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To report a case of endophthalmitis associated with Purpureocillium lilacinum (P. lilacinum) during infliximab treatment for surgically induced necrotizing scleritis, successfully treated with 27-gauge vitrectomy. A single case report. A 71-year-old man who had undergone immunosuppressive therapy, including infliximab, for surgically induced necrotizing scleritis (SINS) in his left eye complained of visual disturbance and eye pain in the eye. He had a past history of surgery for recurrent pterygium: pterygium excision, amnion transplantation with mitomycin C and limbal transplantation. Visual acuity in the left eye was counting fingers at 30 cm, and intraocular pressure was 3.0 mmHg. Slit-lamp examination revealed the presence of anterior chamber cells (3+), and a B-mode ultrasound scan showed a vitreous opacity. We made a diagnosis of endophthalmitis and performed 27-gauge microincision vitrectomy surgery (27GMIVS) with antibiotic perfusion of ceftazidime, vancomycin and voriconazole. Intraoperative findings included a fungus-like ball-shaped opacity in the vitreous, and a close-to-normal retinal appearance. A vitreous body culture identified the presence of P. lilacinum. After 2 months of antibacterial and antifungal therapy, inflammation decreased and visual acuity recovered to 20/100. This is the first report of a case of endophthalmitis associated with P. lilacinum during infliximab treatment for SINS. Scleral thinning due to necrotizing scleritis, especially during immunosuppressive therapy, is a risk factor for endophthalmitis. We found that 27GMIVS was a useful strategy for such a challenging clinical situation.

本稿はケネス・レクスロスの南宋文化を論じた、“Sung Dynasty Culture”（1960 年）と題されたエッセイの翻訳と注釈である。原典は平易な英語で書かれているが、いざ翻訳となると意外にむつかしい。どのような欧文脈であれ、逐語訳にしてしまうと分けがわからなくことは致し方ないのだが、レクスロスのオリジナルな原稿は、インドと中国を西洋文明の光が届かない遅れた国として見下し、収奪の対象としてしかアジアを見ることのできなかった西欧白人社会に向けて書かれたものなのでなおさらである。, This article is basically a Japanese translation of Kenneth Rexroth's essay "Sung Dynasty Culture" (1960) with an introduction and full annotations. The original text is written in plain English, but when it comes to translation, it is surprisingly difficult. It is true that any Japanese translation from European languages is always vague and hard to understand for those who have little knowledge of European history. Rexroth's original manuscript looked down on those Europeans and Americans who are ignorant of the non-Western civilization., 研究ノート

Aim To assess the prevalence, clinical characteristics, treatment, and outcomes of patients who developed ulcerative keratitis (UK) during the course of rheumatoid arthritis (RA) in the modern biologic era. Method We retrospectively reviewed the medical records of 589 patients with RA who visited our department between April 2003 and October 2014, and identified patients who developed UK. We also obtained data about clinical characteristics of RA and UK, complications, treatment, and both visual and life prognoses of these patients. Results Among 589 patients with RA, eight patients (1.4%) were diagnosed with UK. The mean age at the onset of RA was 61.0 years, while the mean age at the onset of UK was 73.0 years. Most of the patients were seropositive and had established RA with a relatively low disease activity. Secondary Sjogren's syndrome was observed in two patients. Peripheral UK occurred as a complication of scleritis, while central UK was not associated with scleritis. Although the mean duration of follow-up was only 3.7 years, visual and life prognoses were both tolerable with therapy, including the use of topical and systemic corticosteroids and calcineurin inhibitors, sometimes combined with biologic disease-modifying antirheumatic drugs (DMARDs) and corneal transplantation. Conclusion This retrospective study demonstrated that the prevalence of UK in patients with RA was 1.4%. Immediate combination therapy, including topical and systemic corticosteroids and calcineurin inhibitors, together with biologic DMARDs and corneal transplantation, was effective for treating RA patients who developed UK in the modern biologic era.

