Guanidinium-carboxylate interactions

The crystal structures of the 1:1 complexes methylguanidinium benzylhydrogenmalonate, (C₂N₃H₈)⁺(C₁₀H₉O₄)^?, MGD.BMAL, and methylguanidinium ethylhydrogenmalonate, (C₂N₃H₈)⁺(C₅H₇O₄)^?, MGD.EMAL, and of the 2:1 complex methylguanidinium sulfate, (C₂N₃H₈)₂SO₄, have been determined from three-dimensional X-ray data. For MGD.BMAL, the complex crystallizes in the triclinic space group P with two formula units in a cell of dimensions a = 6.277(5), b = 8.470(3), c = 13.191(6)Å, α= 91.01(1), β= 99.64(9), γ= 90.83(5)°. The structure has been refined to a final value of R = 0.061 based on 1511 intensities. The MGD.EMAL complex is also triclinic, space group P with two molecules in a cell of dimensions a = 9.254(7), b = 9.625(6), c = 6.778(2) Å, α= 109.6(1), β= 100.8(1), γ= 62.7(1)Å. The crystals of this compound are of low quality, and the final value is R = 0.109 based on 706 intensities. (MGD)₂SO₄ is orthorhombic, space group P2₁2₁2₁, with four molecules in a cell of dimensions a = 7.100(4), b = 12.151(3), c = 13.108(2) Å. Refinement has converged to R = 0.054 based on 907 data. All three crystals exhibit extensive interionic hydrogen bonding. The hydrogen bonding in MGD.BMAL includes a Type B interaction and a Type 1 interaction, the latter being a pairwise interaction from both amino nitrogen atoms on the cation to two carboxylate oxygen atoms from the two different carboxylate groups in an anion. In MGD.EMAL, the anion participates in both a Type A and a Type B pairwise interaction with two neighboring cations. The possible implications of the hydrogen bonding patterns in these two compounds for the role of arginyl side chains in protection of γ-carboxyglutamate residues from decarboxylation are discussed.     

Control of feeding during saline consumption and fluid deprivation in hamsters

Cellular dehydration induced by water deprivation or hypertonic saline injection reduces feeding in a variety of species. Normal feeding in rats is maintained during isotonic saline consumption by increasing the intake of saline compared to the usual intake of water. Hamsters do not show the spontaneous preference for isotonic saline noted in rats, even after adrenalectomy. In the present investigation, feeding by hamsters was depressed during both isotonic and hypertonic saline consumption compared to the usual feeding with water. Saline intakes did not exceed water intakes under similar conditions. When fluid intakes were elevated by prior fluid deprivation, feeding rates increased at all concentrations of saline after a delay proportional to the osmolality of the solution. Positive 24-hr sodium balances were always associated with saline consumption. Water and hypertonic saline injections reduced feeding, and the fluid loads were excreted very slowly. When hamsters were fluid deprived prior to injections, saline totally suppressed feeding, while water increased feeding compared to sham injected controls. It is concluded that cellular dehydration produces a reduction of feeding in hamsters drinking isotonic or hypertonic saline. Reduced feeding with isotonic saline consumption results from the failure of hamsters to increase their ad lib intake of that solution. The prolonged retention of both sodium and fluid after saline consumption or injection suggests that further saline intake may be inhibited by an expansion of the extracellular space.     

Relations between the effect of tetanus toxin on the neuromuscular transmission and histological functional properties of various muscles of the rat

Summary Various doses of tetanus toxin were injected into three hind leg and two fore leg muscles of the rat. The neuromuscular transmission was tested by recording the mass action potential of the muscles elicited by a single electrical stimulus to the motor nerve after strong symptoms of local tetanus had developed. The muscle responses were depressed and blocked at lower toxin doses in the fast tibialis anterior than in the mixed gastrocnemius latemlis, while blocking of the slow soleus required the highest dose. The extensor carpi radialis and the flexor carpi ulnaris muscles showed medium sensitivity. In all five muscles the contraction time was measured and correlated with its individual minimal blocking dose. The more phasic (i.e., the faster) the muscle, the more sensitive its neuromuscular transmission was to tetanus toxin. The proportional distribution of red, white, and intermediate fibres, which are associated with specific end-plate types, was evaluated for the five muscles. The percentage of white fibres in the muscles displayed a very good negative correlation with the blocking dose. The relation between structures of end-plates and effects of tetanus toxin were analysed and it is suggested that the differences in sensitivity to tetanus toxin in the neuromuscular transmission in the five muscles is determined by a differential distribution of endplates with varying sensitivities to this toxin due to structural properties.This study is a part of a doctoral dissertation submitted by one of the authors (H.K.) to the Faculty of Medicine, University of Göttingen. Some of the results were presented at the 48th and 49th Congr. of German Physiol. Soc. (Kretzschmar et al., 1977, 1978) and at the 5th Internat. Conf. on Tetanus (Kretzschmar et al., 1979)     

