Changes in maxillofacial morphology due to improvement of nasal obstruction in rats.

OBJECTIVES: To investigate the effect of release of experimentally introduced nasal obstruction on maxillofacial morphology and percutaneous arterial oxygen saturation (SpO2 ) in rats. MATERIALS AND METHODS: Six-week-old male Wistar rats (n = 36) were divided into a control group (n = 6) and a nasal obstruction group (n = 30). In the nasal obstruction group, the right nostril was occluded with silicon, which was subsequently removed after a given experimental period (days 7, 21, 35, 49 and 63). These animals were then divided into groups D7, D21, D35, D49 and D63 (each n = 6), according to the day at which the obstruction was released. The SpO2 was measured in rats with nasal obstruction at five experimental points. The maxillofacial morphology in rats on the first day and 63 days after the start of the experiment was evaluated by microcomputed tomography. RESULTS: The SpO2 was still lower at 2 weeks after the improvement of the nasal obstruction in the D49 group than in the control group. In addition, the height of the nasal maxillary complex of the D35, D49 and D63 groups was significantly decreased compared with the control group. CONCLUSIONS: The results of this study suggest that long-term unilateral nasal obstruction in growing rats may affect the growth of the nasomaxillary complex and reduce the SpO2 permanently. Therefore, early improvement of nasal obstruction in rats during the growth period may improve the SpO2 and cranial development and promote normal growth and development.

Two dimensional density and its fluctuation measurements by using phase imaging method in GAMMA 10.

Two dimensional (2D) plasma image analysis is useful to study the improvement of plasma confinement in magnetically confined fusion plasmas. We have constructed a 2D interferometer system with phase imaging method for studying 2D plasma density distribution and its fluctuation measurement in the tandem mirror GAMMA 10. 2D profiles of electron density and its fluctuation have been successfully obtained by using this 2D phase imaging system. We show that 2D plasma density and fluctuation profiles clearly depends on the axial confining potential formation with application of plug electron cyclotron heating in GAMMA 10.

Radial density profile measurement by using the multichannel microwave interferometer in GAMMA 10.

Plasma density radial profile measurements are an important study for fusion plasma researches. We reconstructed a multichannel microwave interferometer for radial plasma electron density and density fluctuation measurements with both changing the transmission horn position and using the Teflon lens by only using this system in a single plasma shot. By using this system, we can successfully measure the radial density and density fluctuation spectra in a single plasma shot.

The use of relaxin improves healing in injured muscle.

To improve the functional recovery of injured skeletal muscle, we have focused our efforts on both enhancement of muscle regeneration and prevention of fibrosis. The polypeptide cytokine/growth factor relaxin can inhibit fibrous tissue formation in many tissues. As a member of the insulin-like growth factor family, relaxin also is a potential stimulator of muscle regeneration. In the current experiment, we examined the antifibrotic effect of relaxin in injured skeletal muscle. We also investigated if the injection of relaxin would influence muscle regeneration after injury. Our results demonstrate that relaxin treatment improved histologic and physiologic healing of muscles subjected to traumatic injury.

[Criteria for degrees of a smoothing method applied to mortality rates].

PURPOSE: Age-period-specific mortality rates are smoothed by methods to clarify their characteristics and trends. In the present study a procedure to determine appropriate iteration times for weighted averages was devised, and examined by application to mortality data. MATERIALS AND METHODS: Mortality data for leukemia, solid neoplasms, and traffic accidents caused by cars in Japan were employed. The smoothing method was iterating weighted averages over age-period-specific death rates and their four nearest neighbors in age-period data tables. The primary condition, which the iteration times should satisfy, was that the differences between crude data and smoothed ones were not significantly large, i.e., smoothed mortality data should be within 95% confidence intervals of crude rates. Therefore, the following steps were adopted to determine appropriate times: (1) estimation of 95% confidence intervals for crude age-period-specific mortality rates. (2) determination of iteration times under the primary condition that the proportion of cells left out of the confidence intervals should be around 5%. In cases where the proportion of cells outside of confidence intervals was still below 5% even with 10 or more times of iteration, an index of convergence of smoothing was introduced and used to determine appropriate iteration times. RESULTS: For solid neoplasms, appropriate times of iteration was three. With leukemia, the proportion of cells left out of their confidence intervals remained within 5% with iteration 27 times. Therefore, an index of convergence for the smoothing process was used to avoid over-smoothing. As a result, a value of five was obtained. In the case of car traffic accidents, even only a single smoothing did not fulfill the primary condition, because mortality rates increased very rapidly from age classes of 10-14 to 15-19, the mortality rates for the latter being about ten times larger. To deal with this, the proportion of cells left out of their confidence intervals was calculated excluding these age classes. As a result, five times was found to be appropriate. CONCLUSION: The smoothing method in this study is simple and easy to apply to mortality data. And procedure to determine appropriate iteration times is based on an inferential statistical method. Applying the method to three kinds of mortality data, it was found to be practical and useful for clarification of their characteristics and trends.

