Bioactive compounds and antioxidant activity during ripening of Malpighia glabra fruits.

Malpighia glabra (Malpighiaceae) is a cherry fruit popularly known as acerola or West Indian cherry, with nutraceuticals in each ripening stage. The changes in the phytoconstituents, pigments, sugars, organic acids, and antioxidants were investigated during the fruit ripening and expressed in fresh weight (FW). Gentisic acid was present in the highest concentration in IMGL fruits (11.43 mg/100 g), which was reduced to 0.362 mg/100 g over-ripening. The major flavonoid present was epicatechin, and the concentration increased from 2.11 mg/100 g in immature green large (IMGL) fruits to 19.52 mg/100 g in ripe fruits. Ascorbic acid was the most abundant organic acid present, and the highest concentration was found in the IMGL fruits (2030 mg/100 g). Fructose and galactose were found in the highest concentrations in overripe fruits (2290 mg/100 g and 1460 mg/100 g, respectively). The IMGL fruits showed the highest total antioxidant activity of 5.48% and 5.34% ascorbic acid equivalent in methanolic and aqueous extracts, respectively. Mineral quantification showed that the fruits were rich in potassium and calcium (150.43 and 12.90 mg/100 g, respectively). This study could identify the appropriate stage of acerola fruit for developing functional foods with maximum utilization of phytoconstituents in all stages.

Structure-based screening of natural product libraries in search of potential antiviral drug-leads as first-line treatment to COVID-19 infection.

The study focuses on identifying and screening natural products (NPs) based on their structural similarities with chemical drugs followed by their possible use in first-line treatment to COVID-19 infection. In the present study, the in-house natural product libraries, consisting of 26,311 structures, were screened against potential targets of SARS-CoV-2 based on their structural similarities with the prescribed chemical drugs. The comparison was based on molecular properties, 2 and 3-dimensional structural similarities, activity cliffs, and core fragments of NPs with chemical drugs. The screened NPs were evaluated for their therapeutic effects based on their predicted in-silico pharmacokinetic and pharmacodynamics properties, binding interactions with the appropriate targets, and structural stability of the bound complex using molecular dynamics simulations. The study yielded NPs with significant structural similarities to synthetic drugs currently used to treat COVID-19 infections. The study proposes the probable biological action of the selected NPs as Anti-retroviral protease inhibitors, RNA-dependent RNA polymerase inhibitors, and viral entry inhibitors.

Biosynthesis and regulation of anthocyanin pathway genes.

Anthocyanins are the phenolic compounds responsible for coloring pigments in fruits and vegetables. Anthocyanins offer a wide range of health benefits to human health. Their scope has expanded dramatically in the past decade, making anthocyanin control, influx, and outflow regulation fascinating for many researchers. The main culprit is anthocyanin stability and concentration form, which demands novel ways because these are critical in the food industry. This review aims to examine anthocyanin synthesis via triggering transcription genes that code for anthocyanin-producing enzymes. The balance between production and breakdown determines anthocyanin accumulation. Thus, increasing the anthocyanin content in food requires the stability of molecules in the vacuolar lumen, the pigment fading process, and a better understanding of the mechanism. The promising option is biosynthesis by metabolically engineered microorganisms with a lot of success. This study aims to look into and evaluate the existing literature on anthocyanin production, namely the biosynthesis of anthocyanin pathway genes, production by microbial cell factories, and the regulatory factors that can modulate the production of anthocyanins. Understanding these mechanisms will provide new biotechnological approaches.Key points• Factors affecting the regulation of anthocyanins• Focus on degradation, biosynthesis pathway genes, and alternative systems for the production of anthocyanins• Microbial cell factories can be used to produce large amounts of anthocyanins.

Chemometric evaluation of functional components and anti-quorum sensing activity of mulberry leaves from Indian cultivars: a potential contribution to the food industry.

