A Rapid Micropropagation of nodal explants of Eclipta alba (L.); A Multipurpose Medicinal Herb

An efficient in vitro regeneration protocol was developed for medicinally important plant Eclipta alba. Successful regeneration and multiplication of nodal explants of E. alba were obtained in cytokinin enriched B5 medium. Several cytokinins [6-benzylaminopurine (BAP), kinetin (KIN), thidiazuron (TDZ), gibberellic acid (GA3) and spermidine] were supplemented alone and its combinations for obtaining better results. The best growth frequency response was achieved in the combinations of 1.0 BAP + 0.3 KIN + 1.5 GA3 (mg/L) concentration (7.4 ± 0.9 cm shoot length & 100 % regeneration). Better roots were developed in half-strength B5 medium along with IBA (1.0 mg/L) hormone and exhibits maximum root length (7.0 ± 0.8cm) along with multiple roots (8.8 ± 0.8) at 92 %. The well-developed Plantlets were successfully acclimatized to plastic-cups containing autoclaved sand and garden soil (1:1) and kept undisturbed with plastic cover for maintaining the humidity. The plantlets were watered regularly and maintained at green house

Method Development and Validation of Antiretroviral Drugs in Bulk and Pharmaceutical Dosage Forms

INTRODUCTION: ANALYTICAL CHEMISTRY: Analytical chemistry1 is the branch of chemistry involved in separating, identifying and determining the relative amounts of the components making up a sample of matter. It is mainly involved in the qualitative identification or detection of compounds and the quantitative measurement of the substances present in bulk and pharmaceutical preparation. The newer methods for separating and determining chemical species are known collectively as instrumental methods of analysis. Most of the instrumental methods fit into one of the three following categories viz., spectroscopy, electrochemistry and chromatography. CHROMOGENIC REAGENTS USED IN THE PRESENT INVESTIGATION: Functional groups present in organic drugs determine the way of analyzing them because they are responsible for the properties of substances and determine the identification reaction and the methods of quantitative determination of drugs. Knowing the reactions for detecting functional groups, one can easily analyze any organic drug with a complicated structure. In the present investigation, few visible spectrophotometric methods have been developed for LMV and STV by developing colour in each case with, appropriate reagent. The analytically useful functional groups in the drug have not been exploited completely in developing the new visible spectrophotometric method and so, the drugs have been selected in the present investigation. Different type of reagents like Gibbs reagent, MBTH reagent and BPB reagent were used in the present investigation for developing visible spectrophotometric methods. SUMMARY: Several drugs are available in the form of pharmaceutical formulations to control diseases. Methods of assay for controlling the concentration of these chemicals in the medicine and in the living body are necessary. Pharmaceutical analysis occupies a pivotal role in statutory certification of drugs and their formulations either by the industry or by the regulatory authorities. The complexity of the problem encountered in pharmaceutical analysis coupled with importance of achieving the selectivity, speed, cost, simplicity, precision and accuracy results in new methods of analysis being quickly adopted by pharmaceutical industry. Formulations containing combinations of drugs for potentiating or complementing another in therapy are on the increase. In some cases, no precise analytical methods are reported and quite often the reported procedures need improvements or changes keeping in the view of the advances. Among several instrumental techniques (HPLC, GC, Fluorimetry, NMR, mass spectroscopy covering IR, UV and visible regions) available for assay of drugs, visible spectrophotometric methods depend only on the nature of chemical reaction utilized for colour development and not on sophistication of the equipment. GC method is highly selective and sensitive compared to spectroscopic or other chromatographic methods. GC method is also cost effective as expensive solvents are not required and it is a versatile tool for qualitative and quantitative analysis of drugs and pharmaceuticals. Due to the importance of analysis, present analytical method has been developed for some of the widely used antiretroviral drugs such as lamivudine, zidovudine, stavudine, nevirapine and efavirenz. Hence we planned o develop HPLC, GC and spectrophotometric methods. CONCLUSION: Antiretroviral drugs are medications for the treatment of infection by retroviruses, primarily HIV. When several such drugs, typically three or four, are taken in combination, the approach is known as highly active antiretroviral therapy, or HAART. The American National Institutes of Health and other organizations recommend offering antiretroviral treatment to all patients with AIDS. Although various UV-visible methods have been reported for the estimation of LMV, ZDV and STV it was found that the reagents used in the present study were not used and the methods developed are much sensitive and less time consuming compared to the methods previously develop. The simultaneous estimation of LMV, ZDV and EFZ with UV spectrophotometer using triple point method was not investigated. The work deals with four UV-Visible spectrophotometric methods i.e. oxidation reaction with Cerium (IV) ammonium sulphate, visible spectrophotometric method with BPB, GIBBS reagent and coupling reaction with MBTH reagent. The methods are validated in terms of sensitivity, accuracy and precision. A. Comparative Sensitivity 1> 2 > 3 > 4 B. Comparative accuracy: 3> 4 >2 > 1 C. Comparative precision: 3 > 4 > 1 > 2 Simultaneous estimation of LMV, ZVD and NVP by RP-HPLC was also developed. The retention times of the drugs were less when compared to other methods developed. In terms of sensitivity, accuracy and precision the present developed method has shown good results when compared to the existing method. Lamivudine was quantified by Gas Chromatographic method using Ethyl Chloroformate as a Derivatizing reagent. There were no GC methods reported for lamivudine. LMV analysis was performed after derivatization and the internal standard technique was used for computation. The method development for the assay of LMV was based on its chemical properties. The method developed is very sensitive as the limit of detection is very less. Results of analysis of the pharmaceutical formulations revealed that the proposed methods are suitable for their analysis with no interference from the usual additives. All the methods were found to be linear, precise, accurate, specific and all proved to be sensitive, convenient and effective for the determination of LMV, ZVD, STV, NVP and EFZ in bulk and pharmaceutical dosage forms

