Tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) stimulate osteoclastic bone resorption

As both tissue inhibitor of metalloproteinases-1 (TIMP-1) and TIMP-2 have been reported to inhibit bone resorption, we examined whether TIMP-1 or TIMP-2 in fetal calf serum (FCS), with which culture media were supplemented, affected osteoclastic bone resorption in vitro. Contrary to our expectation, almost complete suppression of osteoclastic bone resorption was observed when both TIMP-1 and TIMP-2 were removed from the FCS. Bone resorption was, however, almost fully restored by the addition of recombinant TIMPs. TIMPs stimulate bone resorption at significantly lower concentrations (∼ng/ml) than those (∼μg/ml) required to inhibit bone resorption. To understand the mechanism of TIMP-dependent bone resorption, we counted and compared the number of tartrate-resistant acid phosphatase-(TRAP-) positive and multinuclear cells in cultures containing either 10% FCS or TIMP-1-free and/or TIMP-2-free FCS. There was essentially no difference in number among these, suggesting that the TIMP role seems to be related to the functional expression of osteoclasts. Metallo-proteinase inhibitors, either BE16627B[l-N-(N-hydroxy-2-isobutylsuccinynamoyl)-seryl-l-valine] or R94138 {N-methyl-(3S)-2-[(2R)-2-hydroxycarbamoylmethylundecanoyl] hexahydropyridazine-3-carboxamide}, could not replace TIMPs, suggesting that the osteoclast-stimulating activity of TIMPs cannot be ascribed to merely their inhibitory effect on matrix metalloproteinases. Received: Oct. 15, 1998 / Accepted: April 5, 1999     

Hypoalbuminemia accelerates erythrocyte membrane lipid peroxidation in chronic hemodialysis patients

BACKGROUND: Persistent hypoalbuminemia is a long-term poor prognostic factor in chronic hemodialysis patients. PATIENTS AND METHODS: We investigated the correlation between the degree of peroxidation of erythrocyte membrane lipids, erythrocyte alpha tocopherol content, erythrocyte glutathione peroxidase activity and serum albumin concentration in twelve patients with uremia not undergoing hemodialysis and fifteen patients on maintenance hemodialysis. RESULTS: The glutathione peroxidase activity in erythrocytes was higher in patients of uremia not undergoing hemodialysis than in chronic hemodialysis patients. A significant negative correlation was observed between the erythrocyte alpha tocopherol content and the degree of erythrocyte membrane lipid peroxidation in chronic hemodialysis patients. There was a statistically significant difference in the degree of erythrocyte membrane lipid peroxidation between patients with chronic hemodialysis-associated hypoalbuminemia and chronic hemodialysis patients having normal serum albumin levels. CONCLUSION: This study suggested that serum albumin inhibits peroxidation of erythrocyte membrane lipids and that hemodialysis induces recovery of serum reductivity. We conclude that persistent hypoalbuminemia worsens the serum antioxidant activity in chronic hemodialysis patients and may contribute to increased oxidative cell damage.     

Molecular-pathological Analysis of a Patient with Three Synchronous Squamous Cell Carcinomas in the Aerodigestive Tract

A case of synchronous squamous cell carcinomas in the soft palate,larynx and esophagus is reported, along with findings of molecular-pathologicalanalysis. A biopsy sample from the aryngeal carcinoma revealedwell differentiated squamous cell carcinoma harboring two pointmutations at codons 144 and 148 of the p53 gene but not at codon299, and more than 50% of the cancer cells showed accumulationof p53 protein immunohistochemically. The esophageal tumor,which was moderately differentiated squamous cell carcinoma,showed immunoreactivity for p53 within the nuclei of 25–50%of cancer cells with a missense mutation at codon 299 but notat codon 144 or 148. This cancer also showed immunoreactivityfor transforming growth factor alpha. On the other hand, thepoorly differentiated squamous cell carcinoma in the soft palateshowed negative immunoreactivity for p53 and no point mutationin exons 5 to 8 of the gene. These results suggest that thethree synchronous squamous cell carcinomas arose as independentevents.     