BACKGROUND: Potential use of many native, easily available vegetal materials for human consumption and value addition is not well recognized. Mulberry, being a traditional industrial crop rich in nutrients and nutraceuticals can be of great importance for the food industry. However, mulberry leaves are mainly being utilized in sericulture and are not exploited for their functional components. Thus, the selection of promising mulberry cultivars, rich in bioactive compounds, like resveratrol and 1-deoxynojirimycin, increase their potential use in functional foods. RESULTS: Chlorogenic acid, myricetin and kaempferol were the major polyphenols present in the nine selected cultivars, in the range 0.001-0.086, 0.003-0.079 and 0.003-0.163 g kg-1 fresh weight (FW), respectively. Protocatechuic acid, epicatechin and rutin were predominantly present in cultivars V-1, G-2 and ML (0.103, 0.080 and 0.121 g kg-1 FW, respectively). Similarly, resveratrol and 1-deoxynojirimycin were highest in cultivars ML and K-2 (0.078 and 0.079 g kg-1 FW, respectively). Leaf extracts of cultivars G-2 and ML were able to effectively inhibit the violacein production with 64.08% and 70.04%, respectively at the concentration of 6 mg mL-1 presumably due to a higher content of polyphenols. Chemometric evaluation of chromatographic data showed the intraspecific variability and secondary metabolite co-existence in different cultivars. CONCLUSIONS: Considering phytoconstituents, cultivars G-2, ML, K-2 and V-1 could contribute efficiently to the rational utilization of mulberry in agro-food industries. Furthermore, cultivars G-2 and ML leaves can be a new source of quorum sensing inhibitory agents. © 2021 Society of Chemical Industry.

Production of small ruminant morbillivirus, rift valley fever virus and lumpy skin disease virus in CelCradle™ -500A bioreactors.

BACKGROUND: Animal vaccination is an important way to stop the spread of diseases causing immense damage to livestock and economic losses and the potential transmission to humans. Therefore effective method for vaccine production using simple and inexpensive bioprocessing solutions is very essential. Conventional culture systems currently in use, tend to be uneconomic in terms of labor and time involved. Besides, they offer a limited surface area for growth of cells. In this study, the CelCradle™-500A was evaluated as an alternative to replace conventional culture systems in use such as Cell factories for the production of viral vaccines against small ruminant morbillivirus (PPR), rift valley fever virus (RVF) and lumpy skin disease virus (LSD). RESULTS: Two types of cells Vero and primary Lamb Testis cells were used to produce these viruses. The study was done in 2 phases as a) optimization of cell growth and b) virus cultivation. Vero cells could be grown to significantly higher cell densities of 3.04 × 109 using the CelCradle™-500A with a shorter doubling time as compared to 9.45 × 108 cells in Cell factories. This represents a 19 fold increase in cell numbers as compared to seeding vs only 3.7 fold in Cell factories. LT cells achieved modestly higher cell densities of 6.7 × 108 as compared to 6.3 × 108 in Cell factories. The fold change in densities for these cells was 3 fold in the CelCradle™-500A vs 2.5 fold in Cell factories. The titers in the conventional system and the bioreactor were not significantly different. However, the Cell-specific virus yield for rift valley fever virus and lumpy skin disease virus are higher (25 virions/cell for rift valley fever virus, and 21.9 virions/cell for lumpy skin disease virus versus 19.9 virions/cell for rift valley fever virus and 10 virions/cell for lumpy skin disease virus). CONCLUSIONS: This work represents a novel study for primary lamb testis cell culture in CellCradle™-500A bioreactors. In addition, on account of the high cell densities obtained and the linear scalability the titers could be further optimized using other culture process such us perfusion.

Identification and Characterization of Polyphenols from Carissa spinarum Fruit and Evaluation of Their Antioxidant and Anti-quorum Sensing Activity.

Quorum sensing controls bacterial virulence through signaling molecules, which plays a vital role in managing foodborne pathogens that cause food spoilage and human infections. Though many synthetic compounds have been used to control infection, antibiotic resistance has become a global issue. Targeting the bacterial quorum sensing using the bioactive compounds could be an alternative strategy to combat their resistance. In this context, polyphenols from an unexplored unripe and ripe fruits of Carissa spinarum were evaluated for anti-quorum sensing activity. The study aimed at determining nutritional, phytochemical composition, and polyphenol profiling to evaluate their antioxidant potential of the fruit. Furthermore, the violacein inhibition, anti-biofilm, and effect on the motility of foodborne pathogens were also studied. The phytochemical content of C. spinarum fruit showed the phenolic and flavonoid content 273.20 mg GAE/100 g and 453.78 mg QE/100 g fresh weight , respectively, in ripe fruit. HPLC characterization of fruit extracts showed high content of syringic acid, resveratrol, and quercetin in ripe, whereas it showed epicatechin and gentisic acid in unripe fruit. The antioxidant activity of the ripe fruit extract exhibited a higher potential to scavenge DPPH radicals with IC50 4.69 mg/mL. Further, the anti-quorum sensing activity was higher in ripe fruit extract at 1.8 mg/mL inhibited with 78.65% violacein production in Chromobacterium violaceum, swimming motility, and biofilm formation in Pseudomonas aeruginosa and Yersinia enterocolitica (66.25% and 59.36% respectively at 1.2 mg/mL). Hence C. spinarum fruit bioactive could be a natural plant source for anti-quorum sensing activity to manage foodborne pathogens over synthetic compounds.

