Collection Of Biological Material Research Articles

Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection

BackgroundParacoccidioidomycosis (PCM) is a neglected systemic mycosis caused by a dimorphic fungus of the Paracoccidioides genus. The standard diagnosis is based on isolation of the fungi in culture, and by microscopic visualization of characteristic multiple budding yeast cells in biological samples. However, in some situations, access to the site of injury prevents the collection of biological material. A variety of immuno-serological techniques has proven useful for allowing inferring diagnosis with a certain degree of certainty, thus optimizing time. The aim of this study was to standardize and validate the Dot-ELISA (DE) assay, comparing it with the serological standard, double immunodiffusion (DI).MethodsIn order to standardize the DE assay, 143 serum samples were used. Out of those, 23 were from apparently healthy patients, 77 were from patients with confirmed PCM and 43 were from patients with other lung infections (tuberculosis, aspergillosis and histoplasmosis). To validate the DE technique, 300 serum samples from patients with PCM clinical suspicion (probable and possible cases) were employed, and these results were compared with those of DI.ResultsThe DE assay showed sensitivity of 91%, specificity of 95.4%, positive predictive value of 96%, negative predictive value of 98.2%, accuracy of 93%, and great precision (k = 0.93). In addition, the nitrocellulose membranes have proved to be viable for using at least 90 days after P. brasiliensis B-339 antigen sensitization.ConclusionDot-ELISA method was found to be an extremely promising tool as serologic screening technique, because of its high sensitivity. Furthermore, Dot-ELISA shows the prospect of being transferred to laboratories of mycoserology including those with fewer resources or even to be used directly in the field. It has an excellent shelf life – membranes coated with antigen can be used for testing without changes in the pattern of reactivity among laboratories – and presents reliable values of sensitivity, specificity, predictive values, accuracy and a high correlation with the serological standard methodology. Based on the present findings, it possible to state that this technique constitutes a remarkable option to be used in routine diagnosis for public health centers.

Maintenance of a Drosophila melanogaster Population Cage.

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life cycle of Drosophila enables collection of large quantities of material from embryos, larvae, pupae and adult flies, in a synchronized way, at a low economic cost. A major strategy for propagating large numbers of flies is the use of a fly population cage. This useful and common tool in the Drososphila community is an efficient way to regularly produce milligrams to tens of grams of embryos, depending on uniformity of developmental stage desired. While a population cage can be time consuming to set up, maintaining a cage over months takes much less time and enables rapid collection of biological material in a short period. This paper describes a detailed and flexible protocol for the maintenance of a Drosophila melanogaster population cage, starting with 1.5 g of harvested material from the previous cycle.

Maintenance of a <em>Drosophila melanogaster</em> Population Cage

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life cycle of Drosophila enables collection of large quantities of material from embryos, larvae, pupae and adult flies, in a synchronized way, at a low economic cost. A major strategy for propagating large numbers of flies is the use of a fly population cage. This useful and common tool in the Drososphila community is an efficient way to regularly produce milligrams to tens of grams of embryos, depending on uniformity of developmental stage desired. While a population cage can be time consuming to set up, maintaining a cage over months takes much less time and enables rapid collection of biological material in a short period. This paper describes a detailed and flexible protocol for the maintenance of a Drosophila melanogaster population cage, starting with 1.5 g of harvested material from the previous cycle.

Restrain of birds with bottles of polyethylene terephthalate, tested in red-browed from the Atlantic Forest

In wild animals, containment is the moment of greatest stress caused by the investigator to the animal due to its natural resistance to the moment of capture, handling, containment and transport, attitudes frankly contrary to his nature. In birds, the restraint must meet certain criteria in order to control the animal's movements, avoiding trauma at the same time that you need to keep your breathing amplitude. The high risk of death during the restraint of these animals raised the need to design a device, from bottles of poly ethylene terephthalate (PET), for containing parrots-browed Amazon (Amazona rhodocorytha), a parrot endemic to the Atlantic Forest in southeastern Brazil, and endangered with extinction, which allowed the observation of respiration, the reduction of handling time of birds for collection of biological material, and consequent reduction of stress and risk of death during the evaluation of several biological data and health of the bird. The PET bottle container can be used as a model for any bird, provided it suits the size of the animal.