Assessment of hepatic graft injury by graft effluent in rodents: N-acetyl-β-glucosaminidase and type III procollagen peptide

We studied the significance of N-acetyl--glucosaminidase (-NAG) and type III procollagen peptide (P-III-P) in the effluent of rodent hepatic grafts. After total hepatectomy, the livers were preserved in chilled, lactated Ringer's solution and then divided into five groups (n=10 each): group 1, 4 h preservation only; group 2, 4 h preservation and rewarming; group 3, 6 h preservation only; group 4, 6 h preservation and rewarming; and group 5, minimal preservation only. The -NAG of groups 2 and 4 was significantly higher than that of groups 1 and 3 (0.98±0.5 U/l vs 0.21±0.12 U/l; P<0.01 and 1.76±0.67 U/l vs 0.38±0.25 U/l, respectively; P<0.01), while that of group 4 was significantly higher than that of group 2 (1.76±0.67 U/l vs 0.98±0.50 U/l; P<0.05). The P-III-P of group 4 was significantly higher than that of group 2 (0.133±0.008 U/ml vs 0.110±0.015 U/ml; P<0.01). We conclude that -NAG is a novel parameter of parenchymal and nonparenchymal cells, while P-III-P reflects the integrity of the hepatic sinusoidal extracellular matrix.     

Cell biological basis for combination radiotherapy using heavy-ion beams and high-energy X-rays.

We investigated the biological effect of combining carbon-beam and X-ray in vitro. The results showed that when we employed Gray equivalent as the indication of therapeutic dose, the effects could be explained with simple additive way in the treatment plan. This fact provides important information about the combined therapy of carbon-beam and X-ray.     

A Question and Answer E-Mail System for Responding to Query from the General Public with Which the System Manager Can Identify Delayed Replies

We constructed a question and answer system for direct delivery of E-mail queries from the general public to the proper hospital representative and with which the system administrator can monitor E-mail replies. The system consists of two sub-systems; one is an E-mail questionnaire sub-system that automatically delivers E-mail queries to the proper person and an E-mail reply monitor sub-system with which the system administrator can monitor the E-mail replies. The monitor system compares the origin address of the E-mail reply and the destination address on a questionnaire filled out at the time the query is made. In a test of the system, 56 queries were received in 31 days, with 37 replied to. The interval from receipt of an E-mail query to reply was calculated. Nineteen replies were submitted within 2 days.     

Comparative studies on patient's and family's basic attributes, psychosocial states and treatment prognoses at three medical institutions for treatment of alcoholism]

A research by questionnaire was conducted at three medical institutions, on basic attributes and psychosocial states of alcoholics' as first time inpatient and respective families. After a year of research, additional study was conducted in regard of abstinence, treatment continuation as outpatient, participation in self-help groups, and family's participation in family groups. Outcomes of researches' statistical analyses showed significant differences concerning medical institutions and basic attributes for the items: treatment histories of alcoholism, treatment or inpatient in other medical departments and alcoholic's family history,; but these differences were considered to be associated with varied courses of referrals. In relation to medical institutions with patients and family's psychosocial states, there were no significant differences on the outcomes of five scales. As for the correlation between medical institutions with a year's prognoses there were no significant differences concerning abstinence and treatment continuation as outpatient. In case of participation in self-help groups and family groups, the prognoses showed significant differences but these differences were considered to be related to varied treatments and different helping ways of medical institutions and varied conditions of local communities for mentioned helps.     

Antiherpetic activities of sulfated polysaccharides from green algae

In order to evaluate the potency of novel antiviral drugs, 11 natural sulfated polysaccharides (SPs) from 10 green algae ( Enteromorpha compressa, Monostroma nitidum, Caulerpa brachypus, C. okamurai, C. scapelliformis, Chaetomorpha crassa, C. spiralis, Codium adhaerens, C. fragille, and C. latum) and 4 synthetic sulfated xylans (SXs) prepared from the beta-(1,3)-xylan of C. brachypus, were assayed for anti-Herpes simplex virus type 1 (HSV-1) activity. Except for one from E. compressa, all SPs showed potent anti-HSV-1 activities with 50 % inhibitory concentrations (IC (50)) of 0.38 - 8.5 microg/mL, while having low cytotoxicities with 50 % inhibitory concentrations of >2900 microg/mL. Anti-HSV-1 activities of SXs were dependent on their degrees of sulfation. To delineate the drug-sensitive phase, 4 polysaccharides, which showed potent anti-HSV-1 activities, were applied to time-of-addition experiments. Among the polysaccharides tested, 3 polysaccharides (SX4, SP4 from C. brachypus, and SP11 from C. latum) showed strong anti-HSV-1 activities with IC (50) of 6.0, 7.5, and 6.9 microg/mL, respectively, even when added to the medium 8 h post-infection. These experiments demonstrated that some sulfated polysaccharides not only inhibited the early stages of HSV-1 replication, such as virus binding to and penetration into host cells, but also interfered with late steps of virus replication. These results revealed that some sulfated polysaccharides from green algae should be promising candidates of antiviral agents which might act on different stages in the virus replication cycle.