Screening of microalgae for biosynthesis and optimization of Ag/AgCl nano hybrids having antibacterial effect

Here we report a facile and novel bio-synthesis technique, using algal extract to reduce silver metal ions into Ag/AgCl nanoparticles. Different concentrations of metallic precursors of silver nitrate (0.1 mM, 0.2 mM, 0.5 mM and 1 mM) were tested with alcoholic extract prepared from biomass of Chlorella sp. for nanoparticle biosynthesis which was screened out of four species namely Chlorella sp., Lyngbya putealis, Oocystis sp. and Scenedesmus vacuolatus. The biomolecules present in the alcoholic extract assisted in the synthesis of nanoparticles by reducing the metallic salt to metal ions and acting as capping agents in order to stabilize the particles. The synthesized particles were characterized for physico-chemical properties. DLS analysis of particles prepared from Chlorella sp. shows the particles with size of 90.6 nm. These biosynthesized nanoparticles show great potential applications in antibacterial activity.

Here we report a facile and novel bio-synthesis technique, using algal extract to reduce silver metal ions into Ag/AgCl nanoparticles.     

Disparities in Influenza Vaccination Coverage Among Women with Live-Born Infants: PRAMS Surveillance During the 2009–2010 Influenza Season

Objectives

Vaccination during pregnancy significantly reduces the risk of influenza illness among pregnant women and their infants up to 6 months of age; however, many women do not get vaccinated. We examined disparities in vaccination coverage among women who delivered a live-born infant during the 2009–2010 influenza season, when two separate influenza vaccinations were recommended.

Methods

Pregnancy Risk Assessment Monitoring System (PRAMS) data from 29 states and New York City, collected during the 2009–2010 influenza season, were used to examine uptake of seasonal (unweighted n=27,153) and pandemic influenza A(H1N1)pdm09 (pH1N1) (n=27,372) vaccination by racially/ethnically diverse women who delivered a live-born infant from September 1, 2009, through May 31, 2010.

Results

PRAMS data showed variation in seasonal and pH1N1 influenza vaccination coverage among women with live-born infants by racial/ethnic group. For seasonal influenza vaccination, coverage was 50.5% for non-Hispanic white, 30.2% for non-Hispanic black, 42.1% for Hispanic, and 48.2% for non-Hispanic other women. For pH1N1, vaccination coverage was 41.4% for non-Hispanic white, 25.5% for non-Hispanic black, 41.1% for Hispanic, and 43.3% for non-Hispanic other women. Compared with non-Hispanic white women, non-Hispanic black women had lower seasonal (crude prevalence ratio [cPR] = 0.60, 95% confidence interval [CI] 0.55, 0.64) and pH1N1 (cPR=0.62, 95% CI 0.57, 0.67) vaccination coverage; these disparities diminished but remained after adjusting for provider recommendation or offer for influenza vaccination, insurance status, and demographic factors (seasonal vaccine: adjusted PR [aPR] = 0.80, 95% CI 0.74, 0.86; and pH1N1 vaccine: aPR=0.75, 95% CI 0.68, 0.82).

Conclusion

To reduce disparities in influenza vaccination uptake by pregnant women, targeted efforts toward providers and interventions focusing on pregnant and postpartum women may be needed.Pregnant women are at increased risk for complications from influenza and are more likely than the general population to be hospitalized due to respiratory illness during influenza season.^1–4 Seasonal vaccination can reduce morbidity and mortality associated with seasonal influenza.^1,2 While the influenza vaccine recommendations for pregnant women date back to the 1960s, it was in 2004 that the American College of Obstetricians and Gynecologists (ACOG) and the Advisory Committee on Immunization Practices (ACIP) recommended that women be vaccinated with the inactivated influenza vaccine any time during pregnancy.^5–7 Historically, the national estimates showed that, of the adult groups recommended to receive seasonal vaccination, pregnant women had the lowest coverage prior to the 2009–2010 influenza season.^7–9 Research has shown that there are racial/ethnic and economic disparities in vaccination coverage among adults. In general, vaccination coverage is lower among non-Hispanic black and Hispanic women than among non-Hispanic white women and women of lower socioeconomic status.^10–16 For pregnant women in particular, coverage prior to the 2009–2010 influenza season was less than 30%.^8,9Because influenza can be especially severe during pregnancy, pregnant women in particular were at increased risk of severe disease and mortality from pandemic influenza A(H1N1)pdm09 (pH1N1) virus infection.¹⁷ During the 2009–2010 influenza season, both the inactivated trivalent seasonal and monovalent pH1N1 vaccinations were recommended for pregnant women.^17,18 Pregnant women were deemed a priority group for the pH1N1 vaccine due to high morbidity and mortality associated with pH1N1 infection within this group. Monovalent pH1N1 vaccine was purchased by the federal government and made available to the public at no cost. Additionally, the monovalent vaccine was made available later than the trivalent seasonal influenza vaccine.Given the recommendation of vaccination for pregnant women and the importance of preventing morbidity and mortality from influenza, we examined disparities in vaccination uptake by pregnant women with recent live-born infants who participated in the Pregnancy Risk Assessment Monitoring System (PRAMS) survey. The rationale for examining data from the 2009–2010 influenza season was to learn about vaccination coverage of a vulnerable population (i.e., pregnant women) during the pandemic.^19,20 Research questions guiding the analysis included (1) What are the differences in vaccination coverage among racial/ethnic groups of pregnant women? and (2) Are there differences by race/ethnicity in the patterns of vaccination uptake for the two influenza vaccinations offered during the 2009–2010 influenza season?     

