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Lipoid proteinosis is a rare multisystem genodermatosis inherited as autosomal recessive trait. We report a case of lipoid proteinosis in a 10-year-old boy born to first-degree consanguineous parents presented with marked hoarseness of voice, accelerated photoaging appearance, enlarged and erythematous tongue with restricted movement and widespread dermatoses. Biopsy of oral mucosa revealed Periodic acid-Schiff (PAS)-positive amorphous eosinophilic hyaline deposits. Mutational analysis revealed a homozygous nonsense mutation with C to T substitution at nucleotide position 1246(c.1246C>T) in exon-8 of the extracellular matrix protein 1 gene leading to a stop codon. Both the parents were unaffected heterozygous carriers. To our knowledge, this is the first case report of lipoid proteinosis with evidence of a novel nonsense genetic mutation from Bangladesh. doi: https://doi.org/10.12669/pjms.39.4.7437 How to cite this: Khan MAH, Reza MA, Sharaf IM, Alam MJ, Rahman MM, Chandra P, et al. Lipoid Proteinosis: Identification of a Novel Nonsense Mutation c.1246C>T:p.R416X in ECM1 gene from Bangladesh. Pak J Med Sci. 2023;39(4):---------. doi: https://doi.org/10.12669/pjms.39.4.7437 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Eco-friendly plasma technology is appeared as promising for the improvement of crop yield and hence it was applied to study the paddy plant growth and yield. Applications of plasma technology in this study are two-fold: (1) paddy seeds (Oryza sativa L.) were treated with low pressure (100 torr) glow air discharge (LPGAD) plasma for duration of 30, 60, 90, 120 and 150 s for finding the highest germination rate for field application, and (2) plasma activated waters (PAWs) were prepared and applied as foliar spray to the plants grown from the treated seeds to investigate the combined effects on plant growth, yield and total soluble protein and sugar concentrations in the produced paddy grains. Seedlings grown from the 90 s LPGAD plasma treated seeds, depending on the results obtained from seed germination test, were transplanted in the field and PAWs were applied 1–5 times during vegetative growth stage of the paddy plants. The results reveal that (a) the maximum paddy seed germination rate of ~ 7% with respect to control was obtained from 90 s treatment duration, out of five treatment durations, with LPGAD plasma, (b) plants growth parameters were enhanced due to the combined effects of plasma seed treatment along with PAW application, (c) defense mechanisms of plants were improved through enhancement of enzymatic activities, (d) concentrations of total soluble protein and sugar were enhanced in the paddy grains, and (e) finally yield was increased by ~ 16.67%.

Citrus macroptera, an indigenous fruit in Bangladesh, is usually used for folk treatment and culinary purposes. Considering its application context, this study was designed to detect and quantify t...

Medicinal plant-derived bioactive compounds have recently gained more interest in biological research as an important source of novel drug candidates. Phyllanthus acidus (L.) is a widely distributed herbal medicinal plant naturally used in Ayurvedic medicine in Bangladesh. The present study focused on exploring the biological potential as well as the inhibitory effect of EAC cell growth with a comparative analysis between Phyllanthus acidus fruit pulp and seed. Crude methanol extract of P. acidus (MEPA) fruit pulp and seed was assessed as DPPH and NO free radical scavengers. While Brine Shrimp lethality bioassay, the standard protocol of phytochemical screening and hemagglutination assay were performed successively to determine the toxic effect on normal cells, the identification of some crucial phytochemicals, and the existence of lectin protein. EAC (Ehrlich’s Ascites Carcinoma) cell growth inhibition was determined by hemocytometer and morphological changes of EAC cells were observed by a fluorescence microscope using Swiss albino mice. The IC50 value of MEPA fruit pulp and seed was obtained as 57.159 µg/ml and 288.743 µg/ml respectively where minimal toxic effects on Brine Shrimp nauplii demonstrates that it is a good source of natural antioxidant compounds. Again, MEPA fruit pulp and seed-mediated effective agglutination of mouse blood erythrocyte strongly support the presence of lectin protein. Furthermore, MEPA fruit pulp and seed extract-treated EAC cells showed 65.71% and 28.57% growth inhibition respectively. The fluorescent microscopic examination of EAC cells treated with MEPA fruit pulp has shown more remarkable structural changes in the nucleus than that of seed. Based on the above findings, the present study reveals that MEPA fruit pulp can be considered as a novel biological candidate for the treatment of fatal diseases shortly.

