Sexual dimorphism in NLR transcripts and its downstream signaling protein IL-1ꞵ in teleost Channa punctata (Bloch, 1793).

Nucleotide-binding oligomerization domain-like receptors (NOD-like receptors or NLRs) are a family of intracellular pattern recognition receptors (PRRs) that initiates as well as regulate inflammatory responses. NLRs are characterized by a centrally located nucleotide binding domain and a leucine rich repeat domain at the C-terminal responsible for the recognition of intracellular microbe-associated molecular patterns (MAMPs) and danger-associated molecular patterns (DAMPs). In the present study in adult spotted snakehead we have investigated the sex-dependent tissue distribution of NLRs known to be associated with inflammation in teleost namely NOD1, NOD2, NLRC3, NLRC5, and NLRX1. Further, the sexual dimorphism in the expression of NLR transcript as well as the pro-inflammatory protein IL-1ß was explored in fish under normal conditions, and in fish exposed to bacterial lipopolysaccharide (LPS). The NLRs show ubiquitous and constitutive expression in all the tissues. Moreover, a prominent disparity between males and females was observed in the basal expression of these genes in various tissues. The sexual dimorphism in NLR expression was also prominent when fish were exposed to LPS. Similarly, IL-1ß exhibited sexual dimorphism in both normal as well as LPS-exposed fish.

Exploring the effect of dihydrotestosterone on nucleotide-binding and oligomerization domain-like receptor expression in spotted snakehead Channa punctata (Bloch 1793).

Sex steroids are known to modulate immune responses and as a result many of the immune parameters in seasonally breeding organisms show reproductive-phase dependent variation. Androgens, the male sex steroids, are largely reported to be immunosuppressive. Together with other pattern recognition receptors, the nucleotide-binding and oligomerization domain-like receptors (NLRs) serve as intracellular sentinels and are essential to defense mechanisms. Interestingly, to date the transcriptional modulation of NLRs by androgens has not been explored. In the present study, we investigated the reproductive-phase dependent expression of NLRs in the male spotted snakehead Channa punctata. Furthermore, the effect of dihydrotestosterone (DHT) on NLR expression was studied. The expression of NLRs was observed to be most pronounced during the spawning phase of the fish, which is marked by the highest testosterone level. In vivo as well as in vitro studies showed the diverse effect of DHT on NLR expression depending on the duration and mode of treatment, as well as the immune tissue studied.

Global impacts of extractive and industrial development projects on Indigenous Peoples' lifeways, lands, and rights.

To what extent do extractive and industrial development pressures affect Indigenous Peoples' lifeways, lands, and rights globally? We analyze 3081 environmental conflicts over development projects to quantify Indigenous Peoples' exposure to 11 reported social-environmental impacts jeopardizing the United Nations Declaration on the Rights of Indigenous Peoples. Indigenous Peoples are affected in at least 34% of all documented environmental conflicts worldwide. More than three-fourths of these conflicts are caused by mining, fossil fuels, dam projects, and the agriculture, forestry, fisheries, and livestock (AFFL) sector. Landscape loss (56% of cases), livelihood loss (52%), and land dispossession (50%) are reported to occur globally most often and are significantly more frequent in the AFFL sector. The resulting burdens jeopardize Indigenous rights and impede the realization of global environmental justice.

Molecular cloning, characterization and 3D modelling of spotted snakehead fbn1 C-terminal region encoding asprosin and expression analysis of fbn1.

