in the interplay of light and ABA signaling regulating plant development and provide an overview of the evolution of these two pathways.

Exploitation of a 1D coordination polymer as a portable kit for an eye-catching fluorometric response towards sensing of trivalent cations.

The development and utilization of coordination polymers (CPs) have drawn interest for potential applications in different fields. Detection of metal ions in efficient and selective manners is an important field of research. It paves the way to protect human health by balancing toxic metal ions and biologically active metal ions in the atmosphere. In this regard, a new one-dimensional (1D) 4-(1-naphthylvinyl)pyridine (4-nvp) based CP [Cd(NCS)2(4-nvp)2]n (1) was synthesized and characterized structurally by single-crystal X-ray diffraction. Interestingly, this 1D CP underwent supramolecular aggregation via π⋯π stacking interactions, which specifically generated an environment for a potent "turn on" response in the presence of trivalent cations (Fe3+, Al3+, and Cr3+) in the nanomolar range but remained silent in the presence of other metal ions. Density functional theory (DFT) computations and X-ray photoelectron spectroscopy (XPS) were performed to establish the sensing phenomena. Fascinatingly, utilizing the sensitivity of 1 in an aqueous medium, a hands-on portable cotton swab kit was developed for instant identification of these three important trivalent metal cations.

Facile construction of an anthracene-decorated highly luminescent coordination polymer for the selective detection of explosive nitroaromatics and the mutagenic pollutant TNP.

Explosive nitroaromatic compounds (epNACs) are a group of chemicals that have caused significant human casualties through terrorist attacks and they also pose health risks. For the benefit of homeland security and environmental health, there is room for advancing research on the precise detection of epNACs. Coordination polymers (CPs) successfully serve this purpose because of their binding abilities and quenching capabilities. In this regard, a one-dimensional (1D) CP [Zn(bdc)(avp)2(H2O)]n (1; H2bdc = 1,4-benzenedicarboxylic acid and avp = 4-[2-(9-anthryl)vinyl]pyridine) was synthesized, which remarkably demonstrated extremely efficient ratiometric and selective sensing capacity toward epNACs and the mutagenic pollutant 2,4,6-trinitrophenol (TNP) with a quick response. Density functional theory (DFT) calculations provided a thorough analysis of the mechanistic routes behind the quenching reaction. Herein, geometrically accessible interaction sites were strategically decorated using anthracene moieties, allowing the quick and precise detection of explosive nitro derivatives and the carcinogenic pollutant TNP with increased sensitivity.

Genomic and Transcriptomic Analyses Illuminates Unique Traits of Elusive Night Flowering Jasmine Parijat (Nyctanthes arbor-tristis).

The night-flowering Jasmine, Nyctanthes arbor-tristis also known as Parijat, is a perennial woody shrub belonging to the family of Oleaceae. It is popular for its fragrant flowers that bloom in the night and is a potent source of secondary metabolites. However, knowledge about its genome and the expression of genes regulating flowering or secondary metabolite accumulation is lacking. In this study, we generated whole genome sequencing data to assemble the first de novo assembly of Parijat and use it for comparative genomics and demographic history reconstruction. The temporal dynamics of effective population size (Ne ) experienced a positive influence of colder climates suggesting the switch to night flowering may have provided an evolutionary advantage. We employed multi-tissue transcriptome sequencing of floral stages/parts to obtain insights into the transcriptional regulation of nocturnal flower development and the production of volatiles/metabolites. Tissue-specific transcripts for mature flowers revealed key players in circadian regulation and flower development, including the auxin pathway and cell wall modifying genes. Furthermore, we identified tissue-specific transcripts responsible for producing numerous secondary metabolites, mainly terpenoids and carotenoids. The diversity and specificity of Terpene Synthase (TPS) and CCDs (Carotenoid Cleavage Deoxygenases) mediate the bio-synthesis of specialised metabolites in Parijat. Our study establishes Parijat as a novel non-model species to understand the molecular mechanisms of nocturnal blooming and secondary metabolite production.