Pleiotropic nature of curcumin in targeting multiple apoptotic-mediated factors and related strategies to treat gastric cancer: A review.

Gastric cancer (GC) is one of the major reasons for cancer-associated death and exhibits the second-highest mortality rate worldwide. Several advanced approaches have been designed to treat GC; however, these strategies possess many innate complications. In view of this, the upcoming research relying on natural products could result in designing potential anticancer agents with fewer side effects. Curcumin, isolated from the rhizomes of Curcuma longa L. has several medicinal properties like antiinflammatory, antioxidant, antiapoptotic, antitumor, and antimetastatic. Such pleiotropic nature of curcumin impedes the invasion and proliferation of GC by targeting several oncogenic factors like p23, human epidermal factor receptor2 including Helicobacter pylori. The side effect of chemotherapy, that is, chemotherapeutic resistance and radiotherapy could be reduced combination therapy of curcumin. Moreover, the photodynamic therapy of curcumin destroys the cancer cells without affecting normal cells. However, further more potential studies are required to establish the potent efficacy of curcumin in the treatment of GC. The current review details the anticancer activities of curcumin and related strategies which could be employed to treat GC with additional focus on its inhibitory properties against viability, proliferation, and migration of GC cells through cell cycle arrest and stimulation by apoptosis-mediated factors.

Physicochemical Composition and Characterization of Bioactive Compounds of Mulberry (Morus indica L.) Fruit During Ontogeny.

Mulberry fruit is well recognized as one of the richest sources of bioactive compounds. We investigated the physicochemical composition and characterized the bioactive compounds during different ripening stages of mulberry (Morus indica) fruit and evaluated their anti-quorum sensing activity on Chromobacterium violaceum. The proximate components such as carbohydrates, proteins and lipids were found to be high in the ripe fruit compared to unripe and mid-ripe fruit. The ripe fruit contained higher content of total phenolics and flavonoids (336.05 and 282.55 mg/100 g fresh weight (FW), respectively). Epicatechin and resveratrol were the major polyphenols detected in the fruit with the range 5.13-19.46 and 4.07-14.45 mg/100 g FW, respectively. Chlorogenic acid and myricetin were predominant in the unripe and mid-ripe fruit (7.14 and 1.84 mg/100 g FW, respectively). The fruit was found to be an excellent source of anti-diabetic compound 1-deoxynojirimycin. The highest content of 1-deoxynojirimycin was present in the mid-ripe fruit, with a content of 2.91 mg/100 g FW. Furthermore, fruit extracts exhibited anti-quorum sensing activity against Chromobacterium violaceum by effectively inhibiting violacein production. Ripe fruit extracts showed the highest activity of 76.30% at 1 mg/mL and thus, could be used as a potent anti-quorum sensing agent. The results could be promising in the selection of appropriate developmental stages for M. indica fruit commercial exploitation in the food formulations rich in potential health components.

Evaluation of Various Drying Methods on Bioactives, Ascorbic Acid and Antioxidant Potentials of Talinum triangulare L., foliage.