Apoptosis plays a pivotal role in the exclusion of abnormal cells without any ruin of surrounding healthy cells. Generally, it occurs through an orderly and autonomously process which is controlled by proper function of various genes. Therefore, the current experiments detect the expression level/pattern of those genes to confirm the involvement of extrinsic and intrinsic pathway using Basella alba leaf (BAL). Several fractions after gel filtration chromatography of BAL extract have been pooled to evaluates its apoptosis induction potentiality on Ehrlich's Ascites Carcinoma (EAC) cells through conducting a number of bio-assays such as cell growth inhibition assay, fluorescence and optical microscopy, DNA fragmentation assay and gene expression analysis etc. The pooled fractions of BAL showed 12-56% inhibitory effect on EAC cell line at the concentration range of 25-400 μg/ml that was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. They also exhibited excellent cell growth inhibition at in vivo and in vitro condition when treated with 10, 20 and 40 mg/kg day. After administration of six consequent days, significant morphological features of apoptosis were observed in EAC cells under both fluorescence and optical microscope which was further supported by DNA fragmentation assay. The polymerase chain reaction amplification of bax, bcl-2 (B-cell lymphoma 2), p53, tumor necrosis factor-α, Fas, NF-kβ (Nuclear factor-Kappa-B), PARP-1 (Poly (ADP-ribose) polymerase), Cyt-c cas-8, cas-9 and cas-3 revealed that the experimental sample able to induce apoptosis in both extrinsic and intrinsic pathways through altering the gene expression. The current findings suggest that sample from BAL occupy wonderful competence to induce cell apoptosis and become an ideal resource for cancer treatment.

We report the draft genome sequence of Klebsiella pneumoniae strain BASUSDALSc45PDB48, isolated from pesticide-contaminated soil; this strain showed the ability to grow in a medium with cypermethrin as the only carbon source. The genome assembly comprised 5,249,704 bp, with 128.17 Ns per 100 kbp, an N 50 value of 5,035,968 bp, a GC content of 57.57%, and 5,349 annotated genes.

This study was conducted to evaluate the toxic effects of an azo dye carmoisine widely used in foods and to investigate its relation to carcinogenicity. Carmoisine administered into mice orally in four different doses as control, low, medium, and high equivalent to 0, 4, 200, and 400 mg/kg bw, respectively, for 120 days. The key toxicological endpoint was observed including animal body weight, organ weights, hematology, biochemistry, and molecular biology assessment. The body weights of medium‐ and high‐dose carmoisine‐treated mice group were significantly decreased as compared to the control mice group. Platelet, white blood cell and monocyte counts of treated group were considerably higher, while Hb and red blood cell counts were drastically lower than the control group. The biochemical parameters such as serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total protein, globulin, urea, and creatinine level were significantly increased, while serum cholesterol level was decreased after treatment as compared to the control. RT‐PCR results showed that expression of Bcl‐x and PARP gene was intensively increased, whereas expression of p53 gene was decreased in the mouse liver tissues treated with carmoisine. This study revealed that high‐dose (400 mg/kg bw) treatment of carmoisine was attributable to renal failure and hepatotoxicity. It also would be suspected as a culprit for liver oncogenesis.

Targeting apoptosis is a promising approach to inhibit the abnormal cell proliferation of cancer progression. Existing anti-apoptotic drugs, many derived from chemical substances, have often failed to combat cancer development and progression. Therefore, identification of apoptosis-inducing anticancer agents from plant-derived sources has become a key aim in cancer research. The present study was designed to explore the regulation of apoptosis by Tabebuia pallida (T. pallida) using an Ehrlich Ascites Carcinoma (EAC) mouse model and compositional analysis by LC-ESI-MS/MS. Dried and powdered T. pallida leaves (TPL), stem bark (TPSB), root bark (TPRB) and flowers (TPF) were extracted with 80% methanol. Using cultured EAC cells and EAC-bearing mice with and without these extracts, anticancer activities were studied by assessing cytotoxicity and tumor cell growth inhibition, changes in life span of mice, and hematological and biochemical parameters. Apoptosis was analyzed by microscopy and expression of selected apoptosis-related genes (Bcl-2, Bcl-xL, NFκ-B, PARP-1, p53, Bax, caspase-3 and -8) using RT-PCR. LC-ESI-MS analysis was performed to identify the major compounds from the most active extracts. In EAC mice compared with untreated controls, the TPL extract exhibited the highest cytotoxicity with significant tumor cell growth inhibition (p< 0.001), reduced ascites by body weight (p< 0.01), increased the life span (pT. pallida possesses significant anticancer activity. The tumor suppressive mechanism is due to apoptosis induced by activation of antioxidant enzymes and caspases and mediated by a change in the balance of pro- and anti-apoptotic genes that promotes cell death.Graphical Abstract

