Some biological effects of Bayluscide on Physa acuta and Helisoma duryi. The fecundity, viability and the size of egg-masses were studied in Physa acuta and Helisoma duryi snails treated with 1 ppm Bayluscide for two hours. Bayluscide application caused significant and highly significant reduction in the fecundity of both Physa acuta and Helisoma duryi. However, an increase in the fecundity was observed for both treated snails in few days after exposure. Moreover, Bayluscide did not lead to stoppage of egg laying at any day of experiment. Bayluscide caused a reduction in the viability of egg-masses produced by treated Physa acuta or Helisoma duryi snails since they produced higher numbers of abnormal eggs containing dead embryo. Moreover, application of 1 ppm Bayluscide for two hours against Physa acuta caused a decrease in the size of egg-masses produced by treated snails. This is expressed by the production of higher percentage of egg-masses containing small numbers of eggs as compared with those masses produced by control snails.


Histo- and immuno-pathological and electron microscopy studies of the splenic red pulps of schistosomiasis mansoni patients. The histo- and immuno-pathological and electron microscopy (EM.) of the splenic red pulps studies were carried out for schistosomiasis mansoni patients and normal controls. The main pathological processes were interstitial haemorrhages, congestion, proliferation and hyperplasia of the red pulps of the schistosomiasis mansoni patients. The IgG showed marked reaction, moderate with IgA and mild with IgM. The E.M. showed an increase in the size of the different types of the reticular cells particularly those of the nuclear membrane with type II, nuclear degeneration, deposition of fibrin threads, marked fibrosis and phagocytosis and formation of pores at the sinusoidal basement membrane. It was concluded that these pathological processes and the alteration in the immunoregulatory system of the spleen of schistosomiasis mansoni patients reflect the role played and perpetrated in the development of splenomegaly.


Immunological response to diphtheria/tetanus vaccine in Schistosomiasis mansoni patients. The cellular and humoral immune responses of patients with S. mansoni infection were evaluated before and one month after each of two intramuscular doses of diphtheria/tetanus toxoid vaccine. Patients were divided into "responder" and "non-responder" groups based on anti-tetanus toxoid (anti-TT) IgG levels after vaccination. The specific anti-TT IgG1 response of the responder group was predominantly in the IgG, subclass. The lymphoproliferative response to PHA was also significantly higher in the responder group; this elevation was detectable before and after each vaccination. The responses to PWM and SPL were comparable in the two groups before vaccination, although the responder group had a higher response to SPL after vaccination. IgG antibodies for schistosome adult worm and egg antigens were significantly lower in the responder group prior to vaccination but not thereafter. Anti-diphtheria IgG antibodies were comparable in the two groups after vaccination at all times. Clinically, the non-responder patients had a higher incidence of splenomegaly (84.6% vs 44.8%) and were significantly older than the responder patients (mean 34.1 yrs vs 18.7 yrs). The cause for the reduced anti-tetanus IgG response in schistosomiasis patients is believed to be multifactorial. T cell or antigen presenting cell dysfunction, high levels of IgG antibodies specific for schistosome antigens, splenomegaly and age are factors that might lead to reduced anti-TT IgG response.


Cyclosporin A in experimental trichinosis scanning electron microscopic study. The surface morphology of adult and larvae of Trichinella spiralis after cyclosporin A treatment was studied by scanning electron microscopy (SEM) of fixed, dried and metal coated specimens. The earliest topographic changes revealed in adults after the drug treatment were in form of disorganization of the cuticle especially between the hypodermal pores which appeared somewhat thickened and irregular. Other changes were in form of bullae formation covering the whole width of the adult surface. Other specimens showed wrinkling of the outer layer that was seen later to peel off. In few worms layer outer was damaged releasing some of the internal organs. Smoothing of the cuticle with obliteration of the grooves between the cuticular ridges were observed as the earliest surface larval changes, ulcer formation which was deeply seated was seen in patchy areas in some specimens. The interaction between the parasite surface and the drug is of significant importance as this can throw light on the possibilities of its mode of action.


Characterization of Egyptian isolates of Trichomonas vaginalis: I. Serotyping. Antitrichomonal hyperimmune sera against T. vaginalis stocks isolated from Egyptian female patients were employed for serological differentiation of somatic and soluble antigens in the Ouchterlony gel double immunodiffusion technique. It was concluded that soluble trichomonal antigens present in association with living flagellates are stock--specific reacting with some, but not all the antitrichomonal hyperimmune sera, while those present in association with dead parasites are common antigens reacting with all the sera. Three stocks, E1, E2 and E3 could be differentiated into two strains using their stock--specific antigens. The somatic antigens of six trichomonal stocks reacted with all the hyperimmune sera denoting common antigenic make up.


Studies on flea fauna in El Fayoum Governorate, Egypt. This paper dealt with the studies of flea fauna as collected on the rodent hosts over one year. The collected rodents (499) in a descending order of abundance in the collection were Rattus rattus, R. norvegicus, Mus musculus, Gerbillus pyramidum and Arvicanthis niloticus. The collected fleas in a descending order in the collection were X. cheopis, P. irritans, N. jasciatus, C. felis, L. segnis and E. gallinacea. The total flea index reached the maximum (0.76) in May and it was zero in December and January. The highest number of fleas was on R. norvegicus and A. niloticus and the lowest was on G. pyramidum. The highest number of rodents and their flea ectoparasites was during the Spring and the lowest was during Autumn. The medical importance of the collected fleas and the whole results were discussed.


On the morphology of the oxyurid nematode Pharyngodon mamillatus (Linstow, 1897) from Chalcides ocellatus from Egypt. The nematode P. mamillatus is described from the lizard Chalcides ocellatus (Forsk.) from Cairo, Egypt. The use of both light and scanning electron microscopy enabled the authors to report new morphological features of the worms, including the cuticular surface, lateral alae and male caudal structures.


Studies on chronic liver diseases in patients with and without Toxoplasma latent infection. Chronic liver diseases mainly due to schistosomiasis is a common clinical feature in Egypt. On the other hand, Toxoplasma latent infection is another common feature. In this paper, it was aimed to study the role of Toxoplasma latent infection in patients with chronic liver diseases mainly due to schistosomiasis. The latent Toxoplasma infection was measured by the positivity of IgG to ELISA and the negativity of IgM to DS-ELISA. The cellular immunity was assessed by the dinitrochlorobenzene (DNCB) skin test. The clinical and the laboratory studies showed that latent Toxoplasma infection had no marked effect on the progression of the chronic liver diseases.


Ultrastructural study of the tegument of Cotugnia polyacantha and Killigrewia streptopelia inhabiting the dove Streptopelia senegalensis aegyptiaca. The ultrastructure of the teguments of the cyclophyllidean cestodes Cotugnia polyacantha and Killigrewia streptopelia naturally infecting the dove Streptopelia senegalensis aegyptiaca, was studied using transmission electron microscopy. The two cestodes inhabit two distinct sites in the small intestine of the same bird. Observations revealed basic similarity between the two cestodes, though, marked differences between the tegument structure details of the two species regarding the structure and dimension of microtriches, types and distribution of vesicles among the regions of the tegumental/perikaryal complex, structure of subtegumental connective tissue and subtegumental muscle layer. The significance of these observations was discussed in possible relation to the different physico-chemical environments known to occur where each cestode resides.


Ultrasonographic findings in the gall bladder in human fascioliasis. This study included forty patients with incubating or open fascioliasis. All were subjected to repeated stool examination, serological and haematological study using IHAT in the former. Ultrasonography was done for upper abdomen stressing on gall bladder and assessing its function. Eighteen patients were diagnosed as incubating fascioliasis by positive IHAT, high eosinophilic count and negative stool examination for Fasciola eggs. No gall bladder changes could be detected by U.S. among those patients. Twenty two patients passed eggs in their stool were diagnosed as open Fasciola. Their gall bladder through U.S. revealed flukes in twelve of them. A weak gall bladder contractility was observed in 64% with open fascioliasis.


The effect of scorpion envenomation on the different organs of albino mice. The effect of sublethal doses of scorpion venom (Buthus quinquestriatus) on the hepatic vascular bed and hepatic parenchyma were studied. The main toxic effect of the venom was primarily on the vascular bed of the liver as manifested by dilatation of branches of the hepatic artery, portal vein together with intravascular thrombi and subcapsular haemorrhages. Apart from mild hydropic degeneration some cells of the hepatic parenchyma showed focal necrosis and Kupffer cells were frequently hypertrophied and contained pigment. In the lung, dilated thrombosed vessels were observed. In the kidney, the vessels showed the same changes. The epithelial and endothelial cells of the glomeruli appeared normal except for slight swelling with intact basement membrane. The tubular cells were swollen with increased granularity and attenuation of the tubular lumen, a picture compatible with hydropic degeneration. The possible mechanisms in inducing such lesions were discussed.


