A review on fisheries and conservation status of Asian horseshoe crabs

Horseshoe crabs are the only extant xiphosurans and are believed to be morphologically unchanged for more than 200 million years. Of the four extant species namely, Limulus polyphemus, Tachypleus tridentatus, Tapinauchenius gigas and Carcinoscorpius rotundicauda, the latter three are found in Asian waters. Recent evidences showed that Asian horseshoe crabs are facing serious threats such as degradation of their spawning grounds and habitat, environmental pollution, overexploitation as a culinary delicacy and biomedical bleeding practices. Baseline data on the distribution and existing population of the wild horseshoe crabs remain poorly known in several Asian regions. Several studies have clearly revealed that pressure due to over-fishing of wild stock has increased tremendously in the last decade. Due to an increase in demand for Tachypleus Amebocyte Lysate (TAL) analogous to Limulus Amebocyte Lysate (LAL) in the United States, there is an urgent need to comprehensively address their fishing and conservation measures in the Asian region. This review addresses the overall studies on three species of Asian horseshoe crabs in relation to their fishing practices, local exploitation of their wild stock either for human consumption (or) by biomedical industries. The authors have structured the discussion on an international scale to address the existing problems in fishing and conservation of horseshoe crabs. Since no specific regulatory force or legislative protection act or a policy to preserve their natural stock are available to this date, this paper strongly recommends representative countries to include horseshoe crabs under their wildlife protection act to avoid further unsustainable exploitation of their wild populations.     

Expression of fibroblast growth factor receptor-1 in rat heart H9c2 myoblasts increases cell proliferation

Aspergillus fumigatus is a highly pathogenic fungus causing a wide spectrum of diseases in immunocompromised as well as immunocompetent hosts. The present work was undertaken to evaluate the cytotoxic nature of fractionated antigens of A. fumigatus against the mammalian cell lines (J774, RAW, CHO and L929). An enriched protein antigenic fraction of A. fumigatus was subjected to con A Sepharose and phenyl Sepharose chromatography. Antigenic fractions, ConAub (conA unbound) and PSC III (fraction III of phenyl Sepharose column) containing low mw antigens showed higher cytotoxicity as compared to other antigenic fractions. PSC III was further purified on HPLC resulting in an 18 kDa homogeneous protein. The purified protein showed high ELISA absorbance values for specific IgG and IgE antibodies in sera of ABPA patients. Monoclonal antibody raised against Asp fl, a major allergen/antigen of A. fumigatus recognised the purified 18 kDa by ELISA and western blot. The 18 kDa allergen/antigen or Asp fl showed similar toxicity towards all the four cell lines (macrophage and fibroblast) with an IC50 of 75 ng/ml or 4.16 nM. Reduction in toxicity of 18 kDa at low temperatures and potentiation in presence of ammonium chloride and monensin indicates mechanism of internalisation of 18 kDa in eukaryotic cells is similar to -sarcin. The present work shows that the 18 kDa allergen/antigen (Asp fl) is a major cytotoxin secreted by A. fumigatus which may play multiple roles in the pathogenesis of Aspergillosis through allergenicity, antigenicity and cytotoxicity. (Mol Cell Biochem 167: 89-97, 1997)     

Re-usable DNA template for the polymerase chain reaction.

DNA covalently bound to an uncharged nylon membrane was used for consecutive amplifications of several different genes by PCR. Successful PCR amplifications were obtained for membrane-bound genomic and plasmid DNA. Membrane-bound genomic DNA templates were re-used at least 15 times for PCR with specific amplification of the desired gene each time. PCR amplifications of specific sequences of p53, p16, CYP1A1, CYP2D6, GSTM1 and GSTM3 were performed independently on the same strips of uncharged nylon membrane containing genomic DNA. PCR products were subjected to restriction fragment length polymorphism analysis, single-strand conformational polymorphism analysis and/or dideoxy sequencing to confirm PCR-amplified gene sequences. We found that PCR fragments obtained by amplification from bound genomic DNA as template were identical in sequence to those of PCR products obtained from free genomic DNA in solution. PCR was performed using as little as 5 ng genomic or 4 fg plasmid DNA bound to membrane. These results suggest that DNA covalently bound to membrane can be re-used for sample-specific PCR amplifications, providing a potentially unlimited source of DNA for PCR.     