Molecular biomarkers and early diagnosis of lung cancer: state of knowledge and future perspectives

Lung cancer is the leading cause of cancer death worldwide and it would be essential to have effective tools to diagnose the disease in its early stages, to identify individuals at highest risk of developing the disease, and to set personalised therapies. The effectiveness of screening for lung cancer with low-dose CT (LDCT) in heavy smokers was demonstrated, in terms of reduction of cause-specific mortality, in recently published data by the study National Lung ScreeningTrial (NLST). In Europe, the introduction of LDCT as screening in individuals at risk is the object of a debate until the results of European randomized trials, expected in 2015-2016, are published. One problem is the high rate of calls for investigations when there is a noncalcified nodule, and it is therefore essential to be able to more accurately identify malignant nodules. The development of specific biomarkers appears to offer promising prospects. Recent advances in genetics and genomics have led to a series of studies aimed at the identification of molecular markers for the diagnosis, the assessment of the risk of developing lung cancer, the molecular characterisation of the different stages of the disease and the personalisation of therapy. Subjects enrolled in trials evaluating LCDT as a test for early detection of lung cancer represent the ideal population in which to study a combined bioinstrumental approach of screening (molecular test and LDCT). This paper reports on the state of knowledge on the possible use of biomarkers in the early detection of lung cancer and molecular analysis conducted within the project ITALUNG, a randomized controlled trial to assess the effect on tumour-specific mortality of LDCT, which provided a collection of biological materials from the subjects enrolled.

The prevalence of mutations in the gene encoding filaggrin in the population of Polish patients with atopic dermatitis.

IntroductionThe genetic background of atopic dermatitis (AD) is complex, involves many genes and their participation varies in varied populations, and depends on the intensity and course of a disease. Changes in the nucleotide sequence of the FLG gene and a reduced number or a deficit of the functional product of processed profilaggrin can be one of risk factors for atopic dermatitis.AimTo determine the prevalence of R501X and 2282del4 mutations of the FLG gene in patients with AD.Material and methodsThe studied group included 60 patients with clinically diagnosed AD, and the control group included 61 healthy volunteers. The study protocol included collection of biological material for tests, DNA isolation and evaluation of its quality and quantity, and PCR amplification of the isolated genetic material.ResultsIn the studied group, both changes in the nucleotide sequence of the FLG gene were detected and in the control group no tested mutations were detected. In 18 (30%) patients with AD, 22 mutations (4 heterozygous and 1 homozygous ones of R501X and 10 heterozygous and 7 homozygous ones of 2282del4) were detected.ConclusionsA high rate of mutations of the FLG gene in patients with clinically diagnosed AD and pathologically dry skin was observed in the studied population. The 2282del4 mutation occurred more often than R501X.

Phenotypic and genotypic aspects of Pseudomonas aeruginosa in chronic wounds: a descriptive study

Wound infections can prolong the inflammatory phase of healing. The bacterium Pseudomonas aeruginosa is commonly found in chronic wounds and contributes to the chronicity. It also presents natural antimicrobial resistance, which complicates treatment, and may vary depending on the product used in the lesions. Objective: To analyze the Pseudomonas aeruginosa strains found in the chronic wounds of outpatients treated with carboxymethylcellulose 2% gel or polyurethane board. Method: a descriptive study with a quantitative approach, through the collection of biological materials from wounds by swab, culture, identification and molecular characterization of the microorganisms found.

Research Concerning the Variability of the Main Characteristics of Lettuce from V.R.D.S. Buzau Germplasm Collection

Lettuce ( Lactuca sativa L.) is the most important crop in the group of leafy vegetables. Lately, the nutritionists recommend more often lettuce consumption because of its high content in vitamins, mineral salts, nutrient substances and low content in carbohydrates. Research made at V.R.D.S. Buzau respond to two major problems: biodiversity conservation and creating new vegetable varieties adapted to nowadays climatic conditions. Since 2011 started the collection of biological material at this species from Buzau vegetable area. In this sense, the lettuce collection from V.R.D.S. Buzau contains over 30 accessions. This study presents the main characteristics of 10 accessions cultivated in open field in 2013 – 2014 periods. In what it concerns accession 14, in 2013 there were registered the highest values concerning plant characteristics (height – 26 cm, diameter – 36 cm, weight – 504.33 g) and eatable part weight (319.33 g), and in 2014 the values of this accession reached or surpassed the mean value of the experience. All accessions presented, had made a head at the consumption maturity, except accession 18. Most variants had green leafs, only 3 accessions (8, 15  and 18) had anthocyanin coloration.