Novel structures in secreting the androgenic gland hormone.

The secretory granules in the androgenic gland of the terrestrial isopod Armadillidium vulgare, which have been indistinct for long time because of vulnerable structures, were revealed by using the rapid-freezing and freeze-substitution method. The fine structure of the androgenic gland is conspicuous by the distribution of numerous particular organelles in the cytoplasm consisting of the endoplasmic reticulum and the Golgi complex, and by having a number of highly organized structures developed between the androgenic gland cells. The structures connect to the intercellular space, which is seen as intercellular canaliculi for exporting the androgenic gland hormone. The plasma membranes near the particular structure of the intercellular canaliculi in the androgenic gland are often specialized to form cellular junctions. The secretory granules including the electron-dense materials, which are supposed to be peptides of androgenic gland hormone, are distributed beside the particular structure of the intercellular canaliculi. Some of the granules are seen to fuse with the plasma membranes. This observation suggests that, in the Armadillidium vulgare, the secretory granules containing androgenic gland hormone are transferred to the extracellular space through the intercellular canaliculi particularly developed for exporting the peptide hormone. This is the first evidence to show the secretory mechanism of the androgenic gland hormone in the Isopoda.

Placental transfer of IgG subclasses in a Japanese population.

BACKGROUND: Maternal immunoglobulin G (IgG), transferred across the placenta to the fetus during intrauterine life, is an important component of the neonatal immunological defence mechanisms against infection. There is controversy with respect to differences in placental transfer of the different IgG subclasses, and no definite data are available on a Japanese population. Therefore, we investigated placental transfer of IgG subclasses in a Japanese population. METHODS: A total of 228 matched pairs of cord and maternal serum samples (20-42 weeks gestation) were assayed for each IgG subclass by an enzyme-linked immunosorbent assay. RESULTS: The mean values and hierarchy of cord/maternal concentration ratios of IgG subclasses at 40 weeks gestation were as follows: IgG1(1.47) > IgG3(1.17) = IgG4(1.15) > IgG2(0.80). The cord/maternal concentration ratios of all IgG subclasses were positively correlated to gestational age. The mean ratios for IgG1 and IgG4 nearly reached a plateau at 39 and 37 weeks gestation, respectively, while those for IgG2 and IgG3 increased until 41 weeks gestation. The ratios of all IgG subclasses for full-term deliveries were reciprocally correlated to the respective maternal IgG subclass serum levels. CONCLUSIONS: The results suggest that although all four IgG subclasses are actively transferred across the placenta, the efficiency of their transfer ranks in the order IgG1 > IgG3 = IgG4 > IgG2. The different results as to placental transfer of IgG subclasses in the literature might be due, at least in part, to different maternal IgG subclass serum levels in the populations studied.

p53 Plasmid Preparation and Techniques for Analysis of Gene Transfer and Expression.

Hepatocellular carcinoma (HCC) is one of the most common causes of cancer death worldwide and is especially prevalent in certain areas of Africa and Asia. The most important etiological factor is infection with the hepatitis B or C virus. Treatment is generally unsatisfactory as the majority of patients are not suitable for surgical resection and chemotherapy is not particularly effective.

[A personal computer system providing various graphs of mortality rates].

PURPOSE: We devised observational methods for showing relations of mortality rates to age and period in various ways. And we contrived a personal computer system to realize the methods. The purpose of this report is to evaluate the methods and the system. METHODS AND MATERIALS: We contrived the system which provides some graphs useful for observing age-period-specific mortality rates. The program was written in Visual Basic on a Windows personal computer. The graphs provided by the system are bird's-eye view, contour map and mortality curves. The system is also made to present any graph desired for any specific required time. A smoothing method is also provided with the system to diminish random error, therefore the characteristics of mortality can be grasped easily. The system was applied to the mortality rates of malignant tumor for the last 45 years (1950-1994) in Japan. RESULTS AND CONCLUSION: Examples which show that one could easily observe the trend and structure of mortality rates by using the method were presented. Because the system can present any of the graphs instantly, so we can observe mortality step-by-step; we can seen the mortality graphs as a whole from a bird's eye view, then observe it in detail by a contour map graph, and furthermore look into the point of interest by mortality curves. Thus, the system will be useful to observe mortality rates.