Purpose To assess the optic nerve head blood flow (ONH BF) response to hyperoxia in glaucoma patients using laser speckle flowgraphy (LSFG), and determine factors influencing vasoreactivity within the ONH. Methods We performed oxygen provocation testing in 15 eyes of 15 primary open-angle glaucoma (POAG) patients and 15 eyes of 15 age-matched control subjects. During the test, LSFG-derived tissue mean blur rate (MBRT) and clinical variables, including blood pressure, were recorded. We evaluated differences in MBRT alteration during systemic hyperoxia between the groups. Additionally, we calculated the mean % change in MBRT against baseline and determined contributing factors. Results Despite similar clinical variables during systemic hyperoxia in both groups, the mean % change in MBRT against baseline was significantly lower in the POAG than control subjects (P < 0.0001). Multiple regression analysis revealed that baseline MBRT and systolic blood pressure (SBP) were contributing factors to mean % change in MBRT (β = 0.44, β = -0.32, respectively). Additionally, baseline MBRT and SBP were strongly correlated to mean % change in MBRT only in the POAG group (r = 0.83, P < 0.0001; r = -0.60, P = 0.02, respectively). Conclusions POAG patients had a weaker vasoreactive response to hyperoxia than controls, and this impaired response was associated with lower basal ONH BF and higher SBP. These findings suggest that pre-existing vasoconstriction in the ONH of eyes with glaucoma might reduce the capacity of the vasoconstrictive response to hyperoxia. Alternatively, the pathways that mediate hyperoxia-induced vasoconstriction could be altered in POAG.

Purpose The assessment of retinal nerve fiber layer defects (RNFLDs) is a useful part of glaucoma care. Here, we obtained en-face images of retinal layers below the inner limiting membrane (ILM) with swept source-optical coherence tomography (SS-OCT), and measured RNFLD angle with new software. Methods This study included 105 eyes of 105 normal tension glaucoma (NTG) patients (age, 59.8 ± 13.2). Exclusion criteria were best-corrected visual acuity 28 mm, non-glaucoma ocular disease, and systemic disease affecting the visual field. We obtained 12 x 9 mm 3D volume scans centered on the macula with SS-OCT (DRI OCT-1, Topcon), and from these scans, created 3 averaged en-face images, each comprising 7 horizontal en-face images (total thickness: 18.2 μm). We labeled these averaged images, according to their depth below the ILM, as en-face images 1 (shallowest), 2 (middle) and 3 (deepest). In each image, a circle was drawn centered on the disc, with a radius of half the distance between the centers of the disc and macula. The investigator marked points where the edge of the RNFLD intersected this circle, and RNFLD angle (RNFLDA) was calculated with new software. Finally, we analyzed the association between RNFLDA, cpRNFLT, weighted RGC count (wrgc) and Humphrey field analyzer (HFA)-measured mean deviation (MD) and hemifield total deviation (TD), both overall and in each hemifield. Results En-face image 2 had the highest interclass reproducibility for measuring RNFLDA (intra-rater intraclass correlation coefficient (ICC): 0.988, inter-rater ICC: 0.962). The correlation coefficients with RNFLDA were: HFA MD, -0.60; superior TD, -0.73; inferior TD, -0.69; overall cpRNFLT, -0.27; superior hemifield cpRNFLT, -0.39; and inferior hemifield cpRNFLT, -0.53 (all p

Purpose To objectively classify the optic discs of open-angle glaucoma (OAG) patients into Nicolela's four disc types, i.e., focal ischemic (FI), myopic (MY), senile sclerotic (SS), and generalized enlargement (GE), with swept-source optical coherence tomography (SS-OCT). Methods This study enrolled 113 eyes of 113 OAG patients (mean age: 62.5 ± 12.6; Humphrey field analyzer-measured mean deviation: -9.4 ± 7.3 dB). Newly developed software was used to quantify a total of 20 optic disc parameters in SS-OCT (DRI OCT-1, TOPCON) images of the optic disc. The most suitable reference plane (RP) above the plane of Bruch’s membrane opening was determined by comparing, at various RP heights, the SS-OCT-measured rim parameters and spectral-domain OCT-measured circumpapillary retinal nerve fiber layer thickness (cpRNFLT), with Pearson's correlation analysis. To obtain a discriminant formula for disc type classification, a training group of 72 eyes of 72 OAG patients and a validation group of 60 eyes of 60 OAG patients were set up. Results Correlation with cpRNFLT differed with disc type and RP height, but overall, a height of 120 μm minimized the influence of disc type. Six parameters were most significant for disc type discrimination: disc angle (horizontal), average cup depth, cup/disc ratio, rim-decentering ratio, average rim/disc ratio (upper and lower nasal). Classifying the validation group with these parameters returned an identification rate of 80.0% and a Cohen’s Kappa of 0.73. Conclusion Our new, objective SS-OCT-based method enabled us to classify glaucomatous optic discs with high reproducibility and accuracy.