The FBN1 gene encodes profibrillin protein that is cleaved by the enzyme furin to release fibrillin-1 and a glucogenic hormone, asprosin. Asprosin is implicated in diverse metabolic functions as well as pathological conditions in mammals. However, till date, there are no studies on asprosin in any non-mammalian vertebrate. In this study, we have retrieved the spotted snakehead Channa punctata fbn1 gene (ss fbn1) from the testicular transcriptome data and validated it. The transcript is predicted to encode 2817 amino acid long putative profibrillin protein. Amino acid sequence alignment of deduced ss profibrillin with human profibrillin revealed that the furin cleavage site in profibrillin is well conserved in C. punctata. Further, differential expression of ss fbn1 was observed in various tissues with the highest expression in gonads. Prominent expression of furin was also observed in the gonads suggesting the possibility of proteolytic cleavage of profibrillin protein and secretion of asprosin in C. punctata. In addition, the C-terminal of the fbn1 gene of C. punctata that codes for asprosin protein has been cloned. Using in silico approach, physicochemical properties of the putative ss asprosin were characterized and post-translational changes were predicted. The putative ss asprosin protein sequence is predicted to consist of 142 amino acid residues, with conserved glycosylation sites. Further, the 3D model of ss asprosin was predicted followed by MD (molecular dynamics) simulation for energy minimization. Thus, the current study, for the first time in non-mammalian vertebrates, predicts and characterizes the novel protein asprosin using in silico approach.

Crosstalk between reproductive and immune systems: the teleostean perspective.

The bidirectional interaction between the hypothalamic-pituitary-gonadal (HPG) axis and the immune system plays a crucial role in the adaptation of an organism to its environment, its survival and the continuance of a species. Nonetheless, very little is known about this interaction among teleost, the largest group of extant vertebrates. Fishes being seasonal breeders, their immune system is exposed to seasonally changing levels of HPG hormones. On the contrary, the presence and infiltration of leukocytes, the expression of pattern recognition receptors as well as cytokines in gonads suggest their key role in teleostean gametogenesis as in the case of mammals. Moreover, the modulation of gametogenesis and steroidogenesis by lipopolysaccharide implicates the pathological significance of inflammation on reproduction. Thus, it is important to engage in the understanding of the interaction between these two important physiological systems, not only from a phylogenetic perspective but also due to the importance of fish as an important economic resource. In view of this, the authors have reviewed the crosstalk between the reproductive and immune systems in teleosts and tried to explore the importance of this interaction in their survival and reproductive fitness.

Teleost NOD-like receptors and their downstream signaling pathways: A brief review.

Nucleotide-binding oligomerization domain-like receptors (NOD-like receptors or NLRs) are key members of the immune system that act as intracellular sentinels. These pathogen recognition receptors are essentially characterized by a central nucleotide binding domain and a C-terminal leucine rich repeat domain responsible for recognition of pathogens. Over the past decade, our understanding of teleosts' NLRs has enhanced significantly although the signaling pathways remain to be elucidated. In this brief review, we have tried to decipher the structural and functional aspects of NLRs in teleost. The review also engages in illustrating the various downstream signaling pathways/molecules reported so far in fishes that enable the NLRs to act as important players in immune responses and defense mechanisms against pathogens. Importantly, we try to explore the lacunae in structural and mechanistic details of NLRs in the teleost that would help in identifying key areas in which research is needed to complete our understanding of NLRs and their structural and functional evolution.

Molecular and functional characterization of spotted snakehead NOD1 with an emphasis on structural insights into iE-DAP binding motifs employing advanced bioinformatic tools.