THE BIOLOGICAL ACTIVITY OF PLANT-DERIVED ANTI-HER2 RECOMBINANT ANTIBODIES

Pharmaceuticals derived from plants, have become one of the leading commercial directions in modern biotechnology. The benefits that offer these technologies, cannot be matched with any other modern technology for producing drugs from recombinant proteins. Main advantages of plant technologies for production of proteins are easy scalability, efficiency, bio-safety, ease of cultivation and collection of biological material. This approach promises to be the most perspective for production of a wide range of drug substances and vaccines. In current investigation we have analyzed in vitro and in vivo biological activity of plant-derived anti-HER2 recombinant antibodies - phytotrastuzumab. Phytotrastuzumab and trastuzumab have similar activity in grows suppression of breast cancer cells overexpressing HER2 in-vitro and were active in suppression of xenografted tumors SK-BR-3 in-vivo.

Analysis of Polymorphic DNA Sequences in the Identification of Individuals and its Possible Use in Biometric Systems

The article presents the achievements in DNA forensic science practice, the latest developments as well as future trends. The article concerns also other than forensic DNA applications as well as expectations, concerns and obstacles. DNA identification technology unlike other biometric techniques requires the collection of biological material and the identification is not performed in real time. DNA utilized in most of forensic identification, is present almost in every living cell in the body. What is more, each cell of the same body has the same DNA molecule which means that it is possible to compare the DNA sampled from different sources, for example saliva with blood or semen from the same person. Rapid development and reliability of DNA technology contribute to the fact that the analysis of polymorphic DNA sequences constitutes a very important evidence used in the court. The unique properties of DNA and rapid development of DNA analytical devices allow to claim that DNA may assume a more important position amongst biometric data in the future.

Biodiversity And Dynamics Of Occurence Of Epigeic Groups In Different Types Of Farming

Abstract The aim of this work was to determine and compare the occurrence of epigeic groups in two methods of farming, ecological one and integrated one. The research was conducte in the locality Nitra – Dolná Malanta in the years 2013 and 2014. The monitoring locality is situated in the south-western part of Slovakia, in altitude 175–180 m on highly productive soils. For the collection of biological material, the earth traps method was applied, used during the vegetation period (from April to October), within both farming systems, at Hordeum sativum, Triticum aestivum and Vicia faba undersowing with Medicago sativa. In canopy of these crops, two soil traps were installed, renewed in monthly intervals. The total of 7,722 exemplars of epigeic groups was obtained, of which 4,355 exemplars were in ecological farming and 3,367 exemplars in integrated farming system. In both treatments, 19 epigeic groups were determined, with dominant abundance of Coleoptera, Collembola, Acarina, Araneae. Also other groups such as Diplopoda, Heteroptera, Chilopoda etc. were observed in lower occurrence. Based on the evaluation of influence of the crop in terms of the occurrence of epigeic groups, the most suitable conditions created Vicia faba with undersowing Medicago sativa (integrated farming) and Triticum aestivum (ecological farming). On the basis of calculated indexes, both farming systems can be evaluated as homeostatically balanced, providing present epigeic groups with topical and trophic conditions.

Diversidad, abundancia y distribución de la macrofauna bentónica de las costas rocosas al norte del Estado Sucre, Venezuela

The rocky intertidal zone is among the most extreme physical environments on Earth. Organisms living in this area are constantly stricken by physico-chemical and biological factors. Due to the ecological importance of these areas, we studied the diversity, abundance and distribution of the rocky coastline benthic macrofauna, from the North coast of Sucre State, Venezuela. We performed bimonthly samplings from November 2008 to September 2009. The collection of biological material in the littoral zone (supra, mid and infralittoral) was done manually with a grid of 0.25m2. Organisms were preserved in 10% formalin for later identification and analysis (ecological parameters and Kruskal-Wallis test to the abundance and diversity). We found a total 19,020 organisms (86 spp.), in 8 phyla, 45 families and 47 genera. Mollusks were the most abundant and diverse (58 spp.), followed by arthropods (12 spp.), annelids (7 spp.), echinoderms (5 spp.), and the less represented cnidarians, sipunculids, nemertinids and urochordates (1 sp.). The zonation found coincided with the universal scheme of zonation. The towns of Rio Boca and Rio Caribe presented the highest values of ecological parameters, and the lowest were found in Playa Grande. Statistical significant differences were found in the abundance and diversity of macrofauna among the three zones. The little information on the composition and distribution of macrobenthic rocky coastline, prevents a better comparison, however the results contribute to the knowledge of the marine biodiversity in this region.