Aspergillus terreus (Trichocomaceae): A Natural, Eco-Friendly Mycoinsecticide for Control of Malaria, Filariasis, Dengue Vectors and Its Toxicity Assessment Against an Aquatic Model Organism Artemia nauplii

Vector-borne diseases like malaria, filariasis, and dengue are transmitted by mosquitoes and they cause global mortality and morbidity due to an increased resistance against commercial insecticides. The present study was aimed to evaluate the neurobehavioral toxicity, knock-down effect, histopathology, ovicidal, adulticidal, and smoke toxicity effect of Aspergillus terreus extract against three mosquito species, namely Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti (Diptera: Culicidae). The isolated fungal strain was identified as A. terreus (GenBank accession no: KX694148.1) through morphological and molecular (phylogenetic) analysis. The morphological changes in the treated fourth instar larvae shown the demelanization of cuticle and shrinkage of the internal cuticle of anal papillae. The time duration of extract exposure against the larvae determines the level of toxicity. The extract treated larvae were displayed excitation, violent vertical and horizontal movements with aggressive anal biting behavior as the toxic effect on the neuromuscular system. The results of the biochemical analysis indicated that a decrease in the level of acetylcholinesterase, α-carboxylesterase, and β-carboxylesterase in extract treated fourth instar larvae of all tested mosquito species. The findings of histopathological investigation shown the disorganization of the abdominal region, mainly in mid, hindgut, and gastric caeca, loss of antenna, lateral hair, caudal hair, upper and lower head hairs in the mycelium extract treated An. stephensi, Cx. quinquefasciatus, and Ae. aegypti. The ovicidal bioassay test results showed the mosquito hatchability percentage was directly related to the concentrations of mycelium extract. Nil hatchability of mosquito eggs was noticed at 500 μg/ml concentration. The adulticidal activity of fungal mycelia ethyl acetate extract resulted in a dose-dependent activity (15 and 30 min recovery periods). The higher concentration of extract (1000 mg/L) acted as a repellent, the adult mosquitoes showed restless movement, uncontrolled/anesthetic flight at last died. The better adulticidal activity was observed in the ethyl acetate extract against An. stephensi, Cx. quinquefasciatus followed by Ae. aegypti with the best score of LD50 and LD90 values and nil mortality was found in the control. The results of smoke toxicity assay of the mycelia extract exhibited significant mortality rate against Ae. aegypti (91%), Cx. quinquefasciatus (89%), and An. stephensi (84%). In addition, the present investigation reported the stability and toxic effects of A. terreus mycelium ethyl acetate extract on Artemia nauplii. The swimming speed (0.88 mm s-1) of A. terreus was reduced with ethyl extract 24 h treatment whereas, the control A. nauplii showed the normal speed of 2.96 mm s-1. Altered behavior and swimming movement were observed in the 8 h A. terreus mycelium extract treated A. nauplii. A pale yellow color substance (metabolites) was found in the mid-gut region of the mycelial extract exposed A. nauplii. The outcome of the present study, suggest that the A. terreus metabolites might serve as an alternative, cost-effective, eco-friendly, and target specific mosquitocidal agent in the future

On the Provable Security of Multi-Receiver Signcryption Schemes

In ATC 2007, an identity based signcryption scheme for multiple receivers was proposed by Yu et al. In this paper, we first show that Yu et al.\u27s signcryption scheme is insecure by demonstrating an universal forgeability attack - anyone can generate a valid signcryption on any message on behalf of any legal user for any set of legal receivers without knowing the secret keys of the legal users. Also, we point out a subtle flaw in the proof of confidentiality given by Yu et al. and show that the scheme does not provide confidentiality. Further, we propose a corrected version of Yu et al.\u27s scheme and formally prove its security (confidentiality and unforgeability) under the existing security model for signcryption.\\ In another direction, Fagen Li et al. have proposed a pairing based multi-recipient signcryption scheme which works in public key infrastructure (PKI). We show that, the scheme proposed by Fagen Li et al. is not adaptive chosen ciphertext secure. We propose a new PKI based multi-receiver signcryption scheme and formally prove confidentiality and unforgeability of the scheme. Since all the previously reported schemes are shown to have flaws either in this paper or else where, the schemes reported in this paper are the only correct and efficient ones (both identity based and PKI based) for multi-receiver signcryption

