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Human cytomegalovirus (HCMV) infection in immunocompromised patients causes considerable morbidity and mortality. Although ganciclovir prophylaxis reduces the incidence of HCMV disease, severe side effects raise serious problems. Thus, the development of new strategies for prophylaxis are clearly needed, and human monoclonal antibodies offer a potential alternative. We describe the cloning, using the phage display system, of a recombinant human Fab fragment against HCMV. A phage display library with 4×106 clones was panned three times against lysates of HCMV-infected cells, and screened by ELISA. Of six antigen-binding clones, one monoclonal antibody reacted strongly to HCMV. In immunostaining analysis, this Fab was able to stain HCMV-infected cells from 24 h post-infection (pi) through to 96 h pi, but not at 6 h pi. In the presence of cytosine arabinoside, HCMV-infected cells were not stained, even at 24 h pi. These results indicate that an HCMV protein that was recognized by the Fab was synthesized in the late phase of infection. In addition, this Fab exhibited neutralizing activity; at 1 μg/ml it reduced HCMV plaque formation by 50%. The Fab was able to neutralize three HCMV strains, but it did not neutralize HSV-1 or -2 infection.

A monoclonal antibody (MAb), MAb ED17 (immunoglobulin G2a [IgG2a]), prepared against trophozoites of Entamoeba dispar SAW1734RclAR cultured monoxenically with Crithidia fasciculata, reacted with 25 of 26 isolates of E. dispar by an indirect fluorescent-antibody test. In contrast, the MAb failed to react with any of 20 isolates of E. histolytica or other enteric protozoan parasites. Western blot (immunoblot) analysis showed that the molecular mass of the E. dispar antigen recognized by the MAb was 160 kDa under reduced conditions. Immunoelectron microscopy revealed that the antigen was mainly located on digested C. fasciculata, but not on undigested organisms. Double staining with a mixture of MAb ED17 and MAb 4G6 (an IgG1 MAb which reacts exclusively with E. histolytica), followed by incubation with a mixture of fluorescein isothiocyanate-labeled anti-mouse IgG2a and tetramethylrhodamine isothiocyanate-labeled anti-mouse IgG1 antibodies, simultaneously identified mixed populations of E. dispar and E. histolytica. This method may prove to be useful for the accurate identification of E. dispar and E. histolytica, even in mixed infections.

Murine monoclonal antibodies (mAbs) were produced against an n-octyl-beta-D-glucopyranoside-extracted fraction of trophozoites of Entamoeba histolytica HM-1:IMSS. Four of the mAbs were reactive with a 150-kDa surface antigen characterized by Western-immunoblot analysis under nonreducing conditions. When the reactivity of the four mAbs with nine reference strains of E. histolytica was examined by an indirect fluorescence antibody test, two of the mAbs (EH3015 and EH3023) were found to react with all nine strains and the other two mAbs (EH3056 and EH3126) reacted with seven strains. The four mAbs did not react with any E. dispar reference strain or with other enteric protozoan parasites. The reactivity of EH3015 and EH3023 with numerous isolates of E. histolytica and E. dispar collected in our laboratories was also examined. The 2 mAbs reacted with all of the 37 E. histolytica isolates tested but did not react with any of the 33 isolates of E. dispar. These results indicate that common antigenic epitopes of E. histolytica are on the 150-kDa surface molecule and that mAbs can distinguish between E. histolytica and E. dispar.

The first successful synthesis of photoresponsive multilayer spiral nanotubes by the introduction of polyfluorinate cationic azobenzene derivative, trans-[2-(2,2,3,3,4,4,4-heptafluorobutylamino)ethyl]-{2-[4-(4-hexyphenylazo)-phenoxy]ethyl}dimethylammonium (abbreviated as C3F7-Azo+), into layered niobate interlayer I by a two-step guest-guest exchange method using the intercalation compound, methyl viologen (MV2+)-K4Nb6O17, as precursor is reported.

Intercalation of tris(2,2′-bipyridine)ruthenium(II), Ru(bpy)32+, into a layered perovskite derived from Aurivillius phase Bi2SrTa2O9 was successfully achieved by a guest ion-exchange method using n...

