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We examined the diet of young Northern Goshawks Accipiter gentilis before and after fledging through the direct observation of prey items delivered by a single pair breeding in central Tokyo. Observations were conducted from 13 May to 14 August 2015. Three hundred prey items primarily belonging to five bird species and two genera of mammals were recorded both before and after fledging. The species contributing most to the total diet were White-cheeked Starling Spodiopsar cineraceus and Feral Pigeon Columba livia, with pigeons contributing most to total biomass followed by White-cheeked Starling. Prey item composition differed between the periods before and after fledging. After fledging, the contribution of White-cheeked Starling to the diet decreased, whereas that of pigeon increased.

Flight is a complicated task at the centimetre scale particularly due to unsteady air fluctuations which are ubiquitous in outdoor flight environments. Flying organisms deal with these difficulties using active and passive control mechanisms to steer their body motion. Body attitudes of flapping organisms are linked with their resultant flight trajectories and performance, yet little is understood about how isolated unsteady aerodynamic phenomena affect the interlaced dynamics of such systems. In this study, we examined freely flying bumblebees subject to a single isolated gust to emulate aerodynamic disturbances encountered in nature. Bumblebees are expert commanders of the aerial domain as they persistently forage within complex terrain elements. By tracking the three-dimensional dynamics of bees flying through gusts, we determined the sequences of motion that permit flight in three disturbance conditions: sideward, upward and downward gusts. Bees executed a series of passive impulsive maneuvers followed by active recovery maneuvers. Impulsive motion was unique in each gust direction, maintaining control by passive manipulation of the body. Bees pitched up and slowed down at the beginning of recovery in every disturbance, followed by corrective maneuvers which brought body attitudes back to their original state. Bees were displaced the most by the sideward gust, displaying large lateral translations and roll deviations. Upward gusts were easier for bees to fly through, causing only minor flight changes and minimal recovery times. Downward gusts severely impaired the control response of bees, inflicting strong adverse forces which sharply upset trajectories. Bees used a variety of control strategies when flying in each disturbance, offering new insights into insect-scale flapping flight and bio-inspired robotic systems. This article has an associated First Person interview with the first author of the paper., Summary: The effect of atmospheric gusts on the flight trajectories of bumblebees, reporting motion of flight influenced by gusts along with flapping-enabled control strategies that could be necessary elements of flight at this scale.

Dolphins are well known as excellent swimmers for being capable of efficient cruising and sharp acceleration. While studies of the thrust production and power consumption of dolphin swimming have been the main subject for decades, time-varying acceleration process during successive fluke beats still remains poorly understood. In this study, we quantified the time-varying kinematics of a dolphin (Lagenorhynchus obliquidens) by directly recording its burst-accelerating swimming before vertical jump in an aquarium with two synchronized high-speed video cameras. We tracked the three-dimensional trajectories of its beak, body sides, and fluke. We found that dolphin could quickly accelerate from 5.0 m s-1 to 8.7 m s-1 merely by 5 strokes (i.e. 2.5 fluke beats) in 0.7 seconds. During the strokes, it was further found that the dolphin demonstrated a great acceleration in downstroke but less acceleration or even a slight deceleration in upstroke. Hydrodynamic forces and thrust power for each stroke were further estimated based on the equation of body motion and a static hydrodynamic model. The drag coefficient of the dolphin was estimated through computational fluid dynamics (CFD) modeling of the steady flows around a realistic geometric model based on 3-D scan data. The thrust and thrust power were then calculated by combining the body kinematics and the drag coefficient, resulting in a maximum stroke-averaged thrust and power-to-mass ratio of 1.3 × 10^3 N and 90 W kg-1 at downstroke, and 3.3 × 10^2 N and 19 W kg-1 at upstroke, respectively. Our results point out the importance of asymmetric kinematics in burst acceleration of dolphin, which may be a useful mechanism for biomimetic design of high-performance underwater robots.

The inhibitory effects of L-type Ca2+ channel antagonists on Na cholate-induced in vitro excystment (CIIE) of Paragonimus ohirai metacercariae were studied. At concentrations of 10 microM, nicardipine and nimodipine inhibited CIIE completely and by approximately 92%, respectively. Nitrendipine and (+/-)-verapamil inhibited CIIE by about one half and one third, respectively. Nifedipine and diltiazem did not inhibit CIIE significantly. At higher concentrations, nitrendipine at 20 microM completely inhibited CIIE, and (+/-)-verapamil at 40 microM inhibited CIIE by 93%. Nifedipine and diltiazem inhibited CIIE only slightly and little, respectively, even at 40 microM. Complete inhibition by nicardipine at 10 microM required preincubation of metacercariae with the antagonist for 15 min. The inhibitory effects of nicardipine and nimodipine were reversible, and most of the nimodipine-treated metacercariae could excyst within 1 h after being washed, but the nicardipine-treated ones started to excyst 1 h after washing. Nicardipine suppressed the active movement of encysted juveniles evoked by Na cholate, whereas nimodipine did not suppress this significantly. These results suggested that L-type Ca2+ channels appeared to be involved in CIIE of P. ohirai metacercariae and that the inhibitory effect of the channels was due primarily to factors other than the inhibition of muscular activity, probably involving the secretion and release of enzymes lytic against the metacercarial cyst wall.

