Selective release of miRNAs via extracellular vesicles is associated with house-dust mite allergen-induced airway inflammation.

BACKGROUND: MicroRNAs (miRNAs) may facilitate cell-to-cell communication via extracellular vesicles (EVs). The biological roles of miRNAs in EVs on allergic airway inflammation are unclear. METHODS: Airway-secreted EVs (AEVs) were isolated from bronchoalveolar lavage fluid (BALF) of control and house-dust mite (HDM) allergen-exposed HDM-sensitized mice. The expression of miRNAs in AEVs or miRNAs and mRNAs in lung tissue was analysed using miRNA microarray. RESULTS: The amount of AEV increased 8.9-fold in BALF from HDM-exposed mice compared with that from sham-control mice. HDM exposure resulted in significant changes in the expression of 139 miRNAs in EVs and 175 miRNAs in lung tissues, with 54 miRNAs being common in both samples. Expression changes of these 54 miRNAs between miRNAs in AEVs and lung tissues after HDM exposure were inversely correlated. Computational analysis revealed that 31 genes, including IL-13 and IL-5Ra, are putative targets of the miRNAs up-regulated in AEVs but down-regulated in lung tissues after HDM exposure. The amount of AEV in BALF after HDM exposure was diminished by treatment with the sphingomyelinase inhibitor GW4869. The treatment with GW4869 also decreased Th2 cytokines and eosinophil counts in BALFs and reduced eosinophil accumulation in airway walls and mucosa. CONCLUSION: These results indicate that selective sorting of miRNA including Th2 inhibitory miRNAs into AEVs and increase release to the airway after HDM exposure would be involved in the pathogenesis of allergic airway inflammation.

Mitochondrial function in Babesia microti and Babesia rodhaini.

The role of mitochondria in the energy metabolism of Babesia microti and Babesia rodhaini was investigated. A variety of mitochondrial inhibitors showed greater sensitivity to B. microti than to B. rodhaini. Additionally, alpha-glycerophosphate- and succinate-cytochrome c reductase activities in the crude mitochondrial fraction from B. microti were substantially higher than those from B. rodhaini. Our results suggest that the mitochondria of these parasites possess a series of "classical" apparati for energy production and their relative functional role may be quantitatively greater in B. microti when compared with B. rodhaini.

Histological observation of large light cells that seem to be surviving hypertrophic chondrocytes in the rat mandibular condyle.

Large light cells (more than 20 microns dia), including some with mitotic figures, appeared to be surviving hypertrophic chondrocytes. Thus at least a few hypertrophic chondrocytes in the rat mandibular condyle may survive, be released into the primary spongiosa, and divide.

A short term in vitro cultivation of Babesia rodhaini and Babesia microti.

In vitro cultivation of Babesia rodhaini (BR) and Babesia microti (BM) was attempted. When RPMI1640 was supplemented with 30 or 40% of non-treated fetal bovine serum (FBS), the gas mixture of 3% CO2-8% O2 best supported the growth of both parasites. Under this optimized condition, the percent parasitized erythrocytes peaked to approximately 4- and 2-times initial values for Br and BM, respectively. The cultivated parasites retained the infectivity to the host mice. BM showed the characteristic feature of division during cultivation. However, the lots of FBS will have to be taken into consideration, since the FBS lots were shown to give large varieties to the results. Selection of the appropriate FBS lot may yield the better growth of these protozoa.

Isoenzyme patterns of glucose-6-phosphate dehydrogenase, malate dehydrogenase and lactate dehydrogenase in Babesia rodhaini and Babesia microti.

Glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), and lactate dehydrogenase (LDH) activities and their isoenzyme patterns in B. rodhaini (BR) and B. microti (BM), the two major causative species of murine babesiosis, were examined. G6PD and LDH activities were higher in BR than those in BM, whereas MDH activity was lower in BR than that in BM. No differences were observed between BR and BM in the mobility of isoenzyme bands of G6PD and MDH. On LDH isoenzyme pattern, at least 5 bands were detected in BR, while only one band in BM. Since each subunit of LDH is known to be coded by different gene, these results suggests that BR and BM are able to be differentiated genetically.

Enzyme activities related to glucose metabolism in Babesia microti and Babesia rodhaini.

