|
| |
|
| |
|
| |
|
| |
|
| |
|
| |
|
| |
|
|
Biopharmaceuticals from the MS point of view
Hubálek, Martin ; Lenčo, J.
The course aims to introduce biotherapeutics, which are becoming an increasingly important part of the pharmacotherapy of serious diseases. Mass spectrometry plays an important role for their quality control in the pharmaceutical industry. We will introduce the audience to biopharmaceuticals, which in this context are peptides, proteins, monoclonal antibodies or oligonucleotides. From the analyst's point of view, we will focus mainly on antibody drugs, looking at the possibilities of their separation and analysis by mass spectrometry, at the level of intact protein molecules, antibody subunits or peptides.
|
|
| |
|
|
Polyploidizační schopnost jeseterů a její vliv na fitness
HUBÁLEK, Martin
Sturgeons (order Acipenseriformes) are considered one of the most ancient groups of fish still present on Earth. The evolution of these 'living fossils' is inherently connected to polyploidization events. Moreover, spontaneously arisen polyploidy has been widely reported in farmed populations of many sturgeon species, and there is also evidence that it randomly occurs in the wild. The persistence of sturgeons for creating polyploid genomes underlines the genome plasticity of these fishes and makes them suitable candidates for polyploid research. However, only a few studies have ever investigated the effects of newly formed polyploidy on fitness or physiology of sturgeons. Broadening the scope of research is of particular interest, since it may shed light on how sturgeon neopolyploids perform in aquaculture and nature, uncover the impact of contemporary polyploidization on sturgeon farming and conservation, and contribute to scientific knowledge regarding the consequences of polyploidy on fish. The second chapter of this thesis introduced an optimized protocol for the creation of biologically triploid populations of sterlet (Acipenser ruthenus) via the second polar body retention (SPBR) achieved by hydrostatic pressure shock. Apart from the presented study, the application of this type of shock in sturgeons has only ever been reported in shortnose sturgeon (Acipenser brevirostrum). Our findings suggested that hydrostatic pressure shock can be more reliable at producing 100% triploids and yield better hatching rates than conventionally used heat shock. Its utilization may thus improve the efficiency in obtaining triploidized progeny of sturgeons. Flow cytometric analysis of relative DNA content in cell nuclei is among the most preferred techniques for the measurement of ploidy levels in sturgeons, but its application has so far been limited to the examination of fresh material. The third chapter of this thesis provided the verified methods for 10-day storage of various types of sturgeon samples - blood, finclip and larva tail tissue. The protocols proposed are easy to use, do not contain centrifugation steps and allow field sampling. The viability and growth of early developmental stages of autopolyploid sterlet kept at both optimal (16 °C) and suboptimal temperatures (10 and 20 °C) were investigated in fourth chapter. The results suggested that irrespective of temperature, biologically triploid sterlet arising from SPBR do not differ from individuals of normal ploidy (normoploids) in terms of survival and growth. The viability of embryos obtained from tetraploidization via the first mitotic division suppression (FMDS) was considerably reduced at optimal temperature and even more so at suboptimal conditions. Biologically tetraploid prelarvae were not able to survive at suboptimal temperatures, assuming their lower thermal tolerance when compared to normoploids or biological triploids. Biological hexaploidy seemed to be lethal, since the individuals obtained from the combined treatment causing both SPBR and FMDS were unable to survive the prelarval period at any temperature. In the fifth chapter of this thesis, the effects of SPBR-induced polyploidy on critical swimming speed, physiology and haematology were investigated using nine-month-old juveniles of two pure sturgeon species, sterlet and Siberian sturgeon (Acipenser baerii), and their reciprocal hybrids. SPBR-induced polyploids of both purebreds and hybrids did not differ from their normoploid counterparts in swimming performance, and the haematological analysis revealed that they were able to fully compensate for having fewer erythrocytes than normoploids in blood haemoglobin and haematocrit. Likewise, the study did not provide any evidence for SPBR-induced polyploidy affecting stress response. These results may indicate that sturgeon polyploids arising from SPBR do not suffer from any substantial physiological limitation and are as fit as their counterparts of normal p
|
guest ::
RSS feed.