To examine cellular changes after diploid-tetraploid transformation in mammalian cells, tetraploid Meth-A cells, which were established from diploid cell line, were compared to the parent diploid cells for serum dependence of growth. Tetraploid Meth-A cells showed a high tolerance for low serum concentrations and could proliferate in a serum-free medium, differing from the parent diploid cells. It was suggested that diploid-tetraploid transition of Meth-A cells resulted in induction of a new cellular function, being that hyperploid cells can proliferate in sever circumstance for nutriments.

Allergens present in the excretory/secretory (ES) products of adult Paragonimus ohirai were biochemically identified. Immunoblot analysis using sera from P. ohirai-infected rats revealed only two allergens to be major proteins in the ES products, with apparent molecular masses (Mr) of 27 and 29 kD. As the ES products contained a high proportion of acidic and neutral cysteine proteinases, we examined whether or not the allergens and the cysteine proteinases were identical. The acidic and neutral cysteine proteinases were biochemically purified from the ES products and showed Mr of 27 and 29 kD, respectively. The two cysteine proteinases had almost identical N-terminal amino acid sequences and were reactive with specific IgE in sera from the infected rats. The allergenicity of the cysteine proteinases was confirmed by 48-hour homologous passive cutaneous anaphylaxis. Immunoblot and immunocapture assays using anti-human IgE monoclonal antibody showed that the proteinase allergens were reactive with specific IgE of patients with paragonimiasis westermani. Also, the cysteine proteinases were reactive with specific IgG of both the infected rats and the patients. Therefore the acidic and neutral cysteine proteinases prepared from the ES products of P. ohirai will be useful allergens and antigens for the immunodiagnosis of paragonimiasis.

Polyploid cells are made by DNA reduplication without cell division, however, it is not easy to establish polyploid mammalian cell lines. It is worth studying the difference in cell character between hyperploid and parent cell lines. Meth-A cells were polyploidized by demecolcine, K-252a, staurosporine and paclitaxel. The cell-cycle responses of highly polyploid Meth-A cells after the removal of the drugs were examined by flow cytometry (FCM). Meth-A cells were highly polyploidized by these drugs. The polyploid Meth-A cells gradually decreased in ploidy after the drug release. A tetraploid Meth-A cell line was established only from the demecolcine-induced polyploid Meth-A cells. The duration of G1, S and G2/M phases of the tetraploid cell line were mostly the same as those of the parent diploid cells, except that the G2/M phase was 1.5 h longer. The chromosome number of tetraploid Meth-A cell line was about twice of the diploidy. A tetraploid Meth-A cell line was established.

An enzyme-linked immunsorbent assay (ELISA) using worm extract antigens from lung flukes of Paragonimus westermani provided good sensitivity to sera from patients with paragonimiasis westermani but high cross-reactivity with sera from most fascioliasis patients and some patients with onchocerciasis or clonorchiasis. To improve the specificity, we tested an ELISA using fluke cysteine proteinases as antigens. Cysteine proteinases were partially purified from the excretory/secretory products of P. westermani by 40-75% ammonium sulfate fractionation, hydrophobic chromatography, and arginine affinity chromatography. An ELISA using the enzyme preparation not only had increased sensitivity to paragonimiasis westermani sera but also reduced cross-reactivity with the fascioliasis, onchocerciasis, and clonorchiasis sera to negligible levels. The reactivity of the ELISA to paragonimiasis miyazakii sera was similar to that of paragonimiasis westermani sera. A proteinase preparation from P. ohirai, which can be obtained easily from infected rats, provided similar results. Therefore, the ELISA using cysteine proteinases of Paragonimus could not distinguish the parasite species with which patients were infected, but it is a valuable assay with which to immunodiagnose paragonimiasis even when the proteinases are prepared from nonhuman species.