In this present study conducted with the LFGD (Low-Frequency Glow Discharge) (Ar + O2) plasma treated maize seeds, to inspect the effect on seed surface modifications, seed germination, growth, development, productivity and nutritional compositions of maize plants. This study reported that LFGD (Ar + O2) plasma treated maize seeds have a potential effect to change its smooth seed surfaces and, it becomes rougher. It also enhances the seed germination rate up to (15.88%), which might help to increase the shoot length (33.42%), root length (10.67%), stem diameter (13.37%), total chlorophyll content (46.93%), total soluble protein (52.48%), total soluble phenol (21.68%) and sugar (1.62%) concentrations in respect controls of our experimental plants. For this reason, the acceptable treatment duration for maize seeds were 30sec, 60sec, 90sec and 120sec. After treatment, the plants exhibited a significant increase in CAT, SOD, APX and GR activities in the leaves and roots, and also significantly changes in H2O2 (208.33 ± 5.87μ molg−1 FW) in the leaves and (61.13 ± 1.72μ molg−1 FW) in the roots, NO was (369.24 ± 213.19μ molg−1FW) and (1094.23 ± 135.44μ molg−1FW) in the leaves and roots. LFGD plasma treatment also contributed to enhancement of productivity (1.27%), nutritional (moisture, ash, fat, and crude fiber) compositions, and iron and zinc micro-nutrition concentrations of maize. From this research, LFGD (Ar + O2) plasma treatment showed a potential impact on the maize cultivation system, which is very effective tools and both in nationally and internationally alter the conventional cultivation system of maize. Because it promotes seed surface modification, improved germination rate, shoot length, root length, chlorophyll content, some of the growths related enzymatic activity, nutrient composition, iron, and zinc micro-nutrients and the productivity of maize., Maize (Zea mays L.); LFGD plasma; Nutrition; Iron; Germination

Highlights • The screening of biological activities of medicinal plants are emphasized due to the adverse effects of synthetic drugs. • The present study demonstrates antioxidant, cytotoxic, and haemagglutination activity as well as EAC cell growth inhibitory effect of methanolic extract of Phyllantus acidus (L.) (MEPA) fruit pulp and seed. • P. acidus is traditionally used as herbal medicine in China, Brazil, and South Asian countries like Bangladesh for the treatment of digestive, respiratory, genitourinary, and skin disorder. • The result of this study suggested that MEPA fruit pulp has a more antioxidant potentiality and EAC cell growth inhibitory property than seed which indicates the presence of effective phytochemicals in MEPA fruit pulp. • Therefore, the MEPA fruit pulp can be considered as a promising biological source to search novel compounds for therapeutic purposes., Medicinal plant-derived bioactive compounds have recently gained more interest in biological research as an important source of novel drug candidates. Phyllanthus acidus (L.) is a widely distributed herbal medicinal plant naturally used in Ayurvedic medicine in Bangladesh. The present study focused on exploring the biological potential as well as the inhibitory effect of EAC cell growth with a comparative analysis between Phyllanthus acidus fruit pulp and seed. Crude methanol extract of P. acidus (MEPA) fruit pulp and seed was assessed as DPPH and NO free radical scavengers. While Brine Shrimp lethality bioassay, the standard protocol of phytochemical screening and hemagglutination assay were performed successively to determine the toxic effect on normal cells, the identification of some crucial phytochemicals, and the existence of lectin protein. EAC (Ehrlich’s Ascites Carcinoma) cell growth inhibition was determined by hemocytometer and morphological changes of EAC cells were observed by a fluorescence microscope using Swiss albino mice. The IC50 value of MEPA fruit pulp and seed was obtained as 57.159 µg/ml and 288.743 µg/ml respectively where minimal toxic effects on Brine Shrimp nauplii demonstrates that it is a good source of natural antioxidant compounds. Again, MEPA fruit pulp and seed-mediated effective agglutination of mouse blood erythrocyte strongly support the presence of lectin protein. Furthermore, MEPA fruit pulp and seed extract-treated EAC cells showed 65.71% and 28.57% growth inhibition respectively. The fluorescent microscopic examination of EAC cells treated with MEPA fruit pulp has shown more remarkable structural changes in the nucleus than that of seed. Based on the above findings, the present study reveals that MEPA fruit pulp can be considered as a novel biological candidate for the treatment of fatal diseases shortly.

