Acyclic diterpene alcohols isolated from four algae of Bryopsidophyceae and their toxicity

Three new acylic diterpenoids belonging to the class of phytol series have been isolated. They were obtained from the ethyl acetate soluble fractions of four siphonaceous green seaweeds, Bryopsis pennata Lamour., Caulerpa taxifolia (Vahl) C. Ag., Codium decorticatum (Woodw.) Howe and Valoniopsis pachynema (Mart.) Børg., collected from Karachi coast of Pakistan. Structures of these compounds were elucidated with the help of spectroscopic methods and confirmed by comparison with the known compounds. Even the known compounds are being reported for the first time from a green algal source. All the compounds were found to display a strong toxicity at all the three concentrations tested in the brine shrimp bioassay

Characterization and evaluation of putative mutant populations of polyembryonic mango genotype Nekkare for dwarfing rootstock traits

Availability of dwarfing rootstocks is an important pre-requisite for improving productivity of mango orchards in India as it facilitates high density planting as well as impart uniformity within an orchard. An attempt was made to induce variability in polyembryonic mango genotype Nekkare for dwarfness by treating kernels with different doses of gamma radiation ranging from 15 to 35 Gy. Irradiation created significant variation in plant height, stem girth, number of nodes, inter-nodal length, number of leaves, leaf blade length and leaf blade width. The highest reduction in seedling height along with highest variation was observed at 35 Gy where the seedling height ranged from 11.50 to 33 cm with a mean of 23.12 cm as compared to mean plant height of 44.55 cm in control ranging from 33.50 to 56 cm. Further, the effect of irradiation on stomatal parameters was also investigated and the highest stomatal length and width was recorded at 15 Gy (63.39 μm) and 20 Gy (63.12 μm) respectively while 30 Gy treatment produced maximum stomatal density (13.85 per μm2). Furthermore, theconcentration of ABA was found to be highest (429.1 ng/gm) in morphologically dwarf (putative mutant) progenies of Nekkare. The results suggest effectiveness of induced mutation for developing dwarfing rootstocks in mango to be used in high density planting

Profil ekspresije receptora hormona rasta (GHR), receptora čimbenika rasta sličnog inzulinu (IGFR) i gena leptina s biokemijskim i endokrinim profilom u jaradi crne bengalske koze (Capra hircus) tijekom različitih razdoblja pretpubertetskog rasta

The aim of this study was to evaluate the expression profile of growth hormone receptor (GHR), insulin like growth factor receptor (IGFR) and leptin genes, together with some blood biochemical (glucose, aspartate aminotransferase, AST; alanine aminotransferase, ALT) and endocrine (growth hormone and cortisol) parameters. For this purpose, blood samples were collected from 25 male and 25 female kids of the Black Bengal goat (Capra hircus), at days 15, 30, 60, 90, 120 and 150. Male kids gained more body weight compared to females. The glucose and cortisol levels significantly (P≤0.01) decreased with the advancement of age. AST, ALT, and growth hormone increased significantly (P≤0.01). In both sexes, plasma glucose level was negatively (P≤0.01) correlated with AST. In males both cortisol and GH levels were positively (P≤0.05) correlated with plasma glucose levels. In female kids only GH showed a positive (P≤0.05) correlation with glucose levels. Female kids had greater expression of the GH receptor, IGFR mRNA and Leptin mRNA on day 30 (P≤0.05) and day 150 (P≤0.01). GHR was only positively (P≤0.05) correlated with IGFR and LEP in female kids. On the basis of the results obtained from our research, we are able to conclude that the lower growth rate in female kids expressed more leptin, which stimulated the somatotrophic axis for the expression of GHR and IGFR during the prepubertal growth stage.Cilj istraživanja bio je procijeniti profil ekspresije receptora hormona rasta (GHR), receptora čimbenika rasta sličnog inzulinu (IGFR) i gena leptina zajedno s krvnim biokemijskim pokazateljima (glukoza, aspartat-aminotransferaza AST, alanin-aminotransferaza ALT) i endokrinim pokazateljima (hormon rasta i kortizol). U tu su svrhu 15., 30., 60., 90., 120. i 150. dan života uzeti su uzorci krvi od 25 muške i 25 ženske jaradi crne bengalske koze (Capra hircus). Muška je jarad imala veći prirast tjelesne mase u usporedbi sa ženskom. Razine glukoze i kortizola s porastom dobi znakovito su se smanjivale (P ≤ 0,01). AST, ALT i hormon rasta znakovito su porasli (P ≤ 0,01). Kod oba je spola razina glukoze u plazmi bila u negativnoj korelaciji (P ≤ 0,01) s AST-om. Kod muške su jaradi kortizol i hormon rasta bili u pozitivnoj korelaciji (P ≤ 0,05) s razinama glukoze u plazmi. Samo je u ženske jaradi hormon rasta pokazao pozitivnu korelaciju (P ≤ 0,05) s razinama glukoze. Ženska je jarad imala veću ekspresiju receptora hormona rasta, mRNA IGFR i mRNA leptina 30. dan (P ≤ 0,05) i 150. dan (P ≤ 0,01). GHR je bio u pozitivnoj korelaciji (P ≤ 0,05) samo s IGFR-om i leptinom u ženske jaradi. Na temelju rezultata ovoga istraživanja možemo zaključiti da niža stopa rasta u ženske jaradi dovela do veće ekspresije leptina, što je stimuliralo somatotropnu os za ekspresiju hormona rasta i IGFR-a u pretpubertetskom razdoblju