An IgM monoclonal antibody (mAb) that reacted with the gut epithelium and luminal contents of adult Paragonimas obirai was prepared from a mouse immunized with the excretory/secretory (ES) products of the parasite. The mAb reacted with the same sites not only of immature worms and newly excysted juveniles of P. ohirai , but also of adult worms of other Paragonimus species, such as P. westermani and P. miyazakii . The mAb did not react with adult antigens prepared from any other helminth parasites. Thus, the mAb is a genus-specific antibody against the gut-associated antigen. Immunoblotting revealed that two antigens with molecular masses (M r s) of 27 and 29 kDa were present in the ES products of adult P. ohirai , whereas four antigens with M r 27, 29, 36, and 38 kDa were detected in adult worm extract. Partially purified fractions containing acidic and neutral cysteine proteinases have 27 and 29 kDa protein molecules reactive with the mAb. The possibilities that the 36 and 38 kDa antigens may be precursors of these proteinases are discussed.

The tegumental glycocalyx of excysted juvenile (EJ) of Paragonimus ohirai was immunobiochemically characterized using a monoclonal antibody (MS-Mab). HPLC gel filtration showed that the antigens detected by two-site ELISA had a molecular weight of greater than or equal to 2 x 10(6) Da (dextran marker). On reduced SDS-PAGE, the glycocalyx antigen retained in the stacking gel was cleaved into several much smaller antigens after pronase treatment. The antigenic activity of the glycocalyx was stable in two-site ELISA to heat and acid treatments, but sensitive to alkali, periodate, base/borohydride, and pronase treatments. Precipitin formation in immunodouble diffusion between MS-Mab and EJ crude antigen was inhibited only by two monosaccharides: galactose and N-acetylgalactosamine. The purified glycocalyx bound strongly to PNA lectin, fairly well to RCA120 lectin, and slightly to SBA lectin, but not to Con A, WGA, UEA-1, DBA, or LFA lectins. Exo-beta-galactosidase treatment increased SBA binding, whereas it decreased PNA binding. PNA was observed to strongly bind to the body surface of living EJ. The antigenic activity of the glycocalyx was remarkably lost by incubation with exo-beta-galactosidase and O-glycanase. The glycocalyx was reactive with sera of P. ohirai-infected rats, and its reactivity was remarkably reduced by O-glycanase treatment. The ELISA level was higher in sera at an early stage of infection than in a late one. These studies show that the EJ tegumental glycocalyx is antigenic in infection, a marked, high molecular weight glycoprotein containing antigenic O-linked sugars, and that the sugar epitope is at the nonreducing terminal of the O-linked sugars and is composed of galactose and N-acetylgalactosamine.

We measured six apolipoproteins (apo AI, AII, B, CII, CIII, and E) by turbidimetric method using an automatic discrete biochemical analyzer and commercially available antisera. The turbidimetric method was compared with the single radial immunodiffusion method. Linearity for serum apolipoprotein assay by the automated turbidimetric method was better than by the single immuno-diffusion method. The linearity by the turbidimetric method was 2.5 G/L for AI, 1.0 G/L for AII, 4.5 G/L for B, 0.12 G/L for CII, 0.3 G/L for CIII, and 0.12 G/L for E. The presence of high concentrations of bilirubin (up to 0.15 G/L) and hemoglobin (up to 50 G/L) interfered with apolipoprotein measurement. Comparison of the immunoturbidimetric and the single radial immunodiffusion (SRID) methods showed excellent coefficients of correlation, r = 0.963, 0.896, 0.846, 0.936, 0.972, and 0.937 for apo AI, AII, B, CII, CIII, and E, respectively. Reference ranges for the six apolipoproteins were determined by using sera from 450 healthy subjects and were 1.4 +/- 0.3 G/L for AI and 0.3 +/- 0.01 for E. The observed levels of AII (P less than 0.001), B (P less than 0.01), and CIII (P less than 0.01) were significantly higher in males. The serum levels of apo B, CII, and E showed a gradual increase with age which was more prominent in females than in males. The levels of apo AI, AII decreased significantly over an 11 day period in 22 patients with myocardial infarction.

Non-specific suppression of the immune response was investigated in Brugia pahangi-infected Lewis rats. The proliferative response of peripheral blood lymphocytes or splenic non-adherent cells to mitogens was significantly reduced by B. pahangi infection. The degree of hyporesponsiveness of splenic non-adherent cells to mitogens was comparable between microfilaremic and non-microfilaremic animals. The suppressed proliferative response of splenic non-adherent cells was restored by blocking with anti-CD8 monoclonal antibody. After separation of T cells into CD4+ and CD8+ subpopulations, only CD8+ T cells from B. pahangi-infected rats suppressed the proliferative response of normal spleen cells to concanavalin A. CD8+ T cells from normal rats had no suppressive effect. On the other hand, the proliferative response of CD4+ T cells to concanavalin A was comparable between normal and infected rats. These results suggest that CD8+ T cells participate in the non-specific suppression of immune response in experimental filariasis.