A cytochemical study of acid phosphatases and esterases during the induction of crown gall in the tomatoLycopersicon esculentum

Synopsis A cytochemical study has been made of acid -glycerophosphatase, acid naphthol-AS-BI-phosphatase and naphthol esterase activities during the induction of crown gall inLycopersicon esculentum byAgrobacterium tumefaciens. There was an increased activity of these enzymes with time after infection, being more marked for the phosphatases than the esterases. It is suggested that the esterase activities may play a part in cell wall development of the maturing gall.     

Modern pollen assemblages from Reasi (Jammu and Kashmir), India: a tool for interpreting fossil pollen records

Twenty five surface samples/moss cushions were collected for palynological analysis from open areas of Reasi District, Jammu and Kashmir (India). These samples were used to investigate the relationships between extant vegetation and modern pollen spectra, which serve as modern analogue for the reliable ecological interpretation of fossil pollen records. The present vegetation in the region comprises tropical dry deciduous forests and subtropical pine forests with scattered stands of oak. The pollen analysis reveals that Pinus sp. (average 69% in the pollen assemblages), amongst the conifers, dominates the pollen rain, which can be attributed to its high pollen productivity and exceptional pollen dispersal efficiency. Cedrus sp. and Podocarpus sp. pollen contribute with an average of 16 and 5% to the total pollen rain. Other conifers such as Picea sp., Abies sp., Juniperus sp. and Tsuga sp., as well as broad-leaved taxa such as Quercus sp., Alnus sp., Betula sp., Carpinus sp., Corylus sp., Juglans sp., Ulmus sp., Salix sp., Elaeocarpus sp., Mallotus sp. and Aesculus sp., have lower averages of 1 to 4.5% in the total pollen rain which could be either due to their poor pollen dispersal efficiency or to the poor preservation in the samples. Tubuliflorae (average 25%), Poaceae (average 6.26%), Cerealia and other crop plants (average 7.68%) are other prominent taxa in the pollen rain. The nearly complete absence of members of tropical dry deciduous forests in the pollen spectra likely is due to the fact that most species in this vegetation type are not wind pollinated.     

Induced Homoeologous Pairing for Transfer of Useful Variability for High Grain Fe and Zn from <Emphasis Type="Italic">Aegilops kotschyi</Emphasis> into Wheat

Biofortification of bread wheat by the transfer of useful variability of high grain Fe and Zn from Aegilops kotschyi through induced homoeologous pairing is the most feasible approach to alleviate micronutrient malnutrition worldwide. Deficiency of chromosome 5B in interspecific hybrids allows homoeologous pairing and recombination of chromosomes of wheat with those of the related species. The interspecific hybrid plants without 5B chromosome showed much higher chromosome pairing than did the plants with 5B. The F₁ plants without 5B chromosome were selected and repeatedly backcrossed with wheat cultivar PBW343. The chromosome number of BC₂F₁ plants ranged from 43 to 60 with several univalents and multivalents. Molecular markers and GISH analysis confirmed the introgression of U/S chromosomes of Ae. kotschyi and their fragments in wheat. The BC₂F₂ plants showed up to 125 % increase in Fe and 158 % increase in Zn compared to PBW343 with Lr24 and Yr36. Induced homoeologous pairing in the absence of 5B was found to be an effective approach for transfer of useful variability for enhanced grain Fe and Zn content for biofortification of wheat for high grain micronutrient content.     

β-Caryophyllene,a phytocannabinoid attenuates oxidative stress,neuroinflammation, glial activation,and salvages dopaminergic neurons in a rat model of Parkinson disease