Nucleotide-binding and oligomerization domain (NOD)-like receptors (NLRs) are cytosolic receptors implicated in recognition of intracellular pathogen associated molecular patterns (PAMPs) and danger associated molecular patterns (DAMPs). Depending upon their effector binding domain (EBD) at the C-terminal, the NLRs are categorized into NLRA, NLRB, NLRC, NLRP and NLRX. NOD1 is a pivotal player in immune responses against bacterial and viral invasions and interacts with pathogens via C-terminal leucine rich repeat (LRR) domain. This study aims at characterizing NOD1 in an economically important teleost of the Indian subcontinent, spotted snakehead Channa punctata. The understanding of pathogen-receptor interaction in teleosts is still obscure. In light of this, combinatorial approach involving protein modeling, docking, MD simulation and binding free energy calculation were employed to identify key motifs involved in binding iE-DAP. In silico analysis revealed that NOD1 consists of 943 amino acids comprising of one caspase recruitment domain (CARD) at N-terminal, one central NACHT domain and nine leucine rich repeat (LRR) regions at C-terminal. Structural dynamics study showed that the C-terminal ß-sheet LRR4-7 region is involved in iE-DAP binding. NOD1 was ubiquitously and constitutively expressed in all tissues studied. Differential expression profile of NOD1 induced by Aeromonas hydrophila infection was also investigated. Lymphoid organs and phagocytes of infected spotted snakehead showed significant downregulation of NOD1 expression. The current study thus gives an insight into structural and functional dynamics of NOD1 which might have future prospect for structure-based drug designing in teleosts.Communicated by Ramaswamy H. Sarma.

Talk renewables, walk coal: The paradox of India's energy transition.

Coal is on the rise in India: despite the devasting impacts of the climate crisis, the awareness for land and forest rights, and political talk of a coal phase-out. In this article, we demonstrate that despite the renewables-led rhetoric, India is in the midst of a transition to (not away from) greater use of coal in its fossil energy system and in the electricity system in particular. We investigate this paradox by combining socio-metabolic and political-ecological analysis of the Indian coal complex. Our framework integrates material and energy flow data as characterizing the Indian fossil energy transition, indicators on the development and structure of the coal industry, and studies of ecological distribution conflicts around coal. The dominant claim to expansive use of coal and the competing counterclaims are indicative of underlying power relations which can also be witnessed in other countries. In India, they extend into the conflicted development of renewable energy including hydropower, in which the land dispossession, exclusion, and injustices associated with the expansion of the coal complex are reproduced. We conclude that the current energy transition - in which coal continues to play a dominant role - is neither sustainable nor just.

Environmental conflicts and defenders: A global overview.

Recent research and policies recognize the importance of environmental defenders for global sustainability and emphasize their need for protection against violence and repression. However, effective support may benefit from a more systematic understanding of the underlying environmental conflicts, as well as from better knowledge on the factors that enable environmental defenders to mobilize successfully. We have created the global Environmental Justice Atlas to address this knowledge gap. Here we present a large-n analysis of 2743 cases that sheds light on the characteristics of environmental conflicts and the environmental defenders involved, as well as on successful mobilization strategies. We find that bottom-up mobilizations for more sustainable and socially just uses of the environment occur worldwide across all income groups, testifying to the global existence of various forms of grassroots environmentalism as a promising force for sustainability. Environmental defenders are frequently members of vulnerable groups who employ largely non-violent protest forms. In 11% of cases globally, they contributed to halt environmentally destructive and socially conflictive projects, defending the environment and livelihoods. Combining strategies of preventive mobilization, protest diversification and litigation can increase this success rate significantly to up to 27%. However, defenders face globally also high rates of criminalization (20% of cases), physical violence (18%), and assassinations (13%), which significantly increase when Indigenous people are involved. Our results call for targeted actions to enhance the conditions enabling successful mobilizations, and for specific support for Indigenous environmental defenders.

Cervical cancer awareness and HPV vaccine acceptability among females in Delhi: A cross-sectional study.

AIMS: The aim of this study is to assess the awareness about cervical cancer and the acceptability of cytological screening and vaccine against human papilloma virus (HPV) among women in Delhi, the national capital of India. MATERIALS AND METHODS: A cross-sectional survey of women was conducted in Delhi to assess the awareness of cervical cancer and acceptability of Papanicolaou (Pap) test and HPV vaccine. The sample size of the population was 450, and a pre-tested questionnaire was administered to them. RESULTS: Majority of the participants (85.11%) were aware of cervical cancer and were willing to undergo diagnosis by Pap test (84.6%). As far as vaccination was concerned, 63.14% found the HPV vaccine acceptable for their daughters. However, very few participants were willing to vaccinate themselves against HPV. CONCLUSION: The high awareness among females in Delhi about cervical cancer and acceptability of screening programs, if done free of cost, shows a positive trend. The only inhibition about HPV vaccine was primarily due to concerns about postvaccination complications. However, inclusion of HPV vaccine in Government-sponsored immunization program would go a long way in increasing the acceptability of the vaccine.

