Purpose We identified calreticulin in human filaria Brugia malayi (BmCRT) that shares 97% homology with Wuchereria bancrofti calreticulin (WbCRT), but only 56% with human calreticulin. We found that BmCRT binds C1q and prevents complement-mediated parasite death; immunization with BmCRT leads to parasite death in a rodent model of the infection. BmCRT could, therefore, be a potential vaccine candidate. In the present study, we determined the levels of BmCRT-reactive IgG and its isotype in bancroftian filarial subjects. Methods Recombinant BmCRT (rBmCRT) was prepared, and the sera of endemic normal subjects (EN), microfilaraemics (Mf+) and chronic amicrofilaraemics (ChMf-) from a bancroftian filaria-endemic area and normal subjects from filaria-non-endemic area (NEN) were probed for IgG and its isotypes reacting with rBmCRT and its domains rN, rP and rC. Results rBmCRT and its rN domain-reactive IgG levels were high in EN and Mf+ groups; rC domain and rP domain showed moderate and very little reactivity, respectively. NEN sera were non-reactive. Moderate levels of rBmCRT-reactive IgG1, IgG3 and IgG4 in EN and Mf+ groups and low levels of IgG2 in Mf+ were found; IgG1 and IgG3 reactivity was found for rBmCRT and its rN domain only, while IgG4 reactivity was moderate for rN domain and low for rP and rC domains. While IgG reactivity was seen in all the endemic subjects, IgG isotype reactivity was found mostly in EN and Mf+ subjects. Conclusions Moderate levels of rBmCRT (and its rN domain)-reactive IgG and its isotypes are present in bancroftian subjects. Preponderance of IgG1 and IgG3 isotypes which bind and activate complement has relevance to vaccine potential of BmCRT.

Aloe barbadensis known for centuries for its herbal remedy belongs to the family of Liliaceae. The aqueous and organic leaf extracts were prepared by using soxhlet apparatus. The extracts were evaluated for qualitative phytochemical analysis and its antioxidant property using DPPH radical scavenging assay, alkaline DMSO method, nitric oxide method, Total antioxidant Assay and Deoxyribose method. The present study showed the presence of alkaloid, flavonoid, reducing sugar, terpenoids and steroids by qualitative phytochemical analysis. The aqueous and organic extracts were prepared by using soxhlet apparatus and screened for anti-oxidant property by different methods. The acetone and methanolic extracts A. barbadensis showed anti-oxidant activity by DPPH radical scavenging assay with IC50 values of 705.2 ± 8.08 and 182.8 ± 1.44 μg/ml respectively. Whereas aqueous and methanolic extracts showed anti-oxidant activity by alkaline DMSO method with IC50 values of 237.5 ± 11.4 and 553.4 ± 3.28 μg/ml respectively. While methanolic extract of A. barbadensis showed activity by nitric oxide method with IC50 value of 43.17 ± 6.29 μg/ml. The methanolic extract of A. barbadensis gave IC50 value of 589.6 ± 5.54 by total anti-oxidant activity and aqueous and methanolic extracts gave IC50 value of 579.8 ± 3.95 and 714.8 ± 7.21 μg/ml respectively by deoxyribose method. The presence of these polyphenols may be responsible for its potent anti-oxidant property. The leaf extracts of Aloe barbadensis showed potent anti-oxidant activity and can be used as a therapeutic agent.

BACKGROUND:Lymphatic Filariasis (LF) is one of the incapacitating and mosquito-borne sicknesses that on progression may prompt a few recognizable types of clutters like extreme lymphedema, hydrocele, and elephantiasis.METHODS:Antigenic preparations of B. malayi adult (BmA), S. cervi adult parasites and microfilariae (mf) total parasite extract were used to analyze the serological reactivity profile with human infectious sera collected from endemic areas of Bancroftian filariasis by performing Western blot and ELISA analysis. Sera from healthy human subjects were also included in the study to determine the variation incurred in the reactivity due to the filariasis infection. Gelelectrophoresis analysis of the crude-extract of BmA revealed seven protein bands while more than ten bands were recognized in S. cervi.RESULTS:our results represent a clear variation in protein patterns among the crude-antigens. ELISA results showed highest prevalence of IgG, IgM and IgG4 antibodies against all antigen preparations when recorded among microfilaraemic chronic infected patients. In both the antigenic preparations, the positive reactions were in the order of microfilaraemic>endemic normal>chronic>acute>nonendemic normal subjects. All sera of Mf+ patients were uniformly positive, while sera of both chronic and endemic normal subjects showed less reactivity.CONCLUSION:In the present study, we endeavoured to establish the extent of cross-reactivity of antigens derived from animal filarial parasites such as B. malayi and S. cervi with W. bancrofti filariasis sera of human patients. Besides, we further analyzed antibody-isotype profile of IgG, IgG4 and IgM in various human infection sera of bancroftian filarial subjects reactive to heterologous parasite antigens derived from adult worms of S. cervi from bovine and B. malayi from bovine and jirds.