AB1056 Development of A Biobank for Research on Rheumatic Diseases Before, during and after Pregnancy

BackgroundInflammatory Rheumatic diseases (IRD) frequently affect women of fertile age. Adverse pregnancy outcomes and recurrent pregnancy complications are increased. The profound immunologic adaptions necessary for maternal tolerance toward the foetus...

Access and benefit sharing of Antarctica's Biological Material

Searching and sampling of Antarctic Biological Material (ABM) is happening with no explicit regulation on access and benefit sharing requirements. Patents already exist on inventions stemming from Antarctic living organisms. The Antarctic Treaty System (ATS) provides mechanisms to ensure that scientific knowledge and data generated from the collection and use of ABM are shared, although commercialization might be a threat to this free exchange of scientific knowledge. Some of the underlying problems regarding the access and benefit sharing of ABM are that under the ATS there are gaps concerning definitions, access to specimens, benefit sharing, commercialization and reporting issues. The Antarctic Treaty Consultative Parties (ATCPs) have decided that the Antarctic Treaty Consultative Meeting (ATCM) is the competent body to discuss the matter, and the ATS is the appropriate framework for managing the collection of biological material in the Antarctic Treaty area and for considering its use. Nevertheless, opinions diverge as to the need for more specific rules on access and benefit sharing other than that already resulting from the obligation to give prior notification and share scientific results.

Large Scale Collections of Biological Material and Ethical First Principles

AbstractCollecting and storing research materials is within the purview of all anthropologists. The establishment of collections, especially the large new biobanks of biological materials, requires an examination of the ways in which ethical first principles of autonomy, beneficence, and justice integrate with the goals and scientific aims of the collection. This article presents an example of the establishment of a repository of material for vervet monkeys (Chlorocebus sensu lato) and the ways in which informed consent, ownership of materials, and data sharing were considered in the establishment of the repository. Concepts of stewardship, derived from museum studies, are provided as an example of the way in which collections are currently regarded and maintained.

Laboratory Monitoring of Individuals with Subclinical Cutaneous Leishmaniasis Infections in Southern Brazil

Background: Cutaneous leishmaniasis (CL) has a high incidence worldwide. Most individuals with CL develop the cutaneous form, but subclinical infections may occur. Objective: The present study used different laboratory methods [polymerase chain reaction (PCR), indirect immunofluorescence (IIF) and enzyme reaction immunoassay (ELISA)] to investigate the incidence, and monitor individuals with subclinical infections. Patients and methods: The six individuals with subclinical infections included residents of a CL endemic area. Collections of biological material were carried out for nearly three years. Results: Two of these individuals were positive for Leishmania, one of them by PCR and the other by PCR, IIF and ELISA after previous negative results. Conclusion: The presence of a subclinical infection suggests the development of an appropriate immune response that can control replication of the parasite and maintain the integrity of the tissue. Individuals with subclinical infections must be correctly diagnosed and monitored using different methods, because of the possibility of the development of the mucosal form, and may serve as reservoirs for the insect vector.

Sleep complaints and cardio-cerebrovascular diseases in the elderly: A 6-year prospective study

