Aggregatibacter actinomycetemcomitans induces detachment and death of human gingival epithelial cells and fibroblasts via elastase release following leukotoxin‐dependent neutrophil lysis

Aggregatibacter actinomycetemcomitans is considered to be associated with periodontitis. Leukotoxin (LtxA), which destroys leukocytes in humans, is one of this bacterium's major virulence factors. Amounts of neutrophil elastase (NE), which is normally localized in the cytoplasm of neutrophils, are reportedly increased in the saliva of patients with periodontitis. However, the mechanism by which NE is released from human neutrophils and the role of NE in periodontitis is unclear. In the present study, it was hypothesized that LtxA induces NE release from human neutrophils, which subsequently causes the breakdown of periodontal tissues. LtxA‐treatment did not induce significant cytotoxicity against human gingival epithelial cells (HGECs) or human gingival fibroblasts (HGFs). However, it did induce significant cytotoxicity against human neutrophils, leading to NE release. Furthermore, NE and the supernatant from LtxA‐treated human neutrophils induced detachment and death of HGECs and HGFs, these effects being inhibited by administration of an NE inhibitor, sivelestat. The present results suggest that LtxA mediates human neutrophil lysis and induces the subsequent release of NE, which eventually results in detachment and death of HGECs and HGFs. Thus, LtxA‐induced release of NE could cause breakdown of periodontal tissue and thereby exacerbate periodontitis.     

Age-related differences in muscle activity, stride frequency and heart rate response during walking in water.

This study aimed to examine whether walking in water produces age-related differences in muscle activity, stride frequency (SF), and heart rate (HR) response. Surface electromyography (EMG) was used to evaluate muscle activities in six older and six young subjects while they walked in water immersed to the level of the xiphoid process. The trials in water utilized the Flowmill which consists of a treadmill at the base of a water flume. The measurement of maximal voluntary contraction (MVC) of each muscle was made prior to the gait analysis. The %MVCs, which refer to the surface EMG measures, from the gastrocnemius of the older subjects were significantly lower than those of the young subjects, in every experimental condition (P<0.05). In contrast, the %MVCs from the rectus femoris (P<0.05) and the biceps femoris (P<0.001) of older subjects were significantly greater than those of young subjects in every experimental condition. Moreover, the SFs of older subjects were also significantly greater than those of young subjects (P<0.05), while the HR responses of older and young subjects were similar. In conclusion, the older subjects had increased hip musculature activity and decreased ankle plantar flexor activity while walking in water, compared with the young subjects.     

Dissociation between plasma and atrial content of atrial natriuretic polypeptide (ANP) following sodium load in rats

To investigate the time course effect of sodium intake on release and synthesis of atrial natriuretic polypeptide (ANP), plasma and atrial content of ANP were measured in rats which had been fed either a high or a low salt diet for 1, 3, 7, 14 and 35 days. Plasma ANP in rats fed the high salt diet for one day was significantly higher than in those fed the low salt diet. However, there were no significant differences between the groups fed either the high or the low salt diet for 3 days or more. In contrast to the direction of change in plasma ANP, atrial content of ANP in rats fed the high salt diet for one day tended to be lower and was significantly lower in those fed for 3 and 7 days than in the low salt diet group, while there were no significant differences between both groups that were fed for 14 and 35 days. These results suggest that ANP is rapidly released into the circulation when sodium is loaded, however, the atrial storage of ANP remains depleted for about one week.     

HPLC-studies on nonmercapt-mercapt conversion of human serum albumin

Human mercaptalbumin (HMA) and nonmercaptalbumin (HNA) could be separated by high-performance liquid chromatography (HPLC) at neutral pH. Using HPLC, the present authors found the nonmercapt-mercapt conversion (HNA----HMA) during hemodialysis and the mercapt-nonmercapt conversion (HMA----HNA) after hemodialysis in chronic renal failure, indicating HMA as the covalent carrier protein for sulfur-containing amino acids.     