The transport pathway of specific dietary carotenoids from the midgut lumen to the silk gland in the silkworm, Bombyx mori, is a model system for selective carotenoid transport because several genetic mutants with defects in parts of this pathway have been identified that manifest altered cocoon pigmentation. In the wild-type silkworm, which has both genes, Yellow blood (Y) and Yellow cocoon (C), lutein is transferred selectively from the hemolymph lipoprotein to the silk gland cells where it is accumulated into the cocoon. The Y gene encodes an intracellular carotenoid-binding protein (CBP) containing a lipid-binding domain known as the steroidogenic acute regulatory protein-related lipid transfer domain. Positional cloning and transgenic rescue experiments revealed that the C gene encodes Cameo2, a transmembrane protein gene belonging to the CD36 family genes, some of which, such as the mammalian SR-BI and the fruit fly ninaD, are reported as lipoprotein receptors or implicated in carotenoid transport for visual system. In C mutant larvae, Cameo2 expression was strongly repressed in the silk gland in a specific manner, resulting in colorless silk glands and white cocoons. The developmental profile of Cameo2 expression, CBP expression, and lutein pigmentation in the silk gland of the yellow cocoon strain were correlated. We hypothesize that selective delivery of lutein to specific tissue requires the combination of two components: 1) CBP as a carotenoid transporter in cytosol and 2) Cameo2 as a transmembrane receptor on the surface of the cells.

In the present study, we purified and sequenced a homolog of the Drosophila imaginal disc growth factor (IDGF) from the hemolymph of Bombyx mori (BmIDGF). Antibodies against BmIDGF were produced and subsequently used in immunoblotting analyses. The immunoblotting analyses demonstrated an extremely high level of BmIDGF in the hemolymph throughout the period of rapid growth of the organs of B. mori. The results of RT-PCR showed that BmIDGF was predominantly expressed in fat bodies. Real-time RT-PCR revealed that BmIDGF transcripts in fat bodies were highly expressed during the feeding stage but significantly suppressed during the molting, wandering, and pupal stages. Starvation brought about a significant decline of BmIDGF mRNAs in the fat bodies and BmIDGF proteins in the hemolymph. After re-feeding, the BmIDGF transcripts in fat bodies and BmIDGF proteins in the hemolymph increased again. In addition, an immunocytochemical study revealed BmIDGF proteins on the surface of wing discs. The present findings suggest that the level of BmIDGF in the hemolymph was modulated by the fat body in response to nutritional conditions and that BmIDGF was transported to target organs through the hemolymph.

In the land application system to purify liquid animal waste, the introduction of vegetation has further increased treatment ability. Although the vegetation beds were covered with zoysiagrass (Zoysia japonica Steud.), we introduced centipedegrass (Eremochloa ophiuroides Hack.) into an experimental bed in order to promote the purification activity of secondary treatment after primary treatment during a midsummer period because centipedegrass thrives even in hotter conditions and to compare its purification ability with that of zoysiagrass. The results showed that the centipedegrass bed had a higher purification ability than zoysiagrass beds in a hot season. The drainage material purified through the centipedegrass bed demonstrated lower values of turbidity, electric conductivity, chemical oxygen demand, total phosphorus, total nitrogen and ammonium-nitrogen, and higher concentrations of nitrate-nitrogen than that purified through the zoysiagrass bed during the middle vegetative stage when the centipedegrass reached the highest biomass. These results suggested that nitrification had occurred vigorously in the centipedegrass plot. It was suggested that the higher nitrogen purification ability of the centipedegrass was linked to both better absorption of nitrogen and nitrification.

The animal waste treatment facility of Azabu University utilizes cultivation beds where liquid animal waste is purified by filtration through soil and vegetation. The beds are presently covered with zoysiagrass (Zoysia japonica Steud.), which thrives in warm weather. However, the purification activities of this grass decrease during winter dormancy. To improve purification activity between autumn and early spring, we introduced redtop (Agrostis alba L.), which thrives in cool conditions, into an experimental bed and compared the purification efficiency with that of zoysiagrass. Drainage purified through the bed of redtop showed significantly lower values of turbidity, chemical oxygen demand, total phosphorus and ammonium-nitrogen concentrations, and significantly higher nitrate-nitrogen concentrations than that through zoysiagrass during the late vegetative stage when redtop had the highest biomass. This suggested vigorous nitrification activity of rhizosphere under aerobic conditions afforded by redtop. The high purification efficiency of redtop appeared to be associated with better essential salts absorption and improved physicochemical properties of soil supporting its growth. The results showed that the purification efficiency of redtop is higher than that of zoysiagrass under seasonal low temperatures.

