Correlation between serum IgE and salivary cortisol levels in subjects with type I allergic disorders.

This study compares the relationship between serum IgE and salivary cortisol levels in 42 normal and in 18 type I allergic subjects. Levels of serum total IgE and salivary cortisol were determined with the UniCAP system and ELISA respectively. In the type I allergic subjects, there was a significant correlation between serum IgE and salivary cortisol levels (p < 0.01). In the normal subjects, on the other hand, no correlation was found. These findings suggest that there may be an association between stress and allergic disorders.

Measurement of salivary adiponectin levels.

We designed a method for measuring salivary adiponectin. In 188 healthy males, salivary adiponectin levels were measured using a commercial enzyme immunoassay kit for plasma with minor modifications. Intra- and inter-assay coefficients of variation for salivary adiponectin ranged from 0.6 to 4.9 and 1.1 to 9.8%, respectively. Salivary adiponectin levels ranged from 0.37 to 6.42 ng/ml, exceeding the kit's detection limit. For the over-43 age group, there was a significant correlation between plasma and salivary adiponectin levels (p<0.000001). These findings suggest the possibilities of salivary adiponectin as a marker of increased risk of non-insulin-dependent diabetes mellitus or cardiovascular disease.

Effect of coffee intake on blood flow and maternal stress during the third trimester of pregnancy.

OBJECTIVE: To investigate the effects of maternal ingestion of an ordinary dose of coffee on maternal stress and placental and fetal blood circulation during the third trimester of pregnancy. METHODS: We performed a Doppler blood flow analysis for 10 women in the third trimester of pregnancy before and after they drank a cup of coffee. Salivary samples were collected from the 10 pregnant women and 14 nonpregnant controls just before coffee intake and 30 min later. Salivary cortisol levels and chromogranin A titers were determined. RESULTS: Coffee intake had no effect on maternal or fetal blood flow. Among the pregnant women, Salivary cortisol levels were significantly reduced after coffee intake but salivary chromogranin A concentration was not significantly different before and after coffee intake. CONCLUSION: The reduced salivary cortisol levels suggest that coffee intake decreases maternal stress during pregnancy.

Thermosensitive polymer-modified liposomes that release contents around physiological temperature.

To obtain temperature-sensitive liposomes which release their contents around the physiological temperature, we designed dioleoylphosphatidylethanolamine liposomes modified with copolymers of N-isopropylacrylamide and acryloylpyrrolidine. Copolymers of acryloylpyrrolidine and N-isopropylacrylamide, which exhibit a lower critical solution temperature around the physiological temperature, were prepared by free radical copolymerization using azobis(isobutyronitrile) as the initiator. The copolymers with anchors to the liposome membrane were obtained by using N, N-didodecylacrylamide as an additional comonomer. The copolymer having the anchor group at the terminal of the polymer chain was also synthesized by copolymerization of these monomers in the presence of 2-aminoethanethiol and subsequent conjugation of N, N-didodecyl succinamic acid to the terminal amino group of the copolymer. Calcein-loaded dioleoylphosphatidylethanolamine liposomes modified with these copolymers were prepared and release of the contents from these liposomes was investigated. It was found that the release from these copolymer-modified liposomes was promoted around and above the lower critical temperature of the copolymer. Also, the liposomes modified with the terminal anchor-type copolymer released the contents more drastically responding to a small temperature change than the liposomes modified with random copolymers containing N,N-didodecylacrylamide units as the anchor.

Temperature-dependent interaction of thermo-sensitive polymer-modified liposomes with CV1 cells.

Egg yolk phosphatidylcholine liposomes modified with a copolymer of N-acryloylpyrrolidine and N-isopropylacrylamide having a lower critical solution temperature at ca. 40 degrees C were prepared and an effect of temperature on their interaction with CV1 cells was investigated. The unmodified liposomes were taken up by the cells approximately to the same extent after 3 h incubation at 37 and 42 degrees C. In contrast, uptake of the polymer-modified liposomes by CV1 cells decreased slightly at 37 degrees C but increased greatly at 42 degrees C, compared to the unmodified liposomes. Proliferation of the cells was partly prohibited by the incubation with the unmodified liposomes encapsulating methotrexate at 37 and 42 degrees C. The treatment with the polymer-modified liposomes containing methotrexate at 37 degrees C hardly effected the cell growth. However, the treatment at 42 degrees C inhibited the cell growth completely. It is considered that the highly hydrated polymer chains attached to the liposome surface suppressed the liposome-cell interaction below the lower critical solution temperature of the polymer but the dehydrated polymer chains enhanced the interaction above this temperature. Because interaction of the polymer-modified liposomes with cells can be controlled by the ambient temperature, these liposomes may have potential usefulness as efficient site-specific drug delivery systems.