Evaluation of insecticidal efficiency of certain new selective formulations against Musca domestica (L). Five new selective formulations were tested for effectiveness, applicability and costs in controlling the housefly Musca domestica. The tested formulations are Alfacron SB, Alfacron 10 WP, Snip IRB (a.i. Azamethiphos, thiophosphorester), Baygon SB and Golden Marlin Results of laboratory studies revealed that Alfacron 10 WP was the most potent compound against the adult fly (100% mortality was achieved after 20 min from exposure). Snip IRB ranked second with 100% mortality after 40 min, followed by Alfacron SB (50 min.) and Golden Malrin (60 min.). On the other hand Baygon SB did not show complete mortality of houseflies since its maximum mortality was 57.5% after 60 min. Laboratory tests also proved that moistening the fly prolonged the efficacy period of the three tested products from 3 to 6 weeks in case of Alfacron SB, from 4 to 7 weeks for Snip IRB and from 7 to 9 weeks in case of Alfacron 10 WP.


Comparative evaluation of antigens used in immunofluorescence (IFAT) antibody test for serodiagnosis of schistosomiasis. The diagnostic value of antigens used in the IFAT for serodiagnosis of schistosomiasis has been evaluated. Cryosections of Schistosoma mansoni adults, formaline fixed cercariae and cryosections of infected liver, from gold hamster, which contained granuloma were used as antigens in this evaluation. The cryosections of adult worm were more reliable and more specific as an antigen for immunofluorescence (Diagnostic efficacy was 95.52%). The cercarial antigens gave the same results as the cryosection of adults. The IFAT using cryosection of infected liver contains granuloma was less reliable due to autofluorescence of eggs in the granuloma with negative sera (Diagnostic efficacy was 49.01).


Definition of receptor binding domains in interferon-alpha. Earlier studies from this laboratory had identified three regions in interferon-alpha (IFN-alpha) that influence the active conformation of the molecule. These domains are associated with the amino acid residues 10-35, 78-107, and 123-166. In this report, we define these domains more accurately by identifying their critical clusters of amino acids. Using a panel of IFN-alpha 2a variants in antiviral, growth inhibitory, and receptor binding studies, we are able to show that these three domains, defined by residues 29-35, 78-95, and 123-140, are likely located on the surface of the molecule, with domains 29-35 and 123-140 in close spatial proximity. We conclude that the 29-35 and 123-140 domains are responsible for IFN-alpha receptor binding interactions and constitute receptor recognition sites in IFN-alpha. Extrapolating from our biological activity data, in the context of a number of predictive algorithms that provide insights into the hydrophobicity/hydrophilicity, surface probability, and flexibility of amino acid clusters, we infer that the residues 29-35 influence the active configuration of IFN-alpha most significantly. This region likely represents a loop structure that is relatively rigid in configuration. The carboxy-terminally located strategic domain, 123-140, is comprised of two clusters of amino acid residues, one that forms part of a rigid alpha-helix, the other a more flexible loop structure. Similarly, the 78-95 domain comprises a portion of an alpha-helical structure that is followed by a loop structure. Close examination of the amino acid sequences in all three regions among the different species of IFN-alpha s and human IFN-beta indicate that the 29-35 and 123-140 domains are most highly conserved, yet some variance is apparent in the 78-95 domain. We propose that the 78-95 region influences species specificity among the murine and human IFN-alpha s and determines the differential specificity of action between human IFN-alpha and human IFN-beta.


Interferon-alpha suppresses the capacity of T cells to help antibody production by human B cells. This study was designed to analyze the effect of interferon-alpha (IFN-alpha) on the potential of T cells to help B-cell differentiation in vitro. Human splenic T cells preactivated via the T-cell receptor (TCR)/CD3 complex, as well as murine EL4 thymoma T cells preactivated with phorbol esters, stimulated human B cells via a species cross-reactive physical interaction to differentiate into antibody-producing cells. If the human or murine T cells were activated in the presence of IFN-alpha, normal proliferation and interleukin-2 (IL-2) production occurred, but the cells did not acquire any B-cell helper potential. Therefore, IFN-alpha modulates the B-cell stimulatory potential of T cells by interfering with the T-cell activation process. In contrast, IFN-alpha neither acted on B cells directly nor on already activated T cells, because it did not suppress B-cell differentiation induced by T cells preactivated in the absence of IFN-alpha. IFN-alpha did not induce the production of inhibitory T-cell factor(s), since T cells preactivated in the presence of IFN-alpha did not inhibit the interaction of B cells with T cells optimally preactivated in the absence of IFN-alpha. Taken together the data indicate that IFN-alpha suppresses the potential of T cells to stimulate B-cell differentiation by interfering with the T-cell activation process, but acts neither on B cells directly nor on already activated T cells.


Constitutive production of interferon-alpha 2 by a human B-lymphoblastoid cell line. Human B-lymphoblastoid cell lines immortalized by Epstein-Barr virus constitutively produce low levels of acid-stable interferon (IFN) that differs in composition from IFN produced by the same cell type after virus induction. We use three approaches to study the constitutive IFN produced by the lymphoblastoid cell line, LuKII: (i) antiviral bioassays with a neutralizing monoclonal antibody; (ii) amino-terminal sequence analysis of the single protein obtained after purification of the IFN by immunoaffinity chromatography and reverse-phase HPLC; (iii) sequence analysis of cDNA clones obtained by screening a LuKII library with an IFN-alpha 2 probe. Our data show that more than 90% of the constitutive IFN results from expression of a single gene coding for IFN-alpha 2. The significance of these results is discussed.


Different interactions of interferon-alpha subtypes at the surface of epithelial and lymphoid cells. The interaction of different interferon (IFN)-alpha subtypes with different cell types was investigated using a unique monoclonal antibody (MAb), I-4-A. This MAb reacts in immunoassays equally with IFN-alpha 2b and IFN-alpha 4a, but does not inhibit the binding of IFN to cell receptors. 125I-labeled I-4-A reacted with IFN-alpha 4a and IFN-alpha 2b bound to receptors on Daudi cells. However, in a "double assay" developed using Daudi cells to measure antiviral and antiproliferative activity, I-4-A neutralized both activities of IFN-alpha 4a, but neither of IFN-alpha 2b. Similarly, in studies on the activation of natural killer (NK) cells, I-4-A neutralized the effect of IFN-alpha 4a but not that of IFN-alpha 2b. In contrast, when cell lines other than lymphoid were studied, e.g., HEp 2 and WISH cells, I-4-A neutralized the antiviral activity of both IFN-alpha subtypes. The neutralization of one IFN-alpha subtype but not another on lymphoid cells suggests a difference either in the receptor-bound form of the subtypes, or in subsequent interactions prerequisite for activation of these cells. Furthermore, the neutralization of a particular IFN subtype, alpha 2b, on epithelial-derived but not lymphoid cells suggests differences in the IFN-receptor complex or the mechanisms of cell activation between these cell types. An implication from these studies is that some IFN-alpha subtypes can exert different functions on lymphoid and epithelial cells.


The effects of interferon-gamma and bacterial lipopolysaccharide on CD14 expression in human monocytes. CD14 has been reported to be the lipopolysaccharide (LPS)-LPS binding protein receptor. The effects of interferon-gamma (IFN-gamma) on CD14 expression have not been clearly established. The purpose of this investigation was to examine the effects of IFN-gamma alone and IFN-gamma followed by bacterial LPS on CD14 expression. Human peripheral blood monocytes were isolated by counterflow centrifugal elutriation (CCE). Monocytes were cultured for 48 h with IFN-gamma alone or for 24 h with IFN-gamma followed by LPS for a second 24 h. IFN-gamma alone caused a down-regulation of CD14 expression, as assessed by flow cytometry, relative to CD14 expression in untreated monocytes. In addition, CD14 expression was even more significantly down-regulated after IFN-gamma pretreatment followed by either Prevotella intermedia or Salmonella typhimurium LPS. Likewise, the percentage of CD14+ monocytes decreased after IFN-gamma alone and even more dramatically after IFN-gamma treatment followed by either LPS. This study clearly demonstrated that IFN-gamma down-regulates CD14 expression and that LPS following IFN-gamma pretreatment potentiates this effect.


