Analysis of capsaicinoid biosynthesis pathway genes expression in callus cultures of Capsicum chinense Jacq. cv. ‘Umorok’

The placental tissue of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’, is used as explants for callus induction. Callus cultures were subcultured after every 32 days and growth curves for a period of six consecutive growth cycles were studied till a stable capsaicinoids producing callus cultures were obtained. The capsaicinoids content in placental tissue explants decreased gradually during the first 2 months of culture as the explants dedifferentiated to form friable callus while the biomass and capsaicinoid content did not show much change in the subsequent growth cycles. The maximum callus biomass of 7.8 g freshweight (FW) or 0.56 g dry weight (DW) per culture were obtained on the 24th day of every growth cycle and the maximum average capsaicinoids content (1.6 mg g^?1 FW capsaicin and 0.78 mg g^?1 FW dihydrocapsaicin) were obtained on the 20th day of every growth cycle. To investigate the underlying dynamics for capsaicinoid biosynthesis during callus formation, comparative gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway were also studied by qRT-PCR analysis. When compared with placental tissue, all the studied genes showed reduced expression during callus formation, especially putative aminotransferase (pAMT) and pungent gene 1 (Pun1), which were extensively down regulated from the 3rd month onwards in the callus cultures. Therefore, the present study revealed that the down-regulated expression of mainly two putative genes in capsaicinoid biosynthetic pathway (pAMT and Pun1) resulted in lower accumulation of capsaicinoids in callus cultures compared to placental tissues of fruits.

     

Length‐weight relationship and relative condition factor of five indigenous fish species from Torsa River,West Bengal,India

Length‐weight relationships (LWRs) for five indigenous fish species under five genera and two families, namely Cyprinion semiplotum (McClelland, 1839), Barilius barna (Hamilton, 1822), Barilius barila (Hamilton, 1822), Gagata sexualis Tilak, 1970 and Pseudolaguvia shawi (Hora, 1921) were studied from Torsa River in West Bengal, India. Sampling was done at quarterly intervals from November, 2015 to December, 2016 and a total of 701 fish specimens were collected using cast nets (mesh size of 15–20 mm), gill nets (mesh size 20–35 mm) and bamboo traps. In the present study, b value ranges from 2.686 to 3.268. LWRs for these fish species (except P. shawi) have not been reported in FishBase. A new maximum length has been recorded for P. Shawi and G. sexualis. The relative condition factor (Kn) values was found to range from 1.004 to 1.028.     

Synthesis of functionalized benzo[<Emphasis Type="Italic">f</Emphasis>]2<Emphasis Type="Italic">H</Emphasis>-chromenes and evaluation of their antimicrobial activities

Knoevenagel cyclocondensations of α-hydroxy naphthaldehyde with β-oxodithioesters and ketene dithioacetals yielded 2H-benzo[f]chromene-2-thiones and 2H-benzo[f]chromen-2-ones, respectively, in high yields. The newly synthesized compounds were evaluated for antifungal and antibacterial activities. Among them, compounds (2-furyl)(3-thioxo-3H-benzo[f]chromen-2-yl)methanone and phenyl(3-oxo-3H-benzo[f]chromen-2-yl)methanone exhibited excellent antifungal activity against tested fungi Curvularia lunata and Fusarium moniliforme. The highest antibacterial activity against the tested bacteria Escherichia coli and Staphylococcus aureus was observed for (4-chlorophenyl)(3-oxo-3H-benzo[f]chromen-2-yl)methanone. The results of antimicrobial screening demonstrate that (2-furyl)(3-thioxo-3H-benzo[f]chromen-2-yl)methanone, phenyl(3-oxo-3H-benzo[f]chromen-2-yl)methanone, and (4-chlorophenyl)(3-oxo-3H-benzo[f]chromen-2-yl)methanone are promising as antimicrobial drugs.     

Cadmium stress-induced oxidative stress and role of nitric oxide in rice (Oryza sativa L.)