Association of ALDH(2) genotypes and alcohol consumption with periodontitis.

There is little information regarding the association between alcohol consumption and periodontitis risk. We assessed whether alcohol consumption and ALDH(2) genotypes were associated with periodontitis. Subjects' lifestyle was examined by a self-administered questionnaire, and the percentage of pocket depths > or = 3.5 mm was used as a periodontal parameter. ALDH(2) genotypes were determined with the use of a PCR/RFLP method. Multiple logistic analyses showed that alcohol consumption was significantly associated with periodontitis, and its odds ratio was 1.98. There was no significant relationship between periodontal status and ALDH(2) genotypes. However, ALDH(2)*1/*2 subjects who consumed > or = 33 g/day of alcohol had a significantly greater percentage of pocket depths > or = 3.5 mm than those whose daily consumption was lower, while there was no significant difference in periodontal status associated with alcohol consumption in ALDH(2)*1/*1 subjects. Our results suggest that alcohol consumption may be a risk indicator for periodontitis in ALDH(2)*1/*2 subjects who consume larger amounts of alcohol.

Secretory IgA in saliva and academic stress.

Several reports have proposed that the concentration of secretory immunoglobulin A (S-IgA) in saliva is an indicator of psychological stress. With this in mind, we decided to examine it in 10 second year medical student volunteers at Kawasaki Medical School course between May 4 and July 13, 2000 and discussed the relationship between S-IbA and the stress from academic examinations. Saliva was collected three times (on rising, at forenoon, and at bedtime) every Thursday. During this period, sporadic academic examinations were held twice and term end examination occurred during the last two weeks. Results showed the concentration of S-IgA significantly higher at the on rising time-point than at the other two time-points. There was also a tendency for the S-IgA level in saliva to be higher on the day before academic examinations and during them and lower on the days between these examinations. In addition, daily variations in the S-IgA concentration sometimes seemed to be disturbed by other academic stress. Therefore it may be possible to use this measurement to monitor psychological stress in students and workers.

Enhanced rectal absorption of [Asu1,7]-eel calcitonin in rats using polyacrylic acid aqueous gel base.

The effects of polyacrylic acid aqueous gel on the absorption of rectally administered [Asu1,7]-eel calcitonin, a calcitonin analogue, were investigated in rats. The [Asu1,7]-eel calcitonin (1 U/kg) was given into the rectal loop in gel bases at various pH (5.5-8.5) and polyacrylic acid concentrations (0.01-1.0% w/v). The maximum hypocalcemic effect was obtained in approximately 30 min after administration of the analogue in a 0.1% w/v polyacrylic acid gel base at pH 5.5. The plasma calcium level decreased by approximately 18% from the initial level. Rectal administration in vehicles such as polyethylene glycol 1000, triglyceride fatty acid mixture base, or saline solution had little or no hypocalcemic effect at a dose of 5 U/kg. The results indicated that a polyacrylic acid aqueous gel base significantly improved the absorption of this analogue. Furthermore, rectal administration in a polyacrylic acid gel base (0.1% w/v; pH 5.5) required a dose 35 times greater than an intravenously administered dose to achieve an equivalent hypocalcemic effect.

Application of surface-coated liposomes for oral delivery of peptide: effects of coating the liposome's surface on the GI transit of insulin.