The role of mast-cell-derived eosinophil chemotactic factor A (ECF-A) on eosinophil-rich periovular granuloma formation was examined by using mast-cell-deficient WBB6F1W/Wv mice infected with Schistosoma japonicum. The average size of granulomas formed around newly deposited eggs in the liver of W/Wv mice was significantly smaller than that observed in control +/+ mice. The number of adult worms recovered or specific IgE titers were comparable between W/Wv and +/+ mice. In contrast, immediate-type hypersensitivity response to specific antigen and dialyzable low-molecular weight ECF in the serum was detectable only in infected +/+ mice. When naive bone marrow eosinophils were incubated with the dialyzable fraction of infected +/+ mice sera, chemotactic reactivity of eosinophils to ECF derived from S. japonicum eggs was significantly enhanced, although that to synthetic ECF-A was depressed. Similar effects were observed when naive eosinophils were treated with synthetic ECF-A. The dialyzable fraction of infected W/Wv mice sera had no such modulating effect on the chemotactic reactivity of eosinophils. These results suggest that an immediate-type hypersensitivity reaction is important in the formation of eosinophilic granulomas around S. japonicum eggs, mainly through the modulating effect of mast-cell-derived ECF-A on the chemotactic reactivity of eosinophils.

The inhibitory effects of various ion channel blockers were examined on in vitro excystment of Paragonimus ohirai metacercariae induced by a bile salt, sodium cholate. At a concentration of 10 microM, bepridil, a non-selective Ca(2+) channel blocker, completely inhibited in vitro excystment, whereas TEA, lidocaine, and R(+)-IAA-94, channel blockers against K(+), Na(+) and Cl(-) ions, respectively, benzamil, an Na(+)/H(+) and Na(+)/Ca(2+) ion exchanger blocker, and R(+)-DIOA, a [K(+), Cl(-)] cotransporter inhibitor, did not. Considering the previous result that Ca(2+) ionophores are also efficient inducing factors for in vitro excystment of P. ohirai metacercariae and the present result, bile salts appear to induce the excystment of P. ohirai metacercariae through evoking the Ca(2+) channels of target cells within the metacercarial juveniles.

Ca2+ ionophores (A23187 and ionomycin) induced in vitro excystment of Paragonimus ohirai metacercariae, whereas Na+ ionophore (monensin) and K+ ionophores (nigericin and valinomycin) did not. The effect of A23187 on in vitro excystment was dependent on the concentration of the ionophore. A high concentration of 1 microM or more induced rapid excystment, but resulted in the death of metacercarial juveniles. The appropriate concentration of 0.2 microM induced in vitro excystment by 5 h after incubation with little or no damage to the juveniles.

We investigated the skeletal muscle voltage-gated chloride channel gene (CLCN1) in two unrelated Japanese patients with Becker's myotonia congenita. The non-myotonic parents of each patient were consanguineous. The proband of each family shares generalized myotonia, transient weakness after rest, and leg muscle hypertrophy. However, the disease severity related to the degree of myotonia differed, even in view of the response to long train nerve stimulation tests. CLCN1 gene analysis revealed a novel Ala659Val missense mutation identified to be homozygous in the more severe patient, while a novel Gln445Stop nonsense mutation was present in the other patient. Both mutations were absent in 90 Japanese normal controls. This is the first report of Japanese cases of Becker's myotonia congenita with CLCN1 gene mutations.

Cultured Meth-A cells always include a small fraction of large cells, which had a DNA content above 4c (polyploid cells). The process from the formation to the disintegration of polyploid Meth-A cells was measured by means of time-lapse videography. Polyploid Meth-A cells arose spontaneously from normal cells (polyploidization), then died by apoptosis. The fraction of polyploid cells gradually increased in seven day-exponential cultures with a low concentration of demecolcine, which is a specific inhibitor of cell division. The results revealed that the polyploid Meth-A cells are generated from normal cells by failing cell division and that they die by apoptosis.

We measured six apolipoproteins (AI, AII, B, CII, CIII, and E) in the serum of patients with several kinds of neural diseases [diabetic neuropathy and neural degenerative disorders (motor neuron degenerative disorders, spinocerebellar degeneration, Parkinson's disease)], comparing them to the age-matched healthy controls using the immunoturbidimetric method. Statistically significant decreases of serum apo-AI, apo-A-II and increases of apo-CIII, apo-E were observed in neural degenerative diseases; and, particularly, higher apo-B and apo-CII concentrations were observed in diabetic neuropathy. Most neural degenerative disease showed lower apo-AII. However, in motor neuron degenerative disorders, higher apo C-II and apo-E were seen. Lower apo-AI was seen in spinocerebellar degeneration, and lower apo-AII was seen in Parkinson's disease. Higher apo-B, CII, and E levels were observed in females with spinocerebellar degeneration and Parkinson's disease, and lower apo-AII was seen in males with spinocerebellar disease.