Post-transcriptional regulation by interferon-alpha of epsilon-globin production in human erythroleukemia K-562 cells. Treatment of human K-562 erythroleukemia cells with interferon-alpha (IFN-alpha) resulted in a six-fold increase in the production of epsilon-globin, a component of embryonic hemoglobin, but in a two-fold or less augmentation of the production of other globin species. There was no comparable increase in the production of epsilon-globin mRNA following IFN treatment. We did, however, observe a shift in the cytoplasmic location of the epsilon-globin mRNA from the messenger ribonucleoprotein (mRNP) to the polysome fraction after IFN-alpha addition. No such shift was noted in the location of the mRNA for A gamma-globin, the major globin product of K-562 cells, which was always present in the polysome fraction. These results are significant with respect to the mechanisms of cellular regulation and differentiation induced by IFNs.


Identification of a complex formed between nuclear proteins and the transcriptional enhancer of interferon-inducible genes that is present in the peripheral blood myeloid cells of CML but not normal individuals. Interferon-alpha (INF-alpha) induces cytogenetic remissions in 20% of chronic myelogenous leukemia (CML) patients. To clarify the mechanisms through which this antiproliferative action of IFN is mediated in the CML cell, a modification of the mobility-shift assay was used to follow the formation of complexes between nuclear proteins and IFN-inducible transcriptional enhancers involved in mediating the cellular effects of IFN-alpha. These studies identified a complex that was present in the myeloid cells of 18/24 (75%) of chronic-phase CML patients tested whose cells contained 100% Philadelphia chromosome positive (Ph+) cells, while the proteins of none of the samples tested from normal peripheral blood samples and only 22% (2/9) of the CML patients in an IFN-induced major cytogenetic remission (less than 30% Ph+ cells) contained these complexes. These studies suggest that the mobility-shift assay detects changes in the CML myeloid cell that distinguish it from the normal myeloid cell.


Double-blind randomized phase I study on the clinical tolerance and biological effects of natural and recombinant interferon-beta. The clinical tolerance of and the effects recombinant human interferon-beta (rHuIFN-beta) obtained from mammalian cells (Chinese hamster ovary cells) exerts on 2',5'-oligoadenyl (2-5A) synthetase activity, human-Mx protein, neopterin, beta 2-microglobulin, interleukin-1 (IL-1) alpha and beta synthesis were compared to those of natural IFN-beta in 12 healthy volunteers. Each subject received a single i.m. injection of 6 x 10(6) IU rHuIFN-beta and natural IFN-beta according to a randomized double-blind cross-over study design. Both were well tolerated and provoked similar changes in clinical indices. Moreover, rHuIFN-beta and natural IFN-beta induced significant and similar increases in 2'-5' adenylates, human Mx protein, and neopterin levels, but neither modulated beta 2-microglobulin, IL-1 alpha or beta synthesis. The sum of these findings indicates that rHuIFN-beta and natural IFN-beta are biologically equivalent. In view of these results, we are of the opinion that these two types of IFN are probably also therapeutically equivalent and, in consequence, that trials to evaluate the response of viral and neoplastic disease patients to rHuIFN-beta are fully justified.


Human interferons-alpha inhibit the production of immunoglobulin M by secreting and nonsecreting lymphoblastoid cell lines. It is well known that human lymphoblastoid interferon-alpha [HuIFN-alpha(ly)] may either increase or decrease antibody production by B cells, in vivo as well as in vitro, depending on the experimental conditions and the system used. We compare here the effect of HuIFN-alpha(ly) and human recombinant interferon-alpha (rHuIFN-alpha) on immunoglobulin M (IgM) production by secreting and nonsecreting human B-like lymphoblastoid cells, respectively, ST8246, and Daudi cells. Under our experimental conditions, Daudi cells were less sensitive to the antiproliferative effect of IFN than previously reported by other authors, and ST8246 cells were insensitive to this antiproliferative effect. In contrast, IgM production was profoundly affected in both cell lines. Thus, we could discriminate between the effect on cell growth from the effect on the immune response. Using high-pressure liquid chromatography (HPLC) separation, mu chains and monomeric and pentameric IgM were distinguished from cytosolic and membrane-associated fractions and from culture medium (extracellular IgM). Pentameric extracellular IgM and monomeric membrane IgM were diminished by HuIFN-alpha(ly) treatment, respectively, in ST8246 cells and in Daudi cells. We conclude that HuIFN-alpha(ly) induces regression of B-like lymphoblastoid cells toward a less mature phenotype.


Induced and down-regulated proteins in the human cultured hairy cell leukemia line JOK-1 and the Burkitt's lymphoma cell line Daudi during incubation with interferon-alpha: a kinetic study. To elucidate the mechanism of action of interferon-alpha (IFN-alpha), the effect on cell proliferation and protein synthesis in the human hairy cell leukemia line JOK-1 and the Burkitt's lymphoma cell line Daudi were investigated. While Daudi cells were inhibited in proliferation and in total protein synthesis, no effect was seen on JOK-1 cells. However, high-resolution two-dimensional gel electrophoresis showed that four polypeptides were induced in JOK-1 cells after IFN-alpha incubation, while an additional 11 were induced and two down-regulated in Daudi cells. Kinetic studies revealed that the changes in JOK-1 cells were only temporary (within 8-16 h) and small to moderate in magnitude (less than four-fold). In Daudi cells, the changes for two of these polypeptides were early (within 2 h), for most of them prolonged (at least 24 h), and for three of them of great magnitude (between 6- and 30-fold). Quantitative analytical assessments indicated that four IFN-alpha-inducible polypeptides, present in low amounts of untreated cells, were highly expressed only in sensitive Daudi cells upon IFN-alpha treatment. This observation might indicate a role for these polypeptides in the inhibition of cell proliferation in Daudi cells. Furthermore, six of the other IFN-alpha-modulated polypeptides were synthesized constitutively in JOK-1 cells at levels comparable to those achieved in IFN-alpha-treated Daudi cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Interferon-alpha activates binding of nuclear factors to a sequence element in the c-fos proto-oncogene 5'-flanking region. Interferon-alpha (IFN-alpha) can regulate the expression of the c-fos proto-oncogene in different cell types. Here we show IFN-alpha-activated binding of murine and human fibroblast nuclear factors to a DNA sequence element located in the 5' upstream region (nucleotides -351/-337) of the c-fos gene. This element, like the conserved enhancer element, the IFN-stimulated response element (ISRE), that mediates transcriptional induction of IFN-alpha-inducible genes, also binds factors in response to platelet-derived growth factor (PDGF) and v-sis-conditioned medium (SCM). The IFN-inducible ISRE shares an 8-bp stretch of sequence homology with the IFN-responsive c-fos SCM element, and competes efficiently for binding of factors to the SCM. Protein-DNA cross-linking experiments with the SCM binding site identified an IFN-modulated nuclear protein of approximately 98 kD. This protein does not appear to be involved in transcription activation, since IFN-alpha failed to stimulate c-fos transcription in nuclear run-off assays, or the c-fos promoter in transient transcription assays of 3T3 fibroblasts. Our data nonetheless suggest the c-fos promoter may be an early target for signal transduction triggered by IFN alpha-receptor interaction.


Opposite sensitivity to the antiproliferative action of interferon-alpha and granulocyte-macrophage colony-stimulating factor in monoblastic U937 cells. Three variants of the human monoblastic cell line U937 with different degrees of sensitivity to the antiproliferative action of interferon-alpha (IFN-alpha were examined for phenotypic differences. The highly IFN-sensitive variant U937-V expressed twice as many IFN-alpha binding sites as both its IFN-alpha-resistant derivative U937-VR and the cell line U937 exhibiting a 20-fold reduction in IFN-alpha sensitivity as compared to U937-V cells. All three variants were IFN-reactive with regard to induction of 2',5'-oligoadenylate (2-5A) synthetase activity and were similarly sensitive to the growth-inhibiting action of IFN-gamma and tumor necrosis factor. Responsiveness to the antiproliferative effect of granulocyte-macrophage colony-stimulating factor (GM-CSF), however, was confined to cell lines U937 and U937-VR. Although expressing a comparable number of GM-CSF receptors, the highly IFN-sensitive variant U937-V was refractory to GM-CSF. Flow cytometry revealed a marked difference in the expression of the antigen CD11b which was detectable on 85% of cells of the U937-V line but only on approximately 25% of cells derived from the U937 and U937-VR lines. Results thus demonstrate opposite sensitivity of U937 cells to the growth-inhibiting action of IFN-alpha and GM-CSF, apparently dependent on the state of U937 differentiation as determined by expression of the CD11b antigen.