Bioremediation through biodegradation is applied for cleaning up several environmental pollutions including petroleum oil spill containing petrol, diesel, mobil, kerosene, lubricating, etc. which have devastated several endangered terrestrial and aquatic ecosystems. Therefore, the current research was aimed to isolate and identify diesel degrading bacteria from the petroleum waste dumping site and determined their degrading efficiency. The bacterial strains were isolated through a minimum salt medium supplemented with 2% diesel as the sole carbon source. The bacteria were identified by morphological, biochemical characterization, and 16S rRNA gene sequencing. The optimized growth pattern was evaluated by utilization of a wide range of temperatures (25, 30, 35, and 40 °C) and pH (5,6,7 and 8) as well as different concentrations of diesel (2, 3, 5and 7%). Finally, the degradation rate was determined by measuring the residual diesel after 7, 14, and 21 days of incubation. The study isolated Enterobacter ludwigii, Enterobacter mori, Acinetobacter baumannii, and Cedecea davisae where all are gram-negative rod-shaped bacilli. All the bacterial strains utilized the diesel at their best at 30 °C and pH 7, among them, A. baumannii and C. davisae exhibited the best degrading efficiency at all applied concentrations. Finally, the determination of degradation rate (%) through gravimetrical analysis has confirmed the potency of A. Baumannii and C. davisae where the degradation rate was around 61 and 52% respectively after 21 days of incubation period with 10% diesel. The study concludes that all of those isolated bacterial consortiums, especially A. baumannii and C. davisae could be allocated as active agents used for bioremediation to detoxify the diesel-containing contaminated sites in a cost-effective and eco-friendly way.

Medicinal plant extract has long been used successfully in ‘unani’ or ‘ayurvedic’ medicine. Medicinal plant extract contains bioactive molecules and activity of these molecules may help to mitigate, eradicate or cure diseases. In the advent for the search for new medicinally important bioactive molecule, the current paper deals with the anti-oxidative, cytotoxic and phytochemical analysis of Scoparia dulcis leaf and root, Curculigo orchioides root, Pandanus fascicularis root and Baccaurea sapida leaf extract. Aqueous and methanolic extracts were made for each of the extracts where they posses significant antioxidative properties. High activities were seen in P. fascicularis and S. dulcis plant extract where IC50 values were 21.87 μg/ml and 173.36 μg/ml respectively. In toxicity test, only P. fascicularis extracts showed lethality in a dose-dependent manner where the LD50 value was 25.64 μg/ml. By the phytochemical analysis, it was found that each of these plant species possesses glycosides, protein, carbohydrates, alkaloid, flavonoid etc. which are pharmacologically active biomolecules. These important properties of those plants showed an indication that these plants can further be tested for the utilization in therapeutic purpose or in cosmetic industry.J. bio-sci. 25: 27-37, 2017