The aim of this study was to assess the efficiency of different drying methods (room, sun, oven, microwave, cross-flow, infra-red, dehumidifier, and freeze-drying) on maximum retention of the nutritional and bioactive compounds profiling of Talinum triangulare, which is a less-known perishable leafy-vegetable. The evaluation of various drying methods is to learn the best appropriate strategy for a post-harvest drying method for retaining all the potential benefits with minor loss. Microwave and freeze-dried samples held the maximum ascorbic acid quantified by HPLC with 1.36 and 1.11 g/100 g DW, respectively. The main carotenoid compounds identified were violaxanthin, lutein, zeaxanthin, ß-carotene isomers, trans-ß-carotene, and cis-ß-carotenes. Gallic, protocatechuic, catechin, para-coumaric, ferulic, rutin, trans-cinnamic, and quercetin are the significant phenolics and flavonoids identified and quantified by liquid chromatography. The efficiency of different solvents on bioactive extractions uncovered that the methanol and 80% aqueous ethanol were good for retention of total phenolics, total flavonoids, and antioxidant compounds, which was affirmed through phosphomolybdate, DPPH, and FRAP assays. Dried T. triangulare foliage could be productively utilized as a promising raw material for food and pharma businesses because of its rich bioactive composition.

Dengue, chikungunya, and scrub typhus are important etiologies of non-malarial febrile illness in Rourkela, Odisha, India.

BACKGROUND: We conducted a diagnostic surveillance study to identify Plasmodium, dengue virus, chikungunya virus, and Orientia tsutsugamushi infections among febrile patients who underwent triage for malaria in the outpatient department at Ispat General Hospital, Rourkela, Odisha, India. METHODS: Febrile patients were enrolled from January 2016-January 2017. Blood smears and small volumes or vacutainers of blood were collected from study participants to carry out diagnostic assays. Malaria was diagnosed using rapid diagnostic tests (RDT), microscopy, and PCR. Dengue, chikungunya, and scrub typhus infections were identified using rapid diagnostic test kits and ELISA. RESULTS: Nine hundred and fifty-four patients were prospectively enrolled in our study. The majority of patients were male (58.4%) and more than 15 years of age (66.4%). All 954 enrollees underwent additional testing for malaria; a subset of enrollees (293/954) that had larger volumes of plasma available was also tested for dengue, chikungunya and scrub typhus by either RDT or ELISA or both tests. Fifty-four of 954 patients (5.7%) were positive for malaria by RDT, or microscopy, or PCR. Seventy-four of 293 patients (25.3%) tested positive for dengue by either RDT or ELISA, and 17 of 293 patients (5.8%) tested positive for chikungunya-specific IgM by either ELISA or RDT. Ten of 287 patients tested (3.5%) were positive for scrub typhus by ELISA specific for scrub typhus IgM. Seventeen patients among 290 (5.9%) with results for ≥3 infections tested positive for more than one infection. Patients with scrub typhus and chikungunya had high rates of co-infection: of the 10 patients positive for scrub typhus, six were positive for dengue (p = 0.009), and five of 17 patients positive for chikungunya (by RDT or ELISA) were also diagnosed with malaria (p < 0.001). CONCLUSIONS: Dengue, chikungunya and scrub typhus are important etiologies of non-malarial febrile illness in Rourkela, Odisha, and comorbidity should be considered. Routine febrile illness surveillance is required to accurately establish the prevalence of these infections in this region, to offer timely treatment, and to implement appropriate methods of control.

Effect of different drying methods on chlorophyll, ascorbic acid and antioxidant compounds retention of leaves of Hibiscus sabdariffa L.

BACKGROUND: Use of the indigenous, easily accessible leafy vegetable roselle (Hibiscus sabdariffa L.) for value addition is gaining impetus as its nutritive and nutraceutical compounds are exposed by investigations. Being a perishable, storage is challenging, hence different methods of drying have been an attractive alternative for its postharvest usage in foods without much compromising its quality and antioxidant potential. RESULTS: Room- and freeze-dried samples were found to have best quality in terms of colour, total flavonoid content (18.53 ± 2.39 and 18.66 ± 1.06 g kg(-1) respectively), total phenolic content (17.76 ± 1.93 and 18.91 ± 0.48 g kg(-1)), chlorophyll content (1.59 ± 0.001 and 1.55 ± 0.001 g kg(-1)) and ascorbic acid content (11.11 ± 1.04 and 8.92 ± 0.94 g kg(-1)) compared with those subjected to infrared, crossflow, microwave, oven or sun drying. Samples treated by room and freeze drying retained maximum antioxidant potential as shown by the phosphomolybdate method and the 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging activity and ferric-reducing antioxidant power assays. Cold water and hot water extracts showed significantly higher total phenolic content and total antioxidant activity owing to the greater solubility of phenolics and destruction of cellular components in polar solvents than in organic solvents. CONCLUSION: The data obtained show the potential for retaining quality parameters of roselle leaf under suitable drying methods.