Neuroleptics Olanzapine is conjugated and oxidized mainly by CYP1A2/CYP2D6. Human CYP1A2 in the 5'-flanking region and intron nucleotide sequence alalysis revealed the existence of a point mutation (-3858) from guanine (wild type; wt) to adenine (mutated type; 1C) caused a significant decrease of enzyme activity, from cytosine (-164) to adenine (mutated; 1F) caused higher inducibility of CYP1A2. Human CYP2D6 (wild type; 1) were also identified; As for the poor metabolizer, nucleotide sequence alalysis reveale the gene deletion (mutated type; 5) caused a deletion of enzyme activity, from cytosine (100) to thymine (mutated; 10) caused a enzyme activity decrease of CYP2D6. A potential side-effects during the treatment with olanzapine is hyperglycemia and newonset diabetes mellitus. There are possible mechanisms by which olanzapine could interfere with glucose metabolism. In this study, we investigate the relationship between olanzapine metabolism by CYP1A2/CYP2D6 and the side effect (fatality from hyperglysemia) to serve as an aid to solve the mechanisms of olanzapine induced abnormal glucose metabolism into diabetes mellitus among 12 Japanese schizophrenic patients. There were 3 patients whose CYP genotypes showed CYP1A2 1C/wt or CYP2D6 10/10, and whose blood olanzapine concentration was higher (about 40.0ng/mL) inspite of within 10mg daily dose. All these 3 patients did not show hyperglysemia, but other 3 patients whose CYP genotypes were normal, showed hyperglycemia. These results suggested that the higher blood olanzapine concentration may be effected by the abnormal metabolism by CYP1A2 and/or CYP2D6, but not always caused hyper glycemia.

Bombyx mori is an excellent model for the study of carotenoid-binding proteins (CBP). In previous papers, we identified and molecularly characterized a CBP from the Y-gene dominant mutants. In the present study, we attempted to correlate and establish lipid metabolism and distribution in these mutants. When [3H]-triolein was fed to the mutants, typical patterns of uptake of labeled fatty acids from midgut to hemolymph and subsequent delivery to fat body and silk glands were obtained in all mutants. Further analysis of lipid and carotenoid profiles revealed that the yellow coloration in the hemolymph associated with lipophorin is not attributed to a difference in lipophorin concentrations among the mutants, nor to its lipid composition, but rather to its carotenoid content. Lipophorin of the Y+I mutant exhibited the highest concentration of total carotenoids of 55.8 microg/mg lipophorin compared to 3.1 microg/mg in the +Y+I mutant, 1.2 microg/mg in the YI mutant and 0.5 microg/mg in the +YI mutant. Characteristic retention time in HPLC of the different classes of carotenoids of lipophorin identified the presence of lutein as the major chromophore (62-77%), followed by beta-carotenes (22-38%). Although lutein and beta-carotene content of mutants' lipophorin differed significantly, the ratio of lutein to beta-carotene of 3:1 was not different among mutants. Similarly, lipid compositions of mutant silk glands were not significantly different, but carotenoid contents were. The significantly high concentration of lutein in the Y+I mutant silk gland represented more than 160-fold increase compared to +Y+I mutant (p

Carotenoids play important and diverse roles in insects. Recently, we purified and partially characterized a carotenoid-binding protein (CBP) from the wild type of Bombyx mori. In this report, we utilized immunoblotting, ELISA and immunocytochemistry to further characterize and localize the expression of CBP in the larval midgut and silk gland obtained from the wild type and four naturally occurring mutants linked to carotenoids transport. CBP was expressed throughout the 5th stadium, with highest expressions on days 4-5 in the silk gland and days 3-5 in the midgut. Immunoblotting analyses demonstrated the presence of CBP along the middle part of the midgut. Microscopic immunocytochemistry demonstrated that the 33 kDa CBP was uniformly expressed along the brush border of columnar cells in the epithelium of the midgut typifying its function in aiding absorption of dietary carotenoids. Similarly, CBP was highly expressed along the distal membrane of the middle part of the silk gland demonstrating its function in uptake of carotenoids from lipophorin. When the middle silk glands and midguts of the four mutants were incubated with rabbit anti-CBP antibody, only proteins of the Y-gene dominant mutants cross reacted with the antibody further accentuating the hypothesis that the CBP is a Y-gene dependent protein.