Cortical Porosity Identifies Women With Osteopenia at Increased Risk for Forearm Fractures

Most fragility fractures arise among the many women with osteopenia, not the smaller number with osteoporosis at high risk for fracture. Thus, most women at risk for fracture assessed only by measuring areal bone mineral density (aBMD) will remain untreated. We measured cortical porosity and trabecular bone volume/total volume (BV/TV) of the ultradistal radius (UDR) using high‐resolution peripheral quantitative computed tomography, aBMD using densitometry, and 10‐year fracture probability using the country‐specific fracture risk assessment tool (FRAX) in 68 postmenopausal women with forearm fractures and 70 age‐matched community controls in Olmsted County, MN, USA. Women with forearm fractures had 0.4 standard deviations (SD) higher cortical porosity and 0.6 SD lower trabecular BV/TV. Compact‐appearing cortical porosity predicted fracture independent of aBMD; odds ratio (OR) = 1.92 (95% confidence interval [CI] 1.10–3.33). In women with osteoporosis at the UDR, cortical porosity did not distinguish those with fractures from those without because high porosity was present in 92% and 86% of each group, respectively. By contrast, in women with osteopenia at the UDR, high porosity of the compact‐appearing cortex conferred an OR for fracture of 4.00 (95% CI 1.15–13.90). In women with osteoporosis, porosity is captured by aBMD, so measuring UDR cortical porosity does not improve diagnostic sensitivity. However, in women with osteopenia, cortical porosity was associated with forearm fractures. © 2014 American Society for Bone and Mineral Research.     

Slovenian population data for five new European Standard Set Short tandem repeat loci and SE33 locus

Aim

To establish the allele distribution and statistical parameters of forensic interest for the D10S1248, D22S1045, D2S441, D1S1656, D12S391, and SE33 loci in Slovenian population and to compare allele frequencies with those from other populations.

Methods

We analyzed blood and buccal swab samples from 333 unrelated, healthy Slovenian individuals. All samples were genotyped using the AmpFlSTR NGM Kit to obtain the allele frequency data for the loci D10S1248, D22S1045, D2S441, D1S1656, and D12S391. Samples from 113 individuals were also analyzed using the PowerPlex ESX 17 system to obtain the allele frequency data for the SE33 locus. Allele frequencies and statistical parameters of forensic interest were determined and frequency profiles compared between Slovenian and other European Caucasian populations using the Arlequin software, version 3.5.1.3.

Results

The investigated short tandem repeat (STR) loci in Slovenian population had a great discriminating potential with a combined discrimination power of 0.99999998. The highest discrimination power and polymorphism information content were observed for the SE33 locus, followed by loci D1S1656, D12S391, D10S1248, D2S441, and D22S1045. When Slovenian allele frequency distribution was compared with other European populations, deviations were found only for Spanish and Italian population for D2S441 and D12S391.

