Chromosome aberrations induced by monomeric acrylamide in bone marrow and germ cells of mice

Acrylamide induces chromatid exchanges and breaks with considerable frequency in spermatogonia of mice with long-term administration (3 weeks), though not, remarkably, with short-term administration (1–2 weeks). At 12 and 24 h after single injections with 50, 100 and 150 mg/kg acrylamide, evaluation of the cytogenetic effect is difficult in the spermatogonia because of an extreme reduction of mitotic cells. Aneuploid and polyploid cells increase with ti,e after treatment in both marrow and spermatogonial cells, while the aberration frequency shows no increase in marrow after both oral-administration and injection. Evidently the spermatogonia are thus rather more sensitive to acrylamide than marrow cells. On the other hand, the SCE frequency is at the control level in treated subjects in marrow and spermatogonia. Acrylamide induces chain quadrivalents, ring quadrivalents, fragments and univalents which are particularly evident in primary spermatocytes in both oral administration and injection, though it is questionable whether these structural changes deal with spermatogonia, or otherwise with the S-phase primary spermatocytes. There is a possibility that the aberrant cells thus produced can develop into spermatozoa carrying a certain type of reciprocal translocation which leads to semi-sterile progeny. In relation to the above problem detailed investigations into this type of rearrangement in primary spermatocytes are needed.     

Identification of a novel member of yeast mitochondrial Hsp70-associated motor and chaperone proteins that facilitates protein translocation across the inner membrane

Here, we report the identification of yeast 15-kD Tim15/Zim17, a new member of mitochondrial Hsp70 (mtHsp70)-associated motor and chaperone (MMC) proteins. The 15-kD MMC protein is a peripheral inner membrane protein with a zinc-finger motif. Depletion of the 15-kD protein led to impaired import of presequence-containing proteins into the matrix in vivo and in vitro. Overexpression of the 15-kD protein rescued the functional defects of mtHsp70 in ssc1-3 cells, and a fusion protein containing the 15-kD protein physically interacts with purified mtHsp70. Tim15/Zim17 therefore cooperates with mtHsp70 to facilitate import of presequence-containing proteins into the matrix.     

One host—one parasitoid system: Seasonal life cycles ofPryeria sinica (Ledipoptera) andAgrothereutes Minousubae (Hymenoptera)

The zygaenid Pryeria sinicaMoore and the ichneumonid Agrothereutes minousubaeNakanishi form a one host—one parasitoid system in nature. Their seasonal life cycles were investigated by laboratory experiments and field observations, and the life-cycle adaptation of the parasitoid to its host was examined. The moth is univoltine. The larva hatches from mid-February to mid-March and feeds on leaf buds and young leaves of ever green Euonymus japonicusThunb . The thermal constants for completing the 1st, 2nd, 3rd, 4th larval instars and prepupal stage were 85.6, 80.5, 85.2, 177,0 and 197.6 degree-days, respectively. The prepupa and pupa vulnerable to the attack by the parasitoid occurred from mid-April to early May and from mid- to late May, respectively. Diapause in the parasitoid is facultative and occurs in the eonymphal stage. The photoperiodic response for this diapause was a long-day type with a critical photoperiod of 13 hr 40 min at 20°C, but it was not expressed at 25°C, most larvae entering diapause irrespective of photoperiod. About 19, 120, 82 and 112 degree-days above 7°C were required to complete the egg, larval, prepupal and pupal development, respectively. These data were superimposed on the photothermograph of Fukuoka, and it is predicted that the 1st adult eclosion would occur in late April and the partial 2nd adult eclosion in early June. The prediction was supported by field observations. The adult eclosion of the parasitoid synchronized well with the apperance of prepupae and pupae of the moth. The parasitoid has two types of seasonal life cycle, one generation and two generations a year. Both types have an extremely long dormant period of 10–11 months due to aestivo-hibernation. This seasonal life cycle enables the parasitoid to maintain its population when the host is in short supply.     

The diversity of receptor recognition in cholesterol‐dependent cytolysins

Cholesterol‐dependent cytolysins (CDCs) are bacterial pore‐forming toxins secreted mainly by pathogenic Gram‐positive bacteria. CDCs generally recognize and bind to membrane cholesterol to create pores and lyse target cells. However, in contrast to typical CDCs such as streptolysin O, several atypical CDCs have been reported. The first of these was intermedilysin, which is secreted by Streptococcus intermedius and has human cell‐specificity, human CD59 (huCD59) being its receptor. In the study reported here, the diversity of receptor recognition among CDCs was investigated and multi‐receptor recognition characteristics were identified within this toxin family. Streptococcus mitis‐derived human platelet aggregation factor (Sm‐hPAF) secreted by S. mitis strain Nm‐65 isolated from a patient with Kawasaki disease was previously shown to hemolyze erythrocytes in a species‐dependent manner, its maximum activity being in human cells. In the present study, it was found that Sm‐hPAF recognizes both membrane cholesterol and huCD59 as receptors for triggering pore‐formation. Moreover, vaginolysin (VLY) of Gardnerella vaginalis showed similar characteristics to Sm‐hPAF regarding receptor recognition. On the basis of the results presented here, the mode of receptor recognition of CDCs can be categorized into the following three groups: (i) Group I, comprising typical CDCs with high affinity to cholesterol and no or very little affinity to huCD59; (ii) Group II, including atypical CDCs such as ILY, with no or very little affinity to cholesterol and high affinity to huCD59; and (iii) Group III, which contains atypical CDCs such as Sm‐hPAF and VLY with affinity to both cholesterol and huCD59.     