Tandem repeat polymers of a critical region of the human interferon-beta promoter exhibit a marked constitutive activity and enhanced responsiveness to transcriptional regulators in transfected HeLa cells. Multiple copy tandem repeats polymers of an authentic 30-bp region of the human interferon-beta (IFN-beta) promoter between positions-91 to -62 relative to the cap site or the hexanucleotide GAAAGT derived from this region, both acted as strong constitutive regulatory elements in transfected HeLa cells. Such polymers were unresponsive to treatment with IFN-alpha despite their considerable homology with the IFN-responsive elements of other genes but were highly responsive to treatment of HeLa cells with IFN-gamma. Virus induction of HeLa cells transfected with polymers of the 30-bp region linked to a CAT gene increased the activity of the reporter gene 500- to 2,000-fold over baseline levels. Treatment with IFN-alpha prior to virus induction did not increase further CAT activity. Cotransfection of HeLa cells with the CAT gene under the control of a 12-element tandem repeat polymer of the human IFN-beta promoter and an expression vector for the IRF-1 transcriptional activator markedly increased CAT activity while cotransfection of HeLa cells with the IFN-beta construct together with an expression vector for the transcriptional regulator IRF-2 markedly decreased CAT activity relative to cells transfected with the IFN-beta polymer alone.


A comparative KAP study of leprosy patients and members of the community in Hlaing and Laung-Lon townships. A KAP study was conducted in the peri-urban Hlaing and rural Laung-Lon Townships in Myanmar. It was found that both the leprosy patients as well as community members were still not sure about the cause of leprosy. Social stigma of leprosy encountered by patients needs to be addressed especially in peri-urban areas. It was also found that the patient's understanding of treatment regularity was still very unsatisfactory, for which health education measures needs to be introduced.


Treatment of paucibacillary leprosy with a regimen containing rifampicin, dapsone and prothionamide. Ninety paucibacillary leprosy patients having indeterminate (I), tuberculoid (TT) and borderline tuberculoid (BT) type of leprosy with bacterial index (BI) of less than two on the Ridley scale were treated with rifampicin (RFM) 600 mg once a month, dapsone (DDS) 100 mg daily and prothionamide (PTH) 250 mg daily. Treatment was stopped at the end of six months. The patients tolerated the drugs fairly well and in only two patients the drugs had to be stopped (in one due to jaundice and in the other due to gastric intolerance). About 6% of patients had early reactions which subsided with additional steroid therapy. The inactivity rate was 60% at six months and this improved to 96% at 12 months. No cases of late reactions and relapses were encountered in the limited follow-up period of six months; and a longer follow-up is necessary for ascertaining the relapse rates. The preliminary results however suggest that the addition of prothionamide to the standard WHO paucibacillary regimen is well-tolerated with increased inactivity rate and fewer instances of late reactions.


Correlation between inhibitory effect of quinolones and mycolic acid metabolism in mycobacteria. Mycolic acids are important components having a significant role in maintaining the rigidity of mycobacterial cell wall. They could also be the barrier for penetration of certain drugs into the bacterial cell. A novel in vitro model system was established for assessing the effect of Ciproflaxacin on mycolic acid metabolism in pathogenic mycobacteria M. Kansasii (which has similar mycolic acid pattern to that from M. leprae) and the effect of norfloxacin in M. intracellulare. These test mycobacteria were exposed in their midlogarithmic phase of growth to 0.5, 1, 2, 3, 4, 5 and 6 micrograms ml of ciprofloxacin and norfloxacin respectively for 1, 2 and 24 hours. Ciprofloxacin completely inhibited the synthesis of mycolates in M. kansasii at 3, 4 and 5 micrograms/ml; whereas norfloxacin exhibited its maximum inhibitory action on mycolic acids in M. intracellulare at 6 micrograms/ml for all the durations of exposure. Inhibition of mycolates directly correlated with bacterial viability which was estimated by colony forming units. The effect of quinolones on mycolic acid metabolism appears to be direct and not secondary to DNA gyrase. The results obtained from this study and our previous findings show that mycolic acid metabolism is affected by various groups of drugs, whose primary sites of activity may be different. The findings of the present study may have significant therapeutic implications in leprosy and other mycobacterial diseases.


Ultrastructural study of mouse dorsal root ganglion cultures infected long term with M. leprae. Ultrastructural changes in the mouse dorsal root ganglion cultures infected long-term with viable M.leprae were studied. Subtle cytomorphological changes and loss of neurites noted in the long-term infected cultures were correlated to early events in the nerve damage.


Leprosy of the tongue.

Leprosy of the tongue. Ten out of the twenty-five lepromatous leprosy patients studied showed clinical evidence of involvement of the tongue, and they presented with various symptoms like loss of taste, stiffness of tongue, bleeding, pain etc. Various types of lesions ranging from small nodule to granuloma formation, ulceration, macroglossia and fissured cracked tongue were noted. The tongue lesions were found to be related to the severity of leprosy.


Leprosy immunopathogenesis and vaccine development. The truly effective immunity against intracellular parasites, including mycobacteria, is mediated by monocyte/macrophages, and in the immunologically responding (resistant) host these phagocytes need minimal antigenic stimulus, specific or non-specific, to become activated and be microbicidal. T-cell mediated delayed hypersensitivity (DTH) causes tissue damage and destruction, which is particularly unwelcome in leprosy because of its nerve-damaging potential. Gamma interferon (INF-gamma), the terminal lymphokine of a DTH response, promotes mycobacterial survival and growth. There are T-cells (TH1 subtypes) that produce DH response either independent of, or, only partly dependent on INF-gamma; this type of DH peaking at 24 hours appears similar to the Jones-Mote type rather than to the tuberculin type of DTH peaking at 48-72 hours and is devoid of the necrotic component of tuberculin type of DTH. M. leprae antigens normally elicit this Jones-Mote type of DH. Suppressor T-cells are associated with a protective immune response, while helper T-cells mediating DTH are harmful. In view of this immunobiology, it would appear that pathogenic mycobacteria that generate a tuberculin type DTH response should not be used as immunogens in leprosy.


Observations on the cultivation of M. leprae and M. tuberculosis in medium 'V' and 'V 1'. Skin scrapings from five different active sites were collected from 14 leprosy patients and inoculated into medium V. Skin scrapings from three leprosy patients were inoculated into medium V 1. All the cultures were incubated at 8-10 degrees C. M. tuberculosis H37Rv, pretreatment isolates and streptomycin resistant strains were inoculated into medium V, with and without antibiotics, and incubated at 8-10 degrees C as well as 37 degrees C. Smears were made from the M. leprae and M. tuberculosis cultures at 0 hours and at different time points. The number of bacilli in the smears were counted. There was no increase in the number of M. leprae or M. tuberculosis in any of the cultures.


Pattern of leprosy deformities among agricultural labourers in an endemic district: a pilot study. A study of 1,338 leprosy affected agricultural labourers in an endemic district revealed that 12% had deformities. The patient's sex, type of disease, duration and educational status seemed to influence pattern of leprosy deformities. The patients continued working despite deformities in order to avoid financial dependence on their family members and loss of dignity.


The CEA family: a system in transitional evolution? The CEA family consists of two structurally and functionally distinct sub-groups; the group including CEA, NCA and CGM-6 which are cell surface-bound by phosphatidyl-inositol (PI) linkages, and the group of BGP splice variants which have trans-membrane and cytoplasmic domains. Although all CEA family members mediate intercellular adhesion in vitro, the PI-linked group show Ca++ and temperature independent adhesion whereas the BGP group show rapidly reversible Ca++ and temperature dependent adhesion. From the close alignment in cDNA nucleotide sequences between family members and between repeated domains in one family member, it is apparent that the CEA family is now rapidly evolving; in fact, analogs of only the trans-membrane BGP group have been found so far in the mouse. The addition of a new group of potent adhesion molecules to complex species at some time after the rodent radiation has strong evolutional implications, which are discussed.


Introduction to the CEA family: structure, function and secretion. Due to the phenomenal progress in the field of tumor immunology that took place during the last twenty years, we dispose today of highly specific and sensitive techniques and reagents like monoclonal antibodies (MAbs). In this context the discovery in human carcinomas of tumor-associated antigens, such as CEA, was of primary importance, especially since the latter was found to have clinical relevance as a tumor marker. Based on animal models, a new in vivo technology for the detection of tumors and metastases was developed in recent years, that uses anti-CEA MAbs, or fragments of them, coupled to radio-isotopes. This technique, called radio-immunodetection (RAID), also paved the way for immunotherapeutic procedures, where again CEA served as the target-antigen. This new technique holds great promise, provided the epitope-specificity of the MAbs is well-controlled: it has been shown that CEA belongs to a large gene-family of at least 22 members, which can be subdivided into two subgroups (i.e., the CEA- and the PSG-subgroup) and which in turn belongs to the immunoglobulin-supergene family. Great structural similarities render the distinction of the various cross-reactive molecules by immunological means rather difficult.