We prepared two kinds of surface-coated liposomes and investigated their potencies as oral dosage forms for peptide drugs by focusing on their effects on the gastrointestinal (GI) transit of drugs. The surface of the liposomes was coated with poly(ethylene glycol) 2000 (PEG-Lip) or the sugar chain of mucin (Mucin-Lip). As a model peptide drug, insulin was encapsulated in these liposomes. Coating the surface with poly(ethylene glycol) was found to reduce the transit rate of liposomes in the small intestine after oral administration to rats in vivo. Mucin-Lip was retained in the stomach longer than PEG-Lip or uncoated liposomes. The effect of surface coating on the intestinal transit of liposomes was determined by means of in situ single pass perfusion in the rat small intestine. Statistical moment analysis was applied to the outflow pattern of both liposomes and encapsulated insulin. The mean transit time (MTT) and deviation of transit time (DTT) in the intestinal tract were calculated. The MTT of PEG-Lip was much longer than those of uncoated liposomes and Mucin-Lip and was significantly shortened after removal of the intestinal mucous layer. These results indicated that PEG-Lip interacts strongly with the intestinal mucous layer, leading to its slow transit in the intestine. In contrast, coating the liposome's surface with mucin did not affect either the MTT or DTT of liposomes in the intestine. This result is in accordance with the in vivo observation that Mucin-Lip was highly retained in the stomach, but not in any region of the small intestine in vivo. Both the MTT and DTT values of insulin encapsulated in PEG-Lip and Mucin-Lip were almost the same as those of liposomes themselves, suggesting that surface-coated liposomes retained insulin in the intestinal tract. However, MTT and DTT of insulin were significantly shorter than those of uncoated liposomes because these liposomes degraded and released significant amounts of insulin during single pass perfusion. The ability of surface-coated liposomes, especially of PEG-Lip, to interact with the mucus layer and slow the transit rate in the GI tract is considered desirable for oral delivery of peptide drugs. Modification of the liposomal surface with appropriate materials, therefore, should be an effective method by which to achieve the oral delivery of peptide drugs.

Single-step purification of F(ab')2 fragments of mouse monoclonal antibodies (immunoglobulins G1) by hydrophobic interaction high performance liquid chromatography using TSKgel Phenyl-5PW.

Hydrophobic interaction high performance liquid chromatography (HPLC) using TSKgel Phenyl-5PW was applicable to single-step purification of F(ab')2 fragments from pepsin digests of mouse monoclonal antibodies of IgG1 class. The digests were applied to the gel equilibrated with phosphate-buffered saline containing 1 M ammonium sulfate. F(ab')2 fragments were adsorbed onto the gel using the same buffer, and eluted by reducing the ammonium sulfate concentration to 0 M. The fraction containing F(ab')2 fragments was homogeneous (purity: higher than 98%) by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration HPLC. The recovery of the antigen binding site was 42-58%. The cycle time of the Phenyl-5PW HPLC was 45 min, and F(ab')2 of up to 2200 mg was purified in a cycle. This method could be useful especially for large scale purification of F(ab')2 fragments.

Single-step purification of F(ab')2 mu fragments of mouse monoclonal antibodies (immunoglobulins M) by hydrophobic interaction high-performance liquid chromatography using TSKgel ether-5PW.

A procedure is described for preparation and single-step purification of F(ab')2 fragments, herein designated as F(ab')2 mu' from mouse monoclonal antibodies of the IgM class. Hydrophobic interaction high-performance liquid chromatography (HPLC) using TSKgel Ether-5PW was well applicable to the purification. The IgM was digested with pepsin at the pepsin-to-IgM ratio of 1:200 (w/w) in 100 mM citrate buffer (pH 4.2) at 37 degrees C for 2 h. The digests were applied to the gel equilibrated with the buffer containing 1 M ammonium sulfate. F(ab')2 mu fragments were adsorbed onto the gel with the same buffer, and eluted by reducing the ammonium sulfate concentration to 0 M. The fraction containing F(ab')2 mu fragments was homogeneous (purity higher than 97%) by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel-filtration HPLC. The recovery of the antigen-binding site was 55-72%. The cycle time of the Ether-5PW HPLC was 40 min, and up to 98 mg F(ab')2 mu fragments. The molecular mass of F(ab')2 mu was estimated to be 144-146 kDa. In comparison with IgM, F(ab')2 mu lost entirely the complement C1q binding activity, and the sugar content was greatly reduced. The binding of IgM with non-specific proteins turned to be negligible, when IgM was converted to F(ab')2 mu, suggesting that the fragments are useful for immunological application.

Enhancement of nasal absorption of insulin and calcitonin using polyacrylic acid gel.