A comparative study was carried out on the glucose metabolism in Babesia microti (BM) and Babesia rodhaini (BR) by analyzing the enzyme activities. The lactate dehydrogenase (LDH) activity in BM showed significantly lower values than that in BR, whereas citrate synthase (CS) and malate dehydrogenase (MDH) activities were remarkably higher in BM. In addition, pyruvate dehydrogenase (PDH), isocitrate dehydrogenase (ICDH), alpha-ketoglutarate dehydrogenase (KGDH), and succinate dehydrogenase (SDH) activities also tended to be higher in BM. Then, the change of enzyme activities related to the proliferation of parasites was examined. In BM infected mice, the parasitemia increased from day 15 to day 19 after inoculation (a.i.). While BM showed decrease of G6PD and LDH activities at day 19 a.i., it showed remarkably increased activities in CS and MDH (368 and 8,842 nmol/min.mg protein, respectively). In addition, PDH, ICDH, KGDH, and SDH activities also tended to increase from day 15 to 19 a.i. In BR infected mice, parasitemia increased from day 9 to day 12 a.i. LDH activity showed a considerable increase at day 12 a.i. (12,920 IU/mg.protein). Although CS and MDH activities also showed a slight increase at day 12 a.i., the activities of PDH, ICDH, KGDH and SDH didn't change from day 9 to 12 a.i. Since these changes observed in the enzyme activities of BM and BR seemed to be correlated with their proliferation, it was suggested that BM and BR depended on aerobic and anaerobic pathways, respectively, for their glucose metabolism.

Tissue residues of sulfadimethoxine following dietary administration to broiler-chickens.

Broilers were divided into four groups and the first group served as the control. The second, third and fourth groups were given feed containing 25, 50 and 100 mg/kg of sulfadimethoxine (SDM), respectively, for 21 days, and thereafter each group received the SDM free feed. On certain days during the experiment period, three broilers in each group were sacrificed and tissues, including blood, heart, liver, spleen, gizzard, thigh muscle, breast muscle and fat, were collected and residual SDM were determined by HPLC. Two days after withdrawal, SDM in each tissue had decreased to below the detection limit of 0.01 microgram/g.

Effects of depletion of T cell subpopulations on the course of infection and anti-parasite delayed type hypersensitivity response in mice infected with Babesia microti and Babesia rodhaini.

To elucidate the role of T cell subpopulations in the protective cell-mediated immune response at the initial phase of infection with Babesia microti (BM) and B. rodhaini (BR), the changes in the course of infection and anti-parasite delayed type hypersensitivity (DTH) response after BM or BR inoculation were investigated in Lyt-2+ T cell or L3T4+ T cell-depleted mice. Depletion of Lyt-2+ T cells strongly enhanced the resistance to BM infection, whereas it increased the susceptibility to BR infection. In contrast, depletion of L3T4+ T cells increased susceptibility to BM infection, while it enhanced resistance to BR infection. The anti-parasite DTH response in BM-infected mice was significantly enhanced by depletion of Lyt-2+ T cells, while significantly reduced by depletion of L3T4+ T cells. No effects of depletion of either Lyt-2+ or L3T4+ cells on DTH response was observed in BR-infected mice. From these results, it was suggested that the roles of Lyt-2+ and L3T4+ T cells in the protective cell-mediated immune response at the initial phase of infection were different between BM- and BR-infected mice, resulting in the difference in their course of infection.

Changes of splenic lymphocyte subpopulation in mice inoculated with Babesia microti and Babesia rodhaini.

Changes of splenic lymphocyte subpopulation after Babesia microti and Babesia rodhaini inoculation in mice were examined by flow cytometric analysis. The B. microti inoculated mice showed a longer period of time from inoculation to the onset of increase or decrease parasitaemia (%), packed cell volume, total spleen cell numbers and surface immunoglobulin positive splenic cell numbers than respective periods in B. rodhaini inoculated mice. The Thy-1 positive cell numbers in B. microti inoculated mice and B. rodhaini inoculated mice pre-immunized with homologous parasites were significantly higher than that of B. rodhaini inoculated mice. The ratio of L3T4 positive cell/Lyt-2 positive cell after inoculation with B. microti was quite similar to that in B. rodhaini mice pre-immunized. However, the ratio in B. rodhaini inoculated mice revealed a lack of an increasing phase. These results suggested that the T-cell dependent early immune response, especially suppressor activity, was closely related to the difference in the course of infection between the non-lethal B. microti and the lethal B. rodhaini infection in mice.

Adjectives that have different meanings in various Latin anatomical names, and expression of these adjectives in Japanese anatomical names.