Ommochrome deficiency in an albino strain of a terrestrial isopod, Armadillidium vulgare.

In order to clarify the cause of ommochrome deficiency in an albino strain of the terrestrial isopod, Armadillidium vulgare, levels of xanthommatin, 3-hydroxykynurenine, 3-hydroxyanthranilic acid and tryptophan in whole body extracts of the albino and the wild type individuals were determined together with enzyme activities of kynurenine-3-hydroxylase, kynureninase and tryptophan-2,3-dioxygenase. Xanthommatin could not be detected in the albinos. The levels of 3-hydroxykynurenine and 3-hydroxyanthranilic acid were determined by high-performance liquid chromatography (HPLC) with electrochemical detection and were markedly low in the albinos compared with the wild type individuals. In contrast to those, the tryptophan levels determined by HPLC with fluorescence detection did not differ significantly between the two phenotypes. In the albino A. vulgare, kynurenine-3-hydroxylase activity was lower and kynureninase activity was higher than in the wild type, although the differences were not statistically significant. Tryptophan-2,3-dioxygenase activity in the albinos was less than 10% that in the wild type. Thus, ommochrome deficiency in the albino A. vulgare is considered to be caused by the extremely low activity of tryptophan-2,3-dioxygenase.

Ommochrome genesis in an albino strain of a terrestrial isopod.

The contents of tryptophan (Trp) metabolites and the activities of the enzymes involved in ommochrome biosynthesis were measured in an albino strain of a terrestrial isopod Armadillidium vulgare. There was little difference between the Trp content in the albino mutant and that in the wild type, although the contents of 3-hydroxykynurenine (3-OH-Kyn), 3-hydroxyanthranilic acid (3-OH-AA) and xanthommatin in the albino were significantly lower than those in the wild type. Tryptophan 2,3-dioxygenase (TDO) activity in the albino was extremely low, while the activities of Kyn-3-hydroxylase and kynureninase did not differ significantly between the two phenotypes. The extremely low activity of TDO is probably one of main reasons why almost no ommochrome pigment is produced in the albino mutant.

Involvement of tryptophan metabolism in the body color of crustacea.

The terrestrial isopod, Armadillidium vulgare is usually grey or black in color, however, red ones are occasionally found in the field. This is caused by the mutation of the ommochrome genesis in the integument. We focused our experiments on the mechanism of pigment genesis in which tryptophan metabolism had been expected to be different from the grey or black wild types. We obtained the result that 3-hydroxyanthranilic acid content was significantly higher in the red phenotype than in the wild type, and kynureninase activity was also higher in the red phenotype.

Involvement of pteridines in the body coloration of the isopod Armadillidium vulgare.

The pteridine content was measured as a function of age in Armadillidium vulgare, and the fine structure of the pteridine-containing granules in the integument was examined in relation to pteridine content. Yellow chromatophores are an essential component of the cream-markings, which are a defining feature of the female A. vulgare. Four kinds of pteridines in the integument including a yellow pigment (sepiapterin) were determined by HPLC. The body color of the red phenotype of A. vulgare varies from dark red to yellowish red and was formerly thought to be due to the quality and quantity of ommochrome pigment. Our analysis of the pteridine content in the integument of this phenotype revealed a significant change in sepiapterin content per body weight with age. Sepiapterin content per body weight decreased gradually with age, while that of biopterin tended to increase with age. Ultrastructural observations of the pigment granules in the yellow chromatophores revealed a corresponding change in the fine structure of pigment granules. In the older adults, some of the electron-dense fibrous materials in the pteridine-containing granules was concentrically arranged, and in the younger adults, most of pteridine-containing granules were electron-lucent. The role of pteridine quality in determining the structure of pteridine-containing granules is discussed.

Splenic adherent cells, stimulated in vitro, induce the reactive formation of lymphoid follicles and germinal centres in draining lymph nodes after subcutaneous transfusion into syngeneic mice.