The effects of polyacrylic acid gel on the nasal absorption of insulin and [Asu1,7]-eel calcitonin were investigated in rats. The nasal administration of insulin (1 IU kg-1) in polyacrylic acid gel at 0.1 and 1% w/v showed maximum hypoglycaemic effects at 30 min and 1 h after administration, respectively. However, the nasal administration of insulin in carboxymethyl cellulose (1% w/v) solution had no hypoglycaemic effect at the same dose. When [Asu1,7]-eel calcitonin (10 U kg-1) was administered nasally in polyacrylic acid gel (0.1% w/v), a prominent hypocalcaemic effect was observed during the first 30 min. Nasal administration of [Asu1,7]-eel calcitonin in saline had no hypocalcaemic effect at the same dose. The results indicate that the polyacrylic acid gel base significantly enhanced the absorption of insulin and [Asu1,7]-eel calcitonin via the nasal cavity.

Effect of non-ionic surfactants in a polyacrylic acid gel base on the rectal absorption of [Asu1,7]-eel calcitonin in rats.

The effect of non-ionic surfactants in a polyacrylic acid gel base on the rectal absorption of [Asu1,7]-eel calcitonin, a calcitonin analogue, was studied in rats. Absorption was enhanced by a microenema which used a polyacrylic acid gel base, but it was reduced by the incorporation of polysorbate 80 (0.1-5% v/v). The incorporation of polyoxyethylene 9 lauryl ether (0.1-5% v/v) in the polyacrylic acid gel base enhanced the absorption. Rectal administration in the base containing 0.5% v/v polyoxyethylene 9 lauryl ether required a dose of the calcitonin 2-3 times greater than an intramuscularly administered dose to achieve an equivalent hypocalcaemic effect.

Design of a polyvinyl alcohol hydrogen containing phospholipid as controlled-release vehicle for rectal administration of (+/-)-propranolol HCl.

Polyvinyl alcohol hydrogels which contained phospholipid, egg yolk lecithin or hydrogenated soya lecithin were designed as a transrectal delivery system for propranolol hydrochloride. The hydrogel preparations containing phospholipid were prepared by a low-temperature crystallization method. The release profile of propranolol from hydrogel preparations containing phospholipid complied with Fickian diffusion (Higuchi model). The release of propranolol from the hydrogel preparation decreased with higher contents of phospholipid (approximately 2% w/w). In rats plasma concentrations of propranolol after rectal administration of hydrogel preparations containing phospholipid (1 and 2% w/w) were prolonged compared with those of rats receiving preparations without phospholipid.

Improved method using a bubble-free adhesion technique for the preparation of semi-serial undecalcified histologic sections containing dental implants.

The preparation technique, with the minimum of artifacts for the semi-serial undecalcified histologic sections containing dental implants, is presented in this study. The sections enabled finer light-microscopic observations to be made. A formalin-fixed tissue block containing a dental implant was dehydrated in ethanol and acetone, and then embedded in polyester resin under 76 cm Hg reduced pressure. The embedded block was trimmed by a cutter and ground by abrasive paper. In a 1.5 Kg f/cm2 pressurized chamber, its polished surface was bonded to a methacrylate slide by means of ethylcyanoacrylate used in an adhesion loading device. This meant that no bubbles could arise in the interface between the slide and the block. The slide-block was then attached to an adsorptive specimen-holder of a hard-tissue cutting machine and cut to a thickness of approximately 50 microns, with use of a diamond blade. The slide-section was ground to 15-40 microns with wet-type abrasive paper and film on a polishing table. Etching with weak acid and surface staining with toluidine blue and methylene blue/basic fuchsin/light green were performed on the section.

Differences between the bony interfaces of titanium and hydroxyapatite-alumina plasma-sprayed titanium blade implants.

Histopathologic study was done to reveal the difference of bony interface in the mandibular bone between titanium blade implants and titanium implants coated with the mixture of hydroxyapatite-alumina by the plasma-spray technique, using the improved undecalcified histologic technique and the image analyzing system. In 4 adult shepherd dogs, the implantation was performed in the molar region of the mandible 2 months after tooth extraction. The coated implants and the non-coated ones were inserted respectively in the mandibles. They were sacrificed on the 5th, 7th, 14th and 28th day postoperatively. In 5-day implant after insertion, granulation tissues or fibrous connective tissues were observed around both implants in bone tissues. In 7-day implant, osteoid tissues regenerated around the apex of both implants, but attached a little to them. In 14-day implant, osteoid tissues or woven bone regenerated around both implants and attached more to the coated implant (approximately 13.7%) than the non-coated one (approximately 6.0%). In 28-day implant, regenerative woven or lamellar bone attached much more to the coated implant (approximately 63.4%) than the non-coated one (approximately 8.8%). Bony interface increased remarkably in the coated implants, chronologically. The results showed that the plasma-sprayed hydroxyapatite-alumina coating was effective for the initial fixation of endosseous dental implants.