Immunohistology of carcinoembryonic antigen (CEA). Specificity of reagents, choice of control tissues and practical application in surgical pathology. Immunohistology of CEA has been used now for many years in research and in diagnostic applications. In this congress paper, the need for specificity checks, correct preservation and quality control of antibodies as well as the choice and preservation of control tissues will be discussed and, in addition, some aspects of the interpretation of CEA immunohistology in surgical pathology are illustrated, based upon prospectively collected data from a routine surgical pathology unit. An approach for collection and analysis is demonstrated and the diagnostic importance of CEA expression together with other antigens in the same tumor.


CEA serum levels in non-neoplastic disease. Several non-neoplastic conditions, i.e., acute and chronic inflammations, benign tumors, renal or hepatic insufficiency, are associated with elevated plasma levels of Carcinoembryonic Antigen (CEA). Usually, CEA elevation in these pathologies is less than 10 ng/ml. CEA values in non-malignant disease can be affected by many factors, which can be classified into five main categories: a) tissular, b) physiological, c) metabolic, d) circulating and e) methodological factors. Synthesis, expression and release are the most important tissular factors. Among the metabolic ones liver, biliary and renal functions play a primary role in the determination of CEA concentrations. In addition, other factors such as the presence of circulating CEA-like substances, autoantibodies and immunocomplexes or the characteristics of the assay method can influence the diagnostic value of the CEA test. All these factors must be carefully considered when the marker is used in clinical practice.


The role of serum carcinoembryonic antigen (CEA) in the management of patients with colorectal carcinoma: the experience of the Istituto Tumori of Milan. CEA determination has attained an important role in the clinical management of patients with tumors of the colorectal tract. In this paper the experience of the Istituto Tumori of Milan is reviewed and the results are discussed. Three hundred and thirty-six patients were followed after curative resection of colorectal carcinoma. The follow-up period was 15 years, from January 1975 to December 1990 (global follow-up 1358 years). In the course of follow-up 136 patients developed recurrent disease. The number of CEA determinations for each patient ranged from 1 to 37 (mean 8, total 3330). CEA levels of presurgical patients were related to the clinical stage. Among patients who developed recurrences 61% showed an increase in CEA serum levels. In 200 patients with a negative follow-up we observed only 15 cases of false-positive results.


CEA and NCA expressed by colon carcinoma cells affect their interaction with and lysability by activated lymphocytes. Heterogeneous lysability by interleukin-2 activated lymphocytes (LAK) and other immune effectors was observed in the human colon-carcinoma lines LoVo/Dx, LoVo/H and HT29. The tumor cells with high susceptibility to LAK (LoVo/Dx, HT29) expressed higher amounts of the adhesion molecules ICAMl, LFA3 and NCA/CEA than cells with low LAK sensitivity (LoVo/H). Monoclonal antibodies against these molecules caused a marked reduction of lysis by LAK of LoVo/Dx and HT29. A pool of these antibodies induced a nearly complete inhibition of the LAK lysis of both lines. Treatment of LoVo/Dx with differentiating agents (dimethylformamide and retinoic acid) led to a decreased expression of the adhesion molecules, including NCA, accompanied by increased resistance to LAK-mediated lysis. Moreover, the presence of CEA soluble antigen drastically inhibited the cytotoxic activity of LAK effectors against HT29 and LoVo/Dx cells, in a dose-dependent manner. These data indicate that sensitivity of colon-carcinoma cells to activated lymphocytes depends on the level of expression of adhesion molecules, including CEA and NCA. Given the role of CEA-related antigens in tumor/lymphocyte interaction, soluble CEA, frequently released by colon-carcinoma, may be involved in immunosuppressive effects induced in vivo by tumor cells.


External quality assurance of the carcinoembryonic antigen (CEA) assay: main findings in six years' experience. In 1984 we initiated a national external quality assessment (EQA) program (supported by the Italian National Research Council, CNR) for the CEA assay; at present, about 200 Italian laboratories are participating in the program. The laboratories assayed the quality control (QC) samples according to their routine procedures and returned the results together with the name of the method/kit they used. The collected results were computer-processed and reports were sent back to the participants. A significant reduction of the CVt (mean between-laboratory agreement) of the CEA assay was observed throughout the EQA survey (from 35% in 1985 to 20-25% in the last cycles). In order to better clarify the differences in variability observed in the first QC cycles against the last ones, we used the ANOVA technique to evaluate the components of variability. The improvement in between-laboratory agreement was mainly due to the reduction of the between-kit component (from 30.5% to 15.2%), rather than to the smaller decrease observed for the within-kit variability (from 18.4% to 14.0%). The results reported for QC samples from different materials showed differences in the between-lab variability and substantial changes of the kit biases, thus suggesting a different specificity of the antibodies used in the various method/kits against different families of CEA molecules. Considerable uncertainty was also encountered in the clinical classification of low pathological samples, which seems mainly due to the variability in cut-off values used by the laboratories for the clinical assessment of the same analytical results. Our data indicate a progressive increase in the reliability of CEA determination during our study and confirm that EQA has improved the reliability of analysis carried out by the participating laboratories, thus stimulating the kit manufacturers to provide more reliable products.


Cost/effectiveness ratio of carcinoembryonic antigen--importance of adequacy of routine requests of tumor markers. Since 1987 we have been evaluating the cost/effectiveness ratio of tumor markers using carcinoembryonic antigen (CEA) as a leading indicator. Preliminary to the evaluation of cost/effectiveness ratio we verified the fitness of CEA requests to the proper clinical problems in order to identify any bias of cost due to inadequate CEA use. 2677 CEA orders were evaluated in 1987. The percentage of inadequate requests was very high (43%). Therefore, it seemed not advisable to carry out the evaluation of cost/effectiveness ratio, while educational actions (divulgation of informative material, service of telephone consultation) were addressed to the physicians of the geographic area of laboratory users. In 1991 the adequacy of CEA requests was reevaluated. The percentage of inadequate requests on 2647 orders was 29.4%. This result, although not yet satisfactory, suggests that proper educational programs may probably improve the fitness of tumor marker requests to correct clinical problems. Additional educational actions are mandatory to further reduce the rate of inadequate tumor marker orders.


Effectiveness of 6-month intermittent administration of natural human interferon-alpha against non-A non-B chronic hepatitis. Interferon (IFN) was administered intermittently for 6 months to the patients with non-A non-B chronic hepatitis (CHNANB), and the effectiveness of the treatment for improving the hepatic function was evaluated. Of 26 patients with CHNANB, 16 received intermittent IFN therapy (IFN group), and 10 were treated by conventional therapies without IFN (non-IFN group). All patients were observed for 1 year. IFN was administered once a day at 3 MU in principle (1 MU in some patients and 6 MU in 1 patient) daily for 1 week immediately after the beginning of the therapy and 3 times a week for the subsequent 6 months at the outpatient clinic. The patients were followed up for at least 6 months after completion of the treatment. In the IFN group, the serum GPT level normalized in 11 (68.8%) of the 16 patients 1 year after the beginning of the treatment. In these 11 patients (normalized group), HCV-RNA was negative or became negative in 3 of the 6 patients in whom the serum HCV-RNA could be examined. Histological grades of inflammation in the liver were also markedly alleviated in the normalized group. The hepatic function did not normalize in any of the 10 patients in the non-IFN group. These findings indicate that IFN therapy is useful for CHNANB.


Cancer imaging with CEA antibodies: historical and current perspectives. This article reviews the history and status of cancer imaging with radiolabeled antibodies against carcinoembryonic antigen (CEA). Although CEA and many other cancer-associated antigens are not distinct for neoplasia, the quantitative increase of these markers in malignant tissues provides a sufficient differential for selective antibody targeting. Animal studies with xenografted human tumors provided the first evidence of the prospects of this technology, followed by initial clinical success with purified goat whole IgG antibodies to CEA, labeled with 131I and with the use of dual-isotope subtraction methods. Subsequently, improved and earlier imaging could be accomplished with monoclonal antibody fragments, which then would permit the use of shorter-lived radionuclides, such as 111In, 123I, and 99mTc. The preferred use of a monoclonal anti-CEA IgG Fab' fragment, labeled with 99mTc by a recently developed, simple and rapid kit, has enabled the detection of small lesions, including those in the liver, within 4 h of injection. By means of SPECT imaging, a high sensitivity and specificity for RAID could be achieved.


