Nanostructure and elastic modulus of single trabecula in bovine cancellous bone

We aimed to investigate the elastic modulus of trabeculae using tensile tests and assess the effects of nanostructure at the hydroxyapatite (HAp) crystal scale on the elastic modulus. In the experiments, 18 trabeculae that were at least 3 mm in length in the proximal epiphysis of three adult bovine femurs were used. Tensile tests were conducted using a small tensile testing device coupled with microscopy under air-dried condition. The c-axis orientation of HAp crystals and the degree of orientation were measured by X-ray diffraction. To observe the deformation behavior of HAp crystals under tensile loading, the same tensile tests were conducted in X-ray diffraction measurements. The mineral content of specimens was evaluated using energy dispersive X-ray spectrometry. The elastic modulus of a single trabecula varied from 4.5 to 23.6 GPa, and the average was 11.5±5.0 GPa. The c-axis of HAp crystals was aligned with the trabecular axis and the crystals were lineally deformed under tensile loading. The ratio of the HAp crystal strain to the tissue strain (strain ratio) had a significant correlation with the elastic modulus (r=0.79; P<0.001). However, the mineral content and the degree of orientation did not vary widely and did not correlate with the elastic modulus in this study. It suggests that the strain ratio may represent the nanostructure of a single trabecula and would determine the elastic modulus as well as mineral content and orientation.     

Normalization of phospholipids concentration of the gastric mucosa was observed in patients with peptic ulcer after eradication of Helicobacter pylori

Background. Phospholipids concentration in the gastric mucosa decreased in patients with Helicobacter pylori infection. The aim of this study is to examine the effects of eradication of H. pylori on decreasing the phospholipids concentration in the gastric mucosa in patients with gastric or duodenal ulcer. Materials and Methods. Phospholipids (phosphatidylcholine, phosphatidylethanolamine, and sphingonomyeline) were measured in biopsy specimens from the antrum and corpus using thin‐layer chromatography. In H. pylori positive patients with gastric ulcer (n = 26) and duodenal ulcer (n = 13), and H. pylori negative controls (n = 20), the biopsy specimens were obtained before and 3 months after eradication. Eradication was performed using lansoprazole, amoxycillin, and clarithromycin. Results. Compared with the H. pylori negative control group, the concentrations of phosphatidylcholine and phosphatidylethanolamine decreased significantly in the gastric ulcer group in both antrum and corpus mucosa, and in the duodenal ulcer group in antrum mucosa. This decrease returned to the control level after eradication. Conclusions. This study demonstrates that the eradication of H. pylori in patients with peptic ulcer normalized the decrease of phosphatidylcholine and phosphatidylethanolamine in the gastric mucosa.     

Possible Mechanism of Action of β-Lactam-Enhancing Factor on Methicillin-Resistant Staphylococcus aureus

We have recently found a factor (Factor T) in aged mixtures of tungstate and phosphate which greatly enhances the antibacterial effects of β-lactams on methicillin-resistant strains of staphylococcal species such as methicillin-resistant Staphylococcus aureus (MRSA), but shows only weak effects on methicillin-susceptible S. aureus and bacterial strains other than staphylococci. Factor T alone did not strongly inhibit cell metabolism and bacterial growth unless an excess amount was added. When Factor T was added to the culture medium beforehand, the growth of MRSA cells was rapidly suppressed just after addition of oxacillin (MPIPC). However, the growth of the cells was inhibited gradually when these two reagents were added in reverse order. For full expression of the enhancing effect, it seemed necessary for cells of MRSA strains to be incubated with Factor T for at least 2–3 hr. When the cells were washed after being sensitized by incubating them for 5 hr with Factor T, it took approximately 1 hr for the cells to recover their resistance to MPIPC. Factor T reduced the amount of penicillin-binding protein-2′ (PBP 2′), and thus sensitized the MRSA strains to β-lactams.     

A method on strain measurement of HAP in cortical bone from diffusive profile of X-ray diffraction

Bone tissue is a composite material composed of hydroxyapatite (HAp) and collagen matrix. As HAp is a crystalline structure, an X-ray diffraction method is available to measure the lattice strain of HAp crystals. However, mineral particles of HAp in bone have much lower crystallinity than usual crystalline materials, which show a diffusive intensity profile of X-ray diffraction. It is not easy to determine quantitatively an infinitesimal strain of HAp from the peak position of diffusive profile. In order to improve the accuracy of strain measurement of HAp in bone tissue and to obtain reproducible results, this paper proposes an X-ray diffraction method applied to a diffusive profile for low crystallinity. This method is to estimate the lattice strain of HAp using not a peak position but a whole diffraction profile. In this experiment, a strip specimen of 28 x 8 x 2 mm was made from bone axial, outside circumferential and cross-sectional circumferential region in the cortical bone of bovine femur. The X-ray diffraction measurements were carried out before and after applying an external load. As a result, the precision of strain measurement was much improved by this method. Although a constant value of macroscopic strain was applied in the specimen, the lattice strain had a lower value than the macroscopic strain and had a different value in each specimen.     

Subchronic stress-induced depressive behavior in ovariectomized mice

AimsMood disorders including depression are more common in women than men, particularly in times of lower estradiol levels. In this study, we investigated the effect of estrogen on emotional behavior in mice in a stress environment.Main methodsFemale mice were divided into four groups: two groups were ovariectomized (OVX) and two were sham-operated. One group each of OVX and sham mice was kept in a normal environment and the other groups were assigned to a daily stress (1 h/day) for 7 days from 5 days after operation. On the 14th day after operation, subjects were measured to assess behavioral specificity, locomotor activity, elevated plus-maze (EPM) behavior, passive avoidance (PA) behavior and forced swimming behavior.Key findingsThe OVX plus stress (OVX + S) group showed a significant prolongation of immobility compared with the other groups. In all the groups there were no changes in locomotor activity, EPM behavior or PA behavior. We further examined the effect of estrogen against depressive behavior in the OVX + S group. The vehicle or 17β-estradiol (E2) was administered s.c. to OVX + S mice for 4 days beginning on post-operative day 11. Subchronic E2 treatment decreased the stress response and improved depressive behavior relative to the vehicle group.SignificanceThese data have important implications regarding the prevention of depression in postmenopausal women undergoing estrogen therapy.     

