How do ionic channel properties depend on the structure of polyene antibiotic molecules?

A study has been made of the properties of ionic channels formed in phospholipid-cholesterol bilayers by polyene antibiotics of various molecular structures. Properties of channels created by natural antibiotics with different structures of the lactone ring (amphotericin B-nystatin-mycoheptin) as well as by some derivatives of amphotericin B modified with respect to the amino and carboxyl groups are compared. Neutralization of one or both charges of the amphotericin B molecule (both by chemical modification and by pH shift) increases the probability of the channel to be in a nonconducting state. An increase of cholesterol concentration in the membrane produces an opposite effect. It is assumed that the electrostatic interaction of the amino group of an antibiotic molecule with the carboxyl group of an adjacent one stabilized the channel. Conductance and selectivity of an open channel are not influenced by changes in the charged groups. These properties strongly depend on the structure of the polar chain of the lactone ring. For example, the appearance of one more carbonyl group in the mycoheptin molecule results in a sharply decreasing anion permeability of channels. An antibiotic concentration which is necessary to observe single channels depends on the polyene chain structure: this is about 10(-7) M for tetraene nystatin and 2.10(-8) M for heptaene amphotericin B an mycoheptin.     

Quantitative protein changes in the hypothalamic neurons of pubertal male rats, castrated neonatally.

The effect of neonatal castration of male rats on the sexual differentiation of the hypothalamus at puberty was studied. Male rats were castrated on days 1, 5 and 7 after birth. Their brains were processed for study on days 83-85. The neurons and cell nuclei of the preoptic area, mediobasal and ventromedial nuclei were assessed for changes in cell and nuclear sizes and dry weight (calculated using interferometric methods). Neonatal castration resulted in size as well as dry weight increase in the neurons of the anterior and mediobasal hypothalamus. The dry weight increased by 34% (P less than 0.001) in the medial preoptic area, by 25% (P less than 0.001) in the arcuate neurons and by 22% (P less than 0.001) in the ventromedial nucleus. The cell nuclei exhibited perceptible weight increase too--in the medial preoptic area 68% (P less than 0.001); 55% in the arcuate neurons (P less than 0.001), and 39% in the ventromedial region. The weight and size increases in neonatally castrated males were equal to those of females of the same age. In rats castrated on day 7, the cell sizes and dry weights of the ventromedial nucleus increased but the cell nuclei exhibited only little change. It is assumed that the changes in the dry weight may be the result of increased synthetic processes in these groups of neurons which are connected with the tonic and cyclic release of gonadotropins. These changes also point to the hypothalamic differentiation shifting to the female type in the absence of the inducing effect of androgens.     

Spectral properties of phosphopyridoxyl-Lys-7(-41)-ribonuclease A.

We have studied the spectral properties of RNAase A containing a phosphopyridoxyl residue at the epsilon-NH2 group of Lys-7 or Lys-14. The overall conformations of the native and modified enzymes were shown to be rather similar. All three proteins have similar circular dichroism spectra within the 220-300-nm region, and similar thermal transition temperatures. All the changes in the RNAase A molecule modified are located in close proximity to the alkylated lysine residue. The phosphopyridoxyl group of (P-Pxy)-epsilon-Lys-41-RNAase A is situated directly at the enzyme active site and is 25% butied in the protein globule. The P-pyridoxyl group of (P-Pxy)-epsilon-Lys-7-RNAase A was shown to be located in the vicinity of the active site and to be more exposed to the solvent. In the pyridoxyl phosphate absorption band, optical activity is induced in both proteins. Study of the pH dependence of the changes occurring in the circular dichroism and absorption spectra has shown that in the modified proteins, the pyridoxyl phosphate chromophore is rather sensitive to the ionic state of the surrounding medium and serves as a "reporter" group when the relationship between structure and function of the RNAase A active site is being investigated.     