The 20/20/20 rule: Practicing pattern and associations with asthenopic symptoms.

Purpose: The present work style and lifestyle have increased the digital device use. Therefore, an increase in digital eyestrain is to be expected. We undertook a survey during coronavirus disease 2019 (COVID-19) pandemic to investigate the practice of 20/20/20 rule and its association with digital device use and asthenopic symptoms. While this rule is commonly advised, little is known about its validity. Methods: An online survey form was disseminated through social media and emails. The questions for eye-related symptoms were similar to the convergence insufficiency symptom survey (CISS). Participants with age ≥5 years were included, with parents completing the survey for children (≤16 years). Results: A total of 432 participants (mean ± standard deviation [SD]: 26.06 ± 13.92 years) were enrolled, of which 125 responses were for children. The 20/20/20 rule was practiced only by 34% of the participants either regularly (n = 38) or occasionally (n = 109). Those who had complaints of burning sensation and headache tended to practice this rule. Among adult participants, more females (47%) practiced this rule when compared to males (23%). Also, adult females significantly (P = 0.04) had more symptoms score when compared to males. In children, no such gender difference was found. Conclusion: Only one-third of participants practice the 20/20/20 rule at least occasionally. More number of adult females being symptomatic and practicing in greater number could be due to higher prevalence of dry eye condition in females. While the symptom of burning sensation could be related to dry eye, that of headache could be related to refractive error or binocular vision dysfunctions.

BBX30/miP1b and BBX31/miP1a form a positive feedback loop with ABI5 to regulate ABA-mediated postgermination seedling growth arrest.

In plants, the switch to autotrophic growth involves germination followed by postgermination seedling establishment. When environmental conditions are not favorable, the stress hormone abscisic acid (ABA) signals plants to postpone seedling establishment by inducing the expression of the transcription factor ABI5. The levels of ABI5 determine the efficiency of the ABA-mediated postgermination developmental growth arrest. The molecular mechanisms regulating the stability and activity of ABI5 during the transition to light are less known. Using genetic, molecular, and biochemical approach, we found that two B-box domain containing proteins BBX31 and BBX30 alongwith ABI5 inhibit postgermination seedling establishment in a partially interdependent manner. BBX31 and BBX30 are also characterized as microProteins miP1a and miP1b, respectively, based on their small size, single domain, and ability to interact with multidomain proteins. miP1a/BBX31 and miP1b/BBX30 physically interact with ABI5 to stabilize it and promote its binding to promoters of downstream genes. ABI5 reciprocally induces the expression of BBX30 and BBX31 by directly binding to their promoter. ABI5 and the two microProteins thereby form a positive feedback loop to promote ABA-mediated developmental arrest of seedlings.

Integration of light and ABA signaling pathways to combat drought stress in plants.

Drought is one of the most critical stresses, which causes an enormous reduction in crop yield. Plants develop various strategies like drought escape, drought avoidance, and drought tolerance to cope with the reduced availability of water during drought. Plants adopt several morphological and biochemical modifications to fine-tune their water-use efficiency to alleviate drought stress. ABA accumulation and signaling plays a crucial role in the response of plants towards drought. Here, we discuss how drought-induced ABA regulates the modifications in stomatal dynamics, root system architecture, and the timing of senescence to counter drought stress. These physiological responses are also regulated by light, indicating the possibility of convergence of light- and drought-induced ABA signaling pathways. In this review, we provide an overview of investigations reporting light-ABA signaling cross talk in Arabidopsis as well as other crop species. We have also tried to describe the potential role of different light components and their respective photoreceptors and downstream factors like HY5, PIFs, BBXs, and COP1 in modulating drought stress responses. Finally, we highlight the possibilities of enhancing the plant drought resilience by fine-tuning light environment or its signaling components in the future.

Plant microProteins: Small but powerful modulators of plant development.