Provably Secure ID-Based Broadcast Signcryption (IBBSC) Scheme

With the advent of mobile and portable devices such as cell phones and PDAs, wireless content distribution has become a major means of communications and entertainment. In such applications, a central authority needs to deliver encrypted data to a large number of recipients in such a way that only a privileged subset of users can decrypt it. A broadcasting news channel may face this problem, for example, when a large number of people subscribe to a daily exclusive news feature. This is exactly the kind of problem that \textit{broadcast encryption} attempts to efficiently solve. On top of this, especially in the current digital era, junk content or spam is a major turn off in almost every Internet application. If all the users who subscribe to the news feed receive meaningless noise or any unwanted content, then the broadcaster is going to lose them. This results in the additional requirement that subscribers have source authentication with respect to their broadcaster. \textit{Broadcast signcryption}, which enables the broadcaster to simultaneously encrypt and sign the content meant for a specific set of users in a single logical step, provides the most efficient solution to the dual problem of confidentiality and authentication. Efficiency is a major concern, because mobile devices have limited memory and computational power and wireless bandwidth is an extremely costly resource. While several alternatives exist in implementing broadcast signcryption schemes, identity-based (ID-based) schemes are arguably the best suited because of the unique advantage that they provide --- any unique, publicly available parameter of a user can be his public key, which eliminates the need for a complex public key infrastructure. In ASIAN 2004, Mu et al. \cite{MSLR04} propose what they call an ID-based authenticated broadcast encryption scheme, which is also a broadcast signcryption scheme, as the security goals are the same. They claim that their scheme provides message authentication and confidentiality and formally prove that the broadcaster\u27s secret is not compromised, but in this paper, we demonstrate that even without knowing the broadcaster\u27s secret, it is possible for a legal user to impersonate the broadcaster. We demonstrate this by mounting a universal forgeability attack --- any valid user, on receiving and decrypting a valid ciphertext from a broadcaster, can generate a valid ciphertext on any message on behalf of that broadcaster for the same set of legal receivers to which the broadcaster signcrypted the earlier message, without knowing any secrets. Following this, we propose a new ID-based broadcast signcryption (IBBSC) scheme, and formally prove its security under the strongest existing security models for broadcast signcryption (IND-CCA2 and EUF-CMA2)

Genome-Wide Linkage Analysis of Global Gene Expression in Loin Muscle Tissue Identifies Candidate Genes in Pigs

BACKGROUND: Nearly 6,000 QTL have been reported for 588 different traits in pigs, more than in any other livestock species. However, this effort has translated into only a few confirmed causative variants. A powerful strategy for revealing candidate genes involves expression QTL (eQTL) mapping, where the mRNA abundance of a set of transcripts is used as the response variable for a QTL scan. METHODOLOGY/PRINCIPAL FINDINGS: We utilized a whole genome expression microarray and an F(2) pig resource population to conduct a global eQTL analysis in loin muscle tissue, and compared results to previously inferred phenotypic QTL (pQTL) from the same experimental cross. We found 62 unique eQTL (FDR <10%) and identified 3 gene networks enriched with genes subject to genetic control involved in lipid metabolism, DNA replication, and cell cycle regulation. We observed strong evidence of local regulation (40 out of 59 eQTL with known genomic position) and compared these eQTL to pQTL to help identify potential candidate genes. Among the interesting associations, we found aldo-keto reductase 7A2 (AKR7A2) and thioredoxin domain containing 12 (TXNDC12) eQTL that are part of a network associated with lipid metabolism and in turn overlap with pQTL regions for marbling, % intramuscular fat (% fat) and loin muscle area on Sus scrofa (SSC) chromosome 6. Additionally, we report 13 genomic regions with overlapping eQTL and pQTL involving 14 local eQTL. CONCLUSIONS/SIGNIFICANCE: Results of this analysis provide novel candidate genes for important complex pig phenotypes

Antibacterial and free radical scavenging activity of a medicinal plant Solanum xanthocarpum

