Vaccination with a non-human random sequence amyloid oligomer mimic results in improved cognitive function and reduced plaque deposition and micro hemorrhage in Tg2576 mice

ABSTRACT: BACKGROUND: It is well established that vaccination of humans and transgenic animals against fibrillar amyloid beta (Abeta) prevents amyloid accumulation in plaques and preserves cognitive function in transgenic mouse models. However, autoimmune side effects have halted the development of vaccines based on full length human Abeta. Further development of an effective vaccine depends on overcoming these side effects while maintaining an effective immune response. RESULTS: We have previously reported that the immune response to amyloid oligomers is largely directed against generic epitopes that are common to amyloid oligomers of many different proteins and independent of a specific amino acid sequence. Here we have examined whether we can exploit this generic immune response to develop a vaccine that targets amyloid oligomers using a non-human random sequence amyloid oligomer. In order to study the effect of vaccination against generic oligomer epitopes, a random sequence oligomer (3A) was selected as it forms oligomers that react with the oligomer specific A11 antibody. Oligomer mimics from 3A peptide, Abeta, islet amyloid polypeptide (IAPP), and Abeta fibrils were used to vaccinate Tg2576 mice, which develop a progressive accumulation of plaques and cognitive impairment. Vaccination with the 3A random sequence antigen was just as effective as vaccination with the other antigens in improving cognitive function and reducing total plaque load (Abeta burden) in the Tg2576 mouse brains. CONCLUSION: These results show that the amyloid Abeta sequence is not necessary to produce a protective immune response that specifically targets generic amyloid oligomers. Using a non-human, random sequence antigen may facilitate the development of a vaccine that avoids autoimmune side effects.     

Development of mycorrhized vitroplants of Jatropha curcas L. at different rooting stages

In this work, we attempted to assess the effects of inoculation of arbuscular mycorrhizal fungus (AMF), Glomus clarum, on the survival and development of micropropagated Jatropha curcas plantlets at different rooting stages. Elongated shoots (average 3?cm) of J. curcas, maintained for 0, 14, or 21?days on rooting medium in the presence or absence of 1?mg?L^?1 indole-3-butyric acid (IBA), were transferred to a sand:soil:vermiculite (1:1/2:1) (v:v:v) substrate. At the time of transplantation, the plantlets were either inoculated or not inoculated with G. clarum that had been monoaxenically produced in Ri-transformed carrot roots. After a 2-week acclimatization period, 100?% of the plants kept for 0 or 14?days in rooting medium survived. However, those that remained for 21?days in rooting medium displayed post-acclimatization survival rates of 93 and 89?% for plants inoculated and non-inoculated with G. clarum, respectively. Colonization rates ranged from 70 to 93?%, and the stimulatory effects of AMF were evidenced by increased phosphorus uptake by plants and increases in all evaluated growth parameters, except plant height. Plants that were not subjected to the rooting stage showed growth similar to or higher than those subjected to the rooting stage, regardless of the addition of IBA. It can be concluded that stimulatory effects of mycorrhizal fungi were observed, and that the inoculation of J. curcas proved effective during the initial period of the acclimatization phase.     

Secondary metabolites of Sophora mollis subsp. griffithii (Stocks) Ali

Two new compounds, (+)-3,5,7-trihydroxy-3-[3′-hydroxy-2′,4′-dimethoxy-5-(3-methyl-2-butenyl)]-phenyl-(3R)-4H-1-benzopyran-4-one (1) and (?)-3-hydroxy-8,9-methylenedioxy-(6aR,11aS)-pterocarpan (2), were isolated from the methanolic extract of Sophora mollis subsp. griffithii. Two known compounds, β-sitosterol (3) and 19βH-lupeol-methyl-ether (4), were also obtained for the first time from this plant. The structures of 1–4 were identified through their spectroscopic data. CD Spectroscopy was also utilized for the structure elucidation of compounds 1 and 2. Compounds 1, 3 and 4 were studied for their effects on immune cells and only 1 was found to be substantially active.     

BAD,a Proapoptotic Protein,Escapes ERK/RSK Phosphorylation in Deguelin and siRNA-Treated HeLa Cells

This study has been undertaken to explore the therapeutic effects of deguelin and specific siRNAs in HeLa cells. The data provided clearly show the silencing of ERK 1/2 with siRNAs and inhibition of ERK1/2 with deguelin treatment in HeLa cells. Additionally, we are providing information that deguelin binds directly to anti-apoptotic Bcl-2, Bcl-xl and Mcl-1 in the hydrophobic grooves, thereby releasing BAD and BAX from dimerization with these proteins. This results in increased apoptotic activity through the intrinsic pathway involved in rupture of mitochondrial membrane and release of cytochrome C. Evidence for inhibition of ERK1/2 by deguelin and escape of BAD phosphorylation at serine 112 through ERK/RSK pathway has been further fortified by obtaining similar results by silencing ERK 1/2 each with specific siRNAs. Increase in BAD after treatment with deguelin or siRNAs has been interpreted to mean that deguelin acts through several alternative pathways and therefore can be used as effective therapeutic agent.     