Parkinson disease (PD) is a neurodegenerative disease characterized by progressive dopaminergic neurodegeneration in the substantia nigra pars compacta (SNc) area. The present study was undertaken to evaluate the neuroprotective effect of β-caryophyllene (BCP) against rotenone-induced oxidative stress and neuroinflammation in a rat model of PD. In the present study, BCP was administered once daily for 4 weeks at a dose of 50 mg/kg body weight prior to a rotenone (2.5 mg/kg body weight) challenge to mimic the progressive neurodegenerative nature of PD. Rotenone administration results in oxidative stress as evidenced by decreased activities of superoxide dismutase, catalase, and depletion of glutathione with a concomitant rise in lipid peroxidation product, malondialdehyde. Rotenone also significantly increased pro-inflammatory cytokines in the midbrain region and elevated the inflammatory mediators such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in the striatum. Further, immunohistochemical analysis revealed loss of dopaminergic neurons in the SNc area and enhanced expression of ionized calcium-binding adaptor molecule-1 (Iba-1) and glial fibrillary acidic protein (GFAP), indicators of microglia activation, and astrocyte hypertrophy, respectively, as an index of inflammation. However, treatment with BCP rescued dopaminergic neurons and decreased microglia and astrocyte activation evidenced by reduced Iba-1 and GFAP expression. BCP in addition to attenuation of pro-inflammatory cytokines and inflammatory mediators such as COX-2 and iNOS, also restored antioxidant enzymes and inhibited lipid peroxidation as well as glutathione depletion. The findings demonstrate that BCP provides neuroprotection against rotenone-induced PD and the neuroprotective effects can be ascribed to its potent antioxidant and anti-inflammatory activities.

     

Targeting CoV-2 spike RBD and ACE-2 interaction with flavonoids of Anatolian propolis by in silico and in vitro studies in terms of possible COVID-19 therapeutics

Propolis is a multi-functional bee product rich in polyphenols. In this study, the inhibitory effect of Anatolian propolis against SARS-coronavirus-2 (SARS-CoV-2) was investigated in vitro and in silico. Raw and commercial propolis samples were used, and both samples were found to be rich in caffeic acid, p-coumaric acid, ferulic acid, t-cinnamic acid, hesperetin, chrysin, pinocembrin, and caffeic acid phenethyl ester (CAPE) at HPLC-UV analysis. Ethanolic propolis extracts (EPE) were used in the ELISA screening test against the spike S1 protein (SARS-CoV-2): ACE-2 interaction for in vitro study. The binding energy values of these polyphenols to the SARS-CoV-2 spike and ACE-2 protein were calculated separately with a molecular docking study using the AutoDock 4.2.6 program. In addition, the pharmacokinetics and drug-likeness properties of these eight polyphenols were calculated according to the SwissADME tool. The binding energy value of pinocembrin was highest in both receptors, followed by chrysin, CAPE, and hesperetin. Based on the in silico modeling and ADME (absorption, distribution, metabolism, and excretion) behaviors of the eight polyphenols, the compounds exhibited the potential ability to act effectively as novel drugs. The findings of both studies showed that propolis has a high inhibitory potential against the Covid-19 virus. However, further studies are now needed.     

The Effects of Dietary Selenium and Vitamin E and their Combination on the Fatty Acids of Erythrocytes,Bone Marrow and Spleen Tissue Lipids of Lambs

The object was to determine the influence of dietary vitamin E, selenium and their combination on the fatty acid con-tent of erythrocytes, bone marrow and spleen lipids of Akkaraman lambs. After supplementation for 15 days, the amount of all fatty acids was slightly higher (p < 0·05) in the vitamin E as compared to the control group, whereas the amount of longer fatty acids was significantly higher (p < 0·01, p < 0·001) in the selenium and combination groups. On the thirtieth day, the amount of all fatty acids was slightly high (p < 0·5) in all the supplemented groups in comparison with the control group. In the bone marrow lipids, the amount of longer fatty acids was decreased (p < 0·05, p < 0·01, p < 0·001) in the vitamin E and combination groups as compared to the control. Although the amount of some fatty acids was high (p < 0·05, p < 0·01) in the selenium group compared to the control, linoleic (18:2), linolenic (18:3) and the polyunsaturated fatty acids (PUFA) were lower (p < 0·05, p < 0·001). In the spleen lipids, the amount of longer fatty acids was slightly decreased (p < 0·05) in the vitamin E group as compared with the control; however the amount of longer fatty acids was significantly higher (p < 0·05, p < 0·01) in the selenium and combination groups in comparison to the control group. Thus dietary supplementation with selenium was more effective than dietary vitamin E supplementation in altering the fatty acid content of the erythrocyte, bone marrow and spleen lipids. © 1997 John Wiley & Sons, Ltd.