Transformations of glycyrrhizic acid. VIII. Synthesis of immunomodulating glycopeptides using tert-butyl amino acid esters

Triterpene glycopeptides, derivatives of glycyrrhizic acid, were synthesized with the use of tert-butyl esters of L-amino acids. The condensation of glycoside with esters was carried out with N-hydroxysuccinimide--N,N-dicyclohexylcarbodiimide or N-hydroxybenzotriazol--N,N'-dicyclohexylcarbodiimide in the presence of a base (triethylamine, N-methyl- or N-ethylmorpholine). The protection groups were removed with the use of trifluoroacetic acid.     

Barriers to school-based health care programs

The purpose of this investigation was to determine the effects of Nd:YAG laser treatment on the permeability of dentine. Forty dentine discs were prepared by horizontal sectioning through the middle coronal third of freshly extracted non-carious third molars. After the removal of the pulp the discs were finished with 600 grit and divided into three test groups and one control group (n = 10). For the test group three different laser power settings were chosen: test group A: 3 x 60 sec, 60 mJ; test group B: 3 x 60 sec, 90 mJ; test group C: 3 x 60 sec, 120 mJ. No laser treatment was performed on the control group. In a two-chamber system the filtration rate of dentine tubules from an exactly defined area of the specimens was measured using a radioactive Ringer solution under a pressure of 30 cm H2O. Permeability measurements were carried out three times prior to lasing, three times immediately following laser treatment and six times after the application of phosphoric acid. Analysis of variance showed a significant influence of the Nd:YAG laser treatment on the permeability of dentine (P < 0.001). The mean quotient of non-treated control vs. lased dentine was 2.19 +/- 0.86 for the 60 mJ beam, 1.49 +/- 0.88 for the 90 mJ beam, and 2.04 +/- 2.17 for the 120 mJ beam. Etching the lased surfaces had a statistically significant influence on the permeability of the dentine only in the 60 mJ group (P < 0.001). The data show that the Nd:YAG laser treatment often increases the permeability of smear layer covered dentine but moderates the increase of permeability after etching the surface with phosphoric acid.     

Interaction of L-glutamic acid with human T-lymphocytes

A specific interaction of [3H]Glu with T lymphocytes from the blood of healthy donors (Kd = 0.236 microM) was revealed and described. It was found that unlabeled quisqualate, a structural analogue of L-glutamic acid, and unlabeled dipeptides Ala-Glu, Glu-Ala, and Glu-Glu competitively inhibit the specific binding of [3H]Glu to T lymphocytes (with Ki 0.19, 2.4, 3.4, and 1.2 microM, respectively). Binding experiments with conjugates of labeled and unlabeled glutamic acid with dextran showed that the receptors of [3H]Glu are localized on the outer surface of the plasma membrane of T lymphocytes.     

The intubating laryngeal mask. II: A preliminary clinical report of a new means of intubating the trachea

To determine the effect of anterior cruciate ligament (ACL) reconstruction on symptoms of pain and instability in patients with chronic ACL insufficiency who had previously undergone meniscectomy, we reviewed a series of 21 symptomatic, previously meniscectomized patients with chronic ACL deficiency (average age, 31 years). Arthroscopically assisted intra-articular ACL reconstruction using a middle, one-third patella-tendon autograft was performed in all cases. All patients had radiographic evidence of degenerative changes before ACL reconstruction. The average time from meniscectomy to ACL reconstruction was 6.6 years. Preoperative and postoperative range of motion, stability, and subjective evaluations were compared. Follow-up averaged 37.4 months (range, 24 to 67 months). Physical examination and postoperative KT-1000 side-to-side measurements revealed three patients (14%) with pathological ligament laxity. One patient had a 2+ Lachman, a 2+ pivot shift, and > 5 mm difference on KT-1000 maximum manual test, and two patients had a 1+ Lachman and a 1+ pivot shift. Range of motion measurements taken at follow-up were not significantly different from preoperative measurements (extension, P = .14; flexion, P = .46). Subjectively, all items on a panel of 15 visual analog scales were improved, but intensity of pain and instability were significantly improved after statistical analysis (P < .05). This review suggests that symptoms of pain and instability in patients with chronic ACL deficiency who have previously undergone meniscectomy can be improved by ACL reconstruction if objective stability is obtained.     

Theoretical considerations on the nature of the pineal 'ageing clock'

The models developed in our laboratory demonstrate that ageing initiates and progresses in the pineal gland. However, the ageing postponing effects of pineal grafting into older recipients cannot be explained by a simple maintenance and/or normalization of the night melatonin synthesis and release. We propose here that the pineal gland monitors and regulates, via its control of neuroendocrine function, the maintenance of 'self-identity' and the capacity of the immune system to recognize and react against any noxious, endogenous or exogenous agent. Senescence is characterized by the extinction of this central pineal function. The progressive decline of the self-recognition capacity distinguishes the typical diseases of ageing expressed as emergence of peripheral desynchronization and autoimmune, anaplastic, neoplastic and degenerative processes. Our approaches aim at a prevention and/or restoration of central pineal functions.     