Excessive use of pesticides in agricultural fields is a matter of great concern for living beings as well as the environment across the world, in particular, the third world countries. Therefore, there is an urgent need to find out an effective way to degrade these hazardous chemicals from the soil in an environment-friendly way. In the current project, a bacterial species were isolated through enrichment culture from carbofuran-supplemented rice-field soil and identified as a carbofuran degrader. The rate of carbofuran degradation by this bacterial species was evaluated using reverse-phase high-performance liquid chromatography (RP-HPLC), which confirmed the ability to utilize as a carbon source up to 4 µg/ml of 99% technical grade carbofuran. The morphological, physiological, biochemical characteristics and phylogenetic analysis of the 16S rRNA sequence showed that this strain belongs to the genus of Enterobacter sp. (sequence accession number LC368285 in DDBJ), and the optimum growth condition for the isolated strain was 37°C at pH 7.0. Moreover, an antibiotic sensitivity test showed that it was susceptible to azithromycin, penicillin, ceftazidime, ciprofloxacin, and gentamycin, and the minimal inhibitory concentration value of gentamycin was 400 μg/ml against the bacteria. It shows beyond doubt from the RP-HPLC quantification that the isolated bacterium has the ability to detoxify carbofuran (99% pure). Finally, the obtained results imply that the isolated strain of Enterobacter can be used as a potential and effective carbofuran degrader for bioremediation of contaminated sites through bioaugmentation.

Cadmium (Cd) is detrimental to crops and the environment. This work examines the natural mechanisms underlying silicon- (Si-)directed Cd detoxification in rice plants. The addition of Si to plants under Cd stress caused significant improvements in morphological parameters, chlorophyll score, Fv /Fm and total soluble protein concentration compared to controls, confirming that Si is able to ameliorate Cd-induced damage in rice plants. This morpho-physiological evidence was correlated with decreased cell death and electrolyte leakage after Si application. The results showed no critical changes in root Cd concentration, while shoot Cd decreased significantly after Si supplementation in comparison with Cd-stressed rice. Additionally, expression of Cd transporters (OsNRAMP5 and OsHMA2) was significantly down-regulated while the concentration of phytochelatin, cysteine and glutathione, together with expression of OsPCS1 (phytochelatin synthase) in roots of Cd-stressed rice was significantly induced when subjected to Si treatment. This confirms that the alleviation of Cd stress is not only limited to the down-regulation of Cd transporters but also closely related to the phytochelatin-driven vacuolar storage of Cd in rice roots. The enzymatic analysis further revealed the role of SOD and GR enzymes in protecting rice plants from Cd-induced oxidative harm. These findings suggest a mechanistic basis in rice plants for Si-mediated mitigation of Cd stress.

A series of programmed cell death that plays a vital role in the exclusion of abnormal cells without any ruin into the surrounding neighboring cells is called Apoptosis. Generally, it occurs in multi-cellular organisms through an orderly and autonomously processes that is controlled by proper function of various genes. In the current studies, cell apoptosis in EAC cells treated with different fractions (BLP-01, BLP-02 and BLP-03) of leaf ex- tract of B. alba was detected by conducting several bio-assays such as cell growth inhibition, fluorescence and optical microscopy, DNA fragmentation and PCR amplification etc. The results of these experiments indicate that the plant extracts are able to inhibit cell growth significantly where morphological features of apoptosis were appeared under both fluorescence and optical microscope. The PCR amplification results showed that the leaf ex- tracts of B. alba were able to cause EAC cell apoptosis in both the extrinsic and intrinsic pathway. An excellent figure of fragmented DNA was found in DNA fragmentation assay when the gel was observed under UV light which confirms the cell apoptosis. The current findings suggest that the samples of this experiment occupy fascinating competence to conduct cell apoptosis and become an ideal resource for cancer research as well as drugs development for cancer treatment.

Tumor necrosis factor-alpha (TNF-α) is a major cytokine for inflammatory response in human body. This is also well linked with obesity and causing different pathophysiological problems in type 2 diabetic mellitus (T2DM) patients because of its pro-inflammatory over expression. However, seemingly harmless nucleotide changes in the promoter region often cause oscillation in expression, results in complications like dyslipidemia and atherosclerosis that ultimately exploit to coronary heart disease (CHD). Therefore, this study was designed to evaluate association between TNF-α (-308G>A) polymorphism at promoter region and CHD in T2DM patients from the northern region (Rajshahi) of Bangladesh. The total number of participants was 96 T2DM patients. TNF-α polymorphism were detected using high resolution melting (HRM) curve analysis. A total of 32 participants were suffering from CHD and 8 polymorphism (3 homozygous and 5 heterozygous) were detected among them. From Fishers Exact test, we found significant (P < 0.05) relationship between TNF-α polymorphism and CHD in T2DM patients. According to Kendalls Tau correlation matrix (r = 0.218), there is a good correlation between polymorphism and CHD. Therefore, the overall results suggest that TNF-α (-308G>A) polymorphism is responsible for inducing CHD in T2DM patients. [ J Adv Biotechnol Exp Ther 2021; 4(1.000): 60-66]