In order to understand the role of Latin adjectives, international anatomical names containing the same adjectives were chosen from among the names of structures in the head and neck: these names were assorted into groups according to the actual meanings of the adjectives. When the Latin adjectives indicated the name of the structure, they signified: belonging to the structure, entering into the formation of the structure, articulating with the structure, transmitting the structure, giving attachment to the structure, or some other relationship to the structure. And, furthermore, the structures that were indicated by the same Latin adjectives might be different. In the Japanese language, those adjectives that have different meanings in other internationally accepted anatomical names were sometimes translated into different words. Because of this, some Japanese anatomical names were more concrete than the corresponding Latin anatomical names. It seemed that compound words and abbreviations, which can be formed easily in the Japanese language, made such expression possible.

The official nomenclature of the articular facets for the rib on the thoracic vertebra: a case for revision.

In Nomina Anatomica 6th ed., the articular facets for the rib on the thoracic vertebra are called Fovea costalis superior, Fovea costalis inferior, and Fovea costalis processus transversi respectively. But, there is a little problem about the names of Fovea costalis superior and Fovea costalis inferior on the body of vertebra, because usually only one facet exists on each side of the body of 10th to 12th (or 9th to 12th) thoracic vertebra. This single facet on the body of vertebra should be called just Fovea costalis.

[Histological observations on structure of bundle bone in area of mylohyoid line].

The purpose of this study was to investigate the structure and the function of the part of the mandible where mylohyoid muscle originates. The bundle bone in the area of mylohyoid line and the tendon fibers on the surface of the bundle bone were observed by light microscopy and scanning and transmission electron microscopy. The conclusions were as follows: 1. It seemed that remodeling occurs frequently in bundle bone in the area of the mylohyoid line. 2. Sharpey's fibers in bundle bone in the area of the mylohyoid line seemed to be flexible. This suggested that Sharpey's fibers can be adapted to the tension from directions that are different to a certain degree. 3. Both directions of Sharpey's fibers in bundle bone in the area of the mylohyoid line and the tendon fibers on the surface of bundle bone were fundamentally the same as that of the muscle fibers of the mylohyoid muscle. But a few existing fibers crossed these fibers. Such Sharpey's fibers and tendon fibers seemed to prevent the exfoliation of the periosteum, and seemed to support main tendon fibers of the mylohyoid muscle when it is contracting.

[Anatomical names of foramina and canales in skeleton].

Latin anatomical names of Foramina and Canales in skeleton were analyzed and compared with Japanese anatomical names for better understanding of the structures of the human body and for possible revision in the future. The conclusions were as follows: 1. In general, short tunnels were called Foramina (singular: Foramen), and long tunnels Canales (singular: Canalis). 2. One end of Canalis was sometimes called Foramen. In this case, Canalis and Foramen were usually modified by the same words. 3. Each name of Foramina contained the word which means form, state, absolute size, region of existence, one of the contents or function of Foramina. 4. Each name of Canales contained the word which means region of existence, one of the contents or function of Canales. 5. Some names of Foramina and Canales that were supposed to mean the region of existence meant one of the contents of the structures. 6. As for Latin anatomical names, the relation between words were relatively clear by the proper use of noun, adjective, nominative, and genitive. 7. Since different Chinese characters were sometimes pronounced similarly in Japanese anatomical names, different structures might be confused. 8. It seemed that some Japanese anatomical names needed partial correction.

[Anatomical names of fossae and foveae in skeleton].

Latin anatomical names of Fossae and Foveae in the skeleton were analyzed and compared with Japanese anatomical names for better understanding of the structures of the human body and for possible revision in the future. The conclusions were as follows: 1. In general, round excavations were called Foveae (singular : Fovea), and nonround excavations were called Fossae (singular : Fossa). Some shallow excavations for articulation and some shallow excavations with the names which indicate their contents were called Foveae even though they were not round. 2. Each name of Fossae contained the word which indicates form, location or content of Fossa, the bone (or osseous structure) which articulates with Fossa, or the muscle which is attached to Fossa. 3. Each name of Foveae contained the word which indicates location, content or articulation of Fovea, the bone (or osseous structure) which articulates with Fovea, or the muscle (or muscular trochlea) which is attached to Fovea. 4. The Japanese name which corresponds to Fossa canina should be changed from Kenshi (canine tooth) = ka (fossa) to Kenshikin (canine muscle) = ka or Koukakukyokin (levator anguli oris muscle) = ka. 5. The Japanese name which corresponds to Fossa pterygopalatina should be changed from Yoku (wing) = kougai (palate) = ka (fossa) to Yokutotsu (pterygoid process) = kougaikotsu (palatine bone) = ka.