Purpose This study aimed to develop a machine learning-based algorithm for objective classification of the optic disc in patients with open-angle glaucoma (OAG), using quantitative parameters obtained from ophthalmic examination instruments. Methods This study enrolled 163 eyes of 105 OAG patients (age: 62.3 ± 12.6, mean deviation of Humphrey field analyzer: -8.9 ± 7.5 dB). The eyes were classified into Nicolela’s 4 optic disc types by 3 glaucoma specialists. Randomly, 114 eyes were selected for training data and 49 for test data. A neural network (NN) was trained with the training data and evaluated with the test data. We used 91 types of quantitative data, including 7 patient background characteristics, 48 quantified OCT (swept-source OCT; DRI OCT Atlantis, Topcon) values, including optic disc topography and circumpapillary retinal nerve fiber layer thickness (cpRNFLT), and 36 blood flow parameters from laser speckle flowgraphy, to build the machine learning classification model. To extract the important features among 91 parameters, minimum redundancy maximum relevance and a genetic feature selection were used. Results The validated accuracy against test data for the NN was 87.8% (Cohen’s Kappa = 0.83). The important features in the NN were horizontal disc angle, spherical equivalent, cup area, age, 6-sector superotemporal cpRNFLT, average cup depth, average nasal rim disc ratio, maximum cup depth, and superior-quadrant cpRNFLT. Conclusion The proposed machine learning system has proved to be good identifiers for different disc types with high accuracy. Additionally, the calculated confidence levels reported here should be very helpful for OAG care.

A clone of a DNA-mediated mobile element (transposon) corresponding to a mariner-like element (MLE) was obtained by carrying out the polymerase chain reaction with genomic DNA of Bombyx mori using a Hyalophora cecropia MLE sequence as a primer. This clone had a size of about 4.2 kb and, after sequencing, was found to contain an RNA-mediated, shorter retrotransposon named L1Bm, which was in turn integrated with a much longer retrotransposon named BMC1. Thus, the mobile elements made a novel tripartite structure. The BMC1 and L1Bm moieties of the composite structure each contained a 63-bp conserved sequence which was subsequently found to be highly conserved in all BMC1 and L1Bm elements registered so far. We propose that the 63-bp stretch may be a recognition site for a retrotransposition mechanism conducted by a reverse transcriptase and an endonuclease complex. On the basis of this inference, we propose a model that predicts how different types of BMC1 and L1Bm elements are dispersed in the genome. In addition, a phylogenetic tree made from the current and extant BMC1 and L1Bm sequences indicated that these elements can be classified into Subfamilies I and II.

The tetramethylarsonium ion has been found in the gill of the clam Meretrix lusoria. We attempted to confirm the conversion of arsenobetaine to the tetramethylarsonium ion in a culture with marine microorganisms occurring in the gill of the clam Meretrix lusoria. Arsenobetaine was aerobically incubated with microorganisms. A garlic-like odor was faintly detected after 5 to 8 days incubation. The mass spectrum of the odorous substance was essentially identical with that of synthetic trimethylarsine. Trimethylarsine oxide (TMAO) was also detected in the culture medium after 3 weeks incubation. Arsenobetaine was biotransformed to TMAO during the incubation in the culture growth by the marine microorganisms. Tetramethylarsonium ion was not detected in the culture medium even after 72 days incubation.