MicroProteins (miPs) are small and single-domain containing proteins of less than 20 kDa. This domain allows microProteins to interact with compatible domains of evolutionary-related proteins and fine-tuning the key physiological pathways in several organisms. Since the first report of a microProtein in mice, numerous microProteins have been identified in plants by computational approaches. However, only a few candidates have been functionally characterized, primarily in Arabidopsis. The recent success of synthetic microProteins in modulating physiological activities in crops makes these proteins interesting candidates for crop engineering. Here, we comprehensively summarise the synthesis, mode of action, and functional roles of microProteins in plants. We also discuss different approaches used to identify plant microProteins. Additionally, we discuss novel approaches to design synthetic microProteins that can be used to target proteins regulating plant growth and development. We finally highlight the prospects and challenges of utilizing microProteins in future crop improvement programs.

Transcription factors BBX11 and HY5 interdependently regulate the molecular and metabolic responses to UV-B.

UV-B radiation acts as a developmental cue and a stress factor for plants, depending on dose. Activation of the transcription factor ELONGATED HYPOCOTYL 5 (HY5) in a UV RESISTANCE LOCUS 8 (UVR8)-dependent manner leads to the induction of a broad set of genes under UV-B. However, the underlying molecular mechanisms regulating this process are less understood. Here, we use molecular, biochemical, genetic, and metabolomic tools to identify the B-BOX transcription factor B-BOX PROTEIN 11 (BBX11) as a component of the molecular response to UV-B in Arabidopsis (Arabidopsis thaliana). BBX11 expression is induced by UV-B in a dose-dependent manner. Under low UV-B, BBX11 regulates hypocotyl growth suppression, whereas it protects plants exposed to high UV-B radiation by promoting the accumulation of photo-protective phenolics and antioxidants, and inducing DNA repair genes. Our genetic studies indicate that BBX11 regulates hypocotyl elongation under UV-B partially dependent on HY5. Overexpression of BBX11 can partially rescue the high UV-B sensitivity of hy5, suggesting that HY5-mediated UV-B stress tolerance is partially dependent on BBX11. HY5 regulates the UV-B-mediated induction of BBX11 by directly binding to its promoter. BBX11 reciprocally regulates the mRNA and protein levels of HY5. We report here the role of a BBX11-HY5 feedback loop in regulating photomorphogenesis and stress tolerance under UV-B.

The evolving role of molecular pathology in the diagnosis of salivary gland tumours with potential pitfalls.

Salivary gland tumors are diagnostically challenging owing to the morphological diversity within any tumor type and overlapping histomorphology and immunohistochemistry amongst different tumours. In past two decades, rapid progress has been made in the field of understanding the pathogenesis of these tumours with the discovery of many tumour specific translocations and rearrangements. This includes CRTC1-MAML2 and CRTC-MAML2 in mucoepidermoid carcinoma, MYBNFIB and MYBL1-NFIB fusions in adenoid cystic carcinoma, PLAG1 and HMGA2 in pleomorphic adenoma, ETV6-NTRK3 in secretory carcinoma, NR4A3 rearrangements in acinic cell carcinoma, PRKD1 mutations in polymorphous adenocarcinoma and EWSR1-ATF1 in clear cell carcinoma. This review is a lens for progress made till date in the molecular pathology of salivary gland tumours with a special focus on their role as diagnostic tools and implications on clinical management of the patient as prognostic and predictive markers.

Comprehensive investigation of fever cases enrolled during 2019 dengue outbreaks from three hyperendemic regions of North 24 Parganas district of West Bengal, India.