[A study of enhancing effects of monocyte culture supernatant on polymorphonuclear neutrophils-chemiluminescence--comparison between normal controls and collagen disease patients]. Priming effect of monocyte culture supernatant on polymorphonuclear neutrophils (PMN)-chemiluminescence (CL) was studied in patients with systemic lupus erythematosus (n = 11) and mixed connective tissue disease (n = 4). In normal controls, N-formyl-methionyl-leucyl-phenylalanine (FMLP) induced PMN-CL was enhanced when PMN were previously incubated for 15 minutes with monocyte culture supernatant (MS) or T lymphocyte culture supernatant (TS) or T lymphocyte culture supernatant (TS). The enhancing effect of MS on PMN-CL was greater than that of TS. This enhancing effect of MS was inhibited by adding of dexamethasone (1 microgram/ml) during the culture. Recombinant human TNF also enhanced PMN-CL as well as MS. When compared the enhancing effects of MS between patients and normal controls, that of patients under corticosteroid therapy (average prednisolone dose 39.5 mg/day) was reduced significantly. Thus, we concluded that the cytokines from monocyte contributed PMN phagocytosis of invading microorganisms, and that this monocyte-mediated PMN phagocytosis was suppressed partly by corticosteroids in collagen disease.


Pilot, multicenter and prospective trials with an anti-CEA antibody. In this paper we summarize the investigations performed by our group utilizing an anti-CEA monoclonal antibody (F023C5) labelled with different radionuclides in humans. Since 1983 radioimmunoscintigraphy (RIS) was performed on 51 patients with 64 localizations of colo-rectal carcinoma (pilot study). A multicenter clinical trial in a large number of patients (509 pts of which 284 with gastrointestinal cancer) was subsequently carried out in collaboration with ten nuclear medicine centres. High sensitivity and specificity values were obtained by these studies and many unsuspected lesions were recorded. In order to better define the clinical role of RIS, a prospective study was performed on 59 patients with suspected local relapses of colo-rectal cancer. A comparative evaluation of RIS, CT scan, US and MRI was done. RIS and MRI had the highest accuracy (86%) followed by CT scan (68%) and US (54%).


Diagnostic applications and therapeutic approaches with different preparations of anti-CEA antibodies. Various preparations of anti-CEA antibodies have shown to detect very sensitively CEA producing tumors. The development of human anti-mouse antibodies (HAMA) prevents from the widespread use of immunoscintigraphy (IS) in the follow-up of patients with colorectal carcinoma. It is, however, not yet clear if genetically reshaped antibodies will solve this problem and it is even less clear if this will change prognosis of these patients. There is certainly room for new therapeutic approaches in colorectal carcinoma. Radioimmunotherapy in combination with other techniques might advantageously complete surgery, chemotherapy and radiotherapy, but needs to be further developed.


CEA determination in the follow-up of extracolorectal neoplasms. CEA was initially described as a tumor and organ specific colorectal antigen, but later found by more sensitive methods in other tumors (stomach, pancreas, lung, breast) and in minor amounts in inflammatory, normal adult and fetal organs of the gastrointestinal tract. The main clinical application of CEA concerns its pretherapeutic and serial determination as circulating antigen in serum and other body fluids by means of CEA-specific, commercially available test kits. By clinical studies a significant correlation has been proven between the pretherapeutic serum CEA level and tumor stages and prognosis. Moreover, serial CEA level changes have been shown a valuable monitor following operation or during radio/chemotherapy anticipating and reflecting the clinical course of disease. In combination with newly established tumor markers, the main clinical indication for CEA determination in addition to colorectal cancer concerns monitoring of patients with stomach (+CA 72-4), lung (+NSE/SCC) and breast cancer (+CA 15-3/MCA).


[A study of nasal carriage of methicillin-resistant Staphylococcus aureus]. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important microorganisms in nosocomial infection. The Hospital staff working in MRSA endemic wards are known to have MRSA in their nasal cavity. The nasal carriage of MRSA was detected in staff members and patients of two hospitals. In Niigata University Hospital, 10 out of 109 nurses and 8 out of 142 doctors were found to be MRSA carriers. On the other hand, in Nagaoka Red Cross Hospital, 25 out of 448 nurses were found to be MRSA carriers, however, no carrier was found in 23 doctors. These strains were also resistant to MCIPC, IPM, TFLX and OFLX, whereas they remained sensitive to VCM. The coagulase types of MRSA isolated from the hospital staff and patients were II, IV and VII, although those of Methicillin-sensitive S. aureus (MSSA) consisted of all types. Elimination of nasal MRSA from the carriers was considered for avoiding hospital outbreaks caused by this potential pathogen. Forty hospital staff and 19 patients, in who's MRSA was found persisting in their nasal cavity, were treated by povidone iodine and chloramphenicol (CP) MRSA disappeared in 44% and 84% of the nasal carriers by povidone iodine and CP, respectively.


Anti-CEA and other antibodies in the study of gastrointestinal tumors. Localization of gastrointestinal tumors by means of labeled monoclonal antibodies is a new, sensitive and suitable technique currently used in several centers. Encouraging results have been documented with several monoclonal antibodies by different authors. This article reviews our experience with radioimmunoscintigraphy in 59 patients with colorectal cancer in follow-up, using 131I and 111In labeled B72.3, and in 16 patients with primary gastrointestinal tumors using 99mTc anti-CEA monoclonal antibody (type F023C5). The sensitivity of both B72.3 and anti-CEA was greater than 70% either for primary tumors and abdominal recurrences or distant metastases except hepatic ones. A significant gradient in antibody uptake was measured on surgical biopsies between tumors and normal tissues allowing a good in vivo contrast for gamma detection. We have defined the impact of some factors affecting in vivo tumor targeting. In fact, pharmacodynamics of MAbs, percentage of injected dose bound to tissues were measured, and in particular antigenic content in tumor nodules was quantified. Furthermore, the results of RIS were compared to those obtained by CT and other imaging modalities.


[Study on lymphocyte-response in early stage of respiratory infection--a view point from experimental Klebsiella pneumoniae pneumonia]. This study was performed to demonstrate the early immuno-responses of lymphocytes in an experimental pneumonia with K. pneumoniae in mature mice (45 week-old) comparing with it in juvenile ones (4 week-old). Acute mice pneumonia was made by inhalation with K. pneumoniae DT-S strains into lung. Changes in lymphocytes including their subpopulation in bronchoalveolar lavage fluid (BALF), peripheral blood, hilar lymphnodes and lung tissue were observed after the inhalation. In addition, lung tissue and hilar lymphnode were examined immunohistologically. The following results were obtained: 1. Total lymphocytes in BALF were more rapidly increased in the mature group than in the juvenile one. But there was no significant change in leukocyte count in peripheral blood between both groups. 2. Such a rapid increase in lymphocytes in BALF in mature group depended on L3T4-Ly6c cells and L3T4-LFA 1 cells. These cells in juvenile group were not accumulated in BALF at initial phase of the infection. But in contrast, they were gradually increased in peripheral blood and in hilar lymphnode. There was significant time-differences in appearance of these cells in BALF between both groups. It might be, that L3T4-Ly6c cells and L3T4-LFA1 cells observed in BALF in mature animals were induced from bronchus-associated lymphoid tissue (BALT) or small lymph tissue in alveolar interstitium, but in juvenile ones were originated in hilar lymphnode. 3. Changes in Ly2-Ly6c cell and Ly2-LFA1 cell were shown the same tendency changes in L3T4-Ly6c cell and L3T4-LFA1 cell. 4. Accumulation of B220:Ly5-LFA1 cell in BALF was not observed in significant difference between the mature group and in the juvenile one. 5. L3T4 cells were markedly accumulation in subcortex area of hilar lymphnode in the juvenile group, but they were only seen scattered in the mature group. 6. It can be concluded that active T lymphocyte begins to response in situ in the early stage of respiratory infection in the mature host and this finding is a characteristic lymphocyte response, that is never seen in the juvenile group.


Application of genetically-engineered anti-CEA antibodies for potential immunotherapy of colorectal cancer. Hitherto anti-CEA monoclonal antibodies (MAbs), normally of mouse origin, have been used primarily for clinical diagnosis of colorectal cancer, either as a tumor marker in serum to monitor tumor recurrence, or latterly as a means to localize in vivo CEA-bearing tumors, and metastases in patients. In vivo diagnosis using mouse anti-CEA MAbs has so far had limited clinical utility because the antibodies elicit a strong anti-mouse immunoglobulin immune response on repeated administration in man. This problem has been addressed by the development of various strategies for "humanization" of mouse anti-CEA MAbs by genetic manipulation of immunoglobulin genes. Such humanized, engineered antibodies markedly attenuate the antigenic response directed against the MAb, such that safe, repeated administration to patients has become feasible. Such humanized anti-CEA antibodies can thus be radioactively-labelled and applied for in vivo monitoring and detection of recurrent malignant disease, or used for therapeutic strategies which similarly take advantage of the ability of the antibodies to target cytotoxic agents selectively to tumor cells. The application of these novel procedures for manipulating MAb structure presents entirely new opportunities for diagnosis and treatment of human colorectal cancer.