Caregivers knowledge, practices about childhood diarrhea and pneumonia and their perceptions of lady health worker program; findings from NIGRAAN implementation research project

Background: Despite 60% coverage by Lady Health Worker (LHW) Program, 30% of child deaths in Pakistan are still due to diarrhea and pneumonia. Caregivers are an important stakeholder yet there is little information on their case management practices and utilization of LHW Program. This study explored caregivers’ knowledge and practices about childhood diarrhea and pneumonia and utility of LHW services before and after a supportive supervision intervention.Methods: Cross sectional surveys were conducted with caregivers’ (mothers) pre and post intervention in project NIGRAAN. The intervention aimed to improve LHSs clinical and supervisory skills of lady health supervisors in order to improve LHW performance and ultimately impact caregiver practices. 4250 households were surveyed. Questionnaire was adapted from PDHS 2012-13. Differences between intervention and control groups were assessed using chi square test. P-value of Results: Comparing baseline to end line, there were significant overall improvements in caregivers’knowledge of loose motion (62 to 84%) and dehydration (12 to 18%) as signs and symptoms of childhood diarrhea. There was also a significant overall increase in caregivers’ knowledge of presenting features of pneumonia- i.e. fever (58 to 86%), cough (51 to 61%) and breathing problems (25 to 57%). The proportion of caregivers seeking advice for diarrhea from public sector significantly improved in intervention arm from 20% to 29%. Private sector however remained overall preferred choice for care seeking. There was significant overall improvement in awareness about LHWs functioning (93 to 99%) and household visits (91 to 98%). Actual care seeking from LHWs however stayed low (≤ 0.3%) Conclusion: In order to improve utility and expand coverage of LHW Program interventions aimed at providing supportive supervision have the potential to improve caregiver practices and utilization of available services and decrease childhood deaths due to preventable illnesses

Electronic-Cigarette Vehicles and Flavoring Affect Lung Function and Immune Responses in a Murine Model

The use of electronic nicotine delivery systems (ENDS), also known as electronic-cigarettes (e-cigs), has raised serious public health concerns, especially in light of the 2019 outbreak of e-cig or vaping product use-associated acute lung injury (EVALI). While these cases have mostly been linked to ENDS that contain vitamin E acetate, there is limited research that has focused on the chronic pulmonary effects of the delivery vehicles (i.e., without nicotine and flavoring). Thus, we investigated lung function and immune responses in a mouse model following exposure to the nearly ubiquitous e-cig delivery vehicles, vegetable glycerin (VG) and propylene glycol (PG), used with a specific 70%/30% ratio, with or without vanilla flavoring. We hypothesized that mice exposed sub-acutely to these e-cig aerosols would exhibit lung inflammation and altered lung function. Adult female C57BL/6 mice (n= 11-12 per group) were exposed to filtered air, 70%/30% VG/PG, or 70%/30% VG/PG with a French vanilla flavoring for 2 h a day for 6 weeks. Prior to sacrifice, lung function was assessed. At sacrifice, broncho-alveolar lavage fluid and lung tissue were collected for lipid mediator analysis, flow cytometry, histopathology, and gene expression analyses. Exposures to VG/PG + vanilla e-cig aerosol increased lung tidal and minute volumes and tissue damping. Immunophenotyping of lung immune cells revealed an increased number of dendritic cells, CD4+ T cells, and CD19+ B cells in the VG/PG-exposed group compared to air, irrespective of the presence of vanilla flavoring. Quantification of bioactive lung lipids demonstrated a \u3e3-fold increase of 2-arachidonoylglycerol (2-AG), an anti-inflammatory mediator, and a 2-fold increase of 12-hydroxyeicosatetraenoic acid (12-HETE), another inflammatory mediator, following VG/PG exposure, with or without vanilla flavoring. This suggests that e-cig aerosol vehicles may affect immunoregulatory molecules. We also found that the two e-cig aerosols dysregulated the expression of lung genes. Ingenuity Pathway Analysis revealed that the gene networks that are dysregulated by the VG/PG e-cig aerosol are associated with metabolism of cellular proteins and lipids. Overall, our findings demonstrate that VG and PG, the main constituents of e-liquid formulations, when aerosolized through an e-cig device, are not harmless to the lungs, since they disrupt immune homeostasis