An intercalation compound of the layered perovskite, K 2 La 2 Ti 3 -O 1 0 , with 5,10,15,20-tetrakis(N-methylpyridinium-4-yl)-21H,23H-porphin (TMPyP 4 + ), as the first example of a large molecule intercalation into this perovskite, was successfully prepared by a guest-exchange method using the n-PrNH 3 + -H x La 2 Ti 3 O 1 0 intercalation compound as an intermediate. Its structural characterization was carried out by means of X-ray diffraction, UV, and polarized spectroscopic investigations. The porphyrin molecules were revealed to be situated with their molecular planes inclined to the (La 2 Ti 3 O 1 0 ) 2 - layers at an angle of 58° because of the high charge density of the host layers.

A unique guest–guest ion exchange method was developed for preparing a thin film of a nano-layered K4Nb6O17·3H2O that possesses both 1) optical transparency and 2) ion-exchangeability, the first ex...

A microfluidic device was successfully fabricated for the rapid serodiagnosis of amebiasis. A micro bead-based immunoassay was fabricated within integrated microfluidic chip to detect the antibody to Entamoeba histolytica in serum samples. In this assay, a recombinant fragment of C terminus of intermediate subunit of galactose and N-acetyl-D-galactosamine-inhibitable lectin of Entamoeba histolytica (C-Igl, aa 603-1088) has been utilized instead of the crude antigen. This device was validated with serum samples from patients with amebiasis and showed great sensitivity. The serodiagnosis can be completed within 20 min with 2 μl sample consumption. The device can be applied for the rapid and cheap diagnosis of other infectious disease, especially for the developing countries with very limited medical facilities.

Amebiasis caused by Entamoeba histolytica at an institution for mentally retarded in Shizuoka Prefecture is reported. Five of the 50 patients showed E. histolytica cysts in their stools and 4 were positive serologically. The polymerase chain reaction and restriction fragment length polymorphism revealed that the isolates were pathogenic-type E. histolytica. Epidemiological analysis revealed that the amebic infection was caused by the abnormal behavior of mentally retarded patients. Administration of diloxanide furoate and metronidazole for cyst-carriers eliminated cysts from the stool and lowered the antibody titer.

It has been established that two distinct species exist within what was originally known as Entamoeba histolytica. These are E. dispar and E. histolytica, for the nonpathogenic and pathogenic forms, respectively. Differentiation of these two organisms is of great clinical importance since they are morphologically indistinguishable and both forms can infect the human intestinal cavity to different degrees. A simple and rapid DNA-extraction method that can be used directly on formalin-fixed stool specimens has been developed. The extracted DNA was used for the identification of the species existing in the stools by polymerase chain reaction (PCR). A total of 72 randomly collected stool samples from the Philippines were analyzed. In all, 19 samples reacted with E. dispar primers, resulting in the expected 101-bp PCR products; however, none reacted with E. histolytica primers. Furthermore, sensitivity assay suggests that genomic DNA from as few as five cysts can be used as a template for PCR. These observations imply that the use of genomic DNA directly extracted from formalin-fixed stool specimens for PCR amplification is a useful tool for obtaining a sensitive and accurate diagnosis that can be applied even in epidemiology studies.

The pathogenicity of 47 strains of Entamoeba histolytica isolated in Pernambuco, Brazil, was examined using the polymerase chain reaction (PCR) followed by restriction-endonuclease digestion. Electrophoretic patterns of PCR products digested with HinfI revealed that all strains were nonpathogenic. The results were entirely in accord with phenotypic properties such as isoenzyme patterns and the failure to bind a pathogenic-isolate-specific monoclonal antibody. When the sensitivity of PCR was examined, amplified products could be detected from template DNA equivalent to five trophozoites. These observations indicate that PCR amplification of genomic DNA and subsequent restriction-enzyme digestion is a useful strategy for obtaining a sensitive and accurate diagnosis. The present study also demonstrates that nonpathogenic strains of E. histolytica predominate in northeastern Brazil.