Monoclonal antibodies against infective third-stage larvae (L3) of Brugia pahangi were generated from mice immunized with L3 antigens. The monoclonal antibodies were L3 stage-specific or stage-nonspecific. A BpG1 monoclonal antibody (IgG1 subclass) showing L3 stage-specificity was examined in detail. BpG1 recognized the surface of B. pahangi L3 and also reacted with the surface of Brugia malayi L3 but not with the surface of filarial worms of other genera, such as Acanthocheilonema viteae and Litomosoides carinii. BpG1 promoted cellular adhesion to the surface of B. pahangi L3. BpG1 bound on living L3 was shed but the shedding rate was relatively slow. The surface antigen recognized by BpG1 had a molecular weight of 58 kDa. It was stable to heat and periodate treatments but sensitive to trypsin digestion and was released from living L3 by SDS but not by Triton X-100 or CTAB. Preincubation of L3 with BpG1 significantly reduced the recovery rate of worms compared with the preincubation with a monoclonal antibody (IgG1 subclass) against the inner tissues of B. pahangi L3 or control supernatant of P3U1 myeloma cells. This result suggests that the antigen containing the BpG1 epitope may be one of the targets of a protective immune response against Brugia infection.

This is the first full report on the genetic linkage between Japanese Huntington's disease and the DNA markers D4S10 and D4S43/S127. With use of the HindIII, BglI, and EcoRI polymorphisms detected at D4S10, and the combination of all these polymorphisms to give composite haplotypes, nine Japanese Huntington's disease families were found to be informative. Three recombinants for D4S10 were detected in these families, giving a maximum lod score of 1.662 at a theta of 0.10. Similarly, when we used the MspI and PvuII polymorphisms detected by D4S43/S127, five families gave informative results. No recombinant was detected in these families, giving a maximum lod score of 3.348 at a theta of 0.00. These results clearly support the view that the Japanese Huntington's disease gene may be identical with the Western gene, in spite of the lower prevalence rate in Japan.

The allergens of the lung fluke Paragonimus westermani were localized by indirect immunostaining in adult fluke sections using pleural exudates from 3 patients with P. westermani. Immunostaining performed by using pleural exudate with the highest level of specific IgE revealed that the P. westermani major allergen (or allergens) was located in the gut epithelium and luminal contents and that minor allergens were in the tegument and parenchyma. The antigens recognized by specific IgG were located at various sites including those recognized by specific IgE. Paragonimus westermani-specific IgE cross-reacted with only the gut of 2 other Paragonimus species, Paragonimus miyazakii and Paragonimus ohirai. The major allergen in the gut also was recognized by the other 2 pleural exudates. These results indicate that the substance present in and secreted from the gut is not only a major allergen but is also a common allergen among Paragonimus species.

Allergen in crude extract of Dirofilaria immitis was purified and separated from IgG-inducing antigens by a combination of DEAE-Sephadex A-50 chromatography, Sephadex G-200 gel filtration and starch gel zone electrophoresis. The purified preparation was proved to be one protein band by sodium dodecyl sulfate polyacrylamide gel (SDS-gel) electrophoresis and one precipitin arc by immunodiffusion. The molecular weight of the purified allergen was estimated to be approximately 20,000 by gel filtration and 15,000 by SDS-gel electrophoresis. The carbohydrate content of the preparation was apparently low, about 2%. The allergen was positively charged, and its determinant group was protein in nature. It was resistant to tryptic, pepsic and chymotryptic digestion, periodate oxidation and DNase and RNase digestion but very sensitive to pronase digestion. Allergen was inclined to aggregate each other in the buffered solution. It was also very resistant to vibration, heat (80°C for 1 h) and acid (pH 2.5) and alkali (pH 11.0) treatments. Rats as well as mice immunized with allergen developed only a reaginic antibody and no hemagglutination antibody.