Cadmium (Cd) is a potential environmental phytotoxicant. The generation of reactive oxygen species (ROS) due to Cd stress is responsible for the induction of oxidative stress in plants. On the other hand, SNP, a NO donor is known to have effect on Cd-induced oxidative stress in plants. We evaluated the effect of NO on the regulation of Cd stress in the rice (Oryza sativa L.) variety MSE-9. Cd treatment was given in the form of 50, 100 and 200 ??M, whereas for interaction study, 100 ??M of Cd and 100 ??M of SNP were used. The result showed that Cd-induced oxidative stress in MSE-9 by generating ROS. However, when SNP was given with Cd stress, it was seen that SNP treatment regulated the stress metabolism in rice seedlings under Cd toxicity by generating NO. It can be said that the SNP in combination with Cd treatment might possess the way to protect rice seedlings under Cd stress.     

Structure-activity relationship of buffalo antibacterial hepcidin analogs

Hepcidin is an anti-microbial peptide expressed predominantly in the liver of many species. Based on the amino acid sequence deduced from buffalo (Bubalus bubalis) hepcidin cDNA (Accession no. EU399814), six peptides Hepc(1-25), Hepc(6-25), Hepc(7-25), Hepc(9-25), Hepc(11-25) and Hepc(15-25) were synthesized using solid-phase fluorenylmethoxycarbonyl (Fmoc) chemistry. CD spectroscopy revealed different spectra of the peptides in different solvents and in all the cases beta-structure was found to be dominant with less alpha-helix as predicted. Quantitation of secondary structure indicated the highest beta-structure for all the six peptides in SDS solution, when used as mimetic for membrane-like environment. The CD spectra of all the peptides taken in water showed that degree of randomness decreased with increase in chain length of the peptide. Out of the six peptides, only Hepc(1-25), Hepc(6-25) and Hepc(7-25) showed antibacterial activity against Staphylococcus aureus (Gram-positive bacteria). The peptides did not show any sensitivity toward E. coli (Gram-negative bacteria). Minimum inhibitory concentration (MIC) showed the lowest value for Hepc(7-25) as an antibacterial agent, followed by Hepc(6-25) and Hepc(1-25). The peptides Hepc(9-25), Hepc(11-25) and Hepc(15-25) with more random structure did not show any antimicrobial activity The study demonstrated that 5 amino acids at N-terminal in buffalo hepcidin can be truncated without loss of antimicrobial activity and further reduction of length of the analog from 20 to 19 amino acids resulted increase in the activity because of increase in beta-structure of the peptide shown by CD spectroscopy.     

Necrosis as an adaptive response to copper toxicity in Ipomoea aquatica Forsk. and its possible application in phytoremediation

Ipomoea aquatica showed symptoms of stem necrosis when exposed to graded concentrations of copper (Cu) in 50 % Hoagland solution. The duration of exposure required to elicit necrotic response was concentration-dependent, where two stages, early necrosis (EN) and advanced necrosis (AN), could be visually distinguished. The apical parts of the exposed plant remained non-necrotic (NN). NN pieces placed in normal Hoagland 50 % solution could grow and produce new roots, nodes and leaves. Comparisons of protein, carbohydrate and copper concentrations among control (CN), EN, AN, NN and regrown and recovered (RC) stem tissues revealed that AN tissue had the lowest protein and carbohydrate concentrations, while NN had the highest. On the contrary, Cu concentration was higher in EN and AN than that in NN and CN. RC tissue had comparable protein, carbohydrate and Cu concentrations to those of CN. Thus I. aquatica could sequester excess Cu in necrotic tissue to keep the apical parts largely free from Cu toxicity. At the same time, more proteins and carbohydrates were synthesized in the apical part, which in turn enabled the plant to survive and grow, even when under Cu stress. This was further aided by the ability of its stem to produce adventitious roots from nodes and give off lateral shoots that bore flowers and leaves. This could be exploited for Cu phytoremediation by growing the plant in Cu-rich medium and then removing the Cu-enriched necrotic portions for safe disposal. The unaffected NN portions could be regrown and reused.     