Effects of black gram (vigna mungo L.cv. Barimash 3) seed treatments with 400 torr dielectric barrier discharge (DBD) air plasma on seed surface morphology, seed germination, seedling growth and antioxidant enzyme activities in the roots, shoots and leaves were investigated. The plasma discharge voltage, frequency, electrode spacing, gas temperature and power were 5kV, 4.5kHz, 60mm, 310K and 45W, respectively. The seeds were treated for the duration ranging from 20 to 180 s. Seed germination rate, seedling growth, total chlorophyll content, total soluble protein and sugar concentrations in the seedlings grown from the treated seeds were found to increase 13.67%, 37.13%, 37.26%,53.60% and 51.71%, respectively, with respect to control. This study reveals that the DBD air plasma was involved in the enhancement of nitrogen complex in the seed coat of black gram which upregulated the protein through nitrogen conversion that was ultimately responsible for the increased seed germination and seedling growth of black gram.

This study investigates the effect and mechanisms of low pressure dielectric barrier discharge (LPDBD) produced with Ar/O2 and Ar/Air technique causing biological stimulation leading to improved germination and growth in wheat. Both plasma treatments caused rougher and chapped seed surface along with noticeable improvement in seed germination in wheat. Beside this, seed H2O2 concentration significantly increased compared to controls subjected to Ar/O2 and Ar/Air while this phenomenon was more pronounced due to Ar/Air plasma. Analysis of plants grown from the plasma treated seeds showed significant improvement in shoot characteristics, iron concentration, total soluble protein and sugar concentration in comparison with the controls more efficiently due to Ar/O2 plasma than that of Ar/Air. Further, none of the plasma treatments caused membrane damage or cell death in root and shoot of wheat. Interestingly, Ar/O2 treated plants showed a significant increase (2-fold) of H2O2 compared to controls in both root and shoot, while Ar/Air plasma caused no changes in H2O2. This phenomenon was supported by the biochemical and molecular evidence of SOD, APX and CAT in wheat plants. Plants derived from Ar/O2 treated seeds demonstrated a significant increase in SOD activity and TaSOD expression in roots of wheat, while APX and CAT activities along with TaCAT and TaAPX expression showed no significant changes. In contrast, Ar/Air plasma caused a significant increase only in APX activity in the shoot. This suggests that Ar/O2 plasma caused a slight induction in H2O2 accumulation without triggering the H2O2 scavengers (APX and CAT) and thus, efficiency affect growth and development in wheat plants. Further, grafting of control and Ar/O2 treated plants showed a significant increase in shoot biomass and H2O2 concentration in grafts having Ar/O2 rootstock regardless of the type scion attached to it. It indicates that signal driving Ar/O2 plasma mediated growth improvement in wheat is possibly originated in roots. Taken together, this paper delivers new insight into the mechanistic basis for growth improvement by LPDBD technique.

BMPR1A (BMP type 1 receptor) is a transmembrane cell-surface receptor also known as ALK3 (activin-like kinases-3) encodes for a type I serine/threonine kinase receptor and a member of the transforming growth-factor β–receptor (TGF-β) super family. The BMPR1A has a significant interaction with BMP-2 for protein activity and also has a low affinity with growth and differentiation factor 5 (GDF5); positively regulates chondrocyte differentiation. The genetic variations can alter the structure and function of the BMPR1A gene that causes several diseases such as juvenile polyposis syndrome or hereditary cancer-predisposing syndrome. The current study was carried out to identify potential deleterious non-synonymous single nucleotide polymorphisms (nsSNPs) in BMPR1A by implementing different computational algorithms such as SIFT, PolyPhen2, SNAP2, PROVEAN, PhD-SNP, SNPs&GO, nsSNPAnalyzer, and P-Mut. From 205 nsSNPs in BMPR1A, 7 nsSNPs (C76Y, C124R, C124Y, C376Y, R443C, R480W, and W487R) were predicted as deleterious in 8 prediction algorithms. The Consurf analysis showed that selected 7 nsSNPs were present in the highly conserved regions. Molecular dynamics simulation analysis also performed to explore conformational changes in the variant structure with respect to its native structure. According to the MDS result, all variants flexibility and rigidity were unbalanced, which may alter the structural and functional behavior of the native protein. Although, three nsSNPs i.e., C124R, C376Y, and R443C have already been reported in patients associated with JPS, but their structural and functional molecular studies remain uncharacterized. Therefore, the findings of this study can provide a better understanding of uncharacterized nsSNPS and to find their association with disease susceptibility and also facilitate to the researchers for designing or developing the target dependent drugs.