世界各地の，いわゆる発展途上国に在留する邦人の数は著しく増加している。それと同時に，各地でA型肝炎， 腸チフスやアメーパ赤痢まど重篤な経口感染症に罹患する邦人の数も増加している。我々は，その主な原因が飲料水の汚染にあると考え，世界各地の発展途上国の飲料水について，特に糞便汚染との関連について調査してきた。今回，飲料水の汚染と在留邦人の腸管内寄生虫感染率との関連について調べたので報告する。我々は，1979年4月から1980年12月までの聞に東南アジア，南西アジア，中近東，東アフリカおよび中南米の種々の国々を訪問し，飲料水を採取し，細菌学的および化学的水質検査を行なった。総数383検水について，まず過マンガン酸カリウム消費量が測定され，ついで一般細菌数および大腸菌群の数が調べられた。その結果，多かれ少なかれ，熱帯発展途上国の飲料水が糞便系汚染にさらされていることが判明した。そこで，水質検査と同時にこれらの飲料水を利用している邦人について，彼らの腸管内寄生虫感染率を調査した。各地に6カ月以上滞在している邦人の成人から小指頭大の糞便を採り，10%ホルマリン加パイアル瓶に入れて日本に持ち帰り， MGL集卵法にて糞便内寄生虫卵を検出した。また邦人の子供達を対象に，セロファン2回法による蟻虫検査を施行した。糞便からは主として蛔虫卵が見出され，その他鞭虫，無鉤条虫，肝吸虫などの卵が検出された。腸管内寄生虫感染率はインドネシア在留邦人に最も高く，次いで南西アジア，東アフリカ，中南米の順にまり，中近東のカタールでは全員陰性とまった。在留邦人のうちで，腸管寄生虫に感染している割合は，同地域の飲料水のうちで大腸菌群が検出された検水の割合と高い正の相聞を示した。すまわち，大腸菌群の検出頻度の高い飲料水を使用している在留邦人ほど，腸管寄生虫に多く感染しており，両者の相関係数はr=0.957であった。しかし，糞便内寄生虫卵陽性率と飲料水の過マンガン酸カリウム消費量との間には相関関係は見出せまかった。また，在留邦人の子供を対象に施行した蟯虫卵の陽性率は飲料水の汚染とは全く無関係であった。, Many Japanese inhabit tropical countries, and some of them are suffering from infectious diseases. In order to know the relationship between the prevalence of these diseases and the degree of faecal pollution of the drinking water, we have examined the drinking water in these countries of the world, and at the same time we have studied the infection rate of intestinal helminths of the Japanese who stayed 6 months or more in these countries. In total over 300 samples of drinking water from countries of South East and South West Asia, Middle East, East Africa and Central and South America were tested with potassium permanganate consumed test, and number of coliform bacilli from the drinking water was counted after cultivation. Relatively high proportion of samples appeared to be polluted by faecal matters. Then, we checked parasite eggs in the stool samples of Japanese who used the water samples in the tropical countries actually. Eggs of Ascaris lumbricoides, Trichuris trichiura, Taenia saginata and Clonorchis sinensis were found among the Japanese. Infection rate of intestinal helminths was proved to be highest among Japanese in Indonesia, and the rate was followed by those of people in South West Asia, in East Africa and in Central and South America. The relationship between the infection rate of intestinal helminths and the degree of pollution of the drinking water was studied. Significant relationship was observed between the infection rate of intestinal parasites of the Japanese inhabitants and the rate of water containing 102 or more/ml of coliform and of total micro-organisms (r=0.957). However, no relationship between the infection rate of Enterobius vermicularis among the Japanese children there and the degree of the pollution of drinking water was found., 日本熱帯医学会雑誌, vol.10(1), pp.41-48; 1982

Rats reinfected with Paragonimus ohirai (P.o.) elicited little secondary IgE response to adult P.o. antigen. In contrast, rats immunized with X-irradiated (2–10 krad) metacercariae elicited not only a marked primary IgE response comparable to that in normally infected rats, but also a marked secondary IgE response after challenge infection. Rats immunized with 2 krad X-irradiated metacercariae yielded a higher secondary IgE response than with 5 or 10 krad irradiated ones, and elicited a secondary response lasting at least 1 year. An IgE response to antigen of newly excysted juvenile parasites (NEJ) was clearly different from the IgE response to adult P.o. antigen. The former IgE response was weak after infection or immunization, but after challenge infection a marked secondary response occurred at a level similar to the IgE response to adult P.o. antigen. The secondary IgE responses to adult P.o. and NEJ antigens were also elicited by reinoculation with X-irradiated metacercariae. Cross-absorption experiments confirmed that there were adult type and NEJ type allergens.

This study was undertaken to prepare monoclonal antibodies which recognized the tegumental antigens of newly excysted metacercaria (NEM) and adult flukes of Paragonimus ohirai. From splenic cells of mice immunized with either the Triton X-100 extract of adult fluke surface or NEM sonicate, monoclonal antibodies were produced by ELISA screening followed by trial immunostaining of fluke sections. We prepared a monoclonal antibody (AS-Mab) recognizing the tegumental antigen of the adult fluke and a monoclonal antibody (MS-Mab) for the NEM tegumental antigen. AS-Mab also bound to the tegument of 1-week-old juveniles but not to that of NEM. MS-Mab bound the tegument from the metacercaria to adult stages, but the degree of immunostaining appeared to decrease with maturation of the flukes. By the double immunodiffusion technique, the two monoclonal antibodies formed crossing precipitin lines against a mixture of the adult surface antigen extract and the NEM antigen extract. MS-Mab and AS-Mab gave precipitin line against the incubation fluid from NEM and adult flukes, respectively.