Lymphatic filariasis (LF) is a chronic and debilitating disease that affects people in tropical and sub-tropical areas of Asia, Africa, and Western Pacific. It is one of the leading community health problems in some of the endemic districts in India including Hardoi district of Uttar Pradesh. The disease is caused by the parasites Wuchereria bancrofti (W. bancrofti), Brugia malayi (B. malayi) and Brugia timori (B. timori), transmitted by the vector Culex, Anopheles and other mosquitoes. This cross-sectional survey study was carried out in rural areas, where its inhabitants vary in socio-economic status, from low to middle-income class. 12 villages of Hardoi district, Uttar Pradesh, India were included. The aim was to see the impact of age and gender on various clinical forms of LF and in estimating its economic and social implications. 260 LF affected people in different parts of Hardoi district were surveyed. The results revealed that the Mass Drug Administration (MDA) coverage reached more than 90%. The overall Microfilaria rate had been reduced, however the prevalence of elephantiasis increased with the progression of age and was found to be highest among people of > 70 years of age, regardless of their gender.

Lymphatic filariasis (LF) is a chronic disease and is caused by the parasites Wuchereria bancrofti (W. bancrofti), Brugia malayi (B. malayi) and Brugia timori (B. timori). In the present study, Setaria cervi (S. cervi), a bovine filarial parasite has been used. Previously, it has been reported that the S. cervi shares some common proteins and antigenic determinants with that of human filarial parasite. The larval stages of filarial species usually cannot be identified by classical morphology. Hence, molecular characterization allows the identification of the parasites throughout all their developmental stages. The genomic DNA of S. cervi adult were isolated and estimated spectrophotometrically for the quantitative presence of DNA content. Screening of DNA sequences from filarial DNA GenBank and Expressed Sequence Tags (EST’s) were performed for homologous sequences and then multiple sequence alignment was executed. The conserved sequences from multiple sequence alignment were used for In Silico primer designing. The successfully designed primers were used further in PCR amplifications. Therefore, in search of a promising diagnostic tool few genes were identified to be conserved in the human and bovine filariasis and these novel primers deigned may help to develop a promising diagnostic tool for identification of lymphatic filariasis.

Unani medicine is now becoming a strong part of medicinal system and it is now popularly practiced by a large segment of the population. This medicinal system originated in Greece and was brought to the subcontinent by Muslim scholars. However, the unani medicine currently practiced in India is vastly different from its Greek roots. Lallemantia royleana (Benth.) is an important folk medicine which is used in number of ailments. This study was conducted to assess the anti-oxidant activity and phyto-chemical analysis of methanolic extracts of L. royleana seeds. The antioxidant activity was determined by using different anti-oxidant methods via 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging assay and hydrogen peroxide scavenging assay. The present study shows that L. royleana seeds possess significant anti-oxidative property therefore, it can be a good remedy for the diseases caused by oxidative stress. The seeds extract contains Alkaloids, Anthraquinones, Flavonoids, Glycosides, Pholobtannins, Tannins and Terpenoids. The antioxidant activity found in DPPH method is more effective than hydrogen peroxide scavenging method. The results of GCMS analysis showed that at least 21 compounds are present in methanolic extract of L. royleana. The compounds which were identified through mass spectrometry (MS) are attached with gas chromatogram (GC). Thus the in-vitro studies clearly indicate that the methanolic extract of L. royleana seeds show significant antioxidant activity and also a better source of natural antioxidant, which might be helpful in preventing the process of various oxidative stresses.