The reactive formation of lymphoid follicles and germinal centres in lymph nodes, induced by subcutaneous transfer of in vitro activated splenic adherent cells into syngeneic mice, were studied. Adherent cells were obtained by incubating spleen cell suspensions for 24 h and activated by incubating for 1 h in the medium containing keyhole limpet haemocyanin (KLH) absorbed onto alumina. Some of the treated adherent cells were irradiated with 10 Gy x-rays, while others were either not stimulated or were stimulated with alumina-KLH but killed by repeated freezing and thawing. Examination of adherent cell smears immunostained with antibodies against, F4/80, Mac-1, Mac-2 and NLDC-145 indicated that many adherent cells displayed macrophage markers but few displayed the interdigitating cell marker. Animals transfused with KLH-treated adherent cells with or without irradiation showed a marked increase in the number of lymphoid follicles and germinal centres in draining lymph nodes, whereas those transfused with adherent cells which had not been KLH-treated or which had been killed after KLH treatment displayed no significant change in the number of follicles. These results were interpreted as indicating that following transfusion, antigen-activated adherent macrophages migrated into the draining lymph nodes and induced the reactive formation of lymphoid follicles and germinal centres outside preexisting follicles.

Stimulatory effects of neopterin on hematopoiesis in vitro are mediated by activation of stromal cell function.

The pteridine neopterin (NP) was shown to be produced by monocytes and is known to be a useful marker of immunological activation, although, its biological activity is still unclear. Recently, we found that intravenous administration of NP increased the numbers of blood leukocytes, and granulocyte-macrophage progenitor cells (CFU-GM) in the bone marrow and spleens of mice. In order to elucidate the mechanism whereby NP stimulates hematopoiesis, the effects of NP on hematopoietic stem cell proliferation and differentiation in vitro were studied using a long-term bone marrow culture (LTMC) system with cloned stromal cell line, MS-5. Adding NP to the LTMC increased the numbers of cells in total, CFU-GM and colony-forming unit in spleen (CFU-S). NP also increased the number of CFU-GM in a soft agar culture system, but it did not enhance CFU-GM colony formation when target bone marrow cells were semi-purified (T, B and adherent cell-depleted bone marrow cells) and cultured in this system, suggesting that NP did not directly affect the proliferation of hematopoietic progenitors. Conditioned medium obtained from NP-treated stromal cells had much greater colony-stimulating activity than that obtained from untreated stromal cells. Furthermore, NP treatment stimulated the production of IL-6 and GM-CSF by stromal cells. All these findings suggest that NP stimulates hematopoietic cell proliferation and differentiation in vitro by activating stromal cell function.

Genetic and biochemical studies on ommochrome genesis in an albino strain of a terrestrial isopod, Armadillidium vulgare.

Genetic studies and quantitative determination of levels of 3-hydroxykynurenine and kynurenine were performed in an albino strain of a terrestrial isopod Armadillidium vulgare. From the results of matings between the albino and the albino, the red, the dark red, or the wild type individuals, the albino A. vulgare seems to be regulated by an autosomal gene(s) recessive to its wild allele. Litter mating of F1 progenies obtained by crossing the albino and the red mutant or the albino and the dark red mutant yielded progenies at a ratio of 3:6:3:4 for the red, the dark red, the wild, and the albino phenotypes, respectively. The albino gene(s) seems not to be allelic but to be epistatic to the red gene(s) with respect to ommochrome biosynthesis. Quantitative determination of 3-hydroxykynurenine carried out by high-performance liquid chromatography with electrochemical detection revealed that the 3-hydroxykynurenine content in the albino was significantly lower than that in the wild or the red type. The whole content of 3-hydroxykynurenine after enzymatic conversion of kynurenine to 3-hydroxykynurenine was still considerably lower than that found in the wild type, even though it increased after the conversion. The albino gene(s) seems to be associated with a blockage at distinct level(s) of ommochrome biosynthesis.

Effect of sublethal X-irradiation on follicular trapping of immune complexes in the mouse lymph node.

In one set of experiments, the effect of sublethal X-irradiation on the 24 h localization of subcutaneously injected immune complexes in the lymph node follicles was studied in mice which were given HRP-anti-HRP complexes into the rear footpad at 1-3 weeks after irradiation and killed 1 day later. The 24 h follicular localization of injected immune complexes in draining popliteal nodes was severely impaired at 7 days after irradiation, at which time residual follicles were markedly depleted of B lymphocytes. In following weeks, residual follicles began to be repopulated, and the 24 h follicular localization of immune complexes became restored. Follicular dendritic cells (FDC), as was detected by the in vitro trapping assay and/or by the immunostaining for complement receptors CR1, were present in lymph nodes at any time after irradiation. Another group of mice were given HRP-anti-HRP complexes at 6 days of X-irradiation and killed from 15 min to 24 h later. Following the injection, complexes localized in residual follicles in draining nodes within 15 min but soon diminished in density and finally disappeared by 24 h after injection. It is obvious that sublethal irradiation affect neither transport of immune complexes to lymph node follicles nor their localization in these follicles. Rather rapid disappearance after temporal localization of immune complexes in residual follicles irradiated mice indicates that persisting FDC were unable effectively to trap immune complexes which were transported and localized in residual follicles. Ineffective trapping by FDC of immune complexes temporally localized in residual follicles is discussed in relation to the depletion of follicular B lymphocytes due to X-irradiation.

