Case Report of a Successful Pregnancy in a Cystic Fibrosis Patient with The c.1521_1523delCTT/c.3718-2477C>t Genotypes.

The aim of this case report was to show the consequences of pregnancy in a cystic fibrosis (CF) patient with a rare mutation. We present a case of a patient with CF, pregnant for the second time, who gave birth to a healthy child. Her mutation status revealed the presence of relatively rare mutation c.3718-2477C>T that is associated with a milder phenotype of the disease. During pregnancy, her vital signs were within normal limits. She had no exacerbations after the third gestational month. Cystic fibrosis is the most common genetic disorder among Caucasians. Over the last few decades, the survival rate and the lifespan of patients with CF have increased progressively. This is why more affected women are choosing to become pregnant. Predictive factors for the pregnancy outcome are basal pulmonary function [measured by forced expiratory volume/1 second (FEV1)], nutritional status [measured by body mass index (BMI)], diabetes and bacterial colonization. The report of our case emphasizes the need for establishing the exact mutations in CF patients who plan to become pregnant in order to predict the possible outcomes of this specific period of life. Moreover, genetic counseling is strongly recommended for the right understanding of the pregnancy risks in such cases.

The functional role of the photosynthetic apparatus in the recovery of Brassica napus plants from pre-emergent metazachlor exposure.

Metazachlor is a chloroacetamide herbicide, frequently used in Brassica napus cultivations around the world. Its primary target is the inhibition of very long chain fatty acid biosynthesis. This study included a morphological and physiological screening of hydroponically grown B. napus, exposed to a concentration range of 0, 0.25, 0.50, 0.75 and 1.0kg metazachlor per hectare. The results indicate that within a month after application, growth and development of B. napus are severely affected by low metazachlor doses. At intermediate metazachlor concentrations, loss of phosphorous and potassium from the plant tissues suggests destabilisation of cellular membranes, which may be a direct consequence of metazachlor application. This membrane instability could be indirectly linked with alterations of electron transport and a reduction of carbon assimilation. At increased metazachlor doses of 0.75kga.i.ha(-1), pigment concentrations are strongly reduced. However, chlorophyll fluorescence parameters seem to remain unaffected at metazachlor doses up to 0.75kga.i.ha(-1). At a metazachlor concentration of 1.0kga.i.ha(-1), negative effects are observed on all tested parameters, resulting in limited survival. The results indicate photosynthesis is assured at intermediate metazachlor concentrations for the cost of growth and development. It is clear that photosynthesis plays a key role in the survival strategy of young plants to overcome initially induced chemical stress.

Morphological, genetic and functional variability of a T-cell hybridoma line.

The variability in the morphology, modal number of chromosomes, TCR expression and functional reactivity of a CII-specific T-cell hybridoma at continuous subcultivation have been investigated. As the number of passages increased, besides the oval semiadherent cells (normal phenotype), fibroblast-like cells (transformed phenotype) were also observed. The two cell subpopulations differed in their karyotype characteristic, as well as in their functional reactivity. The cell population with a normal phenotype was characterized by a tetramodal number of chromosomes (30, 40, 48 and 70) and trisomies of chromosomes 6 and 14, while the cell population with a transformed phenotype was characterized by a trimodal number of chromosomes (11, 68 and 74) and trisomy of chromosome 12. A nullisomy of sex chromosomes was established in both types of cells. In the initial passages of subcultivation, 73.04% of the cells with a normal morphological phenotype expressed TCR-CD3 complexes on their surface and possessed high functional reactivity. After a two-week subcultivation, the values of these indices went down considerably: 46.11% of the cells expressed functional TCR-CD3 complexes, as a result of which their functional reactivity decreased. Only 2.71% of the cells with a transformed morphological phenotype expressed functional TCR-CD3 complexes on their surface. In these cells, a total loss of reactivity towards the specific antigens was established. The achieved results show that at continuous subcultivation the T-cell hybridomas are unstable, and with the increase in the number of passages there appear chromosome rearrangements, leading to loss of their functional reactivity.

Protein extracts for nutritional purposes from fragile strains of Saccharomyces cerevisiae: reduction of the nucleic acid content and applicability of the protein extracts.

92% of the cellular proteins from the osmotically fragile strain Sacch. cerevisiae 211 (MATa srbl srb2 tsl) were extracted by one passage in a high pressure disintegrator at 650 atm. After isoelectric precipitation, 60.1% of the proteins were isolated. The protein extract was rich in proteins (60-70%) but also contained 12-13% nucleic acids and 13-14% carbohydrates. Mild conditions were created for simultaneous reduction of nucleic acids and carbohydrates in the lysate prior to precipitation of proteins. Under these conditions the amount of nucleic acids in the protein extract was reduced to 2%, and that of the carbohydrates to 5.7%. As a result of the whole procedure on a pilot scale, 11 culture of Sacch. cerevisiae 211 grown in molasses based nutritional medium yielded 1 g protein extract. The applicability of this protein extract for nutritional purposes was demonstrated by determination of its amino acid composition, acute toxicity tests with Tetrahymena pyriformis and short-term animal nutrition tests. The results showed that the protein extract is a suitable source of SCP for nutritional purposes.

Protein extracts for nutritional purposes from fragile strains of Saccharomyces cerevisiae: construction of strains and conditions for lysis.

A superfragile strain, Saccharomyces cerevisiae 211, carrying three nonallelic nuclear determinants of cell lysis by osmotic shock was isolated from the haploid progeny of a cross between two laboratory fragile strains VY 1160 and SY 15. The strain Sacch. cerevisiae 211 is prototrophic, grows well in both laboratory and industrial media, only when supplemented with osmotic stabilizers. The average protein content of its biomass after growth in a molasses-based nutritional medium is 48.7%. In laboratory conditions this strain releases about 60% of its cellular proteins by spontaneous lysis in water. However, on a pilot scale the protein yield decreases to about 25%. After a single disintegration step at 650 atm, practically all of the cellular protein--91.5%, is extracted without any additional chemical treatment. This result cannot be reached in any of the wild type strains tested even after triple treatment at 650 atm.

Polyploid fragile strains of Saccharomyces cerevisiae--a novel source of proteins for nutritional purposes.

A series of prototrophic fragile strains of different ploidy (2n, 3n and 4n) has been genetically constructed on the basis of haploid srb1 containing segregants of the fragile Saccharomyces cerevisiae mutant VY 1160. The strains have been characterized by several criteria. In regard to generation time, biomass yield, and nucleic acids content of the cells, the tetraploid srb1 homozygous hybrid is indistinguishable from an industrial strain of S. cerevisiae. However, it is characterized by a higher protein content. Unlike any other laboratory or industrial strains, the original mutant and these hybrids possess an ability for lysis upon suspension in hypotonic solutions. The dependence of the percentage of lysed cells on the growth phase and concentration of osmotic stabilizer in the medium has been investigated. The quantity of proteins in the soluble and insoluble fractions obtained after lysis of these strains by osmotic shock has been determined. These hybrids can be considered as a potential industrial source of proteins for nutritional purposes.