Conclusion

Slovenian population does not differ significantly from other European populations in terms of allele frequency distributions for the six analyzed STR loci. Based on forensic efficiency values, SE33 may be considered the most informative locus, which makes it especially useful in forensic investigations.In their recommendations for autosomal short tandem repeat (STR) DNA typing in forensic casework, the European Network of Forensic Institutes (ENFSI) and the European DNA Profiling (EDNAP) Group proposed the use of additional five STRs, three mini-STRs (D2S441, D10S1248, D22S1045), and two highly polymorphic STRs (D1S1656 and D12S391). The increased number of the European Standard Set (ESS) loci resulted in improved discrimination power, sensitivity, and reproducibility for the analysis of minute amounts of DNA (1,2). In the last few years, several commercial STR typing kits have been released, with five new ESS loci. The new kits have been shown to be robust enough to successfully genotype even degraded DNA from old bone material (3-5). These kits also include the AmpFlSTR NGM^TM PCR Amplification Kit (Applied Biosystems, Foster City, CA, USA) and the PowerPlex ESX 17 System (Promega, Madison, WI, USA), which we used for our population study. Allele frequencies for autosomal STRs (6,7), Y-chromosomal STRs (8), and mitochondrial DNA (9) were already determined for Slovenian population and the aim of this study was to apply new genetic markers in routine forensic casework to achieve higher evidential value of STR typing and to increase the number of short STR loci, which are better preserved in degraded samples. Some European population studies have already investigated the new ESS loci (D10S1248, D22S1045, D2S441, D1S1656, D12S391) and we compared Slovenian allele frequencies with them (10-17). Beside the analysis of 5 new ESS loci we also analyzed SE33 locus and compared allele frequencies with Austrian (15), Italian (16), German (18), and Spanish population (19).     

Outpatient follow-up appointments for patients having curative treatment for cancer of the head and neck: are the current arrangements in need of change?

Although there are recommendations, there is little evidence about the rationale for the frequency and duration of review appointments for patients with cancer of the head and neck. We have recorded the pattern of follow-up in a tertiary cancer centre and its association with survival and recurrent disease. We used clinical letters and a prospectively maintained database to obtain details on 297 patients who were treated curatively for squamous cell carcinoma (SCC) of the oral cavity between 2005 and 2008. Mean (SD) age was 63 (12) years and 58% (n = 171) were male. Most patients were seen about 6 times in year one, 3 times in year 2, twice in year 3, twice in year 4, once or twice in year 5, and once yearly beyond year 5. Fewer clinics were scheduled for and attended by patients over 75 years of age, those with overall clinical grades 0-1, and those treated by operation alone in contrast to those who also had adjuvant radiotherapy. Patients were usually seen about 15 times over the 5 years. Taking into account the stage of the tumour and overall mortality, the number and timing of follow-up visits is adequate for the needs of patients with stage II-IV disease. Those with stage I disease may be considered for discharge after the third year if they are told about the risk factors, and signs and symptoms of recurrent disease, and surveillance in primary care.     

Utility of B-cell epitopes based peptides of RD1 and RD2 antigens for immunodiagnosis of pulmonary tuberculosis

Tuberculosis (TB) continues to be a major health problem due to lack of accurate, rapid, and cost-effective diagnostic tests. Serodiagnostic tests incorporating highly specific region of difference (RD) antigens (early secretory antigenic target 6 [ESAT-6], culture filtrate protein 10 [CFP-10], culture filtrate protein 21 [CFP-21], and mycobacterial protein from species tuberculosis 64 [MPT-64]) have recently been shown to be promising for specific diagnosis of TB in our lab. However, only few studies have reported the use of synthetic peptides of RD antigens, and none has used them to differentiate TB from sarcoidosis, a close mimic of smear-negative pulmonary TB (PTB) with entirely different management. The present study was conducted with an aim to study the utility of B-cell epitopes based peptides of RD1 (ESAT-6, CFP-10) and RD2 (CFP-21, MPT-64) antigens for immunodiagnosis of PTB for which sputum smear-positive PTB patients, sputum smear-negative PTB patients, sarcoidosis patients, and healthy controls (n = 24/group) were recruited. Bioinformatic software Bcepred was used to predict linear B-cell epitopes, using physico-chemical properties on a non-redundant dataset. Seven peptides as representative B-cell epitopes of ESAT-6, CFP-10, CFP-21, and MPT-64 were evaluated as targets of the antibody responses in TB patients and controls by enzyme-linked immunosorbent assay (ELISA). The current study showed sensitivity with individual peptides ranging from 37.5% to 83.3% for smear positive, 25% to 58.3% for smear negative as compared to 4.16% to 20.8% for sarcoidosis. Four out of 7 peptides that showed higher reactivity with TB patients and better discrimination from sarcoidosis patients representing ESAT-6, CFP-10, CFP-21, and MPT-64 were selected for multiepitope ELISA. The combination of peptides yielded 83.3% sensitivity for smear positive, 62.5% for smear negative, and only 4.16% for sarcoidosis. The specificity, however, for all the peptides/combination was 100%. Combination of peptides has proven to be better than individual peptides as per the latest criteria of the World Health Organization according to which a test that can replace smear microscopy with sensitivity of >90% for smear-positive patients and >65% for smear-negative TB patients with a specificity >95%, and thus, the present study suggests that a test based on combination of peptides selected from mycobacterial RD1 and RD2 antigens could be important for promoting an early diagnosis and management of otherwise difficult to diagnose smear-negative PTB patients. Moreover, it can also be used to discriminate sarcoidosis from PTB, thus preventing the misdiagnosis and mismanagement.     