乙型肝炎表面抗原对BALB/c小鼠细胞免疫应答的影响

目的研究不同来源乙肝表面抗原(HBsAg)对BALB/c小鼠的某些细胞免疫应答的影响.方法采用血源、CHO和酵母HBsAg分别免疫BALB/c小鼠,于免疫后不同时间制备脾脏单个核细胞(MNC),测定MNC对刀豆蛋白(ConA)和细菌脂多糖(LPS)的增殖反应性;采用绵羊红细胞(SRBC)作为致敏原,测定不同HBsAg免疫小鼠后迟发性超敏反应(DTH)的发生程度.结果不同来源HBsAg对ConA或LPS刺激的反应指数(SI)表现出不同的时间曲线,一般于免疫20?d和25?d时显著升高,其中血源HBsAg较高,酵母较低.H.M.-HBsAg在免疫5?d时即显出较高的SI值.CHO-HBsAg对SRBC诱导的DTH反应具有显著的抑制作用(P＜0.05).结论不同来源HBsAg诱导细胞免疫反应的类型和程度存在差异,CHO细胞来源的HBsAg对TH1细胞介导的DTH反应有抑制作用.     

Restriction site polymorphism at the LPA (Lp(a) apoliprotein; apoliprotein(a)) locus

A restriction site polymorphism in the Lp(a) apolipoprotein gene (the LPA gene) is reported. The basis for the polymorphism is presence or absence of an MspI restriction site that appears to be 3' to the last kringle IV structure of the gene. The "1" gene (presence of the restriction site) has a frequency of 0.316 and the "2" gene (absence of the restriction site) has a frequency of 0.684. Both members of each of 67 monozygotic (MZ) twin pairs had the same genotype and there was Mendelian segregation of the DNA variants in 40 families with a total of 75 children. There was a lower proportion of people with genotype 1-1 in the top quartile than in the 3 bottom quartiles of the population distribution of Lp(a) lipoprotein levels but the difference did not reach statistical significance.     

Interaction of sensory and cognitive processes during visual recognition: The role of the associative areas of the cerebral cortex

The first part of the present study used a model of Alzheimers disease in two groups of animals (three monkeys in each), given injections of neurotoxins (monkeys of group I) and physiological saline (monkeys of group II). Before injections, all monkeys were trained to discriminate stimuli containing different types of information (spatial frequency grids and geometrical figures of different colors and with different spatial relationships between objects) and to perform spatial selection. The dynamics of impairments in the characteristics of working memory were identified using delayed differentiation tasks in monkeys of both groups before injections and every two months after injections. Quantitative measures of impairments were made using the entropy of visual recognition, which characterizes uncertainty in decision-taking. The development of Alzheimers disease in rhesus macaques was characterized by a deficit of working memory, resulting from lesions to the two component processes of memory. Impairments of the first of these in monkeys of group I were manifest as a significant increase in entropy, which is associated with correct decision-taking. The magnitude of the increase depended on the type of visual information. Impairments of the second component were characterized by increases in entropy associated with refusals to take decisions and were independent of the delay duration and the type of visual information. Monkeys given injections of physiological saline showed no significant changes in these characteristics. The features of working memory were also studied in the second part of the investigation, using four groups of Rhesus macaques: intact, those with bilateral extirpation of the sulcus principalis or field 7 or both: degradation again identified two components. Entropy associated with this was increased significantly for most of the stimuli tested on monkeys of all extirpation groups as compared with intact animals. Significant differences were found in these characteristics for a number of stimuli, which depended on the location of the structures removed. The characteristics of impairments of the components of working memory resulting in the development of Alzheimers disease showed that the cholinergic mechanisms responsible for sensory processing differ from those involved in decision-taking. The structural-functional organization of the interaction of sensory and cognitive processes controlled by the motivation and attention systems is discussed, as is the role of the associative areas of the cortex.Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 89, No. 10, pp. 1226–1239, October, 2003.     