Genome-wide identification of MATE, functional analysis and molecular dynamics of DcMATE21 involved in anthocyanin accumulation in Daucus carota.

Anthocyanins are a subclass of flavonoids that are synthesized in the endoplasmic reticulum and then transported to the vacuole in plants. Multidrug and toxic compound extrusion transporters (MATE) is a family of membrane transporters that transport ions and secondary metabolites, such as anthocyanins, in plants. Although various studies on MATE transporters have been carried out on different plant species, this is the first comprehensive report to mine the Daucus carota genome to identify the MATE gene family. Our study identified 45 DcMATEs through genome-wide analysis and detected five segmental and six tandem duplications from the genome. The chromosome distribution, phylogenetic analysis, and cis-regulatory elements revealed the structural diversity and numerous functions associated with the DcMATEs. In addition, we analyzed RNA-seq data obtained from the European Nucleotide Archive to screen for the expression of DcMATEs involved in anthocyanin biosynthesis. Among the identified DcMATEs, DcMATE21 correlated with anthocyanin content in the different D. carota varieties. In addition, the expression of DcMATE21 and anthocyanin biosynthesis genes was correlated under abscisic acid, methyl jasmonate, sodium nitroprusside, salicylic acid, and phenylalanine treatments, which were substantiated by anthocyanin accumulation in the in vitro cultures. Further molecular membrane dynamics of DcMATE21 with anthocyanin (cyanidin-3-glucoside) identified the binding pocket, showing extensive H-bond interactions with 10 crucial amino acids present in the transmembrane helix of 7, 8, and 10 of DcMATE21. The current investigation, using RNA-seq, in vitro cultures, and molecular dynamics studies revealed the involvement of DcMATE21 in anthocyanin accumulation in vitro cultures of D. carota.

Evolutionary selectivity of amino acid is inspired from the enhanced structural stability and flexibility of the folded protein.

AIM: The present study attempts to decipher the site-specific amino acid alterations at certain positions experiencing preferential selectivity and their effect on proteins' stability and flexibility. The study examines the selection preferences by considering pair-wise non-bonded interaction energies of adjacent and interacting amino acids present at the interacting site, along with their evolutionary history. MATERIALS AND METHODS: For the study, variations in the interacting residues of spike protein (S-Protein) receptor-binding domain (RBD) of different coronaviruses were examined. The MD simulation trajectory analysis revealed that, though all the variants studied were structurally stable at their native and bound confirmations, the RBD of 2019-nCoV/SARS-CoV-2 was found to be more flexible and more dynamic. Furthermore, a noticeable change observed in the non-bonded interaction energies of the amino acids interacting with the receptor corroborated their selection at respective positions. KEY FINDINGS: The conformational changes exerted by the altered amino acids could be the reason for a broader range of interacting receptors among the selected proteins. SIGNIFICANCE: The results envisage a strong indication that the residue selection at certain positions is governed by a well-orchestrated feedback mechanism, which follows increased stability and flexibility in the folded structure compared to its evolutionary predecessor.

Salt Stress-Induced Anthocyanin Biosynthesis Genes and MATE Transporter Involved in Anthocyanin Accumulation in Daucus carota Cell Culture.

Anthocyanins biosynthesis is a well-studied biosynthesis pathway in Daucus carota. However, the scale-up production at the bioreactor level and transporter involved in accumulation is poorly understood. To increase anthocyanin content and elucidate the molecular mechanism involved in accumulation, we examined D. carota cell culture in flask and bioreactor for 18 days under salt stress (20.0 mM NH4NO3/37.6 mM KNO3) at 3 day intervals. The expression of anthocyanin biosynthesis and putative MATE (multidrug and toxic compound extrusion) transporter expression was analyzed by qRT-PCR. It was observed that there was a significant enhancement of anthocyanin in the bioreactor compared to the control culture. A correlation was observed between the expression of MATE and the anthocyanin biosynthesis genes (CHS, C4H, LDOX, and UFGT) on the 9th day in a bioreactor, where maximum anthocyanin accumulation and expression was detected. We hypothesize the involvement of MATE in transporting anthocyanin to tonoplast in D. carota culture under salt stress.