Ir gene influence on DNP-specific B-memory-cell formation

It is well known that Ir genes control the antibody production. To investigate whether they also influence B-memory-cell generation, CBA mice (nonresponders) were primed with dinitrophenyl-poly-(L-tyr,L-glu)-poly-(D,L-ala)-poly-(L-lys)-DNP-TGAL conjugate. At the same time animals were injected with ovalbumin (OVA) to activate OVA-specific T helpers. Two to four weeks later animals were challenged with DNP-OVA conjugate and the number of IgG-producing B cells was determined. The data presented indicate that carrier-specific MHC-restricted T helpers are not required for B-memory-cell generation. It is concluded that the defect of IgG response to DNP-TGAL in CBA mice is caused by a block in the maturation of memory cells to antibody-producing cells.     

俄罗斯北部平原(VYCHEGDA河盆地)全新世植物群和植被

对Vychegda河流域靠近Baika村(61°16'N,46°44'E)洪积平原的一个剖面进行了孢粉学研究.化石点位于泰加林带中部,靠近欧洲云杉林和亚乌拉尔-西伯利亚云、冷杉-西伯利亚松林界限附近.该河流盆地森林覆盖率高达98%.根据放射性碳年代测定,沉积涵盖了全新世大约自9000 yrs B.P.以来的大部分地层.主要运用化石植物群组成来恢复这一地区全新世以来的植被和气候变化,通过某些化石植物群的全部植物种类的现代地理分布的分析,可以发现与过去植被最接近的现代对应植被类型的位置,通过确定所有植物种现代生长地区的景观和气候来推测与古环境最接近的景观和气候.整个孢粉组合序列中云杉、松和桦占统治地位,但是植物群的成分和植被有变化.暖温带树种,如椴、榆及栎等在森林中的含量有变化,在全新世最暖期,相当于中晚大西洋期(6000-4500 yrs B.P.)达到最高.这些变化通过对应的现代植物群在俄罗斯平原的"迁移"反映出来,从中、南乌拉尔向西至Sukhona河盆地然后再返回到研究区的Vychegda河下游.运用古植物群的方法定量恢复的主要气候指标表明在大西洋期7月份月均温比现代高2.5℃,1月份月均温比现在高出1℃,年降雨量和现在接近.这段时间可以认为是本地区全新世的气候最适宜期.     

Conformational variability of recombination R-triplex formed by the mammalian telomeric sequence

Alignment of three nucleic acids strands, in which the third strand is identical to one of the DNA duplex strands, occurs in various cellular systems. In the case of telomeric t-loops, recognition between the DNA duplex and the homologous single strand is likely to be mediated by proteins through formation of the transient recombination-type R-triplex. Earlier, using 2-aminopurine as a fluorescent reporting base, we evaluated the thermodynamic characteristics of intramolecular R-triplex formed by a mixed nucleotide sequence. Here, we used this approach to explore a propensity of the telomeric TTAGGG repeat to form the R-triplex. The circular dichroism spectral changes detected upon formation of the R-triplex suggest that this process is accompanied by specific conformational changes in DNA, including a local destabilization of the target duplex next to a GGG run revealed by the fluorescence of the reporting 2-aminopurine base. Surprisingly, stability of the R-triplex formed by telomeric sequence depends strikingly on the counter ion, being higher for Na⁺ than for Li⁺. Taken together these findings indicate a significant conformational variability of telomeric DNA in the context of recombination-type R-triplex, a phenomenon of possible biological relevance.     

Dynamic changes in the response of molluscan neurons to cyclic AMP with repeated injections

Microiontophoretic injection of cAMP (but not of non-cyclic AMP) into Helix snail neurons induces rapid reversible membrane depolarization. The reversa potential of the cAMP response varies between +10 and -30 mV in different cells. The amplitude of the responses gradually increases (15 experiments), decreases (4 experiments) or remains unchanged (40 experiments) when the interval between the injections is less than 5 minutes. The identified neurons of the different preparations exhibit different types of dynamic changes in the amplitude of the responses to repeated cAMP injections. The possible role of the retaining current in the origin of the potentiation of cAMP responses was investigated. Potentiation of cAMP responses was preserved after the retaining current was switched off only in cases with no membrane depolarization due to spontaneous leakage of the substance from the microelectrode. It is suggested that potentiation or depression of cell responses to repeated cAMP injections is caused by dynamic changes in the cell cAMP system.