Loci controlling lymphocyte production of interferon γ after alloantigen stimulation in vitro and their co-localization with genes controlling lymphocyte infiltration of tumors and tumor susceptibility

Low infiltration of lymphocytes into cancers is associated with poor prognosis, but the reasons why some patients exhibit a low and others a high infiltration of tumors are unknown. Previously we mapped four loci (Lynf1–Lynf4) controlling lymphocyte infiltration of mouse lung tumors. These loci do not encode any of the molecules that are involved in traffic of lymphocytes. Here we report a genetic relationship between these loci and the control of production of IFNγ in allogeneic mixed lymphocyte cultures (MLC). We found that IFNγ production by lymphocytes of O20/A mice is lower than by lymphocytes of OcB-9/Dem mice (both H2 ^pz ) stimulated in MLC by irradiated splenocytes of C57BL/10SnPh (H2 ^b ) or BALB/cHeA (H2 ^d ) mice, or by ConA. IFNγ production in MLCs of individual (O20 × OcB-9)F₂ mice stimulated by irradiated C57BL/10 splenocytes and genotyped for microsatellite markers revealed four IFNγ-controlling loci (Cypr4-Cypr7), each of which is closely linked with one of the four Lynf loci and with a cluster of susceptibility genes for different tumors. This suggests that inherited differences in certain lymphocyte responses may modify their propensity to infiltrate tumors and their capacity to affect tumor growth.     

External factors involved in the regulation of an extracellular proteinase synthesis in Bacillus megaterium

Summary A mathematical model was formulated to describe the kinetics and stoichiometry of growth and proteinase production in Bacillus megaterium. Synthesis of the extracellular proteinase in a batch culture is repressed by amino acids. The specific rate of formation of the enzyme (r _E) can be described by the formula {ie373-1}, where k ₂ and k ₃ stand for the non-repressible and repressible part of enzyme synthesis respectively, k _{S 2} is a repression coefficient and S ₂ indicates the concentration of amono acids; the values of k ₂ and k _{S 2} depend on the composition of the mixture of amino acids. Even in a high concentration, a single amino acid is less effective than a mixture of amino acids. The dependence of the proteinase repression on the concentration of an external amino acid (leucine) follows the same course as its rate of incorporation into proteins, approaching saturation at concentrations higher than 50 M (half saturation approximately 10 M). However, the total uptake of leucine did not exhibit any saturation even at 500 M external concentration.Symbols X biomass concentration, g/l - E proteinase concentration, unit/l - t time, h - S ₁ concentration of glucose, g/l - S ₂ concentration of amino acids, g/l - specific growth rate, l/h - rE specific rate of enzyme production, unit/g/h - k ₁ growth kinetic constant, l/h - k ₂ product formation kinetic constant (for non-repressible part of enzyme synthesis), unit/g - k ₃ product formation kinetic constant (for repressible portion of enzyme synthesis), unit/g - k _{S 1} saturation constant, g/l - k _{S 2} repression coefficient for a certain amino acid or amino acids mixture, g/l     

Contribution of leaves of different ages to plant carbon balance as affected by potassium supply and water stress

The carbon balances of whole, 21-d old French bean plants (Phaseolus vulgaris L.) grown in standard nutrient solution (1K) and its modifications without (OK) or surplus (2K) potassium were calculated from the daily photosynthetic carbon inputs of individual leaves, and the daily respiratory carbon losses by individual leaves, stalks and petioles, and roots. Under the three K concentrations, maximum net photosynthetic rates (P_n) were found in the 2nd or in the 3rd trifoliate leaves, maximum respiratory rates (R_d) in the youngest, 4th trifoliate leaves; the P_n/R_d ratio decreased with leaf age. In all leaves of 2K plants, leaf dry masses and thicknesses, P_n, P_n/P_d ratios, and stomatal and intracellular conductances were lower than in OK and IK plants. Daily whole-plant net carbon gain was highest in IK plants, whereas in OK and 2K plants it was 98.0 and 81.3 % of IK, respectively. Similar values were found in the parameters of growth analysis, namely in net assimilation rates and relative growth rates. No differences were found in water potential (Ψ _w ) or water saturation deficit (W_sat) in the OK, 1K and 2K plants sufficiently supplied with water or during wilting and resaturation. The decrease in Ψ_w to −0.97 MPa was associated with a 19.9 %, 31.4 % and 23.4 % decrease in P_n of OK, 1K and 2K plants, respectively, but no effect on R_d was found. In the three variants, the short-time effect of mild water stress was fully reversible.     