Ikeda T. , Oikawa Y. and Fujita K. 1982. Kinetics and localization of parasite-specific IgE in Paragonimus ohirai -infected rats. International Journal for Parasitology 12: 395–398. In Wistar rats infected with Paragonimus ohirai (P.O.), P.O. -specific IgE responses of the mesenteric and mediastinal lymph nodes and spleen were determined by homologous adoptive cutaneous anaphylaxis (ACA) assay since P.o. -specific IgGa was not detected by either 2 h or 4 h PCA. Intraperitoneal (i.p.) infection with metacereariae elicited similar patterns of ACA response in the three lymphoid tissues examined, with the mediastinal lymph node giving the highest response. ACA positive cells were detected 2 weeks after infection, peaked at 3 weeks and then declined. These kinetics of ACA responses nearly paralleled the kinetics of serum P.o. -specific IgE titre. In intrapleural infection with metacereariae, on the other hand, the mediastinal lymph node gave a high ACA response comparable to the lymph node in i.p. infection, but the mesenteric lymph node and spleen gave negligible ACA responses. In infection established by i.p. transplantation of 4–5-week-old worms, only the mediastinal lymph node of the three lymphoid tissues responded and its response was at a low ACA level. The level of serum P.O. -specific IgE was much lower in the above two infections than in i.p. infection with metacercariae.

In Guatemala, an indirect hemagglutination (IHA) test for onchocerciasis, performed with a crude saline extract of Onchocerca volvulus and blood samples taken on filter paper, showed a high level of sensitivity and specificity. IHA titers of blood samples from the ear lobe taken on filter paper and of sera obtained by venipuncture showed a high correlation.

During the development of a cell aggregate of Dictystelium discoideum into a fruiting body, an antigenic acid mucopolysaccharide is synthesized only in the prespore cells of a cell mass. In this study, the subcellular distributions of UCPgalactose: polysaccharide transferase and UDPglucose pyrophosphorylase involved in biosynthesis of the mucopolysaccharide were determined. The transferase was specifically localized in the smaller vesicles with lighter density than the prespore-specific vacuoles identifiable electronmicroscopically. In contrast to the enzyme, the antigenic mucopolysaccharide was exclusively localized in the prespore-specific vacuoles. Unlike the transferase, UDPglucose pyrophosphorylase was confined to the soluble fraction. The sucrose gradient profiles of the transferase activity in the 5000 × g supernatant gave two main peaks. When the profiles were compared among standing and migrating slugs and culminating cell mass, the difference in the profiles closely reflected the state of biosynthesis of the acid mucopolysaccharide in eac developmental stage.

An epidemiological survey for onchocerciasis was carried out in the San Vicente Pacaya area of Guatemala. A total of 2,153 inhabitants were examined by a single skin snip, and 664 (30.8%) were positive for microfilariae. Slit lamp examination of the anterior chamber of the eye revealed microfilariae in 6.2%; the positive rate for microfilariae in the anterior chamber was proportional to the microfilarial density in the skin. The altitude-dependent character of onchocercal infection was shown in this mountainous area, suggesting that transmission occurs principally between 600 and 1,300 m. Of 1,217 persons examined simultaneously by skin snip and by palpation for nodules, 587 were positive by one or both methods; 101 infections were detected by onchocercal nodules only and these were usually in children or persons living in areas of low endemicity. It was concluded that both the skin snip and a search for nodules are required for accurately determining the prevalence of onchocerciasis in Guatemala., American Journal of Tropical Medicine and Hygiene, 28(1), pp.67-71; 1979

A method was described for isolation of the prespore specific vacuole (PSV) from slugs of the cellular slime mold, D. discoideum. A cellular component, which was fractionated in accordance with immunohistochemical staining using heteroplastic antispore serum, was found to consist of only the PSV. It was thus concluded that the PSV is identical with the cytoplasmic granule which has been shown by the antiserum to be specifically present in the prespore cell, and hence that the PSV is the only structure which contains the prespore specific substance (antigenic mucopolysaccharide). The isolated PSV contained polysaccharide equivalent to 14% of its protein content, and antigenic mucopolysaccharide constitutes about 60% of the total polysaccharide.

In seven patients with paragonimiasis westermani, parasite-specific IgE and IgG levels in sera and pleural effusion were determined by an enzyme-linked immunosorbent assay (ELISA). The sensitivity to adult excretory-secretory (E-S) antigen was compared with the sensitivity to whole worm extract antigen, and the former was more sensitive in both an IgE-ELISA and IgG-ELISA. Both parasite-specific IgE and IgG could be detected by ELISA at levels much higher than those in control subjects using E-S antigen. When specific IgE and IgG levels in sera and pleural effusion of individual patients were compared, the latter had higher values. The difference between levels of specific IgE in pleural effusion and serum did not correlate with that of specific IgG. These results indicate that specific IgE and IgG antibodies form locally, i.e., in the lung, and that pleural effusions from patients with paragonimiasis are more suitable than serum for immunodiagnosis.