For the past several decades, dengue fever has been emerging in epidemic proportions in several regions of the world. During August-September 2019, an increasing number of fever cases were being reported from some areas of North 24 Parganas district of West Bengal, India. Accordingly, outbreak investigation of fever cases from these affected areas of Bongoan, Barasat, and Habra was carried out. To characterize clinical and biochemical features of fever cases as well as to investigate the utility of CRP as a Dengue severity marker in resource-limited settings. We systematically enrolled 108 patients from the affected region of North 24 Parganas. Standard diagnostic assays along with routine serological and biochemical parameters were performed. Of the 108 patients, 77 (71%) were confirmed with Dengue infection followed by 22 (20%) DENV seronegative and 9 (8%) coinfected DENV cases. Among the 77 confirmed Dengue patients, 53 (69%) had primary infection while 24 (31%) had secondary infection. Among the DENV clinical symptoms, fever (r = 0.50; p = 0.004), headache (r = 0.40; p = 0.03) and abdominal pain (r = -0.40; p = 0.02) were found to bear significant correlation with DENV viral load. The predominant circulating serotype was found to be DENV2. CRP Dengue severity cut-off level of 10.15 mg/L (AUC: 0.85; 86% sensitivity, 77% specificity) was obtained. CRP had correlation with viral load (r = 0.4, p = 0.05) within febrile phase of infection. The performance of biomarkers can be influenced by local epidemiology, geography, and several patient factors, therefore, CRP Dengue severity cut-off value may be region-specific. This study for the first time attempts to estimate CRP Dengue severity cut-off value based on routine immunoturbidometric evaluation from Dengue Hyperendemic zones of North 24 Parganas, West Bengal, Eastern India.

B-box protein BBX32 integrates light and brassinosteroid signals to inhibit cotyledon opening.

Cotyledon opening is a key morphological change that occurs in seedlings during de-etiolation. Brassinosteroids (BRs) inhibit the opening of cotyledons in darkness while light promotes cotyledon opening. The molecular regulation of the interplay between light and BR to regulate cotyledon opening is not well understood. Here, we show the B-box protein BBX32 negatively regulates light signaling and promotes BR signaling to inhibit cotyledon opening in Arabidopsis (Arabidopsis thaliana). BBX32 is highly expressed in the cotyledons of seedlings during de-etiolation. bbx32 and 35S:BBX32 seedlings exhibit enhanced and reduced cotyledon opening, respectively, in response to both light and brassinazole treatment in dark, suggesting that BBX32 mediates cotyledon opening through both light and BR signaling pathways. BBX32 expression is induced by exogenous BR and is upregulated in bzr1-1D (BRASSINAZOLE RESISTANT1-1D). Our in vitro and in vivo interaction studies suggest that BBX32 physically interacts with BZR1. Further, we found that PHYTOCHROME-INTERACTING FACTOR 3 (PIF3) interacts with BBX32 and promotes BR-mediated cotyledon closure. BBX32, BZR1, and PIF3 regulate the expression of common target genes that modulate the opening and closing of cotyledons. Our work suggests BBX32 integrates light and BR signals to regulate cotyledon opening during de-etiolation.

Quantification of gaze reaction time in infants with Pediatric Perimeter.

PURPOSE: We quantified the eye/head (gaze) reaction time in infants to establish a normative database for the Pediatric Perimeter device. Additionally, we tested the hypothesis that gaze reaction time will reduce with age. METHODS: A cross-sectional study was conducted. Healthy infants between 3 to 10 months of age were recruited. Peripheral visual field stimuli (hemifield and quadrant stimuli) were presented in the Pediatric Perimeter device. Infant's gaze to these stimuli was observed, documented in real time, and video recorded for offline analysis. RESULTS: A total of 121 infants were tested in three age group bins [3-5 months, n = 44; >5-7 months, n = 30 and >7-10 months, n = 47]. Overall, 3-5 months old had longer reaction time when compared to the older infants particularly for stimuli presented in the quadrants (Kruskal-Wallis, p<0.038). A significantly asymmetric difference (p = 0.025) in reaction time was observed between the upper (median = 820ms, IQR = 659-1093ms) and lower quadrants (median = 601ms, IQR = 540-1052ms) only for the 3-5 months old infants. CONCLUSION: This study provides the normative gaze reaction time of healthy infants. With increase in age, there is reduction in reaction time and disappearance of reaction time asymmetry in quadrant stimuli. The longer reaction time for upward gaze could be due to delayed maturation of neural mechanisms and/or decreased visual attention.