Biological defense factors show diurnal variations in their expression levels or activities. These variations can induce the different sensitivity to external toxicants of a day. We reported earlier that mice showed clear diurnal variation of cadmium (Cd)-induced toxicity, i.e., chronotoxicity. In this report, we investigated additional new evidences for the cadmium (Cd)-induced chronotoxicity, and considered the mechanisms contributed to this chronotoxicity. Male C57BL/6J mice were injected with CdCl₂ (6.4 mg/kg, one shot) intraperitoneally at 6 different time points of a day (zeitgeber time (ZT); ZT2, ZT6, ZT10, ZT14, ZT18 or ZT22) followed by monitoring the mortality until 14 days after the injection. We observed extreme difference in survival numbers: surprisingly, all mice died at ZT2 injection while all mice survived at ZT18 injection. Moreover, in non-lethal dose of Cd (4.5 mg/kg), the values of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) used as indexes of hepatotoxicity markedly increased at ZT6 injection while mostly unchanged at ZT18 injection. To consider the mechanisms of this extreme diurnal variation, we examined biochemical studies and concluded that the diurnal variation was not caused by the differences in hepatic Cd level, basal hepatic metallothionein (MT) level, and induction level or induction speed of hepatic MT. We suggested that one of the candidate determination factors was glutathione. We believe that the "chronotoxicology" for metal toxicity may be classic, yet new viewpoint in modern toxicology field.

Cadmium (Cd) is one of the endocrine disrupter and is a well-known testicular toxicant. Recently, we reported that Cd-induced mortality was markedly different by injection timing. In this report, we investigated whether severity of testicular toxicity was affected by injection timing of Cd. C57BL/6J mice (male, 7 w) were received single intraperitoneal injection of CdCl(2) (4.5 mg/kg) at zeitgeber time 6 (ZT6) or ZT18; these injection timings showed highest (ZT6) or lowest (ZT18) mortality in our previous study (Miura, 2012). After one week of the injection, several parameters for testicular toxicity such as epididymal sperm motility and numbers of sperm head both in cauda epididymidis and testis were measured. At ZT6 injection group, all parameters examined were significantly reduced compared to the control group. However, very interestingly, no significant changes were observed at ZT18 injection group. We obtained similar results by another experiment in which mice were received single subcutaneous injection of CdCl(2) (4 or 6 mg/kg) followed by measuring the parameters ten days after the injection. This diurnal variation was not contradictory to the result of the lethal toxicity which we showed earlier. Therefore, our results indicate that the testicular toxicity of Cd is also influenced by the injection timing.

The presence of long repetitive sequences is demonstrated in the genome of the silkworm, Bombyx mori. Members of this BMC1 family reveal several features typical of the L1 (long interspersed sequence one) family of mammals, except for species specific elements. The number of BMC1 elements is estimated to be approximately 3500 per haploid genome. Elements containing the full length unit of 5.1 kb are dispersed throughout the genome and their restriction sites are conserved, although most members are preferentially truncated to varying extents at their 5' ends. DNA sequencing indicates that this element contains six tandem repeats of 15 bp CpG-rich sequence in the 5' proximal region. It terminates with a 3' oligo(A) stretch, and is flanked at both ends by a 7-10 bp target sequence duplication. In addition, there is significant evidence for amino acid sequence homology with reverse transcriptase domains of other L1 families, especially F, Doc and Jockey of Drosophila melanogaster. No large open reading frame is present. The BMC1 element is suggested to be dispersed in the genome by a transposition mechanism involving RNA intermediates.

To the Editor,A 66-year-old woman was admitted to our department for the evaluation and treatment of palpable purpura and skin ulcers of the bilateral lower extremities (Figure 1A) and thrombocytop...

We examined the expression of apolipophorin-III (apoLp-III) during embryonic development of the silkworm Bombyx mori. ApoLp-III mRNA was first expressed 24 h after oviposition, which corresponds to the time of germ band formation. The amount of apoLp-III in the eggs increased from day 2, peaked on day 4, and then gradually decreased until hatching (on day 9.5). ApoLp-III was apparently synthesized during early embryogenesis, as radioactive amino acids were incorporated into newly synthesized apoLp-III in three-day-old eggs. Moreover, radioactive apoLp-III was found only in the embryo and not in the extraembryonic tissue. KBr density gradient ultracentrifugation of egg homogenates showed that apoLp-III was associated with low-density lipophorin (LDLp). These results suggest that LDLp is required for the delivery of lipids for organogenesis during embryogenesis.

Since high levels of prions, the causative agent of bovine spongiform encephalopathy (BSE), accumulate in the brain and spinal cord, contamination of beef carcasses with central nervous system tissue (CNST) may occur at post-slaughter process. In this study, we investigated CNST contamination on the surface of beef carcasses using glial fibrillary acidic protein as a marker after splitting and evaluated the effects of washing procedures on contamination removal. High levels of CNST contamination was detected immediately after splitting, especially in the area close to the spinal column. This suggests that spinal cord fragments are attached to carcasses at the time of splitting even though the spinal cords have been removed by vacuum before splitting. Steam cleaning or manually washing with normal pressure water around the spinal column, performed prior to washing with high-pressure water, was found to be effective for reducing the level of CNST contamination. Furthermore, manually washing with high-pressure water could reduce CNST contamination to almost negligible levels. These results are useful for preparation of appropriate sanitation standard operating procedures to reduce the risk of CNST contamination of carcasses for prevention of exposure to BSE prion via the food chain.