Serendipitous Discovery of a Benign Obturator Nerve Schwannoma: Case report with a brief clinical review.

Schwannomas are typically benign tumours of the peripheral nerves. However, they seldom arise from the obturator nerve. We report a case of an uncommon swelling (2.5 × 3.5 cm) in a 65-year-old male cadaver, found during a routine dissection session for first Bachelor of Medicine and Surgery students in the Department of Anatomy, Kasturba Medical College, Manipal, India, in 2019. It was seen originating from the left obturator nerve in the pelvis at the level of the sacral promontory. Histopathological investigation revealed a schwannoma. The hypocellular tumour was arranged in a sweeping fascicle pattern with patches of myxoid degeneration. Obturator schwannomas, though rare, can exist in cadavers, as seen in the present case. Hence, it should be considered as a differential diagnosis for clinical cases of pelvic masses and eliminated only after thorough radiological examination. Knowledge about the existence of such schwannomas is therefore essential.

Micro and nanoemulsions of Carissa spinarum fruit polyphenols, enhances anthocyanin stability and anti-quorum sensing activity: Comparison of degradation kinetics.

The low stability of anthocyanins is a constraint in the food industry. The present work has been carried out to overcome this low stability by encapsulating fruit concentrate of underutilized plant Carissa spinarum (CS) with polyphenols in microemulsions (CSME) and nanoemulsions (CSNE). Increasing the amount of CS reduced the particle size from 1154 to 70-300 nm whereas addition of Tween 80 reduced it optimally to 5-25 nm. Degradation of anthocyanins in control and ME/NE proceeded with zero- and first-order reaction rates, respectively, at 28 °C (half-life 6, 25 and 40 days, respectively). The degradation kinetics of phenolics-flavonoids were also studied. CSNE exhibited higher anti-quorum sensing (QS) activity than CSME against Chromobacterium violaceum (73.7%); it inhibited biofilm formation by 70.1 and 64.4% in Pseudomonas aeruginosa, and Yersinia enterocolitica, respectively. This is the first report of using the more stable ME/NE to study anti-QS activity, an alternative to conventional antibiotics.

Exogenous Serotonin and Melatonin Regulate Dietary Isoflavones Profoundly through Ethylene Biosynthesis in Soybean [Glycine max (L.) Merr.].

Serotonin and melatonin are important signaling and stress mitigating molecules. However, their role and molecular mechanism in the accumulation of isoflavones are not clearly defined. To elucidate their functions, serotonin and melatonin were applied to in vitro cultures of soybean at different concentrations and analyzed to assess the accumulation of isoflavone content followed by transcript levels of biosynthesis genes at different time intervals. Increased total phenolics, total flavonoids, and different forms of isoflavone content were observed in the treatments. Expression levels of critical genes in isoflavone, ethylene, jasmonic acid, abscisic acid, and melatonin biosynthesis and related transcription factor were quantified. A correlation was observed between the expression of ethylene biosynthesis genes (S-adenosylmethionine synthase and 1-aminocyclopropane-1-carboxylate oxidase) and isoflavone biosynthesis genes (chalcone synthase, chalcone reductase, and isoflavone synthase). We hypothesize that, under serotonin and melatonin treatments, ethylene biosynthesis may play a role in the increase/decrease in isoflavone content in soybean culture.

Suppression of N-glycan processing enzymes by deoxynojirimycin in tomato (Solanum lycopersicum) fruit.

The present study investigated the potential of a small molecule inhibitor, 1-deoxynojirimycin (DNJ), to extend the shelf life of tomatoes. The optimum concentration of DNJ and the proper ripening stage for treatment were standardized using response surface methodology, following a central composite design. The concentration of DNJ used for the analysis was 0.15 mM, and 0.30 mM and the ripening stages of the tomato fruit analysed were immature green, mature green, breaker, ripen and over-ripen. Analysis of the influence of the DNJ treatment of the fruit using quadratic multiple regression models considering the factors colour, texture, and free sugars revealed significant responses. A DNJ concentration of 0.30 mM and fruit-ripening stage of mature green was found to be optimal for the treatment. DNJ-treatment maintained fruit firmness throughout ripening with a significant reduction in reducing sugar formation. Enzyme activity of the N-glycan processing enzymes involved in cell wall softening, α-mannosidase and ß-d-N-acetylhexosaminidase revealed a significant reduction in their activity by 2 and 3.5-fold, respectively. Down-regulation of expression of important ripening-related and softening process-associated genes, aminocyclopropane carboxylic synthase-4, aminocyclopropane carboxylic oxidase, polygalacturonase and pectin methylesterases at 4, 5, 6 and 5-fold, respectively, was also observed. The present results showed that the treatment of mature green tomato fruit with DNJ at a concentration of 0.30 mM can delay the ripening of the tomato fruit by inhibiting cell wall and N-glycan processing enzymes.