Light and abscisic acid interplay in early seedling development.

Abscisic acid (ABA) plays a crucial role in plant development, regulating germination, seedling development and stomatal movements, especially under adverse conditions. Light interacts with the ABA signalling pathway to fine tune these processes. Here, we provide an overview of the recent investigations on ABA-light interplay during early plant development after germination. We discuss the multilayered and reciprocal interactions between ABA signalling components and several light signalling modulators, including photoreceptors, transcription factors and posttranslational modifiers. ABSCISIC ACID INSENSITIVE5 acts as a central convergence point for these interactions during postgermination seedling development. ABA also regulates the adaptation of seedlings to challenging light environments. Furthermore, we enlist the role of ABA-light cross-talk in regulating seedling establishment in crops and highlight open questions for future investigations.

The role of solute transporters in aluminum toxicity and tolerance.

The prevalence of aluminum ions (Al3+ ) under acidic soil conditions inhibits primary root elongation and hinders plant growth and productivity. Al3+ alters the membrane potential, displaces critical ions in the apoplast and disrupts intracellular ionic concentrations by targeting membrane-localized solute transporters. Here, we provide an overview of how Al3+ affects the activities of several solute transporters especially in the root. High Al3+ level impairs the functions of potassium (K+ ), calcium (Ca2+ ), magnesium (Mg2+ ), nitrate (NO3 - ) and ammonium (NH4 + ) transporters. We further discuss the role of some key transporters in mediating Al tolerance either by exclusion or sequestration. Anion channels responsible for organic acid efflux modulate the sensitivity to Al3+ . The ALUMINUM ACTIVATED MALATE TRANSPORTER (ALMT) and MULTIDRUG AND TOXIC COMPOUND EXTRUSION (MATE) family of transporters exude malate and citrate, respectively, to the rhizosphere to alleviate Al toxicity by Al exclusion. The ABC transporters, aquaporins and H+ -ATPases perform vacuolar sequestration of Al3+ , leading to aluminum tolerance in plants. Targeting these solute transporters in crop plants can help generating aluminum-tolerant crops in future.

PIF3/HY5 module regulates BBX11 to suppress protochlorophyllide levels in dark and promote photomorphogenesis in light.

Greening of cotyledons during de-etiolation is critical for harvesting light energy and sustaining plant growth. PIF3 and HY5 antagonistically regulate protochlorophyllide synthesis in the dark. However, the mechanism by which the PIF3/HY5 module regulates genes involved in protochlorophyllide synthesis is not clear. Using genetic, molecular and biochemical techniques we identified that the B-BOX protein BBX11 acts directly downstream of PIF3 and HY5 to transcriptionally modulate genes involved in protochlorophyllide synthesis. Dark-grown bbx11 and 35S:BBX11 seedlings exhibit an enhanced and reduced ability to green, respectively, when exposed to light. Transcript levels of HEMA1 and CHLH are upregulated in 35S:BBX11 seedlings that accumulate high levels of protochlorophyllide in the dark and undergo photobleaching upon illumination. PIF3 inhibits BBX11 in the dark by directly binding to its promoter. bbx11 suppresses the cotyledon greening defect of pif3 after prolonged dark, indicating that the PIF3-mediated regulation of greening is dependent on BBX11. The enhanced greening of hy5 is also suppressed in hy5 lines overexpressing BBX11. In light, HY5 directly binds to the promoter of BBX11 and activates its expression to regulate BBX11-mediated hypocotyl inhibition. We show that a PIF3/HY5 module regulates BBX11 expression in opposite ways to optimise protochlorophyllide accumulation in the dark and promote photomorphogenesis in light.