[Biochemical analysis of lipopolysaccharides from respiratory pathogenic Branhamella catarrhalis strains and the role of anti-LPS antibodies in Branhamella respiratory infections]. We characterized lipopolysaccharides (LPSs) from respiratory pathogenic Branhamella catarrhalis (BC) strains, and evaluated the protective property of anti-BC LPS antibody in BC respiratory infections. LPSs from four strains of BC were lipooligosaccharide having no O-side chain and a M(r) of 3 KDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). All of them produced different patterns, showing two to four bands on SDS-PAGE. We found high level of anti-BC IgG antibody in convalescent sera from a patient with BC respiratory tract infection by ELISA. This IgG antibody recognized BC LPSs on Western blots. Two respiratory pathogens of BC (strains; 87-122, 88-23) were tested in a bactericidal assay employing a convalescent sera. 87-122 strain was susceptible to antibody-dependent, complement-mediated killing, while 88-23 strain was resistant. The killing of 87-122 strain was inhibited by addition of the homologous BC LPS to the convalescent sera in a dose-dependent manner. These data support that anti-BC LPS antibody may mediate complement-lysis of some strains of BC, and play a protective role in BC respiratory infections.


[Antigen detection in the murine models of systemic candidiasis]. We compared two latex agglutination tests for serum candidal antigen in murine models of systemic infection and gastrointestinal colonization. In the intravascular-infection model, mannan detection test was positive at 2 hours after inoculation, but Cand-Tec was less sensitive than mannan detection test in this model. In the gastrointestinal colonization model followed by systemic infection in neutropenic mice, the mannan antigen test became positive after 3 weeks of colonization with increased number of Candida of the stool, and Cand-Tec became positive after 5 weeks in neutropenic mice.


[Study on the bacteriological examination of sputum and bronchoscopy specimens from 31 cases with pneumonia due to Chlamydia psittaci]. We carried out the bacteriological examination of sputum and bronchoscopy specimens from 31 cases with pneumonia due to C. psittaci. The results obtained were as follows: 1. The positive culture of sputum and bronchoscopy specimens were 38.7% (12/31). 2. The organisms detected from them were 13 strains of gram-negative bacilli, 2 of gram-positive cocci and one gram-positive bacillus. 3. Significant differences were observed in the white blood cell count between the cases of positive culture and those of normal upper respiratory tract flora (p less than 0.05). From the results we conclude that it would be better that we add the proper antimicrobial drugs to chlamydial antibiotics in the treatment of patients with leukocytosis.


[Two cases of diffuse panbronchiolitis receiving long-term erythromycin (EM) therapy with acute exacerbation due to EM-resistant pneumococcus]. The first case was a 73-year-old woman with chief complaints of fever, cough, purulent sputum and dyspnea. EM therapy was begun in December 1983 due to a diagnosis of diffuse panbronchiolitis (DPB). Subsequently, P. aeruginosa was persistently detected, while in February 1991 at the time of an acute exacerbation of the DPB P. aeruginosa and S. pneumoniae were detected by TTA. The second case was a 65-year-old man with chief complaints of fever, cough and purulent sputum. DPB was diagnosed and EM therapy was begun in December 1985. In January 1991, pneumonia developed, at the time when S. pneumoniae was detected by TTA. In both cases, rapid disappearance of S. pneumoniae from the sputum and alleviation of symptoms were obtained with carbapenem antibiotic administration. Both strains were resistant to EM, Tetracycline (TC), Minocycline (MINO) and Clindamycin (CLDM). Particularly, S. pneumoniae of case 2 showed low sensitivity to Ampicillin (ABPC), Cefotiam (CTM) and Cefoxitin (CFX) as well. These cases showed acute exacerbations due to EM-resistant pneumococcus during long-term therapy with EM, and are of interest in that they may shed light on the relation between long-term EM therapy and the emergence of resistant pneumococcus.


[Epidemiological and bacteriological study on gonococcal infections]. Epidemiological and bacteriological studies on Neisseria gonorrhoeae isolated in Sapporo, Japan, in 1980 and 1991 performed and the following results were obtained. 1. The range of age in the patients infected with Neisseria gonorrhoeae tended to be younger than those in the whole country. 2. Male patients in the early 20s or younger with gonococcal urethritis were often infected by bon-professional females but those in their late 20s or older were often infected from professional females, for example prostitutes and hostesses. 3. The rate of professional females who were positive to gonococci reached 17.4% and young females in their teens with cervicitis had the highest morbidity rate of gonococci than those in the older females. 4. The latent period in gonococcal infections tended to become longer gradually. 5. The isolation rate of penicillinase producing Neisseria gonorrhoeae (PPNG) showed a peak of 23.9% (61/255) in 1985, but gradually declined thereafter and it was 3.7% (1/27) in 1991. 6. An investigation on auxotype showed a decline of proto and Pro-strains and an increase of AHU-strains in non-PPNG. And most of the PPNG belonged to proto or Pro-strains. 7. With the relationship between auxotype and sensitivity to AMPC, AHU-strains were more sensitive than proto or Pro-strains.


[An outbreak of municipal water-associated food poisoning caused by Salmonella enteritidis]. An outbreak of municipal water-associated food poisoning by Salmonella Enteritidis occurred in Takatoh Town. Nagano prefecture during September 4 to 19, 1989. The major symptoms observed in the 680 patients consisted of diarrhea (70.9%), abdominal pain (51.2%), fever (44.6%), headache (27.9%), nausea (5.9%) and vomiting (5.7%). In the outbreak, S. Enteritidis was the only suspected etiological agent isolated from both patients and municipal water supply. All of the 21 isolates were lysine decarboxylase activity negative. In other respects, the isolates conformed to the general characteristics of Salmonella. All of them were further characterized by phagovar 8 type, uniform drug susceptibilities and carrying 2.7 kb plasmid. Inspite of the evidence that the infection caused by a peculiar strain linked to the strain in municipal water supply used by the patients, it was impossible to determine the exact source of contamination in the water. The outbreak above seems to be the most large-scale among those of Salmonella found poisoning associated with drinking water in Japan.


[Food poisoning caused by enteroinvasive Escherichia coli (O164:H-)--a case in which the causative agent was identified]. Food poisoning due to "Godofu (Sasayuki tofu)" as a main causative foodstuff which broke out on July 14, 1988. There were 670 out of 918 persons who ingested this food who became ill (incidence 73.0%). The main symptoms were diarrhea (93.4%), fever (77.5%), abdominal pain (64.5%), and vomiting (19.9%). A high degree of fever and watery diarrhea were characteristic of this poisoning. The average latent period was 35 hours with a range of one to 156. The O164:H- strains of enteroinvasive Escherichia coli (EIEC) were detected from 22 of the 32 fecal samples collected from the patients, five of ten samples collected from workers engaged in tofu making, and one sample of left-over Godofu. The virulence of EIEC strains isolated from the patients, workers, and left-over food was confirmed by invasion into HeLa and HEp-2 cells, Sereny test, and ELISA test to detect invasive plasmid-derived protein of the organism (conducted at Tokyo Metropolitan Research Laboratory of Public Health). These EIEC strains were sensitive (less than or equal to 0.19 to 6.25 micrograms/ml) to GM, ABPC, CBPC, CER, CET, NA, PB, MINO, TC and CP as well as KM and OFLX which were used for treatment. However, their susceptibility to FOM varied to some extent (6.25 to 25.0 micrograms/ml) and one strain isolated from a tofu worker was resistant to MINO, TC, FOM and CP (25 to greater than or equal to 100 micrograms/ml). Since investigation revealed that Godofu was left at room temperature about 29 degrees C until ingested, we did a experiment to check the bacterial growth in Godofu under similar conditions at the time of outbreak.(ABSTRACT TRUNCATED AT 250 WORDS)


[Mechanism of acquired resistance against Legionella--blastogenic responses of murine spleen lymphoid cells following the stimulation with Legionella antigens]. We have tried to characterize the blastogenic responses of murine spleen lymphoid cells from BALB/c mice immunized with Legionella pneumophila serogroup (SG) 1 (Philadelphia 1 strain) and non-treated mice. Lymphoid cells from immunized mice showed stronger blastogenic responses following stimulation with concanavalin A or formalin-treated L. pneumophila SG1 whole cell antigen than those showed by lymphoid cells from non-treated mice. These cells from immunized mice also responded strongly when stimulated in vitro with other SGs of L. pneumophila, while these responded weakly when stimulated with other species of Legionella. Serum antibody titers of immunized mice against each SG of L. pneumophila were examined and the cross reactions were also recognized. However, the relatedness of serum antibody titers and the blastogenic responses against each serogroup of L. pneumophila was small. The epitopes recognized by the cellular immunity might be different in part from those recognized by serum antibodies, and investigations should be made on what the cellular immunity recognizes and how it works.