The present study aimed to examine anti-microbial, anti-oxidant and cytotoxicity in leaf extract of Ricinus communis extract (in different solvent). The leaf powder of R. communis was extracted using different solvents. The anti-bacterial activity of the extracts was determined by agar well and disc diffusion method. The extracts were also subjected to phytochemical analysis. The anti-oxidant activity of the extracts was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH), alkaline DMSO, deoxyribose and nitric oxide scavenging assay. The cytotoxicity of the extracts was estimated using MTT cell proliferation assay. The photochemical qualitative analysis of methanollic plant extracts revealed the presence of alkaloid, flavonoid, tannins, glycoside, reducing sugar, anthraquinones and saponins. The methanollic extract showed zone of inhibition of 15 mm each against Bacillus subtilis, Staphylococcus epidermis and Saccharomyces cereviceae by using well diffusion method, whereas S. cereviceae gave 12 mm zone of inhibition by disc diffusion at a concentration of 40 mg/mL. The anti-oxidant activity by different methods gave IC50 value of 102.1 ± 4.16, 30.27 ± 3.85 and 382.6 ± 3.30 µg/mL in aqueous, benzene and ethyl acetate extract respectively by using DPPH method. The acetone extract gave IC50 value of 357.1 ± 4.96 µg/mL by nitric oxide method. The aqueous and acetone extract gave IC50 value of 860.1 ± 7.73 and 626.7 ± 2.25 µg/mL, respectively by deoxyribose method. The chloroform and ethyl acetate extract showed cytotoxicity in A549 cell line having IC50 value of 687 ± 3.92 and 957 ± 4.46 µg/mL respectively by MTT cell proliferation assay whereas, aqueous extract in Jurkat cell line gave IC50 value of 918 ± 2.05 µg/mL. This study demonstrates that the R. communis extracts are potential source for anti-microbial, anti-oxidant and anti-cancer agent. Further study is needed to identify the specific bioactive compounds, their mode of action and their non-toxic nature in in vivo condition.   Key words: Ricinus communis, (3-(4, 5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide) (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH).

The scientists have identified number of herbs from different parts of India having medicinal properties.The aim of the study was to examine antimicrobial, anti-oxidant and cytotoxicity in fruit extract of Myristica fragrans (aqueous, cyclohexane, benzene, chloroform, acetone, ethyl acetate, ethanol and methanol).The phytochemical analysis revealed the presence of alkaloid, flavonoid, tannins, glycoside, reducing sugar, steroids and terpenoids that may be responsible for its anti-microbial, anti-oxidant and cytotoxicity in M. fragrans extracts.The methanolic extract showed considerable anti-microbial activity against S. aureus and the zone of inhibition was found to be 16 mm at the concentration of 40 mg/ml of plant extract by well and disc diffusion method.The anti-oxidant activity by different methods was screened and the highest activity was shown by methanol extract and gave IC50 value of 56.83 ± 5.67 µg/ml by DPPH method.The superoxide radicals were scavenged strongly by M. fragrans acetone extract which showed IC50 value of 321.2 ± 12.75 µg/ml.The nitric oxide radicals were scavenged strongly by M. fragrans methanol extract demonstrated IC50 value of 45 ± 1.55 µg/ml.The hydroxyl radicals were strongly scavenged by M. fragrans benzene extract and showed IC50 value of 181.3 ± 4.04 µg/ml.The extracts of M. fragrans were found cytotoxic to cancerous cell lines by MTT cell proliferation assay.The chloroform extract of M. fragrans showed cytotoxicity in A549 cell line with IC50 value of 392 ± 6.51 µg/ml.

Lymphatic filariasis, a neglected parasitic disease caused by tissue dwelling human filarial nematodes such as Wuchereria bancrofti, Brugia malayi and B. timori is considered to be a major complication for the socio-economic development in developing countries. A number of inflammatory responses are associated with the diseases such as adeno-lymphangitis, lymphoedema, hydrocele and elephantiasis. The present study is directed towards the identification and amplification of Hha 1 gene for diagnosis of B. malayi. The specific primer Hha1 specific to Brugia malayi was used for detecting the parasites and was found to give optimum yield in the positive control samples. The results were confirmed from the amplified fragment having size of 322 bp of B. malayi. Using this primer as a diagnostic tool for the detection of filariasis might be the most promising aspect of the study and offers scope for detection of both the parasites even at low levels of infection.