New insight in staphylococcin research: bacteriocin and/or bacteriocin-like inhibitory substance(s) produced by S. aureus AB188

Summary Staphylococcus aureus AB188 (a clinical isolate from wound pus) has been found to produce bacteriocins and/or bacteriocin-like inhibitory substance(s) tentatively termed staphylococcin Bac188. It has a broad activity spectrum against many Gram-positive (e.g. B. subtilis, S. aureus, E. faecalis), Gram-negative bacteria (e.g. E. coli, S. typhi and S. dysenteriae), and dermatophytes including Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton longi and Trichophyton rubrum. Interestingly, staphylococcin Bac188 also showed very potent activity against many clinical isolates of Mycobacterium tuberculosis. Staphylococcin Bac188 showed wide thermostability and remained stable over the wide pH range (pH 2–14). It was also resistant to treatment with chloroform, catalase, lipase and lysozyme, but showed sensitivity to proteinase K, trypsin and α-chymotrypsin suggesting its proteinaceous nature. Staphylococcin Bac188 revealed bactericidal effects on the S. aureus SS-1 sensitive strain as well as on E. coli and S. typhi, suggesting a similar mode of action on both Gram-negative and Gram-positive organisms. The antimicrobial, antidermatophytic and antimycobacterial activities expressed by S. aureus AB188 correlate with the production of a bacteriocin and/or bacteriocin-like inhibitory substance with properties similar to other staphylococcins reported earlier. To our knowledge, this is the first report describing such wide possible clinical applications of a bacteriocin and/or bacteriocin-like inhibitory substance produced by S. aureus AB188, suggesting further investigation for potential therapeutic development.     

Prevalence of ESBL and MBL encoding genes in Acinetobacter baumannii strains isolated from patients of intensive care units (ICU)

The aim of this study was to investigate the prevalence of ESBL and MBL encoding genes among A. baumannii isolates. In this cross sectional study, 100 A. baumannii strains were isolated from ICU wards of 3 educational hospitals of Hamadan City, Iran in 2011. Phenotypic identification of the production of ESBLs and MBLs has been carried out by using E-test and DDST methods, respectively. PCR technique was used for amplification of the ESBL and MBL encoding genes, namely: CTX-M, SHV, TEM, OXA-51, VIM-Family, IMP-Family, SPM-1, SIM-1, and GIM-1. Eighty seven (87%), 95 (95%), 98 (98%) and 95 (95%) out of 100 A. baumannii isolates were resistant to imipenem, meropenem, ceftazidime and cefotaxime, respectively. Also, 99% and 7% of the isolates were MBLs and ESBLs produced phenotypically. Thirty (30%), 20 (20%) and 58 (58%) out of 100 A. baumannii isolates have been confirmed to harbor the bla_VIM-family, TEM and SHV genes, respectively. Our results show no significant relationship between the detected gens with production of MBLs and ESBLs in spite of high prevalence of MBL encoding and drug resistant A. baumannii. Probably some other genes rather than what we studied are involved in phenotypic production of MBLs and ESBLs and subsequent drug resistance in Hamadan area, Iran.     

Assessment of the role of general,biochemical and family history characteristics in kidney stone formation

Aim

The main objective of the study was to determine the urinary risk factors involved in kidney stone formation.

Method

In this study a total number of 101 patients (64 males and 37 females) between the age group 2 and 70 years were selected. Personal characteristics like age, family history, clinical sign and symptoms, education, monthly income, living style, smoking or tobacco chewing habit, dietary intake and daily amount of drinking water were recorded.

Results

The study showed that the risk of kidney stone formation was high in the median age group (16–25 years) both in male and female population. The most important factors associated with this were lack of drinking clean water, over weight and obesity as well as family history (37.5% and 27.02% in men and women, respectively).

Conclusion

Our study has confirmed that lack of drinking sufficient amount of water, increasing weight and obesity and family history are some major factors contributing to the increased risk of kidney stone formation. Therefore it is very important to live a healthy life, drink clean water and control weight to prevent such diseases.     

Molecular analysis of V617F mutation in Janus kinase 2 gene of breast cancer patients

BackgroundBreast cancer is a multifactorial disease with the highest frequency in females. Genetic and environmental factors can cause mutation in several genes like tyrosine kinase, JAK2 gene which may initiate cancer. Molecular analysis of mutations in the JAK2 gene along with determination of environmental, clinical and haematological risk factors associated with breast cancer patients is need of hour to improve patient's healthcare. Somatic JAK2 valine-to-phenylalanine (617 codon) mutation is one of the widely prevalent mutations.MethodsBlood was collected from seventy breast cancer patients after their consent. The questionnaire included risk factors, age group, locality, number of children, tumor type, family history, time of initial diagnosis, no of cycles/month, water conditions and exposure to radiations. Molecular analysis were carried out from genomic DNA using Sanger sequencing and allele-specific PCR to check the V617F point mutation.ResultsThe breast cancer risk factors includes unfiltered water (68.57%), urban (58.57%), menopause (55.71%), family history of cancer (18.57%), tumor grades (II, 37.14% and III, 35.71%), consanguineous marriages (44.28%) and having more than 3–4 children (45.71%). Prevalence of breast cancer was higher after the age of 35 and maximum at 35–50. In allele-specific PCR of 70 patients, 25 patients were wild type (229 bp), 25 patients were with partially deleted gene (200 bp), and 20 patient had shown no or less than 40 bp size fragments. In Sanger's sequencing of 70 BC cases, 18% were found to be positive for V617F point mutation, including 6 homozygous (T/T) and 7 heterozygous (G/T) mutations at nucleotide position 1849 in exon 14 of the JAK2 gene.ConclusionsEnvironmental and clinical risk factors were associated with breast cancer which can be overcome by improving awareness of associated risks, health facilities and reducing stress.