Subcellular organization of Streptomyces aureofaciens and overproduction of chlortetracycline

Biosynthesis of chlortetracycline is localized differently under low- and high-production conditions (standard low-production strain and its high-production variant). The experimental evidence was based on the assay of anhydrotetracycline oxygenase in subcellular fractions, ultracytochemical localization and electron-probe X-ray microanalysis of the product in the mycelium. Overproduction of chlortetracycline is closely associated with compartmentation of biosynthetic enzymes and with an efficient export of the antibiotic out of the cell.     

Molecular characterization of β-thalassemia in Czechoslovakia

Summary We have identified different -thalassemia mutations in 93 members of 34 families of Czech or Slovakian descent using gene amplification, hybridization with specific ³²P-labeled oligonucleotide probes, sequencing of amplified DNA, and gene mapping. The GA mutation at IVS-I-1 was found in 18 families; other Mediterranean mutations were IVS-II-1 (GA), IVS-II-745 (CG), IVS-I-110 (GA), and codon 39 (CT); these were present in 9 additional families. The GT mutation at codon 121, known to cause Heinzbody -thalassemia, was present in 3 families, and the frameshift at codons 82/83 (-G), first described in the Azerbaijanian population, in 2 families. A newly discovered allele was a frameshift at codons 38/39 (-C). One -thalassemia allele was incompletely characterized. We observed in 2 families a TC mutation at position +96 UTR (untranslated region) relative to the termination codon; this mutation likely is a rare polymorphism, -Thalassemia was rare; only one person carried the -^3.7 heterozygosity, and one other had a yet to be identified -thalassemia-1, while seven had the ^{anti 3.7} triplication.     

Timing of laparoscopic aspiration of preovulatory oocytes in heifers

Twenty-one heifers were synchronized with PGF(2) alpha and 22 heifers were stimulated with FSH-P in decreasing doses and synchronized with PGF(2) alpha. The beginning of LH rise was observed to be 47.6+/-14.0 h and 35.9+/-1.3 h (P < 0.05) and the peak of LH rise was observed at 53.8 +/- 13.4 h and 40.14+/-1.4 h (P < 0.05) after the luteolyticum administration in the synchronized and superovulated group respectively. The beginning of LH rise was observed 6.9+/-6.7 h and 4.8+/-2.6 h (P < 0.05) and the LH peak was observed 11.8+/-7.7 h and 8.3+/-3.2 h (P < 0.05) after the frist symptoms of oestrus in the synchronized and superovulated group respectively. Ovulation was not observed in stimulated heifers in the period of 23-25 h after the preovulatory LH rise. Compact cumulus oophorus was seen at 30.5%, expanded at 67.0 and partial at 2.5% during this interval of 23-25h. Within this same interval 26.9%, 51.3% and 21.8% oocytes without perivitelline space, with perivitelline space and with extruded first polar body were aspirated respectively. It may be concluded from the reported results that to recover fully mature oocytes for in vitro fertilization, it will be necessary to monitor the preovulatory period of the donor cow in great detail.     

An improved method to isolate lichen algae by gel filtration

Photobiont cells of the lichen Evernia prunastri have completely been separated from their fungal partner by filtration through a bed of Sepharose 2B. Both mannitol and ribitol have been quantified by gas-liquid chromatography in the different steps of the isolation procedure. Absence of mannitol, which is exclusively produced by the mycobiont, has been used as the best probe to monitor isolation.     

Invertase immobilization via its carbohydrate moiety

After periodate oxidation of its glycosidic component, invertase was covalently bound onto three types of modified solid supports: glycidyl methacrylate, styrene-divinylbenzene copolymers, and bead cellulose. Direct reaction of the invertase aldehyde groups that were formed with amino groups of the support and use of the modified Ugi reaction have been employed as immobilization procedures. Apart from binding methods, the important effects of the buffer, support, conditions of periodate oxidation, and the length of the spacer on the activity of the enzyme conjugate have been investigated. Superior conjugate activity was obtained, via modified Ugi reaction, by the immobilization of a suitably oxidized invertase to a styrene-divinylbenzene copolymer having free amino groups.