Summary. Background: The evolution of structurally and functionally similar proteins with highly diverse physiological roles within a single organism is of great interest. Australian elapid snakes offer an excellent opportunity to study the molecular evolution of prothrombin activators. Venom from Pseudonaja textilis contains pseutarin C, a group C prothrombin activator. Its enzymatic subunit is structurally and functionally similar to mammalian factor (F) Xa, whereas its non-enzymatic subunit is similar to FVa. As vertebrates, the snakes also contain a system to activate prothrombin in their own blood during injury. These hemostatic factors are produced in the liver. Results: Here we describe the presence of two molecular forms of FX expressed in the liver of P. textilis. Both isoforms have molecular signatures and domain architecture of FX. However, one isoform shows ∼94% sequence identity with the snake FX from Tropidechis carinatus, whereas the other is much closer (90% identity) to the catalytic subunit of pseutarin C (PCCS). Real-time polymerase chain reaction reveals that the latter isoform is expressed ∼56 000 times lower in the liver of P. textilis. However, the isoforms are not expressed in the venom gland. Conclusion: A detailed analysis of deletions and insertions along with the sequence indicates that the second isoform is an intermediate caught in the evolution of venom prothrombin activator from the blood coagulation FX. Thus, this isoform represents a ‘molecular fossil’ and reveals the likely evolutionary path of recruitment of FX in the venom gland.

Anticancer activities of p-menth-1-ene-4,7-diol (EC-1) isolated from Eucalyptus camaldulensis Dhnh. were studied on Ehrlich ascites carcinoma (EAC) cells by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. Anticancer activities also analyzed in EAC-bearing mice by assessment of cancer growth inhibition, changes in cancer volume, changes in life span, and hematological parameters. Apoptosis was analyzed by fluorescence microscope, DNA fragmentation assay, and flow cytometry. The expression of apoptosis-related genes, Bcl-2, Bcl-X, PARP-1, p53, and Bax, were analyzed using polymerase chain reaction (PCR). EC-1 significantly inhibited proliferation of EAC cells in vivo and restored the altered hematological parameters of EAC-bearing mice. Cytological observation by fluorescence microscope showed apoptosis of EAC cells upon treatment with EC-1. Also, DNA fragmentation assay revealed EAC cells' apoptosis following EC-1 treatment. Increased mRNA expressions of p53 and Bax genes and negative expressions of Bcl-2 and Bcl-X were observed in cells treated with EC-1. These findings confirmed the induction of apoptosis by EC-1. In addition, MTT assay showed dose-dependent anticancer activity of EC-1 against EAC cell. Cell cycle analysis revealed that EC-1 treatment caused suppression of EAC cells at S phase. To conclude, EC-1 is a novel anticancer compound and showed antiproliferative and apoptotic activities in cellular and mice models.

This work reports the DC characteristics of dual gated large area graphene metal oxide semiconductor field effect transistor (MOSFET). The sheet charge density dependent quantum capacitance is obtained self-consistently with considering the impurities concentration of the gate oxide layer. The potential profile as well as sheet charge density of graphene channel is calculated. The C-V and I-V characteristics are illustrated here. Finally, the velocity-field relation is shown.