Genomic and non-genomic effect of cortisol on phagocytosis in freshwater teleost Channa punctatus: an in vitro study.

The non-genomic action of glucocorticoid, besides classical genomic action, is recently implicated in regulation of phagocyte activities in mammals. With regard to the non-mammalian vertebrates, this study in the teleost, Channa punctatus, for the first time demonstrates the regulation of innate immunity by cortisol following non-genomic pathway. Cortisol suppressed the phagocytic activity of splenic phagocytes in a time- and concentration-dependent manner. Intriguingly, it impeded the phagocytosis within 15 min which is too short for conventional genomic action. The cortisol-induced rapid inhibition could not be altered by transcription and translation inhibitors, suggesting the involvement of non-genomic pathway. Since membrane impermeable BSA-cortisol mimicked the rapid inhibitory effect of cortisol at 15 min, we speculated that cortisol exerted its non-genomic effect on phagocytosis by acting at membrane site. These membrane-bound glucocorticoid receptors seem similar to cytosolic GR, as rapid inhibitory effect of cortisol was blocked by the cytoplasmic glucocorticoid receptor blocker RU-486. Using inhibitors for adenylate cyclase/protein kinase A (PKA) and estimating intracellular cAMP, adenylate cyclase-PKA pathway was seen involved in mediating the rapid non-genomic action of cortisol in phagocytes of C. punctatus. In contrast to the rapid effect, inhibitory effect of cortisol on phagocytosis after 1h was blocked by protein synthesis inhibitors, thus implicating genomic regulation. An overview of our data suggests that cortisol regulates phagocytosis in C. punctatus via genomic as well as non-genomic mechanisms. Further, the non-genomic action of cortisol is mediated via membrane-bound GR coupled to cAMP-PKA system.

Diurnal variation in phagocytic activity of splenic phagocytes in freshwater teleost Channa punctatus: melatonin and its signaling mechanism.

The aim of the present study was to understand the rhythmic changes in innate immune response in freshwater fish Channa punctatus. Furthermore, the putative role of melatonin as the zeitgeber was explored. The phagocytic activity of splenic phagocytes assessed at 6-h intervals showed higher phagocytic activity during light phase than dark phase. The increased phagocytic activity during light phase was diminished by melatonin administration at 09:00 h. Implication of melatonin in control of diurnal variation in phagocytic activity was substantiated by administering irreversible tryptophan hydroxylase inhibitor, para-chlorophenylalanine (pCPA) at 18:00 h. pCPA abrogated the decrease of phagocytosis observed during dark phase, and the same was restored after melatonin administration. The direct involvement of melatonin in modulation of phagocytosis was demonstrated following in vitro experiments. Melatonin suppressed the phagocytic activity in a concentration-dependent manner without affecting the viability of phagocytes. The existence of functional membrane-bound melatonin receptors on fish phagocytes was pharmacologically demonstrated. Luzindole, melatonin membrane receptor antagonist, completely blocked the inhibitory effect of melatonin on phagocytosis. Further receptor-coupled adenylate cyclase-protein kinase A (PKA) pathway was implicated in transducing the melatonin effect as both adenylate cyclase and PKA inhibitor completely nullified the melatonin-induced suppression. An increased intracellular cAMP level in response to melatonin ascertained the second messenger status of cAMP for downstream signaling. However, manipulation of phospholipase C/PKC failed to influence the effect of melatonin on phagocytic activity. These observations in C. punctatus evidenced the diurnal rhythmicity in phagocytic activity that is regulated by melatonin following membrane-bound receptor-coupled cAMP-PKA pathway.