The amino acid analysis of the purified allergen from Dirofilaria immitis revealed that it had a total number of 98 amino acid residues. The allergen was remarkably rich on glutamic acid (15 residues), lysine (14 residues) and aspartic acid (12 residues). No half-cystine was obtained. The ratio of the acidic residues to the basic ones was 1.4. The weak positive charge of the allergen is contributed by the fewer number of basic residues relative to acidic residues.

Parasite-specific IgE response in rats infected with metacercariae of the lung fluke, Paragonimus ohirai, was enhanced about 8 times by Bordetella pertussis adjuvant. In rats infected by intraperitoneal transplantation of adult worms, the adjuvant markedly increased the usually low or absent parasite-specific IgE response. The most effective time of the adjuvant administration was different between the two modes of infections.

Once organisms entered the alveolar macrophages the growth of these organisms was similar, independent of the animal species. To our knowledge it has not been determined whether adult rats are more susceptible to Toxoplasma after intranasal and pulmonary inoculation than after intraperitoneal inoculation. Differences in alveolar and peritoneal macrophages have been observed also in mice; alveolar macrophages may exert a stronger antibacterial action than the peritoneal exudate macrophages (Walker, 1976. In Immunobiology of the macrophages, D. S. Nelson (ed.). Academic Press, pp. 91-111). It would be of interest to determine whether the rat peritoneal macrophages could protect rat alveolar macrophages. It is not clear whether the low multiplication of Toxoplasma in peritoneal cells of rats is associated with killing or whether this is merely an inhibition of rate or multiplication. Ultrastructural studies may clarify this problem. This work was supported in part by the Vicerrectoria de Investigaci6n, Universidad de Costa Rica, and the Consejo Nacional de Investigaciones Cientificas y Tecnolo6gicas (CONICIT). We thank Aleida Quir6s and Zaida Umafia for their excellent secretarial assistance and Fabio Camacho for his concerned care of the laboratory animals.

A sero-epidemiological study of onchocerciasis was carried out with the IHA test in Guate- mala. In the endemic area, 94.8% of 191 subjects with microfilariae, 82.1% of 172 cases with onchocercal nodules, and 22.5% of236 cases without either microfilariae or nodules gave positive reactions in the IHA test. On the other hand, the positive rate was 3.4% and 2.0% in each of 2 nonendemic populations. A follow-up study of reactors in the IHA with neither microfilariae nor nodules showed that 11 (42%) of 26 cases were microfilaria positive 6-7 mo after the first examination. A clear correlation was found between the results of the IHA test and skin biopsy in surveyed communities. The age distribution curve of the IHA test was closely associated with skin biopsy although the former was higher. IHA titers rose in proportion to microfilarial density. A sex-related difference was evident in the IHA positive rate in the subjects from medium and low endemic areas, but little difference was shown in individuals from a higher endemic area. Although onchocerciasis can be diagnosed by detecting microfilariae in the skin, skin biopsy is not sensitive, especially in light or early stages of infection. Therefore, a stan- dardized immunological test may offer a pref- erable method of diagnosis and for epidemi- ological studies. The immunodiagnosis of onchocerciasis has been attempted by various workers using the skin test and complement fixation test (as reviewed by Kagan, 1963), im- munofluorescent antibody test (Lucasse, 1962; Lucasse and Hoeppli, 1963; Woodruff and

In rats infected with Paragonimus ohirai (P.o.), IgE response against the parasite was influenced by both the site of metacercarial inoculation and the age of the inoculated worms, whereas the IHA response was not influenced by these variables. Intraperitoneal infection and oral infection with 10 metacercariae induced a high level of P.o.-specific IgE (maximum PCA titer of 1:128 and 1:111, respectively). In contrast, intrapleural infection induced little IgE (1:496, maximum titer). Thus migration of the parasite in the peritoneal cavity and the liver, or the former alone, was an important event in the production of IgE, because intrapleurally inoculated metacercariae normally develop into mature worms without migrating into the abdominal cavity. Intraperitoneal transplantation of five immature or adult worms produced significantly lower IgE levels and correlated with the age of the transplanted worms. Four-week-old worms were observed to induce negligible or no P.o.-specific, IgE antibody. The stage-specific, IgE resonse observed in infections established by transplantation appears to be partly dependent on migration of the parasite in the abdominal cavity. Worms in the pleural cavity may have suppressed IgE production.