Localization of spectrin isoforms in the adult mouse heart

The distribution of two isoforms of spectrin in the adult mouse heart was investigated by Western blotting and immunocytochemistry by use of monospecific antibodies to erythrocyte spectrin and nonerythroid brain spectrin (240/235). Western blotting revealed proteins analogous to both isoforms of -spectrin in adult heart. Light-microscopic immunocytochemistry indicated that erythroid spectrin was distributed throughout the myocardium, with immunofluorescence localized to plasma membranes, Z-lines, and intercalated discs. Antibodies to brain spectrin (240/235) exhibited staining throughout the heart, with a generally diffuse distribution except for the prominent immunoreactivity associated with the intercalated discs. Nonerythroid spectrin immunofluorescence was detected in the endothelial cells of the endocardium and the mesothelial cell lining of the epicardium. Erythrocyte spectrin was not detected in the endocardium or the epicardium. The identification and localization of spectrin isoforms in the mammalian heart suggest the importance of spectrin proteins in the structural integrity and proper function of cardiac cells and tissues. This is the first demonstration of two different -spectrin subunits in the mammalian heart.     

Switching dynamics and the transient memory storage in a model enzyme network involving Ca2+/calmodulin-dependent protein kinase II in synapses

 The signal processing through a chain of phosphorylation-dephosphorylations mediated by a pair of enzymes, Ca²⁺/calmodulin-dependent protein kinase II and the associated phosphatase, is formulated as a non-autonomous dynamical system in the framework of non-autocatalytic, intraholoenzyme reaction dynamics. A classification of switching characteristics of the system is made in the parameter space comprising the three controllable system parameters: an input-pulse intensity and initial concentrations of the two associated enzymes. It is found that a region of parameter space exists termed the transition zone, that exhibits a quasi-switching behaviour characterized by a signal storage time being prolonged by more than several orders of magnitude (10⁴ times in certain cases) for the increase of two orders of magnitude in the input signal intensity. The effect of alterations of certain rate constants on the quasi-switching property is explored. It is numerically demonstrated that the Ca²⁺/calmodulin-dependent kinase II-related phosphatase is the most important key enzyme for regulating the signal storage time. Received: 25 April 1994/Accepted in revised form: 16 December 1994     

A Simple Method for Making a Vibrating Probe System

A simple and easy method for making the vibrating probe systemfirst developed by Jaffe and Nuccitelli [(1974) J. Cell Biol.63: 614] is described. The best performance of the probe insea water was 30.7 µA cm^-2 per one µV output, 0.4µA cm^-2 peak-to-peak output-noise and negligible output-driftat 30-µm vibrating amplitude and 3-s amplifier time constant.Sample applications were made for both freshwater and marinealgae. (Received March 22, 1984; Accepted May 28, 1984)     

Effect of pH on preferential antibacterial-activity of ethylenediaminetetraacetic acid (EDTA)]

Ethylenediaminetetraacetic acid (EDTA), a chelating agent, was examined for the antibacterial activity against 15 species of bacteria by treating with a 10mM solution at pH adjusted to 5.0, 7.0 or 9.0. All bacterial species tested were classified into three groups; tentatively named the pH5 EDTA-sensitive group comprising Vibrio cholerae and Staphylococcus aureus, the pH9 EDTA-sensitive group comprising Escherichia coli and Pseudomonas aeruginosa and the EDTA-nonsensitive group comprising Proteus mirabilis. The EDTA-sensitivity grouping may be used as a tool for preferential decontamination of certain bacteria in live edible fishes, although further experiments are needed to characterize more strains and also species of bacteria.     

Expression of the pS2 gene in human gastric cancer cells derived from poorly differentiated adenocarcinoma

NH2-terminal amino acid sequence of the pS2 protein produced and secreted by human gastric cancer cells, MKN-45, was determined to be identical to that of MCF-7 cells. A clone encoding pS2 protein was isolated from the cDNA library constructed from MKN-45 cells. The nucleotide sequence was identical to that of pS2 cDNA previously isolated from human breast cancer cells, MCF-7, except for one nucleotide in the 3' untranslated region. Thus, in this cell line, the pS2 gene product is translated and secreted as in MCF-7 cells. RNA blot hybridization analysis revealed that pS2 gene was expressed well in two (MKN-45 and KATO-III; derived from poorly differentiated adenocarcinoma) but not in three cell lines (MKN-1, MKN-28 and MKN-74; from well differentiated adenocarcinoma), suggesting that expression of the pS2 gene depends on the state of cell differentiation. These results suggest that pS2 is expressed in human gastric cancer cells in an estrogen-independent manner and is possibly associated with the malignant state of cells.