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RESEARCH AND PATENTS

Halotech DNA SL, in collaboration with researchers affiliated with the Department of Biology of the Universidad Autónoma de Madrid [Autonomous University of Madrid], the Ministry of Science and Innovation and the European Union , has developed, and continuously develops the following R&D projects:

  1.  “DNA fragmentation analysis systems for the evaluation of sperm quality and the generation of antibiograms” carried out between 2008 and 2010 as part of the NEOTEC grant of CDTI (Centro para el Desarrollo Tecnológico Industrial [Centre for Industrial Technology Development]) (IDI-20080728). Funded by the Ministry of Science and Innovation.
  2. “New methodology for the analysis of human sperm DNA quality” for the year 2008 and 2009 as part of the Programa Nacional de Investigación Aplicada (National Applied Research Programme). Applied collaborative research sub-programme in the form of projects conducted in science and technology parks. (PPT-010000-2008-012).

    a) Improve existing systems for the analysis of human sperm DNA quality, attempting to optimise the times spent on the process.

    b) Implement new strategies and processes adapted to other species with commercial value based on the knowledge acquired in humans.

  3. “Magic Bullet -Optimisation of treatment with off-patent antimicrobial agents of ventilator-assisted pneumonia” FP7-HEALTH-2011-TWO-STAGE.

    The objectives of the project are:

    a) Development of a basic system to create antibiograms that do not exceed a time limit of 60 minutes after isolation of the responsible bacteria. The methodology is based on technical variants of those developed for human spermatozoa. Currently, and through a European project that ends this year, bacterial variants associated with Ventilator-Assisted Pneumonia are being investigated.

SCD TECHNIQUE:

The method is based on SCD (Sperm Chromatin Dispersion) technology (Fernández et al., J. Androl 24: 59-66, 2003; Fertil Steril 84: 833-842, 2005). Spermatozoa from fresh, frozen, or diluted samples are immersed in an inert agarose microgel on a pretreated slide. After, they are subjected to an initial acid treatment that denatures the DNA in those sperm with fragmented DNA. Following this, they are treated with a lysis solution that eliminates most nuclear proteins. After this treatment, spermatozoa with fragmented DNA do not show a halo, and those with intact DNA show large halos around the nucleoid.

Our patent for DNA fragmentation has been widely tested in universities, clinics, and hospitals in over 50 countries. The highly reproducible and reliable technique is based on Halotech's principles: simplicity, speed, and accessibility. To learn more about our solutions, you will have access to results and publications backed by the major scientific journals, which lend credence to the optimal and beneficial use of our kits

DETECTION OF OXIDATIVE STRESS:

The presence of destructive reactive oxygen species (ROS), a concept associated with oxidative stress, results when natural antioxidant defences are incapable of blocking ROS activity. The net result is that cellular damage occurs at different levels. Somatic and germ cell lines appear to be targets of ROS: when they affect germ cell lines, excess oxidative stress has a direct impact on male fertility (Aitken and De Iuliis, 2010).

Several trials are currently in progress, focused on the evaluation of the various levels of ROS and the molecules responsible for producing oxidative stress. However, they are not used very often in clinical andrology laboratories. Oxisperm provides an established, easy, and reliable assay to the clinic to measure possible excesses of anions associated with oxidative stress (O2). This test is based on the nitro blue tetrazolium (NBT) assay, in the form of a reactive gel (RB) in the Oxisperm kit. The NBT assay is based on the ability of water-soluble tetrazolium salt to be converted into a blue formazan crystal due to the action of superoxide anions (Baehner et al., 1976).

In sperm, the products of this reaction adhere to sperm membranes and can be easily visualised in a bright field microscope (Image 1). These crystals produce an increase in the intensity of the colour in the RG (from yellow through to different levels of purple-blue, as seen in the diagram), that can be easily quantified and compared using a colour scale. Alternatively, a colourimeter can be used to measure the absorbance of the wavelengths, which range from 530 nm to 630 nm. The intensity of the colour is closely tied to the level of oxidative stress (an excess of superoxide ions) in the sample (see the colour palette).