Glutathione-S-transferase is a major phase-II detoxification enzyme in parasitic helminthes. Previous research highlights the importance of GSTs in the establishment of chronic infections in cytotoxic microenvironments. Filarial nematodes depend on these detoxification enzymes for their survival in the host. GST plays an important role in filariasis and other diseases. GST from W. bancrofti and B. malayi are very much different from human GST. This structural difference makes GST potential chemotherapeutic targets for antifilarial treatment. In this study we have checked the efficacy of some well known antifilarial compounds against GST from B. malayi and W. bancrofti. The structure of BmGST was modeled using modeller9v10 and was submitted to PMDB. Molecular docking study reveals arbindazole to be the most potent compounds against GST from both the filarial parasites. Role of some residues playing important role in the binding of compounds within the active site of GST has also been revealed in the present study. The BmGST and WbGST structural information and docking studies could aid in screening new antifilarials or selective inhibitors for chemotherapy against filariasis.

Medicinal plants are the best source to obtain a variety of herbal drugs. The use of plant extracts and photochemical both with known anti-microbial properties can be of great importance in therapeutic treatments. The plants have provided a good source of anti-infective agents and many of them remain highly effective in the fight against microbial infections. Therefore in the present study seven medicinal plants that are Emblica officinalis, Ficus bengalensis, Myristica fragrans, Acacia arabica, Aloe barbadensis, Ricinus communis and Zizyphus jujuba were screened for potential anti-bacterial activity against medically important bacterial strains, such as Pseudomonas aurogenosa, Proteus vulgaris, Staphylococcus aureus and Streptococcus cereviceae. The anti-microbial activity was determined in methanolic extracts using agar well diffusion method. Streptococcus cereviceae showed resistance against the plant extracts. Emblica officinalis and Aloe barbadensis showed strong anti-bacterial activity against all the tested bacterial strains. Hence, this plant extract can be used to evaluate any bioactive natural products that may serve as leads in the development of new pharmaceuticals that can address the unmet therapeutic needs.

Crude antigenic preparations from heterologous filarial parasites gave false positive results because of complex nature of these antigens and their cross-reactivity with other helminth parasites. In the present study, efforts have been made to isolate and characterize the antigens from Setaria cervi important for diagnostic purposes. The fractionation of S. cervi somatic antigenic preparation on Sephacryl S-200 resulted in separation of three major antigenic peak fractions. Crossed immunoelectrophoretic analysis, using immune rabbit serum, revealed 13-14 antigens in SFP-I pool fraction, which showed high reactivity with filarial patients sera as compared to other two pool fractions. This SFP-I fraction was further purified by DEAE-Cellulose column chromatography. Out of the 4 antigen pool fractions, DFP-IV fraction showed high ELISA reactivity with filarial patient serum pool (Wuchereria bancrofti and Brugia malayi) as compared to other fractions. The SDS-PAGE analysis of DFP-IV fraction revealed 2 major and 1 minor protein bands (mol. wt. range 65-70 kDa). Crossed immunoelectrophoresis also showed the presence of 3 antigenic peaks in DFP-IV fraction. The purified DFP-IV fraction showed high reactivity with filarial patients sera but did not cross-react with sera from ascaris and hookworm infections thereby suggesting the filaria-specificity and potential for immunodiagnosis of human filariasis.

Medicinal plan ts are the best source to obtain a variety of herbal drugs. The use of plant extracts and photochemical both with known anti - microbial properties can be of great importance in therapeutic treatments. The plants have provided a good source of anti - infective agents and many of them remain highly effective in the fight against microbial infections. Therefore in the present study s even medicinal plants that are Emblica officinalis, Ficus bengalensis, Myristica fragrans, Acacia arabica, Al oe barbadensis, Ricinus communi s and Zizyphus jujuba were screened for potential anti - bacterial activity against medically important bacterial strains, such as Pseudomonas aurogenosa , Proteus vulgaris , Staphylococcus aureus and Streptococcus cereviceae . The anti - microbial activi ty was determined in methano lic extracts using agar well diffusion method. Streptococcus cereviceae showed resistance against the plant extracts . Emblica officinalis and Aloe barbadensis showed strong anti - bacterial activity against all the tested bacteria l strains. Hence, this plant extract can be used to evaluate any bioactive natural products that may serve as leads in the development of new pharmaceuticals that can address the unmet therapeutic needs.