The present study describes the phytochemical profiles, antibacterial, and antioxidant properties of different solvent extracts of Solanum xanthocarpum leaves. Phytochemical analysis results revealed the presence of terpenoids, tannins, steroids, and phenols. Methanolic extract of plant had a maximum quantity of phenol (28.3 ± 2.0 mg) and flavonoids (25.2 ± 1.2 mg) than others. Similarly, the methanolic extract showed excellent antibacterial activity and exhibited the highest inhibitory effect against Pseudomonas aeruginosa (12 ± 0.5 mm), Salmonella typhi (10 ± 0.6 mm), Staphylococcus aureus (9 ± 1.0 mm), and Escherichia coli (7 ± 1.3 mm). The average minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were reported in the range of 3.2 to 6.9 µg/ml (for MIC) and 6.0 to 14.5 µg/ml (for MBC), respectively. The remarkable antioxidant activity was observed in chloroform and methanol extract on the DPPH radical scavenging activity with the lowest IC50 value of 197.245 μg/ml (chloroform) and 201.04 μg/ml (methanol) and compared with control (ascorbic acid 239.36 μg/ml). GC-MS analysis revealed the presence of six major bioactive compounds as follows; 2,Octylcyclopropene-1-Heptanol (42.81%), Hexadecanoic acid (26.63%), 1-methylene-2b Hydroxymethyl-3,3-Dimethyl-4b-(3Methylbut-2enyl)-C (9.3%), Phytol (7.5%), (1,3,3-Trimethyl-2-Hydroxymethyl-3,3-Dimethyl-4–3-Methylbut-2-Enyl)-C (7.2%). 3,7,11,15-Tetramethyl-2-Hexadecen-1-Ol (6.3%). The FT-IR spectrum reflected the presence of the twelve peaks at the range of 3746.38 cm-1 (O-H stretch alcohols), 3424.18 cm-1 (O-H stretch phenols), 2926.01 cm-1 (C-H stretch alkanes), 2857.55 cm-1 (C-H stretch alkanes), 2084.04 cm-1 (-C = C stretch alkynes), 1595.76 cm-1 (N-H bend primary amines), 1402.49 cm-1 (C-C stretch in ring aromatics) and others. This study suggests S. xanthocarpum as a potential candidature for having better antibacterial and antioxidant property and identified several bioactive compounds by GC-MS analysis

Larvicidal, Histopathological Efficacy of Penicillium daleae against Larvae of Culex quinquefasciatus and Aedes aegypti Plus Biotoxicity on Artemia nauplii a Non-target Aquatic Organism

Mosquitoes can transmit the terrible diseases to human beings. Soil-borne fungal products act as potential source for low-cost chemicals, used for developing eco-friendly control agents against mosquito-vector borne diseases. The prime aim of study was to check the larvicidal potential of fungus mycelia (by ethyl acetate solvent) extract from Penicillium daleae (KX387370) against Culex quinquefasciatus and Aedes aegypti and to test the toxicity of brine shrimp Artemia nauplii, by observing the physiological activity. The ethyl acetate extract of P. daleae mycelia (after 15 days) from Potato dextrose broth (PDB) medium revealed better result with least LC50 and LC90 values of I-IV instars larvae of Cx. quinquefasciatus (LC50 = 127.441, 129.087, 108.683, and 93.521; LC90 = 152.758, 158.169, 139.091, and 125.918 μg/ml) and Ae. aegypti (LC50 = 105.077, 83.943, 97.158, and 76.513; LC90 = 128.035, 106.869, 125.640, and 104.606 μg/ml) respectively. At higher concentration (1000 μg/ml) of extracts, mortality begins at 18 h of exposure and attained 100% mortality after 48 h exposure. Overall, the activity was depends on the dose and time of exposure to the extracts. The stereomicroscopic and histopathological analysis of Ae. aegypti and Cx. quinquefasciatus larvae treated with mycelium ethyl acetate extract showed complete disintegration of abdominal region, particularly the midgut and caeca, loss of cuticular parts and caudal hairs. Morphological characterization of the fungi was performed and taxonomically identified through 5.8s rDNA technique. The phylogenetic analysis of rDNA sequence was carried out to find out the taxonomic and the evolutionary sketch of isolate in relation to earlier described genus Penicillium. Behavior and swimming speed alteration was analyzed together with mortality. The results of the experiment indicates that swimming behavior recorder (SBR) is a appropriate tool to detect individual swimming speed of the A. nauplii organisms, since the values have been obtained in accordance with control monitored results showed the 2.75 mm s-1 and after 24 h treated found to be 0.72 mm s-1, respectively. The extract-exposed to A. nauplii showed changes in body structures, i.e., intestine enlargement, eye formation, outer shell malformations and loss of antennae. In the present study, we aimed to investigate the toxicity of the ethyl acetate extract of P. daleae on A. nauplii larvae by performing the mortality, behavior and alterations in swimming responses. This is the first time report on the larvicidal efficacy of P. daleae ethyl acetate extract against Cx. quinquefasciatus and Ae. aegypti larvae