The objective of this study was to characterize the transfer of diacylglycerol (DAG) and cholesterol from larval Bombyx mori lipophorin to ovarioles. Transfer studies were carried out by incubating pupal ovarioles (5-day) with [3H]-cholesterol and [3H]-DAG-labeled lipophorin under different conditions. Transfer of both cholesterol and DAG exhibited hyperbolic dependency on lipophorin concentration with apparent Km values of 0.83 ± 0.17 mg/ml and 0.74 ± 0.16 mg/ml, respectively. Pretreatment of ovarioles with anti-lipid transfer particle (LTP) IgG significantly inhibited transfer of labeled DAG to ovarioles (75%) and not cholesterol. Injection of B. mori pupae (day 4) with anti-LTP IgG significantly affected the weight (65%), number of eggs (49%), amount of lipid (74%), and protein (65%) of the adult ovaries. Matured eggs had a very faint yellow color and deformed shape compared to controls. The inhibitory effect demonstrates the active role LTP plays in growth of ovaries, development, and oogenesis. The effect on vitellogenin shortage on egg development and maturation was determined by implanting ovaries in male recipients that lack vitellogenin. An 80% decline in egg production was observed. However, the mature eggs were normal in shape, color, and lipid content. Thus, restricting lipid or protein delivery to developing ovaries would dramatically affect choriogenesis.

We established basic conditions for transient gene expression and selection of antibiotics in the cultured cell line of silkworm, Bombyx mori, by use of the promoter of the heat shock protein (hsp70) gene of Drosophila melanogaster. The control promoter (hsp70) promoted the expression of chloramphenicol acetyltransferase (CAT) gene ligated at the downstream, dependent on the orientation of the promoter in the silkworm cell. The cell line is able to be supplied for the promoter assay of the silkworm genes. The concentration for the drug selection was determined as 0.75 mg/ml on neomycin analog, G418 (geneticin).

1. 1. The transcribed region of Bombyx mandarina rDNA was identical to that of B. mori rDNA when the restriction maps and partial nucleotide sequences were compared. The result supports the assumption that Bombyx mandarina is an ancestor of Bombyx mori and that the two were subdivided very recently. 2. 2. Non-transcribed spacer (NTS) of four clones derived from the two insects was slightly different from one another, which seemed to be due to the difference in the number of repeated sequences distributed in the spacer. 3. 3. The five clones from B. mandarina had the type I insertion sequence (IS) homologous to that in Bombyx mori 28S rDNA. There was micro-heterogeneity in the structure of IS.

Tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (TFPB) anion was highly lipophilic, practically insoluble in water, and durable against acid and oxidants. Partition equilibria of alkali TFPB between water and organic solvents and the stability in acid media are described and compared with the properties of the parent tetraphenylborate and of some fluorine-containing homologues.

Two types (R a1 and R a2) of nucleolus organizers were identified in the genome of Bombyx mandarina (Japan) which occurs in Japan. Genetical analysis of a hybrid with B. mori suggested that the loci of both nucleolus organizers are allelic and correspond to the R 0 locus of B. mori. These nucleolus organizers segregated and were inherited by the progeny in a Mendelian fashion. The majority of the R a1 rDNA units were 10.6 kb in length and had an additional EcoRI site in the transcribed spacer region when compared with the same size unit of R 0. On the other hand, the KpnI site present in the non-transcribed spacer region of the R 0 rDNA was not detected in the R a1 unit. The 15.1 kb unit observed in the R a2 locus was the same as the unit with the type II intron of R 0. The four major components of R a2 rDNA, with lengths of 10.6, 15.1, 15.7 and 20.8 kb, were also found in the R 0 locus, and thus the R a2 and the R 0 loci were considered to be closely related. Usually the functional unit of rDNA in the nucleolus organizers of homologous or non-homologous chromosomes cannot be easily distinguished. However, in the case of B. mandarina (Japan), distinct functional 10.6 kb units were observed in the allelic R a1 and R a2 loci. Therefore the existence of the two distinct units suggest the possibility of introducing the chromosomes of the interspecies in the genus Bombyx.