Despite the medical importance of Entamoeba histolytica , neither an effective vaccine nor chemoprophylaxis to prevent amebiasis has yet been developed. On the other hand, it has been reported that passive immunization with murine monoclonal antibodies to several E. histolytica antigens is effective in preventing amebic liver abscess formation in animal models (1,2). Therefore, if human antibodies with such specificity can be obtained, it is possible that mortality from amebiasis could be reduced by passive immunization. However, hybridoma technology has not been very successful for the production of human monoclonal antibodies. Recently, new technologies have been established to generate human monoclonal antibodies. We have developed a vector for the expression of Fab fragments in Escherichia coli (3). Four recombinant human monoclonal antibody Fab fragments specific for E. histolytica have been prepared from the peripheral blood of a patient with an amebic liver abscess (4). However, these Fab fragments did not react with the surface of trophozoites, and did not inhibit amebic adherence to human erythrocytes. We report here the successful production of a recombinant human monoclonal antibody that inhibits in vitro adherence of E. histolytica trophozoites.

We designed a medium (YIGADHA-S) (1) for the trial of axenic cultivation of Entamoeba dispar based on the caseinfree YI-S medium (YI-S) (2,3). YIGADHA-S is different from YI-S in that glucose is replaced by gluconic acid and dihydroxyacetone, and its sterilization is carried out by filtration. Consequently, one strain of E. dispar (CYNO16:TPC) was adapted to this YIGADHA-S and grew stably. However, in this YIGADHA-S culture system, other strains of E. dispar still required a culture associate, such as 10% formalin fixed or autoclaved Pseudomonas aeruginosa or Crithidia fasciculata. Moreover, we recently found that over 18 different cells of animal or plant origin, which were autoclaved (121 8 C, 15 min), could also support the growth of E. dispar. These studies led us to search for a property common to every cell examined (e.g., protozoan, mammalian, and plant cells) for supporting the growth of E. dispar in YIGADHA-S. These cells had either mitochondria, hydrogenosome, or mitochondria and chloroplast. On the other hand, some protozoan cells [ E. histolytica , E. dispar , E. moshkovskii (Laredo), E. invadens (IP-1), Giardia lamblia (Portland I)], and human erythrocytes, which lacked mitochondria or mitochondria-like organelles, did not support the growth of E. dispar in this medium. Based on these findings, we speculated that some substance existing commonly in mitochondria, hydrogenosomes, chloroplasts, and bacteria might support the growth of E. dispar. The present study was attempted to test the growth-promoting effect of a plant nonheme iron–sulfur protein (ferredoxin) on E. dispar in culture. Materials and Methods

A lambda gt11 cDNA library was constructed from the poly(A)+ RNA of trophozoites of Entamoeba histolytica HM-1:IMSS strain. The library was immunologically screened with monoclonal antibody 4G6, which is specific for the 30,000-Mr antigen of pathogenic isolates. A 0.7-kb clone was isolated, and its nucleotide sequence was determined. To examine whether this gene was specific for pathogenic isolates, a polymerase chain reaction was performed by using four sets of primers and the genomic DNA of pathogenic and nonpathogenic isolates as templates. Amplified DNAs were detected not only in pathogenic isolates but also in nonpathogenic isolates. However, when sequences of amplified DNA of these isolates were compared, minor differences were observed. By considering the presence or absence of recognition sites of some endonucleases, it was possible to distinguish between the pathogenic and nonpathogenic isolates. When various isolates with different zymodemes were examined by polymerase chain reaction and enzyme digestion, the results of typing were entirely in accord with those of zymodeme analysis. These results indicate that there is dimorphism in the genomic DNA coding the 30,000-Mr antigen of E. histolytica and that the combined use of the polymerase chain reaction and enzyme digestion is a useful strategy for identification of species and determination of pathogenicity.

(1991). The validity of serodiagnosis using a monoclonal antibody against Trypanosoma cruzi-specific Mr 25 000 antigen for chagasic patients without cardiomyopathy. Annals of Tropical Medicine & Parasitology: Vol. 85, No. 2, pp. 275-276.

The seroprevalence of Entamoeba histolytica infection in the residents of seven provinces in China was examined by using an enzyme-linked immunosorbent assay with a crude antigen and a recombinant surface antigen, C-Igl, of the parasites. A total of 1,312 serum samples were investigated. The positivity rates for these two antigens were 11.05% and 6.25%, respectively. There was no significant difference in the seropositivity to E. histolytica between men and women. We used a logistic regression model and maximal-likelihood methods to estimate the prevalence of E. histolytica infection from sequential serologic data. Seropositivity in Sichuan, Guizhou, and Sinkiang Provinces was higher than that in Beijing, Shanghai, and Qinghai Provinces. The present study provides an overview of seropositivity to E. histolytica infection in seven provinces in China and use the logistic regression model estimation method to achieve a more accurate measure of amebiasis prevalence.