Epidemiological and Ecological Studies of Japanese Encephalitis in Okinawa,Subtropical Area in Japan. I. Investigations on Antibody Levels to Japanese Encephalitis Virus in Swine Sera and Vector Mosquito in Okinawa,Miyako and Ishigaki Islands

From 1985 to 1989, serum specimens of swine raised in the northern, central and southern areas in Okinawa island were examined for Japanese encephalitis (JE) virus antibody by ELISA and hemagglutination-inhibition test. The antibody positive rate was found to be higher in the north and central than in the south. The 2-mercaptoethanol sensitive antibody to JE was detected mostly in June and July, and occasionally in other months except February and March. There was no month when all specimens from three areas turned antibody-negative simultaneously, indicating that JE virus transmission to swine lasted longer in Okinawa island than in other temperate areas in Japan. From 1986 to 1991, the vector mosquitoes (Culex tritaeniorhynchus) were collected in a pig farm in the south of Okinawa island. A total of 153 strains of JE virus was isolated from the vector mosquitoes mainly in June. In Miyako and Ishigaki islands, the antibody positive rate in swine sera was found to be extremely low, compared with that in Okinawa island. In Miyako island, where no paddy rice field is cultivated, a few adults as well as larvae of the vector mosquito were collected, while in Ishigaki island, where there are many watered rice fields, a lot of adults as well as larvae were collected. Although the environmental situation is quite different between the two islands, JE virus transmission appeared to be very low in both islands.     

Locoregional immunotherapy of head and neck cancers utilizing allogeneic spleen cells — a report of 2 cases

Frozen-stored human spleen cells (SC) cultured with streptococcus preparation OK-432 acquired direct cytotoxicity to autologous as well as allogeneic tumor cells. The activated cells started to produce cytocidal cytokine TGIF, which is distinct from previously known cytokines.We examined the possibility of allogeneic adoptive immunotherapy (AIT) using these OK-432-stimulated spleen cells (OK-SC) in two cancer patients. Rapid necrosis of cancer tissue and remarkable decreases of tumor markers in tumor effusion were observed. There were no severe side effects.     

Inhibition of tumor cell growth by a human B-cell line

An Adenocarcinoma cell line (Breast-M) and an Epstein-Barr virus (EBV)-infected B-cell line (Hairy-BM) were established from breast tumor tissue. The Hairy-BM was CD20⁺, CD25 (Tac)+ and surface immunoglobulin (sIg)+. Hairy-BM suppressed the in vitro proliferation of Breast-M in a time and a dose-dependent manner. The suppression was also found in 5 different human tumor targets showing tumor-Hairy-BM binding, but not; in 2 murine tumor targets showing no significant tumor-Hairy-BM binding. Lytic activity of Hairy-BM was found only against Breast-M.Abbreviations sIg Surface immunoglobulin - CTL Cytotoxic T-cells - NK Natural killer - IL2 Interleukin 2 - LAK Lymphokine activated killer - CSN Culture supernatant - MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoolium bromide - PCR Polymerase chain reaction - TIL Tumor-infiltrating lymphocytes - HCL Hairy cell leukemia - TNF Tumor necrosis factor     

A retrospective serological study of Japanese who contracted dengue fever in Thailand

Between September and November 1981, some members of a survey team from Japan suffered from a febrile illness diagnosed clinically as dengue fever during their stay in a village in Khon-Kaen Province, in the north-eastern part of Thailand. The morbidity rate in the team was as high as 69% (11/16). Blood samples were taken from 12 of the 16 members of the team in February, 1982 in Japan and the serum specimens were examined for antibodies to dengue (DEN), Japanese encephalitis (JE) and yellow fever (YF) viruses respectively. The results of the tests indicated that all 8 members who had had symptoms had been infected with DEN type 1 virus. No case of inapparent infection with dengue viruses was found. Of these 8 persons, seven had had neutralizing (N) antibody to JE virus before infection, but their clinical manifestations had been similar to those of an individual without N antibody to JE virus and were typical symptoms of dengue fever, such as leukopenia and "saddle-back" fever, without hemorrhagic manifestations, as seen from platelet counts and hematocrit values.     

The histochemistry of complex carbohydrates in the ovarian follicles of adult mice

Summary In the ovarian follicles of adult mice, complex carbohydrate-containing structures have been studied by means of light microscopic histochemical methods. In the ovarian follicles, the zona pellucida of oocytes, follicular fluid and intercellular matrix of the granulosa layer are found to exhibit positive reactions for complex carbohydrates with 1,2-glycol and acidic groups and -D-mannosyl and -D-glucosyl residues. The present histochemical analyses have revealed that the complex carbohydrates common to the three types of the structures are hyaluronic acid, sulfated glycosaminoglycans other than isomeric chondroitin sulfates and neutral glycoproteins and that sialic acid is a particular moiety of the zona pellucida, whereas isomeric chondroitin sulfates being that of the follicular fluid and intercellular matrix of the granulosa layer. The histophysiological activities of the carbohydrate-containing structures have been discussed with special reference to their histochemical properties determined in the present study.