Screening,Purification and Properties of a Specific Antiphage Substance,AFS, against Filamentous Phage of Escherichia coli

We have made screening for antiphage antibiotics using an assay system of a male-specific filamentous phage, fl, and Escherichia coli as its host, and found a proteinous active substance named AFS (anti-filamentous phage substance) which was produced by an Actinomyces belonging to the group of Streptomyces lavendulae. It was purified from the filtered broth of the strain by ammonium sulfate precipitation and chromatographies of SP-Sephadex, CM-Sephadex and Sephadex G–50. The purified preparation was judged to be homogeneous by several column chromatographies and electrophoresis. Its molcular weight was about 5100 and isoelectric point was pH 9.9. Fourty two residues of amino acids, rich in the hydrophobic ones, were contained in one mole of AFS, but no carbohydrate was detected.     

The effect of 17-N substituents on the activity of the opioid κ receptor in nalfurafine derivatives

We have previously reported the essential structure of the opioid κ receptor agonist nalfurafine hydrochloride (TRK-820) for binding to the κ receptor. In the course of this study, we focused on the effect of the substituent at 17-N in nalfurafine on the binding affinity for the κ receptor. The exchange of the 17-N substituent in nalfurafine from cyclopropylmethyl to fluoro-substituted alkyl groups, which are strong electron withdrawing substituents, almost completely diminished the binding affinities for the μ and δ opioid receptors, but the binding affinity for the κ receptor was still maintained. As a result, nalfurafine derivatives with 17-fluoro-substituted alkyl groups showed higher selectivities for the κ receptor than did nalfurafine itself. With regard to the κ agonistic activities, the conversion of the 17-N substituent in nalfurafine from cyclopropylmethyl to fluoro-substituted alkyl groups led to the gradual decrease of the agonistic activities in the order corresponding to their binding affinities for the κ receptor. In contrast, the derivative with the bulky 17-isobutyl group showed lower affinity and agonistic activity for the κ receptor than the derivatives with the smaller functional groups. This research suggested that both the electronic property and the steric characteristics of the 17-N substituent would have a great influence on the binding property for the κ receptor.     

Relationship between the lifestyle of a Siboglinid (Pogonophoran) polychaete, Oligobrachia mashikoi, and the total sulfide and nitrogen levels in its habitat

A gutless polychaete of the family Siboglinidae, Oligobrachia mashikoi, known in the past as a beard worm of the group Pogonophora, inhabits Tsukumo Bay of the Noto Peninsula in the Sea of Japan. Photographs were taken of this polychaete projecting about one third of the length of its tentacles outside of its tube. The tube protruded several mm from the sea bottom. These are the first field photographs of beard worms. The trophosome of this beard worm harbors sulfur-oxidizing bacteria. In fact, the muddy sediment where this worm inhabits smells slightly of hydrogen sulfide. Total sulfide levels, which can be an indicator of the generation of hydrogen sulfide gas, were measured at 10 locations in the bay. Furthermore, at the location which this species inhabits, the total sulfide levels in the vertical direction were determined. In addition, the total nitrogen levels, which can indicate the quantity of organic substances, were measured. The sediment inhabited by this worm was determined to have total sulfide levels of 0.24-0.39 mg/g dry mud, measured in the form of acid-volatile sulfide-sulfur. The total nitrogen levels were 1.0-1.5 microg/mg dry mud. These values suggest that the bottom of Tsukumo Bay has not been deteriorated by eutrophication. The levels were, however, highest in the surface layer of the sediment. These results suggest that hydrogen sulfide is generated in the surface of the sediment by sulfate-reducing bacteria, and that O. mashikoi appears to able to live in an environment that contains a slight amount of sulfide.     

Reevaluation of erythropoietin production by the nephron

Erythropoietin production has been reported to occur in the peritubular interstitial fibroblasts in the kidney. Since the erythropoietin production in the nephron is controversial, we reevaluated the erythropoietin production in the kidney. We examined mRNA expressions of erythropoietin and HIF PHD2 using high-sensitive in situ hybridization system (ISH) and protein expression of HIF PHD2 using immunohistochemistry in the kidney. We further investigated the mechanism of erythropoietin production by hypoxia in vitro using human liver hepatocell (HepG2) and rat intercalated cell line (IN-IC cells). ISH in mice showed mRNA expression of erythropoietin in proximal convoluted tubules (PCTs), distal convoluted tubules (DCTs) and cortical collecting ducts (CCDs) but not in the peritubular cells under normal conditions. Hypoxia induced mRNA expression of erythropoietin largely in peritubular cells and slightly in PCTs, DCTs, and CCDs. Double staining with AQP3 or AE1 indicated that erythropoietin mRNA expresses mainly in β-intercalated or non α/non β-intercalated cells of the collecting ducts. Immunohistochemistry in rat showed the expression of HIF PHD2 in the collecting ducts and peritubular cells and its increase by anemia in peritubular cells. In IN-IC cells, hypoxia increased mRNA expression of erythropoietin, erythropoietin concentration in the medium and protein expression of HIF PHD2. These data suggest that erythropoietin is produced by the cortical nephrons mainly in the intercalated cells, but not in the peritubular cells, in normal hematopoietic condition and by mainly peritubular cells in hypoxia, suggesting the different regulation mechanism between the nephrons and peritubular cells.