Interactions of pluronic block copolymers with brain microvessel endothelial cells: evidence of two potential pathways for drug absorption

The effect of stimulus size on the pattern onset-offset visual evoked potential elicited with stimuli of two different wavelengths is studied under intensive yellow adaptation: (1) The onset response obtained with a 460 nm pattern is of negative polarity (N1) and saturates in amplitude with a stimulus radius of 7 deg. The onset response obtained with a 550 nm pattern is of positive polarity and continues to increase up to the maximum size (32.2 deg). (2) The peak time of N1 (460 nm) decreases with increasing stimulus size, that of P1 (550 nm) remains constant. These results are discussed as reflecting either varying retinal and brain anatomy, or cone activity, color-opponent activity, or luminance contrast activity.     

131I dose-dependent thyroid autoimmune disorders in children living around Chernobyl

The nucleotide sequence of 35,400 bp at approximately 10 kb from the right telomere of chromosome VII was determined. The segment contains the MAL1 locus, one of the five unlinked loci sufficient for maltose utilization. Until now, each of these loci was considered to contain three genes (for regulator, permease and alpha-glucosidase), but a fourth gene, presumably an extra alpha-glucosidase gene, was found at MAL1 adjacent to the usual cluster of three genes. The two glucosidase genes are present in opposite orientation, forming an inverted repeat structure. In addition to the four genes at MAL1, there are 11 complete, non-overlapping open reading frames (ORFs) longer than 300 bp in the sequence presented here. A new ABC transporter gene (YGR281w), required for oligomycin resistance was found (YOR1; Katzman et al., 1995), and the previously sequenced BGL2 (YGR282c), ZUO1 (YGR285c) and BIO2 (YGR286c) genes were located. The sequence of BIO2, a biotin synthetase gene, required substantial correction and the size of Bio2p is 375, rather than 356, amino acids. Two ORFs show rather weak similarities to animal genes: YGR278w to an unknown ORF of Caenorhabditis elegans and YGR284c to the murine Surf-4, a member of a cluster of at least four housekeeping genes. The remaining five ORFs do not encode known functions, but three of these show weak to high similarities to other ORFs in the Saccharomyces cerevisiae genome and one (YGR280c) codes for a particularly lysine-rich protein.     

Expression of the brain creatine kinase gene in rat RT4 peripheral neurotumor cell lines and its modulation by cell confluence

Creatine kinases (CK) catalyze the reversible transfer of a high energy phosphate group between creatine phosphate and ADP to regenerate ATP in cell types where the requirements for ATP are extensive and/or sudden. Previously, we have shown in primary rat brain cell cultures that brain CK (CKB) mRNA levels are highest in astrocytes and oligodendrocytes and much lower in neuronal cells. However, little is known of the factors which regulate CKB expression in the central nervous system and peripheral nervous system. To begin to investigate these factors, we asked in this report (1) if this pattern of CKB expression was also characteristic of some established glial and neuronal cell lines derived from the PNS; (2) whether CKB expression could be rapidly modulated by culture conditions, and (3) if CKB is expressed in cells with characteristics of glial cell progenitors. In subconfluent cells, CKB mRNA and enzyme activity were found to be high in both the rat RT4 peripheral neurotumor stem cell RT4-AC36A and its glial cell derivative RT4-D6. Conversely, CKB mRNA and activity were 5- and 8-fold lower, respectively, in the neuronal derivative RT4-E5 and, more dramatically, CKB was undetectable in neuronal RT4-B8 cells. Maintaining RT4-D6 glial cells at confluence rapidly increased CKB enzyme activity by 7-fold, such that D6 cells contained about 25% of the CKB level in lysates prepared from either whole adult rat brain or primary cultures of rat brain astrocytes. The levels of CKB mRNA and immunoreactive protein were also correspondingly increased in confluent D6 cells. These confluence-mediated increases in CKB appeared to be due to cell-cell contact and not the depletion of serum growth factors or an increase in intracellular cAMP. This study indicates that CKB expression is highest in cells displaying glial properties and can be rapidly modulated by appropriate culture conditions. The results are discussed in relation to the factors which may regulate CKB expression in vivo.     

Use of fluorescent dyes for measurement and localization of organelles associated with Ca2+ store release in human neutrophils

Fura-2 and its lipid analogue, FFP-18, were used to measure changes in cytosolic free Ca2+ concentration within human neutrophils. Whereas fura-2 was employed to monitor cytosolic Ca2+ increases throughout the cytosol, FFP-18 was used to monitor Ca2+ changes only near the membrane. This latter probe was incorporated into the plasma membrane as its acetoxymethyl ester (FFP-18-AM) but as de-esterification was catalysed by cytosolic esterases, the Ca(2+)-sensing probe (FFP-18 acid) accumulated on the inner face of membrane. The fluorescence of esterified probe on the extracellularly facing membrane leaflet was quenched by the membrane-impermeant ion Ni2+. Under these conditions, near membrane Ca2+ changes which resulted from the release of Ca2+ from intracellular stores was possible by conventional ratio fluorescence measurement of FFP-18. From the timing of arrival of Ca2+ at the plasma membrane, it was proposed that there were two Ca2+ storage sites, liberated by different stimuli, one close to the plasma membrane and the other more distant. In order to discover whether organelles within the neutrophil had distributions which correlate with the Ca2+ release sites, fluorescent dyes for structures within the cytosol were employed. We have previously shown that the location of the intracellular membrane stain, DiOC6 (3) corresponds to the distant Ca2+ release site. Here a second stain, BODIPY-C5 ceramide, has also been used and is shown to stain a peripheral region of the neutrophil, in a similar pattern to the near membrane Ca2+ storage site. These data therefore raise the question of whether these stains mark the organelles in neutrophils which are the two Ca2+ storage and release sites.