Centella asiatica extract selectively decreases amyloid β levels in hippocampus of Alzheimer's disease animal model

PSAPP mice expressing the ‘Swedish’ amyloid precursor protein and the M146L presenilin 1 mutations are a well‐characterized model for spontaneous amyloid β plaque formation. Centella asiatica has a long history of use in India as a memory enhancing drug in Ayurvedic literature. The study investigated whether Centella asiatica extract (CaE) can alter the amyloid pathology in PSAPP mice by administering CaE (2.5 or 5.0 g/kg/day) starting at 2 months of age prior to the onset of detectable amyloid deposition and continued for either 2 months or 8 months. A significant decrease in amyloid β 1–40 and 1–42 was detectable by ELISA following an 8 month treatment with 2.5 mg/kg of CaE. A reduction in Congo Red stained fibrillar amyloid plaques was detected with the 5.0 mg/kg CaE dose and long‐term treatment regimen. It was also confirmed that CaE functions as an antioxidant in vitro, scavenging free radicals, reducing lipid peroxidation and protecting against DNA damage. The data indicate that CaE can impact the amyloid cascade altering amyloid β pathology in the brains of PSAPP mice and modulating components of the oxidative stress response that has been implicated in the neurodegenerative changes that occur with Alzheimer's disease. Copyright © 2008 John Wiley & Sons, Ltd.     

Modified formulation of CPDA for storage of whole blood, and of SAGM for storage of red blood cells, to maintain the concentration of 2,3-diphosphoglycerate

BACKGROUND AND OBJECTIVES: A dramatic decrease in the level of 2,3-diphosphoglycerate (2,3-DPG) takes place during the storage of whole blood (WB) in CPDA (citrate-phosphate-dextrose-adenine) and a similar decrease occurs during the storage of red blood cells (RBCs) in SAGM (saline-adenine-glucose-mannitol). The aim of the present study was to prevent this decrease by modifying CPDA and SAGM. MATERIALS AND METHODS: The pH of WB anticoagulant or RBC preservative solution was maintained at 7.6 by autoclaving the dextrose solution separately, by incorporating ascorbic acid and nicotinic acid into both CPDA and SAGM (to produce modified CPDA and SAGM solutions), and by reducing the concentration of adenine and adding citrate to the modified SAGM solution. The concentration of 2,3-DPG in WB after 28 days of storage in modified CPDA, and in RBCs stored in modified SAGM, was compared with that in WB or RBCs stored in unmodified solutions. RESULTS: The initial 2,3-DPG levels were maintained after 28 days in the modified formulations [10.63 +/- 2.58 microM/g of haemoglobin (Hb) in the case of modified CPDA and 12.07 +/- 1.47 microM/g of Hb in the case of modified SAGM], whereas in standard CPDA and SAGM solutions, the concentration of 2,3-DPG decreased to very low levels (0.86 +/- 0.97 microM/g Hb for CPDA and 0.12 +/- 0.008 for SAGM). CONCLUSIONS: Our modification in the formulation of CPDA or SAGM is effective in arresting the dramatic decrease in the level of 2,3-DPG that occurs during storage of WB and RBCs in unmodified solutions.     

Design,Synthesis, and Biological Evaluation of New Azulene-Containing Chalcones

Azulene-containing chalcones have been synthesized via Claisen–Schmidt condensation reaction. Their chemical structure has been established by spectroscopic methods where the ¹H-NMR spectra suggested that the title chalcones were geometrically pure and configured trans (J = 15 Hz). The influence of functional groups from azulene-containing chalcones on the biological activity of the 2-propen-1-one unit was investigated for the first time. This study presents optical and fluorescent investigations, QSAR studies, and biological activity of 10 novel compounds. These chalcones were evaluated for their antimicrobial activity against Gram-positive and Gram-negative bacteria. The results revealed that most of the synthesized compounds showed inhibition against Gram-negative microorganisms, independent of the substitution of azulene scaffold. Instead, all azulene-containing chalcones exhibited good antifungal activity against Candida parapsilosis, with MIC values ranging between 0.156 and 0.312 mg/mL. The most active compound was chalcone containing azulene moieties on both sides of the 2-propene-1-one bond, exhibiting good activity against both bacteria-type strains and good antifungal activity. This antifungal activity combined with low toxicity makes azulene-containing chalcones a new class of bioorganic compounds.