Malaria in Sundargarh district, Odisha, India: Epidemiological and behavioral aspects from surveys.

To characterize malaria and assist in prevention efforts, we conducted a series of epidemiological studies in Sundargarh district, India, as part of an NIH-funded International Center of Excellence for Malaria Research. In a published survey around Rourkela in 2013-2014 (N = 1307), malaria prevalence was found to be 8.3%. Using these data, villages were divided into low (<2%), medium (2-10%) and high (>10%) malaria prevalence, and risk factors assessed by type of village. In the six low malaria villages, four persons were positive by PCR; in the four medium malaria villages, prevalence was 7% (35 infections, 7 P. vivax); and in the three high malaria villages, prevalence was 21% (62 infections, 10 P. vivax and 5 mixed with P. vivax and P. falciparum). A total of 30.6% infections were submicroscopic and 40.6% were asymptomatic. Our analyses showed that the rainy season and male gender were risk factors for malaria; in high malaria villages, young age was an additional risk factor, and indoor and outdoor spraying was protective compared to no spraying. We undertook a subsequent behavioral survey in four of the medium and high malaria villages in 2017 to investigate the behavioral aspects of malaria risk. Among 500 participants in 237 households, adult men (15+ years) were more likely to be outside in the evening (34.5% vs. 7.9% among adult women 15+ years and 0.7% among children, p < 0.001), or to sleep outside (7.5% vs. 0.5% and 0%, respectively, p < 0.001). Although women were more likely to get up before 6 a.m. (86.6%, vs. 70.5% among men, 50.7% among children, p < 0.001), men were more likely to be outside in the early morning (77.6% among men, 11.2% among women, and 11.1% among children, p < 0.001). More children used insecticide treated nets the previous night (73.4%) than men (45.6%) or women (39.6%), and repellents were used by 29.5% of 234 households (insecticide creams were not used at all). Malaria control and elimination in India will need local approaches, and the promotion of repellent cream use by at-risk groups could be further explored in addition to mass-screen or treat programs in high-risk villages.

Effects of beta-1,3-glucan from Septoria tritici on structural defence responses in wheat.

The accumulation of the pathogenesis-related (PR) proteins beta-1,3-glucanase and chitinase and structural defence responses were studied in leaves of wheat either resistant or susceptible to the hemibiotrophic pathogen Septoria tritici. Resistance was associated with an early accumulation of beta-1,3-glucanase and chitinase transcripts followed by a subsequent reduction in level. Resistance was also associated with high activity of beta-1,3-glucanase, especially in the apoplastic fluid, in accordance with the biotrophic/endophytic lifestyle of the pathogen in the apoplastic spaces, thus showing the highly localized accumulation of defence proteins in the vicinity of the pathogen. Isoform analysis of beta-1,3-glucanase from the apoplastic fluid revealed that resistance was associated with the accumulation of an endo-beta-1,3-glucanase, previously implicated in defence against pathogens, and a protein with identity to ADPG pyrophosphatase (92%) and germin-like proteins (93%), which may be involved in cell wall reinforcement. In accordance with this, glycoproteins like extensin were released into the apoplast and callose accumulated to a greater extent in cell walls, whereas lignin and polyphenolics were not found to correlate with defence. Treatment of a susceptible wheat cultivar with purified beta-1,3-glucan fragments from cell walls of S. tritici gave complete protection against disease and this was accompanied by increased gene expression of beta-1,3-glucanase and the deposition of callose. Collectively, these data indicate that resistance is dependent on a fast, initial recognition of the pathogen, probably due to beta-1,3-glucan in the fungal cell walls, and this results in the accumulation of beta-1,3-glucanase and structural defence responses, which may directly inhibit the pathogen and protect the host against fungal enzymes and toxins.