Yellow marking and pteridine contents in the integument of albino Armadillidium vulgare.

The albino mutant strain in the woodlice, Armadillidium vulgare, was investigated with respect to the yellow patterns on the dorsal integument. Pigment cells were observed with electron microscope in order to determine the cell types of yellow markings. Quantitative analyses of pteridines in the albino were carried out by HPLC. The result indicated that the albino integument contain sepiapterin, biopterin, pterin, isoxanthopterin as in the wild type and the red mutant strain. The total amount of the four pteridines in the albino was about half as much as that in the red phenotype for both males and females, respectively. Males and females showed almost the same totals and ratios of the four pteridines in the albino and red phenotypes. Therefore, pteridine contents in both phenotypes of A. vulgare may not be related to the activity of androgenic gland hormone. Yellow chromatophores of the albino and red phenotypes were morphologically identical, emitting a yellow fluorescence. These cells contained numerous electron-lucent pigment organelles which were similar to pteridine granules of the wild type.

The induction of cardioangitis by Lactobacillus casei cell wall in mice. I. The cytokine production from murine macrophages by Lactobacillus casei cell wall extract.

A single intraperitoneal injection of the water-soluble cell wall extract from Lactobacillus casei (LCWE) in mice can induce acute inflammatory heart involvement such as cardiac arteritis, similar to the heart lesions in Kawasaki disease. To clarify whether inflammatory cytokines participate in pathogenic mechanisms of LCWE-induced cardioangitis in mice, we studied in vitro cytokine production (IL-1, TNF alpha, and IL-6) from peritoneal macrophages (M phi) stimulated with LCWE in comparison with that of lipopolysaccharide (LPS). LCWE significantly activated M phi of BALB/c in production of IL-1 and TNF alpha compared with LPS. However, the magnitude of IL-6 synthesis by LCWE was approximately similar to that of LPS. These results demonstrate that the pathogenesis of LCWE- induced cardiac lesions may be associated with increased production of inflammatory cytokines.

[Pharmacokinetic, bacteriological, and clinical studies on SY5555 in children].

Pharmacokinetic, bacteriological and clinical studies on SY5555 were performed in children. The results were as follows: 1. A total of 15 patients considered to have bacterial infections were treated with SY5555. Each dose, 5 mg/kg, was orally administered 3 times daily, for 4-11 days. Clinical efficacies of SY5555 in 13 patients with bacterial infections (1 with pneumonia, 2 with bronchitis, each 1 with maxillary sinusitis, 2 with otitis media, 5 with pharyngitis, 1 each with gastroenteritis and pyelonephritis) were evaluated as excellent in 10 patients and as good in 3 patients with an efficacy rate of 100%. Two patients with viral infection and malignant lymphoma were not evaluated. Thirteen causative strains in 7 species were found in 10 patients. Streptococcus pneumoniae in 1/3, Haemophilus influenzae in 2/2, Streptococcus pyogenes 4/4, Salmonella spp. in 1/1, Escherichia coli in 1/1 were eradicated. Only one patient developed mild diarrhea as an adverse reaction. Another patient showed elevated GPT (glutamate pyruvate transaminase). The abnormality was mild and the patient recovered after the cessation of SY5555 administration without specific treatment. 2. MICs of SY5555 were examined against 33 clinical isolates. SY5555 has low MICs against Enterococcus faecalis and other Gram-positive cocci. 3. Pharmacokinetic studies Peak plasma concentrations of SY5555 was 1.15 micrograms/ml at a dose level of 4.9 mg/kg orally administered at fasting. Based on the above results and the broad spectrum of the anti-bacterial activities, SY5555 appears to be a promising antibiotics that is usable as a single agent for the primary therapy of respiratory tract infections, skin soft tissue infections and urinary tract infections in children.