A novel photofunctional material composed of polyfluorinated cationic azobenzene and layered potassium titanoniobate was synthesized and its photochemical behavior was investigated. Although polyfluorinated cationic azobenzene could not be intercalated into the interlayer region of layered potassium titanoniobate directly, the intercalation compound was obtained by guest–guest-exchange with the hexylammonium-TiNbO5 intercalation compound. X-Ray diffraction, TGA, IR, TEM, UV-visible spectroscopy and elemental analysis results indicated that the polyfluorinated cationic azobenzene was intercalated into the interlayer spaces of the potassium titanoniobate. The AFM image for the intercalated compound with a basal spacing of 3.90 nm was consistent with the X-ray diffraction data. The spectral properties as well as X-ray diffraction results have revealed that the adsorbed polyfluorinated cationic azobenzene molecules form J-like aggregates with bi-layers in the interlayer space of the potassium titanoniobate. The intercalation compound exhibited an excellent reversible cis–transphotoisomerization by successive illumination with UV light at 365 nm and visible light at 458 nm. The basal spacing changed reversibly upon photoisomerization of the intercalated polyfluorinated cationic azobenzene.

The cytotoxic activity of stearylamine-bearing liposomes against Toxoplasma gondii (RH strain) was examined. When tachyzoites were treated in vitro with liposomes consisting of 20 mol% stearylamine and 80 mol% phosphatidylcholine (130 ,g/ml total lipids), more than 95% of the parasites were killed within 90 min. Intraperitoneal injection of 10 mg of 30 mol% stearylamine/70 mol% phosphatidylcholine-liposomes in mice shortly before or after T. gondii challenge afforded protection from death for more than 30 days to 70- 80% of the treated mice, whereas all untreated mice succumbed within 9 days. The liposome-injected mice that survived remained symptom-free and behaved normally. Recently, liposomes have been shown to be a potent vehicle with which to carry and deliver anti-protozoal drugs (Alving, 1986). Antimoni- als and primaquine were encapsulated in lipo- somes that were used for the therapy of experi- mental leishmaniasis and malaria, respectively. When the therapeutic effects of the encapsulated

We have developed a new LCD controller incorporating a Compression Feedforward Driving, which will be second generation Feedforward Driving. In spite of having only one SDRAM, this LCD controller can offer nearly ideal performance, improving the response time through utilizing image compression technology to reduce the frame memory requirement.

The results of an epidemiological survey in a 190-patient institution for mentally retarded were reported. Twenty percent of the patients had either cysts or trophozoites of Entamoeba histolytica in their stools, and 38% were positive serologically. The amebic outbreak revealed a sex-independent but age-dependent distribution; younger patients had more serious symptoms in cases invasive amebiasis. A high prevalence of amebic infection was found in the heavily retarded patients, and the positive cases tended to concentrate in certain training classes. Further demographic analysis suggests that the amebic infection was possibly caused by abnormal behavior of heavily retarded patients.

The results of the 13 fractures of the distal end of the femur were examined. Two cases of epiphysiolysis and one monocondylar fracture obtained good results by the cancellous screw fixation. In ten supracondylar and intercondylar fractures, six were treated by AO condylar plate and four by the other methods. All cases treated by the other method and intercondylar fractures treated by AO condylar plate showed unsatisfactory results. Surgical treatment of the distal end of the femur should be treated under technical familiarity and full equipment.

We reported our surgical method, currently adopted for duplicated proximal phalanx type.The operation was started with extracting the phalanx out of radial thumb making filet flap and, after advancement suture was done as to thenar muscle and capsule, aiming to expand the diameter and rirth, transfered filet flap to the remained thumb. By drawing the proximal phalanx of ulnar thumb up to the radial aspects, we could secure the abducted position of the thumb when suturing the filet flap. Our new contrivance in this method is to secure the advancement suture of thenar muscle by maintaining the ulnar thumb to the abducted position with radial flap.