Probing the SAM Binding Site of SARS-CoV-2 nsp14 in vitro Using SAM Competitive Inhibitors Guides Developing Selective bi-substrate Inhibitors

The COVID-19 pandemic has clearly brought the healthcare systems world-wide to a breaking point along with devastating socioeconomic consequences. The SARS-CoV-2 virus which causes the disease uses RNA capping to evade the human immune system. Non-structural protein (nsp) 14 is one of the 16 nsps in SARS-CoV-2 and catalyzes the methylation of the viral RNA at N7-guanosine in the cap formation process. To discover small molecule inhibitors of nsp14 methyltransferase (MT) activity, we developed and employed a radiometric MT assay to screen a library of 161 in house synthesized S-adenosylmethionine (SAM) competitive methyltransferase inhibitors and SAM analogs. Among seven identified screening hits, SS148 inhibited nsp14 MT activity with an IC50 value of 70 ± 6 nM and was selective against 20 human protein lysine methyltransferases indicating significant differences in SAM binding sites. Interestingly, DS0464 with IC50 value of 1.1 ± 0.2 μM showed a bi-substrate competitive inhibitor mechanism of action. Modeling the binding of this compound to nsp14 suggests that the terminal phenyl group extends into the RNA binding site. DS0464 was also selective against 28 out of 33 RNA, DNA, and protein methyltransferases. The structure-activity relationship provided by these compounds should guide the optimization of selective bi-substrate nsp14 inhibitors and may provide a path towards a novel class of antivirals against COVID-19, and possibly other coronaviruses

Human dendritic cells express the complement receptor immunoglobulin which regulates T cell responses

The B7 family-related protein V-set and Ig containing 4 (VSIG4), also known as Z39Ig and Complement Immunoglobulin Receptor (CRIg), is the most recent of the complement receptors to be identified, with substantially distinct properties from the classical complement receptors. The receptor displays both phagocytosis–promoting and anti-inflammatory properties. The receptor has been reported to be exclusively expressed in macrophages. We now present evidence, that CRIg is also expressed in human monocyte-derived dendritic cells (MDDC), including on the cell surface, implicating its role in adaptive immunity. Three CRIg transcripts were detected and by Western blotting analysis both the known Long (L) and Short (S) forms were prominent but we also identified another form running between these two. Cytokines regulated the expression of CRIg on dendritic cells, leading to its up- or down regulation. Furthermore, the steroid dexamethasone markedly upregulated CRIg expression, and in co-culture experiments, the dexamethasone conditioned dendritic cells caused significant inhibition of the phytohemagglutinin-induced and alloantigen-induced T cell proliferation responses. In the alloantigen-induced response the production of IFNγ, TNF-α, IL-13, IL-4, and TGF-β1, were also significantly reduced in cultures with dexamethasone-treated DCs. Under these conditions dexamethasone conditioned DCs did not increase the percentage of regulatory T cells (Treg). Interestingly, this suppression could be overcome by the addition of an anti-CRIg monoclonal antibody to the cultures. Thus, CRIg expression may be a control point in dendritic cell function through which drugs and inflammatory mediators may exert their tolerogenic- or immunogenic-promoting effects on dendritic cells.Usma Munawara, Khalida Perveen, Annabelle G. Small, Trishni Putty, Alex Quach, Nick N. Gorgani, Charles S. Hii, Catherine A. Abbott and Antonio Ferrant

Brassinosteroids as promoters of seedling growth and antioxidant activity under heavy metal zinc stress in mung bean (Vigna radiata L.)

