Correlation between cell survival,clonogenic activity and micronuclei induction in DMBA-OC-1R cells treated with immunospecific albumin microspheres containing cisplatin and 5-fluorouracil

An ideal chemotherapeutic strategy would be to deliver a high concentration of drug that would be released in sustained small amounts from targeted microspheres to effectively kill only the tumour cells and thus reduce toxicity to normal tissue. Clonogenic and cell survival growth curve assays, as well as the micronucleus assay, were used to determine the feasibility of employing targeted immunomicrospheres in the treatment of cancer. Cells of a rodent ovarian carcinoma cell line, were exposed to cisplatin and 5-fluorouracil, either as free drug or encapsulated in albumin microspheres that were either conjugated to monoclonal antibodies or not. In cell survival growth curve assays, cell survival was reduced to 1.2% of the control when cells were treated with drug-containing immunomicrospheres. 3.2-fold more micronuclei were found in those cells that had been exposed to the drugs in immunomicrospheres than in those subjected to untargeted microspheres. All three assays demonstrated that the targeted immunomicrospheres were more effective in delivering cisplatin and 5-fluorouracil directly to the cells than the unconjugated microspheres, thus suggesting that targeted chemotherapy might be a more effective option in the treatment of cancer.     

Cupriavidus alkaliphilus sp. nov., a new species associated with agricultural plants that grow in alkaline soils

A group of 20 bacterial strains was isolated from the rhizosphere of different agricultural plants growing in alkaline soils in the northeast of Mexico. The phylogenetic analysis of the 16S rRNA gene sequence from four strains showed that this novel group belonged to the Cupriavidus genus, with C. taiwanensis (～98.9%) and C. necator (～98.8%) as the closest species. However, DNA-DNA reassociation values were less than 20%. The novel group did not fix nitrogen and lacked nifH and nodA genes, unlike C. taiwanensis. Whole-cell protein patterns were highly similar among the 20 strains but different from the closest Cupriavidus species. BOX-PCR patterns were distinct among the 20 strains but also differed from other Cupriavidus type species. The major cellular fatty acids from strains ASC-732(T) and SLV-2362 were C(16:0), C(18:1) ω7c/12t/9t and C(16:1) ω7c and/or C(15:0) iso 2OH. The major polar lipids consisted of phosphatidylglycerol, cardiolipin, phosphatidylethanolamine, 2-hydroxylated-phosphatidylethanolamine and an unknown aminolipid. The DNA G+C content of strain ASC-732(T) was 66.8mol%. All 20 strains grew in the presence of 5-10mgmL(-1) arsenic, 1mgmL(-1) zinc, and 0.1mgmL(-1) copper. Consequently, the group of strains was considered to represent a novel species for which the name Cupriavidus alkaliphilus sp. nov. is proposed. The type strain is ASC-732(T) (=LMG 26294(T)=CIP 110330(T)).     

Salivary immunity in elderly individuals presented with Candida-related denture stomatitis

doi: 10.1111/j.1741‐2358.2011.00476.x
Salivary immunity in elderly individuals presented with Candida‐related denture stomatitis Objectives: Elderly individuals with Candida‐related denture stomatitis (DS) present with a reduced defence against Candida albicans. This study evaluated levels of antimicrobial mediators in the elderly DS saliva and salivary neutrophils’ activation characteristics compared with elderly and young without DS. Methods: Salivary peroxidases (SPO) and elastase activities (ELA), nitric oxide (NO), transforming growth factor beta (TGF‐β), IL‐6 and CCL3 production were determined in saliva from elderly with or without DS, and young control individuals. TLR4, CXCR1, CD11b, CD16 and CD32 expression on salivary neutrophils were evaluated. Correlations between number and apoptosis rate of salivary neutrophils, enzymatic activities and cytokine levels were determined. Results: Elderly DS individuals exhibited the lowest SPO and ELA activities. Also, the activity of both enzymes was low in elderly without DS. Although both elderly groups showed higher salivary NO and TGF‐β levels compared to young control groups, elderly DS presented the highest salivary NO, TGF‐β, IL‐6 and CCL3 levels. Decreased percentages of salivary TLR4⁺ and CD16⁺ neutrophils were detected in both elderly groups. Although these damages could influence the establishment and persistence of DS, the highest levels of salivary IL‐6 and CCL3 in elderly DS could be preventing more serious complications.     