1. 1. We characterized several small RNAs in various tissues and strains of the silkworm, Bombyx mori by the in vitro capping and in vitro transcription. 2. 2. Three subspecies of 7SL RNA, which is believed to be involved in protein secretion, were detected in the commercial strains. Structural differences among the three molecules were due to the nucleotide substitution at two positions or more. 3. 3. A very AU-rich RNA, which is about 500 bases long, was always present in any tissue of Bombyx mori while 55 base RNA was only observed in a substantial amount in the fat body and culture cells. 4. 4. These small RNAs were also observed among the in vitro transcripts of total Bombyx DNA in the cell-free extracts from the silkgland and fat body.

A new photometric method was proposed to determine a minute concentration of tetraarylborate anions in aqueous solutions. Tetraphenylborate as low as 10-5 - 10-6 mol dm-3 could be determined by use of the spectral change in the visible absorption peak of rhodamine 6G at 527 nm. The absorption band of crystal violet at 590 nm could be also useful. The photometric sensitivities for the organic borates were as high as 5 - 7 × 104 absorbance unit per mol dm-3 of the borate at 10-5 mol dm-3 of the dye for 1 - 5 × 10-6 mol dm-3 range of the borates. Analytical interferences from ordinary electrolytes and organic solvents supposedly being present in sample solutions were examined to be assured.

The structure of eremophilenolide is confirmed to be (1) by X-ray analysis. The cis-fused decalin system takes all chair “steroid-like conformation.”

To investigate the behaviour of strontium and calcium at the gill absorption of fishes, gill-perfusing experiments were conducted using the fresh water or sea water adapted eel, Anguilla japonica and following results were obtained. 1) Discrimination against strontium in favour of calcium was observed at the gill absorption of these minerals. 2) The strontium-calcium discrimination factor (DFgill absorptive) expressing this discriminating rate was 0.74 in case of fresh water eel. In this case, the fresh water of external medium contained 100 mg. per 1000 ml. of medium. 3) In marine eels, on the other hand, this DFgill absorptive was found as 0.67 and this heigher rate of discrimination than that observed in fresh water eels seems to be caused by the pretty high concentration of calcium and strontium in sea water. In this case the sea water of the external medium contained 498 mg. Ca and 12 mg. Sr per 1000 ml. of medium. The authors wish to express their cordial thanks to Assoc. Prof. NAKAMURA and Mr. S. KASAHARA, Fisheries Experimental Station of Tokyo Univ. and members of Atsumi Fish-breeding Co. Ltd., for the facilities they gave to authors during the collection of marine eels. They are also indebted to Mr. Y. ENOMOTO, National Institute of Radiological Sciences, for his technical assistance.

Using 294 goldfish, serum calcium and magnesium in various degree of gonadal maturation and effect of estradiol on serum calcium and magnesium were investigated, and the following results were obtained.1. Conspicuous elevation of total calcium level was observed in the serum of goldfish injected with estradiol. The elevation of total magnesium level was detected also in the serum of these fish.2. Relationship was observed between the total calcium level in blood serum and ovarian maturation of goldfish. Matured female goldfish showed hypercalcemia. On the contrary, no distinctive relationship was observed between the calcium level in the serum of male goldfish and testicular maturation. Total calcium in the serum of male goldfish was 3.17±0.12 mEq/L on the average.3. Total serum magnesium of male and female goldfish was 1.63±0.12 and 1.50 ±0.07 mEq/L on the average, respectively.

Effects of estradiol, Solu-Cortef and parathyroid extract on the urinary and serum calcium levels of fish were investigated, using the snake-head fish, Channa argus and the following results were obtained. 1. Conspicuous elevation of total calcium level was observed in the serum of fish injected with estradiol. The elevation of total calcium level was observed also in the urine of these fish. 2. No remarkable changes were detected on the urinary and serum calcium level of fish after the injection of Solu-Cortef. 3. Slight, but statistically significant elevation of urinary and blood calcium levels was found in the fish injected with parathyroid extract.

The handy mill (Freezer-Mill) is useful for a DNA extraction from tissues and organs. In the present experiment, we prepared a high molecular weight DNA (100 to 140kb) from the posterior silk gland of Bombyx mori by use of the mill.