The escalation of harmful pollutants, including heavy metals, due to industrialization and urbanization has become a global concern. To mitigate the negative impacts of heavy metal stress on germination and early plant development, growth regulators have been employed. This study aimed to evaluate the response of mung bean (Vigna radiata L.) to zinc stress in the presence of brassinosteroids, focusing on seedling growth and antioxidant potential. Mung bean seedlings were treated with three concentrations of 24-epibrassinolide (EBL) (0.1, 0.2, and 0.4 PPM) with or without zinc. Results demonstrated that the application of brassinosteroids, combined with zinc stress, significantly enhanced germination percentage (about 47.06, 63.64, and 120%), speed of germination (about 39.13, 50, and 100%), seedling growth (about 38% in case of treatment combined 0.4 PPM 24-EBL and 1.5 mM ZnSO4) and seedling vigor index (204% in case of treatment combined 0.4 PPM 24-EBL and 1.5 mM ZnSO4) compared to zinc-treated seedlings alone after 24 h. The activities of antioxidative enzymes (catalase, ascorbate peroxidase, polyphenol oxidase, and peroxidase) and total soluble protein content decreased, while lipid peroxidation and proline content exhibited a significant increase (p ≤ 0.05) when compared to the control. However, the negative effects induced by heavy metal stress on these parameters were significantly mitigated by EBL application. Notably, the most effective concentration of EBL in overcoming zinc stress was found to be 0.4 PPM. These findings underscore the potential of exogenously applied brassinosteroids as a valuable tool in phytoremediation projects by ameliorating heavy metal stress

Genetic variation between hybrid (Labeo rohita ♂, Cirrhinus mrigala ♀ and Labeo rohita) by RAPD marker

Abstract The present work was to study the genetic variability between the major carps Labeo rohita and Cirrhinus mrigala and their hybrids of L. rohita (male♂) and C. mrigala (female♀). Genetic variability was studied by employing RAPD molecular markers. 25 samples of each target species having different sizes with the same age group for the determination of interspecific variation were collected. The morphometric parameters such as body weight, total length, tail length, and lengths of dorsal and anal fins of each individual were recorded and results showed that wet body weight, total length, dorsal fin, anal fin, and tail fin length are positively correlated and then the DNA was extracted using the inorganic salt-based method and conformed by Gel electrophoresis. Twenty-four arbitrary decamer primers were used to get species-specific RAPD analysis Distinct and highly reproducible RAPD profiles with significant genetic variability was detected among species. Only five primers showed amplification. The RAPAD primer OPB-05 produced a total of seven bands out of these 5 monomorphic and 2 polymorphic, so in this case, the percentage polymorphism was 28.57%. The Hybrid show more than a 50% difference from the Labeo rohita. This shows that the Hybrid more resembles C.mrigala. Phylogenetic analysis demonstrated that hybrid (L. rohita ♂ X Cirrhinus mrigala ♀) is the closest to C. mrigala and the farthest from L. rohita. Overall data are presented concerning the applications of RAPD markers for hybrid identification, genetic diversity assessment, and studying taxonomic relationships at a molecular level

Comparative transcriptome analysis provides novel insights into molecular response of salt-tolerant and sensitive polyembryonic mango genotypes to salinity stress at seedling stage

IntroductionIncreased soil salinity in the recent years has adversely affected the productivity of mango globally. Extending the cultivation of mango in salt affected regions warrants the use of salinity tolerant/resistant rootstocks. However, the lack of sufficient genomic and transcriptomic information impedes comprehensive research at the molecular level. MethodWe employed RNA sequencing-based transcriptome analysis to gain insight into molecular response to salt stress by using two polyembryonic mango genotypes with contrasting response to salt stress viz., salt tolerant Turpentine and salt susceptible Mylepelian.ResultsRNA sequencing by Novaseq6000 resulted in a total of 2795088, 17535948, 7813704 and 5544894 clean reads in Mylepelian treated (MT), Mylepelian control (MC), Turpentine treated (TT) and Turpentine control (TC) respectively. In total, 7169 unigenes annotated against all the five public databases, including NR, NT, PFAM, KOG, Swissport, KEGG and GO. Further, maximum number of differentially expressed genes were found between MT and MC (2106) followed by MT vs TT (1158) and TT and TC (587). The differentially expressed genes under different treatment levels included transcription factors (bZIP, NAC, bHLH), genes involved in Calcium-dependent protein kinases (CDPKs), ABA biosynthesis, Photosynthesis etc. Expression of few of these genes was experimentally validated through quantitative real-time PCR (qRT-PCR) and contrasting expression pattern of Auxin Response Factor 2 (ARF2), Late Embryogenesis Abundant (LEA) and CDPK genes were observed between Turpentine and Mylepelian.DiscussionThe results of this study will be useful in understanding the molecular mechanism underlying salt tolerance in mango which can serve as valuable baseline information to generate new targets in mango breeding for salt tolerance