Pea ( Pisum sativum L.) lectin is known to have interesting pharmacological activities and of great interest on biomedical research. In the current research pea lectin was purified followed by ion exchange chromatography on DEAE column and affinity chromatography on glucose-sepharose column. The lectin shown 11.7–84% inhibitory effect against Ehrlich ascites carcinoma (EAC) cells at the concentration range of 8–120 μg/ml in RPMI 1640 medium as determined by MTT assay. Pea lectin was also shown 63% and 44% growth inhibition against EAC cells in vivo in mice when administered 2.8 mg/kg/day and 1.4 mg/kg/day (i.p.) respectively for five consequent days. When Pea lectin injected into the EAC bearing mice for 10 days its significantly increased the hemoglobin and RBC with the decreased of WBC levels toward the normal. Apoptotic cell morphological change of the treated EAC cells of mice was determined by fluorescence and optical microscope. Interestingly, cell growth inhibition of the lectin was significantly reduced in the presence of caspase inhibitors. Treatment with the lectin caused the cell cycle arrest at G 2 /M phase of EAC cells which was determined by flow cytometry. The expression of apoptosis-related genes, Bcl-2 , Bcl-X and Bax was evaluated by reverse transcriptase polymerase chain reaction (RT-PCR). Intensive increase of Bax gene expression and totally despaired of Bcl-2 and Bcl-X gene expression were observed in the cells treated with Pea lectin for five consecutive days.

Snake venom is a mixture of various proteins and peptides with distinct pharmacological properties. They have the unique aptitude to attack specific physiological systems of the prey with remarkable specificity and thereby obstruct the natural function culminating in death and debilitation of the victim. The origin of snake venom toxins is one of the most interesting question that intrigued scientists for a long time. In this chapter, we will discuss the origin of a particular group of venom proteins, namely prothrombin activators. The results from our recent studies provide molecular evidence that this class of deadly toxins have evolved by gene duplication and recruitment of blood coagulation factor gene. Thus a new toxin has evolved after recruitment from a simple body protein that is involved in its own lifesaving haemostasis.

Summary. Background: Proteins with new function originate through gene duplication followed by divergence. In nature, occurrence of structurally and functionally similar proteins performing highly diverse physiological roles within an organism is rare. Several Australian elapid snakes have two parallel prothrombin activating systems with distinct physiological roles. For example, in Tropidechis carinatus, trocarin D exists in the venom and acts as toxin, whereas coagulation factor (F) X exists in plasma and plays a role in hemostasis. Results: Here, we show that FX and the trocarin D genes are expressed in a highly tissue-specific manner in T. carinatus. To understand their origin, recruitment and evolution, we determined the complete structure of their genes. Both genes have eight exons with identical exon–intron boundaries. All the introns are 92–99% identical with the exception of intron 1, indicating a recent gene duplication event. The first intron of the trocarin D gene is also nearly identical to that of the FX gene, except for two deletions (255 and 1406 bp) and three insertions (214, 1975, and 2174 bp). The third insertion has a potential scaffold/matrix attached region. The putative promoter of the snake FX gene shares similar cis-elements compared with those of human and murine FX genes. Interestingly, the trocarin D promoter has a 264-bp insertion with core promoter sequences and cis-elements that are known to induce high-level expression. This insertion might be responsible for switching from constitutive expression of the FX gene to inducible expression of trocarin D. Thus, we named this segment as VERSE (Venom Recruitment/Switch Element). Conclusion: To our knowledge, this is the first molecular evidence for the recruitment of a duplicated gene for expression in venom glands by a simple insertion.

SummaryIt is uncommon for similar pathways/systems to be involved in highly divergent functions within single organisms. Earlier, we have shown that trocarin D, a venom prothrombin activator, from the Australian rough-scaled snake Tropidechis carinatus, is structurally and functionally similar to the blood coagulation factor Xa (FXa). The presence of a haemostatic system in these snakes implies that they have two parallel prothrombin activating systems: one in the plasma, that participates in the life saving process of blood clotting and the other in their venom, where it acts as a toxin. Here, we report the complete cDNA sequence encoding the blood coagulation factor X (FX) from the liver of T. carinatus. Deduced T. carinatus FX sequence shows ~80% identity with trocarin D but ~50% identity with the mammalian FX. Our present study confirms the presence of two separate genes – one each for FX and trocarin D, that code for similar proteins in T. carinatus snake. These two genes have different expression sites and divergent uses suggesting that snake venom prothrombin activators have probably evolved by the duplication of the liver FX gene and subsequently marked for tissue-specific expression in the venom gland.