Influence of different mucosal resiliency and denture reline on stress distribution in peri-implant bone tissue during osseointegration. A three-dimensional finite element analysis

doi: 10.1111/j.1741‐2358.2011.00569.x Influence of different mucosal resiliency and denture reline on stress distribution in peri‐implant bone tissue during osseointegration. A three‐dimensional finite element analysis Objective: The aim of this study was to evaluate the influence of mucosal properties and relining material on the stress distribution in peri‐implant bone tissue during masticatory function with a conventional complete denture during the healing period through finite element analysis. Materials and Methods: Three‐dimensional models of a severely resorbed mandible with two recently placed implants in the anterior region were created and divided into the following situations: (i) conventional complete denture and (ii) relined denture with soft lining material. The mucosal tissue properties were divided into soft, resilient and hard. The models were exported to mechanical simulation software; two simulations were carried out with a load at the lower right canine (35 N) and the lower right first molar (50 N). Data were qualitatively evaluated using Maximum Principal Stress, in MPa, given by the software. Results: All models showed stress concentrations in the cortical bone corresponding to the cervical part of the implant. The mucosal properties influenced the stress in peri‐implant bone tissue showing a different performance according to the denture base material. The simulations with relined dentures showed lower values of stress concentration than conventional ones. Conclusions: It seems that the mucosal properties and denture reline have a high influence on the stress distribution in the peri‐implant bone during the healing period.     

Fast molecular evolution associated with high active metabolic rates in poison frogs

Molecular evolution is simultaneously paced by mutation rate, genetic drift, and natural selection. Life history traits also affect the speed of accumulation of nucleotide changes. For instance, small body size, rapid generation time, production of reactive oxygen species (ROS), and high resting metabolic rate (RMR) are suggested to be associated with faster rates of molecular evolution. However, phylogenetic correlation analyses failed to support a relationship between RMR and molecular evolution in ectotherms. In addition, RMR might underestimate the metabolic budget (e.g., digestion, reproduction, or escaping predation). An alternative is to test other metabolic rates, such as active metabolic rate (AMR), and their association with molecular evolution. Here, I present comparative analyses of the associations between life history traits (i.e., AMR, RMR, body mass, and fecundity) with rates of molecular evolution of and mitochondrial loci from a large ectotherm clade, the poison frogs (Dendrobatidae). My results support a strong positive association between mass-specific AMR and rates of molecular evolution for both mitochondrial and nuclear loci. In addition, I found weaker and genome-specific covariates such as body mass and fecundity for mitochondrial and nuclear loci, respectively. No direct association was found between mass-specific RMR and rates of molecular evolution. Thus, I provide a mechanistic hypothesis of the link between AMRs and the rate of molecular evolution based on an increase in ROS within germ line cells during periodic bouts of hypoxia/hyperoxia related to aerobic exercise. Finally, I propose a multifactorial model that includes AMR as a predictor of the rate of molecular evolution in ectothermic lineages.     

Realism in systematics through biogeographical consilience

There is an overlooked gap between any phylogenetic hypothesis and the natural world shaped by historical evolutionary processes, since the main concern during phylogenetic analyses is solely the search for congruence among characters under a defined criterion. Given a scientific realistic view, however, phylogenetic hypotheses are scientific theories that try to depict the historical series of cladogenetic events among biological entities. In this sense, the challenge is to establish a form of measuring the degree of truthfulness of our phylogenetic hypotheses. Here, we advocate the use of consilient biogeographical hypotheses to recognize the biological meaning of a phylogenetic inference apart from its instrumentalist value. Our proposal is based on the assumption that robust biogeographical hypotheses allows us to be close to the real evolutionary history of taxa. © The Willi Hennig Society 2011.     