Quantitative study on the interface between bone tissue and Blade-vent implants using the image processing system.

Undecalcified histopathologic sections of two titanium blade-vent implants (which had survived for five years in a dog) were made, and image analyses of the bone-implant interface were determined. One implant was inserted conventionally in the right mandible, and the other was inserted shallower than usual in the left mandible, so that the shoulder was located at the same level as the residual bone ridge. Using an image analysis technique, we observed, on the deeper implant, a 0-56% rate of bony contact around the implant and a 71-84% rate of bone-vent occupancy in the vents. With the shallower implant, a 0-46% rate of bony contact and a 26-60% rate of bone-vent occupancy were observed. When the two implants were compared, the deeper implant showed relatively higher bony contact and bone-vent occupancy than its less-submerged control in the five-year study.

[The anti-mutagenic effects of human saliva investigated by umu-test. Part 1. Effects of filtration and storage at low temperatures].

The anti-mutagenic effects of human saliva were examined by the umu-test. The SOS-inducible DNA-damaging activity of AF-2 was markedly decreased by pre-treatment with human fresh saliva. The anti-mutagenic activity of human saliva was retained even after the filtration (0.45 microns). This activity was also observed after 24 hr-storage at 10 degrees C or -12 degrees C. These results were consistent with previous reports using Ames tests. Significant differences in the activity, however, were observed between sample donors and between experiments with the same sample donor. Further analyses of the effects of the factors contained in saliva as well as the effects of life-styles, including dietary-habits of sample donors will be necessary.

Efficacy of a new quinolone, levofloxacin in patients with surgical infections.

The pharmacokinetics and clinical efficacy of levofloxacin (LVFX, DR-3355), an optically pure S(-)-enantiomer of ofloxacin, were studied in patients after surgery. In the pharmacokinetic study, 4 patients undergoing bile drainage were given 2 100-mg tablets by mouth. Peak levels of LVFX were from 2.22 to 4.02 micrograms/ml of plasma at 2-4 hours after the oral administration, and from 7.5 to 11.3 micrograms/ml of bile at 2-6 hours. Forty-three patients with surgical infections, including 16 skin and soft-tissue infections and 12 wound infections, were treated with LVFX. Twenty-eight (70%) of the 40 patients whose results could be evaluated had excellent or good results; 42 (93%) of the 45 causative organisms identified were eradicated. An episode of diarrhea with chills and fever occurred in a 38-year-old man. The results suggested that LVFX has satisfactory antimicrobial effects in surgical infections.

[A false aneurysm caused by rupture of a double velour Dacron graft. A case report].

A 71-year-old man was operated for a complete occlusion of the left common iliac artery caused by arteriosclerosis obliterans. Cross-over bypass was performed from the right external iliac artery to the left superficial femoral artery in June 1978, using a Double Velour Dacron graft (Cooley) 8 mm in diameter. In February 1985, the patient noticed the pulsatile mass in his left groin. Angiography showed a false aneurysm at the distal anastomosis of the graft. On operation, 2 aneurysmal dilatations were found on the distal anastomosis. After resection of the aneurysms, the reconstruction was established by EPTFE graft successfully. The resected specimen showed that one of the aneurysms was caused by the anastomotic failure of the distal portion of the graft, but that the other was the false aneurysm caused by rupture of the implanted graft. Two ruptured holes were confirmed to be 20 X 25 mm, 10 X 15 mm in size respectively, 10 mm proximal to the anastomotic aneurysm. Rupture of the Dacron arterial prosthesis is a rare complication. Only 23 cases have been reported in the United States. But some literatures show that the incidence of the failure of the Dacron prosthesis was about 3%. Therefore, we should make a long-term follow-up to the patients implanted Dacron arterial prostheses.