Transforming growth factor-ß1 in the cerebrospinal fluid of patients with distinct neurodegenerative diseases.

A chronic inflammatory condition may underlie neurodegenerative disorders, including Parkinson's disease (PD) and Alzheimer's disease (AD). For example, both PD and AD patients show an increase in transforming growth factor-ß1 (TGF-ß1) levels in their cerebrospinal fluid (CSF). TGF-ß1 is a cytokine that inhibits inflammation. In the present study, using an enzyme-linked immunosorbent assay, we tested the hypothesis that the level of TGF-ß1 in the CSF of patients with amyotrophic lateral sclerosis (ALS), spinocerebellar degeneration (SCD), or multiple system atrophy-cerebellar subtype (MSA-C) would be elevated compared with that of normal controls. We found that TGF-ß1 levels in the CSF were not significantly different between these patients and normal controls. Our data suggest that the level of TGF-ß1 in the CSF is an unreliable biomarker of ALS, SCD, and MSA-C.

Genetic screening of 104 patients with congenitally malformed hearts revealed a fresh mutation of GATA4 in those with atrial septal defects.

We analysed the GATA binding protein 4 gene, or GATA4, along with the NK2 transcription factor related, locus 5 gene, or NKX2.5, to determine their genetic contribution to 104 sporadic patients in Indonesia with congenitally malformed hearts, 76 cases having atrial septal defect and 28 tetralogy of Fallot. We found only 1 novel mutation of GATA4 in those with atrial septal defects. Analysis of the genetic background of the parents of the patient showed for the first time that a new mutation of GATA4 can cause sporadic atrial septal defects. We failed to discover any other mutations of either the GATA4 or NKX2-5 genes, supporting the marked genetic heterogeneity of human congenital cardiac defects.

Spatiotemporal characteristics and mechanisms of intracellular Ca(2+) increases at fertilization in eggs of jellyfish (Phylum Cnidaria, Class Hydrozoa).

We have clarified, for the first time, the spatiotemporal patterns of intracellular Ca(2+) increases at fertilization and the Ca(2+)-mobilizing mechanisms in eggs of hydrozoan jellyfish, which belong to the evolutionarily old diploblastic phylum, Cnidaria. An initial Ca(2+) increase just after fertilization took the form of a Ca(2+) wave starting from one cortical region of the egg and propagating to its antipode in all of four hydrozoan species tested: Cytaeis uchidae, Cladonema pacificum, Clytia sp., and Gonionema vertens. The initiation site of the Ca(2+) wave was restricted to the animal pole, which is known to be the only area of sperm-egg fusion in hydrozoan eggs, and the wave propagating velocity was estimated to be 4.2-5.9 mum/s. After a Ca(2+) peak had been attained by the initial Ca(2+) wave, the elevated Ca(2+) gradually declined and returned nearly to the resting value at 7-10 min following fertilization. Injection of inositol 1,4,5-trisphosphate (IP(3)), an agonist of IP(3) receptors (IP(3)R), was highly effective in inducing a Ca(2+) increase in unfertilized eggs; IP(3) at a final intracellular concentration of 12-60 nM produced a fully propagating Ca(2+) wave equivalent to that observed at fertilization. In contrast, a higher concentration of cyclic ADP-ribose (cADPR), an agonist of ryanodine receptors (RyR), only generated a localized Ca(2+) increase that did not propagate in the egg. In addition, caffeine, another stimulator of RyR, was completely without effect. Sperm-induced Ca(2+) increases in Gonionema eggs were severely affected by preinjection of heparin, an inhibitor of Ca(2+) release from IP(3)R. These results strongly suggest that there is a well-developed IP(3)R-, but not RyR-mediated Ca(2+) release mechanism in hydrozoan eggs and that the former system primarily functions at fertilization. Our present data also demonstrate that the spatial characteristics and mechanisms of Ca(2+) increases at fertilization in hydrozoan eggs resemble those reported in higher triploblastic animals.

Levels of t-PA and PAI-2 in gingival crevicular fluid during orthodontic tooth movement in adults.

BACKGROUND: The regulation of plasminogen activation is a key element in controlling proteolytic events in the extracellular matrix. OBJECTIVE: The aim of this study was to investigate gingival crevicular fluid (GCF) levels of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI -2) during orthodontic tooth movement in adults. METHODS: Five male subjects (Mean age: 22.5 +/- 2.8 years) and five female subjects (Mean age: 23.4 +/- 3.9 years) were used. Each subject had one upper canine retracted into an extraction space. The contralateral and opposing canines, which were not moved, served as controls. GCF was collected at the distal cervical margins of the experimental and control teeth 0, 1, 24, and 168 hours after a retracting force was placed. GCF levels of t-PA and PAI-2 were determined by commercially available ELISA kits. RESULTS: After 24 hours of tooth movement the levels of t-PA and PAI-2 in the GCF were significantly higher from the experimental canines compared with the control teeth. There were no significant experimental-control differences at 0, 1, and 168 hours. There were no differences in the total protein levels up to 168 hours after orthodontic tooth movement. CONCLUSIONS: These results indicate that the amounts of t-PA and PAI-2 in the GCF increase with orthodontic tooth movement, and suggest that such increases may be involved in extracellular matrix degradation in response to mechanical stress. Failure to detect elevated levels of t-PA and PAI-2 at 168 hours was attributed to decay of the force retracting the canines.

MexAB-OprM specific efflux pump inhibitors in Pseudomonas aeruginosa. Part 4: Addressing the problem of poor stability due to photoisomerization of an acrylic acid moiety.

Exchange of the ethylene tether in a series of pyridopyrimidine-based MexAB-OprM specific efflux pump inhibitors to an amide bond stabilized the olefin of the acrylic acid moiety, preventing facile photoisomerization to the Z-isomer. Furthermore, the activity was drastically improved in the amide tether variants, providing extremely potent acrylic acid and vinyl tetrazole analogues.