Endophytic fungi from Combretum leprosum with potential anticancer and antifungal activity

The recent increase in human diseases and cancers requires new drugs to combat them. Sources have been found in rare microorganisms, those from extreme habitats, and from endophytes. In this study, the biological activity of endophytic fungi associated with the Brazilian medicinal plant Combretum leprosum was assessed. Cytotoxic and antiproliferative effects were evaluated using seven human cancer cells lines (HeLa, ECV304, B16F10, J744, P388, Jurkat and k562). In addition the minimum inhibitory concentration (MIC) against pathogenic human fungal was determined using four Candida species and the filamentous fungi Cryptococcus neoformans and Trichophyton rubrum. A compound from extracts of phylotype Aspergillus oryzae CFE108 exhibited the most significant cytotoxicity effect against histiocytic sarcoma J774 (IC₅₀ of 0.80 μg?mL^?1), leukemia Jurkat (IC₅₀ of 0.89 μg?mL^?1), bladder carcinoma ECV304 (IC₅₀ of 3.08 μg?mL^?1) and cervical cancer HeLa (IC₅₀ of 2.97 μg?mL^?1). The extract from phylotypes Fusarium oxysporum CFE177 displayed antifungal activity and inhibited the growth of Candida glabrata (4 μg?mL^?1) as well as that of C. neoformans and T. rubrum with the lowest MIC being 62.5 μg?mL^?1. In addition, the fractions from A. oryzae CFE108 showed marked morphological activity (rounding up) on endothelial cells (tEnd.1 cells), which is indicative of potential antivascular activity. Our results indicate that the endophytes associated with this medicinal plant may be a source of novel drugs.     

Floristic relationships of the woody component in rocky outcrops savanna areas in Central Brazil

Rocky outcrops usually have a peculiar vegetation which differs from that of the surrounding vegetation matrix. In Brazil, a savanna type (cerrado rupestre) is particularly characterized by rocky outcrops and is one of eleven vegetation types which occur throughout Cerrado biome. It has been scarcely studied in detail. Therefore, the aim of this study was to investigate the phytogeographical patterns of the woody species in ten areas of the rocky outcrop savanna in the Brazilian Central Plateau. The difference in spatial distribution of the species was assessed through indicator species and CCA ordination analyses. Ten indicator species showed significantly different distribution. The first axis of the ordination showed association with the edaphic gradient, based on maximum temperature, pH and physical properties of the soils (eigenvalue 0.63). The second axis showed an eigenvalue 0.46, associated with poor rainfall and land declivity. It appears that the woody flora of the rocky outcrop savanna areas of the Brazilian Central Plateau is relatively homogeneous, with low β diversity. Apparently, the size of the woody species populations is what distinguishes the ten investigated areas. Therefore, initiatives and actions for the Cerrado biome conservation must consider not only the size of the protected areas but also that of the populations to be protected, especially the woody species on rocky outcrops.     

Adsorption of HSA, IgG and laminin-1 on model hydroxyapatite surfaces--effects of surface characteristics

Ellipsometry and mechanically assisted sodium dodecyl sulphate elution was utilized to study the adsorption of human serum albumin (HSA), human immunoglobulin G (IgG), and laminin-1, as well as competitive adsorption from a mixture of these proteins on spin-coated and sintered hydroxyapatite (HA) surfaces, respectively. The HA surfaces were characterized with respect to wettability and roughness by means of water contact angles and atomic force microscopy, respectively. Both surface types were hydrophilic, and the average roughness (Sa) and surface enlargement (Sdr) were lower for the sintered compared to the spin-coated HA surfaces. The adsorbed amounts on the sintered HA increased as follows: HSA < laminin-1 < IgG < the protein mixture. For the competitive adsorption experiments, the adsorbed fractions increased accordingly: HSA < laminin-1 < IgG on both types of HA substratum. However, a higher relative amount of HSA and laminin-1 and a lower relative amount of IgG was found on the spin-coated surfaces compared to the sintered surfaces. The effects observed could be ascribed to differences in surface roughness and chemical composition between the two types of HA substratum, and could have an influence on selection of future implant surface coatings.