Role of adrenoceptor-coupled second messenger system in sympatho-adrenomedullary modulation of splenic macrophage functions in live fish Channa punctatus.

In order to understand the role of sympatho-adrenomedullary (SAM) system in mediating stress effect on non-specific immune responses in fishes, the splenic macrophage phagocytic and respiratory burst activities of normal and chemically sympathectomized Channa punctatus under restraint stress were studied. Chemical sympathectomy abrogated the differential effects of acute stress on diverse functions of macrophages. The SAM regulation of macrophage activities was substantiated by in vitro experiments with catecholamines, the end product of SAM system. Further, for the first time in fishes, different adrenoceptors and their precise second messenger system regulating diverse functions of macrophages by catecholamines were demonstrated. Norepinephrine (NE)/epinephrine (E) decreased the phagocytosis through beta-adrenergic receptor as only propranolol, the beta-adrenergic receptor antagonist, blocked the suppressive effect of NE/E. However, dopamine (DA) regulates phagocytosis solely via the dopaminergic receptor. The DA effect was mimicked by DA receptor agonists, apomorphine and bromocryptine. Adenylate cyclase system linked to beta-adrenoceptor/dopaminergic receptor seems to be involved in mediating the effect of catecholamine on phagocytosis since db cAMP inhibited the phagocytosis in a dose-dependent manner. In case of superoxide production, only phenoxybenzamine, an alpha-adrenergic receptor antagonist, was seen effective in blocking the stimulatory effect of NE/E. Further, Ca2+ as second messenger system coupled to alpha1-adrenergic receptor was shown to mediate this effect since phospholipase C (PLC) inhibitor, U73122 and intracellular calcium chelating agent, BAPTA-AM downregulated the NE/E-induced superoxide production. The role of calcium in modulation of superoxide production was also emphasized using calcium ionophore A23187.

Dual mode of catecholamine action on splenic macrophage phagocytosis in wall lizard, Hemidactylus flaviviridis.

In the present study, in vitro concentration-related effect of catecholamines, dopamine (DA), norepinephrine (NE), and epinephrine (E) was observed on phagocytic activity of splenic macrophages to understand the impact of sympatho-adrenal-medullary (SAM) activation on innate immunity in wall lizard Hemidactylus flaviviridis under stress condition. Restraint stress for 1 h resulted in marked suppression of macrophage phagocytosis, suggesting that supra-physiological level of catecholamines in response to SAM activation under stress suppressed phagocytosis. This interpretation was reinforced since all the catecholamines considerably reduced phagocytosis at high concentrations ranging from 10(-7) to 10(-5)M. On the contrary, DA, NE, and E at low concentrations considerably stimulated phagocytosis, which increased with the decrease of concentrations ranging from 10(-11) to 10(-15)M. Further, effect of NE and E was blocked by beta-adrenergic blocker suggesting the beta-adrenoceptor-dependent regulating mechanism of NE and E. DA acts through both beta-adrenoceptor-dependent and D1/D2 class receptor-dependent mechanism, since beta-adrenergic blocker could partially block the DA effect. beta-Adrenoceptor-linked adenylate cyclase-mediated cAMP action in modulation of phagocytic activity was evident as 3-isobutyl-1-methyl-xanthine suppressed phagocytosis. Further, to delineate the mode of dual effect of catecholamines through beta-adrenergic receptors, in vitro concentration-related effect of cAMP was investigated on macrophage phagocytosis. cAMP depending on concentration had opposite effect on phagocytosis, and its stimulatory effect at low concentrations was reversed by actinomycin D and cycloheximide, whereas these transcription and translation inhibitors, respectively, failed to alter the inhibitory effect of cAMP at high concentrations. This suggests the concentration-related two different pathways of catecholamine action, classical non-genomic at high concentration while genomic pathway at low concentration.