Introduction While sleep disturbances may contribute to the development of cardio- cerebrovascular diseases (CVD), few studies have examined the association of sleep complaints (insomnia complaints and excessive daytime sleepiness (EDS)) with history of CVD and incident CVD in persons over 65. Materials and methods CVD was assessed at baseline and at the two, four, and six-year follow-ups in 5494 non-demented subjects. Self-reported insomnia complaints (poor sleep quality, difficulty in initiating sleep, difficulty in maintaining sleep, and early morning awakening), EDS and sleep medication use were evaluated at baseline. Logistic regression models and Cox proportional hazard models, with delayed entry and age of participants as the time scale, were adjusted for socio-demographic, lifestyle and clinical variables. Results At baseline, 748 participants had a past-history of CVD which was associated with EDS (OR = 1.28 95% CI = [1.05–1.57]) and the number of insomnia complaints (OR = 1.26 95% CI = [1.03–1.55] for 1–2 insomnia complaints; OR = 1.32 95% CI = [1.03–1.71] for >2 complaints). In longitudinal analyses, neither the four components of insomnia nor the number of insomnia complaints were significantly associated with first or recurrent CVD events (n = 391 events). EDS was independently associated with future CVD events even after adjusting for prescribed sleep medication and past-history of CVD (HR = 1.35 95% CI = [1.06–1.71]). Conclusion Our results suggest a complex relationship between sleep complaints and CVD. Insomnia complaints are more likely to be a consequence of CVD, whereas EDS appears to be a determinant of CVD independently of past-history of CVD. EDS screening may thus constitute a means of detecting persons at high risk of CVD. Acknowledgements The 3C Study is conducted under a partnership agreement between Inserm, the Bordeaux II University and Sanofi- Synthelabo and was supported by the FRM, the CNMTS, DGS, MGEN, Institut de la Longevite, AFSSPS, the Regional Governments of Aquitaine, Bourgogne and Languedoc-Roussillon and, the Fondation de France, the Ministry of Research-Inserm Programme Cohorts and collection of biological material). The Lille received a grant from Eisai. Part of this project is financed by two grants from the ANR.

CHAMACOS: A Birth Cohort Study in an Immigrant Population

Background. Epigenetic mechanisms may provide a link between environmental exposures and effects on children’s health and provide clues for possible interventions. Birth cohorts with repeated collection of biological material can allow for an evaluation of epigenetic changes from thein utero period through childhood. Aims.To describe the CHAMACOS birth cohort study and the use of measures of DNA methylation to link environmental exposures and child health. Methods.We enrolled 601 pregnant women primarily of Mexican origin and have followed their children (currently 12 years old). We measuredin uteroand child exposure to numerous environmental agents in stored biological samples. In blood samples collected from the cord (fetal blood) and from the child at age 9, we measured global and site-specific DNA methylation in by Illumina Infinium Human Methylation 450K BeadChips. Results. We have previously shown that exposure to various environmental agents have been related to fetal growth, neurodevelopment and obesity. We have now begun to examine whether epigenetic changes may mediate these associations. We observed that approximately 15.5% of all investigated CpG sites were differentially methylated between the fetal period and age 9. We also found a significant difference in methylation by sex. Markers of DNA methylation will be examined in relation to prenatal exposure to environmental pollutants and as mediators of children’s health. Conclusions.Birth cohort studies can allow for the investigation of the role of epigenetics in mediating the relationship of environmental exposures and health of children. Acknowledgements: This work was supported by PO1 ES009605 and R01 ES017054 from the NIEHS and R82670901, RD83171001, and RD83451301 from the U.S. EPA.

Beta-glucan feeding effect on biochemical and immune responses in vaccinated and non-vaccinated piglets against proliferative enteropathy

The aim of this study was to compare the results of active non-specific immuno-modulation in form of feeding food additive based on beta-glucan to pregnant sows and consecutive specific immuno-modulation by vaccination of their sucklings. Experimental sows were fed feedstuff with preparation on basis of beta-glucan at a 5% concentration from day 14 before parturition until the weaning of piglets; control sows were fed standard feedstuff only. Sucklings were vaccinated with a single dose of 2 ml of oral vaccine Enterisol® Ileitis one week before weaning. Collection of biological material was done 3 × in sows and 5 × in sucklings. Post-vaccination examination of blood serum for antibodies against L. intracellularis in sucklings in experimental and control groups was negative, probably due to insufficiently long period of rejection of antibiotics-medicated feedstuff and by chlorinated water. With the exception of immunological profile, no essential changes were recorded in the dynamics of other indices of examined profiles in groups of sows in relation to immune-modulation. In groups of sucklings from sows fed beta-glucan supplemented feedstuff significant changes were determined for various indices compared with sucklings from sows fed standard foodstuff. This is the first similar study in pig herds in Slovakia.

Open Data Sharing in the Context of Bioresources

Recently many international initiatives have been developed to improve access to scientific information and to promote open data sharing. In the complex field of bioresources, the BRIF (Bioresource Research Impact Factor) project aims to create suitable methods to recognise and measure the use and impact of biological resources in scientific/academic work, in order to maximize access by researchers to collections of biological materials and attached databases, and to recognize efforts involved in their maintenance. The lack of a proper recognition of scientific contribution is in fact a major obstacle which impedes bioresource sharing. In this context, the BRIF initiative can be considered as a tool to facilitate research resource sharing.