Cytolytic activity of liposomes consisting of stearylamine and phosphatidylcholine (SA/PC-liposomes) was examined in vitro against the bloodstream forms of Trypanosoma brucei gambiense. More than 99% of the cells (2 X 10(6)/ml) were killed within 30 min by treatment with 15 mol% SA/PC-liposomes (100 microM total lipids). As few as 1.2 X 10(12) liposomes per ml (equivalent to 2 nM liposome) showed trypanocidal activity. Fluorescence microscopy of cells treated with the dansylated SA/PC-liposomes suggested that the liposomes bound to and accumulated on the cell surface, eventually damaging the plasma membrane. SA/PC-liposomes showed no significant hemolysis when incubated with human and mouse erythrocytes under conditions that killed greater than 99.9% of the T. b. gambiense trypomastigotes. Human leukocytes were also shown to be less susceptible to SA/PC-liposomes than T. b. gambiense. These results may point to a new direction in strategy for therapy of African trypanosomiasis.

Augmentation de la resistance mecanique et de la tenacite de la fonte a graphite spheroidal par addition de 2 a 4% de nickel et traitement thermique donnant une matrice ferrite-bainite. Comparaison avec des echantillons de fonte malleable a structure austenitique. Etude microfractographique

The case of a seven-year-old boy with acute osteomyelitis of the distal femoral epiphysis is reported. Acute hematogenous osteomyelitis begins commonly in the metaphysis and in children the epiphyseal plate bars the spread to the epiphysis and the adjacent joint. Epiphyseal osteomyelitis is very rare, frequently spreading into the adjacent joint and being more latent than metaphysial osteomyelitis.

A mouse monoclonal antibody TCF87, prepared previously, was reactive with Trypanosoma cruzi-specific Mr 25,000 antigen regardless of strain. The Mr 25,000 antigen was recognized by all sera from chagasic patients living in different areas of South America, when examined by Western immunoblotting analysis. Although many antigens of T. cruzi epimastigotes were also recognized by sera from patients with leishmaniasis, the Mr 25,000 antigen of T. cruzi did not react with leishmaniasic sera. These results indicate that Mr 25,000 antigen recognized by TCF87 is valuable as a diagnostic antigen for Chagas' disease. When a competition enzyme-linked immunosorbent assay using TCF87 was carried out, all sera from Chagas' disease patients showed positive inhibition. By contrast, all patients with leishmaniasis or other parasitic diseases were scored as seronegative. The present study suggests that competition enzyme-linked immunosorbent assay using monoclonal antibody against the Mr 25,000 antigen of T. cruzi will be useful for serodiagnosis of Chagas' disease in areas where leishmaniasis is co-endemic.

Stress roentgenograms before and after treatment of lateral ligament tears of the 15 ankles treated by cast immobilization were analyzed.Eleven out of 15 ankles recovered roentgenographic stability satisfactorily.To the above cases and chronic cases, we asked questionnaires based on Seligson scale by letter.Results showed a tendency that the greater the roentgenographic instability of the ankle is, the lower the functional rating of the ankle is.

Two monoclonal antibodies reacted with the Trypanosoma cruzi-specific antigen of an apparent Mr 25,000 from all developmental forms (Tachibana et al. 1986). This T. cruzi-specific antigen was found at the plasma membrane by immunoperoxidase electron microscopy using the monoclonal antibodies TCF48 and TCF87. The TCF48 and TCF87-treated cells showed stain deposits at the plasma membrane clearly distinguishable from those in cells treated with a monoclonal antibody against a surface antigen. This suggests that the epitope(s) of the Mr 25,000 antigen is located on the inner surface or in the matrix of the plasma membrane. TCF48 and TCF87 also reacted with an antigen on the microtubules of the axoneme, but not with the subpellicular microtubules. These results suggest that the T. cruzi-specific Mr 25,000 antigen is common to both the plasma membrane and axoneme but it is not located at the subpellicular microtubules. Its identity and that of the surface antigen, Gp25 (Scharfstein et al. 1983) as well as its role in the pathogenicity of the parasite are discussed.

If the feed contains micro-components and reflux ratio is extremely high, distillation problems sometimes become impossible to solve because micro-components behave in a complicated manner. The acetic acid purification problem is an example of these problems. No conventional single algorithm could solve this problem. It can be solved by a combination algorithm : to make a favorable initial profile, the successive iteration algorithm precedes the Newton-Raphson one. A detailed consideration of convergence is given in the text.A novel column profile of micro-components is observed : it shows three extremes in column concentration of formic acid, and two extremes for water, which we had never seen. Failure of the single algorithm comes from this novel type of profile.