Length–weight relationships for Eutropiichthys murius (Hamilton, 1822), Coilia reynaldi Valenciennes, 1848 and Johnius gangeticus Talwar, 1991 from lower stretch of the River Ganga,India

Length‐weight relationships (LWRs) for three fish species from the River Ganga (India) is presented. Sampling was conducted from the lower stretch of the River Ganga (Patna: 25°36′51.66″N & 85°12′7.02″E to Freserganj: 21°35′40.58″N & 88°15′28.92″E) during April, June and September and December of 2017. Specimens were caught using gill nets (18 nos.; mesh 18–32 mm), and bag nets (3 nos.; mesh 14–22 mm). The values a and b from LWRs ‐were found to be 0.007 and 2.977 for Eutropiichthys murius; 0.003 and 3.001 for Coilia reynaldi; 0.009 and 3.010 for Johnius gangeticus.     

Evidence for the High Importance of Co-Morbid Factors in HFE C282Y/H63D Patients Cared by Phlebotomies: Results from an Observational Prospective Study

Despite type I haemochromatosis (HC) is mainly associated with the HFE C282Y/C282Y genotype, a second genotype -C282Y/H63D- has mostly been described in other patients. Its association with HC, apart from any associated co-morbid factors, remains unclear and complex to interpret for physicians. This study assesses the weight of this genotype and the role of co-morbid factors in the occurrence of iron overload. This prospective study included the C282Y/C282Y (n = 172) and C282Y/H63D (n = 58) patients enrolled in a phlebotomy program between 2004 and 2007 in a blood centre of western Brittany (Brest, France), where HC is frequent. We compared prevalence of these two genotypes, as well as patients’ profile regarding degree of iron overload and prevalence of co-morbid factors. First, we confirmed the obvious deficit of C282Y/H63D compound heterozygotes among patients cared by phlebotomies. This genotype was 3.0 times less frequent than the C282Y/C282Y genotype among those patients (18.9% vs. 56.0%) whereas it was 4.9 times more frequent in the general population (4.3% vs. 0.9%; p<0.0001). Despite a similar level of hyperferritinaemia, the C282Y/H63D patients who came to medical attention had a milder plasma iron overload, reflected by a lower transferrin saturation median (52.0% vs. 84.0%; p<0.0001). They also exhibited more frequently co-morbid factors, as heavy drinking (26.0% vs. 13.9%; p = 0.0454), overweight (66.7% vs. 39.4%; p = 0.0005) or both (21.3% vs. 2.6%; p<0.0001). Ultimately, they required a lower amount of iron removed to reach depletion (2.1 vs. 3.4 g; p<0.0001), clearly reflecting their lower tissue iron. This study confirms that H63D is a discrete genetic susceptibility factor whose expression is most visible in association with other co-factors. It highlights the importance of searching for co-morbidities in these diagnostic situations and of providing lifestyle and dietary advice.     

MicroRNA‐29b modulates Japanese encephalitis virus‐induced microglia activation by targeting tumor necrosis factor alpha‐induced protein 3

Japanese encephalitis virus (JEV), a single‐stranded RNA (ssRNA) virus, is the leading cause of encephalitis in Asia. Microglial activation is one of the key events in JEV‐induced neuroinflammation. Although the various microRNAs (miRNAs) has been shown to regulate microglia activation during pathological conditions including neuroviral infections, till date, the involvement of miRNAs in JEV infection has not been evaluated. Hence, we sought to evaluate the possible role of miRNAs in mediating JEV‐induced microglia activation. Initial screening revealed significant up‐regulation of miR‐29b in JEV‐infected mouse microglial cell line (BV‐2) and primary microglial cells. Furthermore, using bioinformatics tools, we identified tumor necrosis factor alpha‐induced protein 3, a negative regulator of nuclear factor‐kappa B signaling as a potential target of miR‐29b. Interestingly, in vitro knockdown of miR‐29b resulted in significant over‐expression of tumor necrosis factor alpha‐induced protein 3, and subsequent decrease in nuclear translocation of pNF‐κB. JEV infection in BV‐2 cell line elevated inducible nitric oxide synthase, cyclooxygenase‐2, and pro‐inflammatory cytokine expression levels, which diminished after miR‐29b knockdown. Collectively, our study demonstrates involvement of miR‐29b in regulating JEV‐ induced microglial activation.