乳酸菌Z222产胞外多糖(EPSⅠ)对免疫细胞功能的影响

目的　检测乳酸菌Z2 2 2 产胞外多糖EPSⅠ对体外免疫细胞功能的调节作用。方法　不同浓度的EPSⅠ作用于小鼠的腹腔巨噬细胞和脾细胞 ,分别用中性红染色法检测巨噬细胞的吞噬能力 ,用MTT法检测淋巴细胞在体外的转化能力、NK细胞杀伤肿瘤细胞Yac 1的能力和产细胞因子IL 2的活性。结果　 (1)EPSⅠ单独作用小鼠脾细胞就能促进体外淋巴细胞增殖 ,且表现出剂量依赖关系。EPSⅠ亦可促进ConA诱导的体外小鼠淋巴细胞转化。 (2 )EPSⅠ体外作用于小鼠腹腔巨噬细胞均可提高MΦ的吞噬能力 ,与对照组比较差异都有显著性。 (3)与对照组相比EPSⅠ能增加体外NK细胞的活性 ,杀伤率最大值达 6 2 .4 1%± 2 .5 4 %。 (4 )EPSⅠ使小鼠脾细胞产IL 2的能力与对照组比较 ,差异有显著性。结论　乳酸菌多糖EPSⅠ对小鼠的免疫功能有一定的调节作用。     

Glomerular expression of cell-cycle-regulatory proteins in human crescentic glomerulonephritis

To elucidate the mechanism underlying crescentic formation, we assessed the phenotypic characterization and cell-cycle protein expression in human crescentic glomerulonephritis (CRGN). Kidney tissue specimens taken from CRGN patients (10 patients with pauci-immune type rapidly progressive glomerulonephritis (RPGN), 2 patients with Henoch-Schönlein purpura nephritis, and 1 patient with IgA nephropathy) were examined immunohistochemically. Most of the cellular components of the crescents expressed cytokeratin, whereas few cells expressed PHM-5. CD68-positive cells were minor components of cellular crescents, indicating that the major principal cellular component of the crescents is made up of cells with the parietal glomerular epithelial cell (PEC) phenotype. Additionally, serial section analysis revealed that Ki-67-positive cells in the crescents were frequently cyclin-A positive and Bcl-2 positive, but seldom cyclin-B₁ positive. Moreover, the expression of cyclin-dependent kinase inhibitor p27^Kip1 was low in the cellular crescents, despite being exclusively positive in podocytes within the same section. We concluded that the major component of the cellular crescents is made up of PECs and that apparent expression of cyclins and Bcl-2 and restrained expression of p27^Kip1 may be synergistically associated with the development of cellular crescents in human CRGN.     

Diminished neo-antigen response to keyhole limpet hemocyanin (KLH) vaccines in patients after treatment with chemotherapy or hematopoietic cell transplantation

Relapse is the most common cause of treatment failure for advanced cancer, even those treated with autologous hematopoietic cell transplantation (HCT). Effective tumor-specific immunotherapy may decrease relapse, however, this will fail if the immune system is unable to respond. We developed a strategy to test immune responses with a single injection of the bona fide neo-antigen KLH. The model was first tested in 37 normal volunteers using three KLH vaccines: Intracel KLH, Biosyn KLH, and Biosyn KLH + adjuvant. Despite finding the immunogenic epitope conserved in both products, intact Intracel KLH induced a better response compared to a purified 350/390 kDA subunit of KLH contained in the Biosyn KLH product. Addition of a synthetic oil adjuvant (Montanide ISA51) restored the response to a single injection of Biosyn KLH. A quantitative readout measured by a KLH-specific cellular and humoral response with isotype switching 1 month after KLH vaccination was established. To test the integrity of the adaptive immune response in cancer patients, we vaccinated 14 patients post-HCT and 19 patients with advanced cancer with KLH vaccines that elicited a 100% response rate in normal volunteers. In marked contrast to normal subjects, both responses were significantly impaired up to 16 months after autologous HCT with an intermediate response in advanced cancer patients. KLH vaccines are safe and require only a single injection to test neo-antigen responses providing an optimal platform for definitive testing of strategies to improve diminished immune recovery after chemotherapy or post-HCT.