[Detection of Chlamydia trachomatis in first-voided urine sediments from male urethritis by polymerase chain reaction]. Chlamydia trachomatis was detected from first-voided urine sediments of 97 male patients with urethritis by polymerase chain reaction (PCR). Since urine and urinary sediments only treated with proteinase K inhibited DNA amplification by PCR, DNA was further purified by phenol extraction and concentrated. Two oligonucleotides based on sequences within the major outer membrane gene from C. trachomatis serovar L2 were used as primers. A DNA fragment of 242 bp specific for C. trachomatis was amplified by PCR and detected by agarose gel electrophoresis. The DNA fragment was amplified by PCR in all specimens of urine sediments from 50 patients with Chlamydiazyme-positive urethral swab. In 38 specimens of urine sediments from 47 patients with Chlamydiazyme-negative urethral swab, PCR was negative. The overall coincidence rate between the PCR for detecting C. trachomatis in first-voided urine sediments and Clamydiazyme in urethral swab was 90.7% (88/97). Detection of C. trachomatis from first-voided urine sediments by PCR was considered to be noninvasive and useful for the diagnosis of male urethritis due to C. trachomatis.


[Dose finding study of cefepime for chronic respiratory infections]. In order to determine the optimal dose of cefepime (CFPM) for respiratory tract infections, a dose finding study was conducted in patients with chronic respiratory tract infections, and the clinical properties of the drugs were compared with those of ceftazidime (CAZ). Each drug was administered by intravenous drip infusion at the dose of 2 g/day of CFPM, 4 g/day of CFPM or 2 g/day of CAZ for 14 days. 1. The overall efficacy rates evaluated by the committee were 82.6% (19/23) for the CFPM 2 g/day group, 85.0% (17/20) for the 4 g/day group and 79.3% (23/29) for the CAZ 2 g/day group, with no statistically significant difference among the three groups. 2. The overall efficacy rates evaluated by the attending physicians were 91.3% (21/23) for the CFPM 2 g/day group, 78.9% (15/19) for the CFPM 4 g/day group and 75.9% (22/29) for the CAZ 2 g/day group, with no significant difference among the three groups. 3. Bacteriological eradication rates were 88.2% (15/17) for the CFPM 2 g/day group, 68.8% (11/16) for the CFPM 4 g/day group and 63.2% (12/19) for the CAZ 2 g/day group, with no statistically significant difference among the three groups. 4. The incidences of adverse reactions were 3.8% (1/26) for the CFPM 2 g/day group, 9.1% (2/22) for the CFPM 4 g/day group and 3.4% (1/29) for the CAZ 2 g/day group, with no significant difference among the three groups. The incidences of abnormal laboratory findings were 37.5% (9/24) for the CFPM 2 g/day group, 15.0% (3/20) for the CFPM 4 g/day group and 3.4% (1/29) for the CAZ 2 g/day group. There was a significant difference among the three groups. 5. Utility rates assessed by the committee were 81.8% (18/22) for the CFPM 2 g/day group, 76.2% (16/21) for the CFPM 4 g/day group and 75.9% (22/29) for the CAZ 2 g/day group. Utility rates assessed by attending physicians were 90.9% (20/22), 78.9% (15/19) and 72.4% (21/29), respectively. There was no significant difference among the three groups. From the above results, it is concluded that the optimal dosage of CFPM is 2 g/day for chronic respiratory tract infections.


[A comparative study of cefepime for bacterial pneumonia]. The efficacy, safety and usefulness of cefepime (CFPM), a new cephem antibiotic, in bacterial pneumonia, were evaluated in a comparative study against ceftazidime (CAZ). Each drug was administered by intravenous drip infusion at a dose of 1.0 g (nominal potency) twice daily for 14 days, and the following results were obtained. 1. A total of 183 cases were enrolled in this study. Efficacy rates ("good" or better responses) as evaluated by the subcommittee were 90.3% (65/72) in the CFPM group and 94.0% (63/67) in the CAZ group, with no significant difference between the 2 groups. 2. Efficacy rates ("good" or better responses), as evaluated by attending physicians, (in the same bacterial pneumonia cases which were subjected to evaluation by the subcommittee) were 87.5% (63/72) in the CFPM group and 89.6% (60/67) in the CAZ group, with no significant difference between the 2 groups. 3. Bacteriologically, eradication rates were 96.9% (31/32) in the CFPM group and 96.7% (29/30) in the CAZ group with no significant difference between the 2 groups. 4. The incidence of side effects was 5.9% (5/85) in the CFPM group and 4.8% (4/84) in the CAZ group, with no significant difference between the 2 groups. No significant difference was also found between the 2 groups in the incidence of abnormal laboratory findings; 28.4% (23/81) of the case in the CFPM group and 34.1% (28/82) in the CAZ group. 5. As for overall usefulness of the drug in bacterial pneumonia cases, utility rates ("useful" or better evaluations) as evaluated by the subcommittee were 88.9% (64/72) in the CFPM group and 92.5% (62/67) in the CAZ group. The rates as evaluated by investigators (in cases judged as evaluable by the subcommittee) were 87.5% (63/72) and 85.1% (57/67), respectively. There were no significant differences between the 2 groups. These results indicated that CFPM is very useful for the treatment of bacterial pneumonia.


[A comparative study of cefepime for chronic respiratory tract infections]. The clinical efficacy, safety and usefulness of Cefepime (CFPM), a new cephem antibiotics, in chronic respiratory infections were evaluated in a comparative study against Ceftazidime (CAZ). Each drug was administered by intravenous drip infusion at a dose of 1.0 g (nominal potency), twice daily for 14 days, and the following results were obtained: 1. A total of 170 cases were enrolled in this study. Efficacy rates ("good" or better responses) as evaluated by the subcommittee were 86.2% (56/65) in the CFPM group and 84.5% (60/71) in the CAZ group, with no significant difference between the two groups. 2. Efficacy rates ("good" or better responses) as evaluated by attending physicians were 83.3% (55/66) in the CFPM group and 84.5% (60/71) in the CAZ group with no significant difference between the two groups. 3. Bacteriologically, eradication rates were 83.3% (40/48) in the CFPM group and 88.2% (45/51) in the CAZ group, with no significant difference between the two groups. 4. Side effects occurred in none of the patients in the CAZ group and in 4 of the 66 patients in the CFPM group. There was a significant difference between the two groups (Fisher's test p = 0.0489). The incidence of abnormal laboratory findings were 17.6% (12/68) in the CFPM group and 21.1% (16/76) in the CAZ group. There was no significant difference between the two groups. 5. The utility rates evaluated by the subcommittee were 81.8% (54/66) in the CFPM group and 84.5% (60/71) in the CAZ group with no significant difference between the two groups. Only in the incidence of side effects, there was a significant difference between the two groups (Fisher's test p = 0.0489), but there was no significant difference in other items of efficacy, safety and usefulness between the two groups. These results indicate that CFPM is useful for the treatment of chronic respiratory tract infections.


[Growth inhibitory activity of clinical isolates of Pseudomonas aeruginosa against Staphylococcus aureus (MRSA and MSSA)]. Anti staphylococcal activity by clinical isolates of Pseudomonas aeruginosa was tested by the reversed agar plate and the filter paper stamp methods. Almost 40% of Pseudomonas aeruginosa inhibited the growth of both Methicillin resistant Staphylococcus aureus (MRSA) and Methicillin sensitive Staphylococcus aureus (MSSA). Green pigment (Pyocyanin) produced strains showed a strong inhibitory effect against MRSA and MSSA respectively. But some other pigment (Yellow, Red) strains also showed anti staphylococcal activity. These data suggest the colonization of Pseudomonas aeruginosa with anti staphylococcal activity may not be eradicated by the anti pseudomonic antibiotics.


[Effect of gentiana violet against methicillin-resistant Staphylococcus aureus (MRSA)]. The bactericidal effect of gentiana violet against MRSA isolated from clinical specimens was studied both in vitro and in vivo. The results obtained are as follows: 1) Minimum bactericidal concentration (MBC) of gentiana violet to MRSA was between 0.0025% and 0.08% and the MBC was not influenced even if 25% human whole serum exists in the medium. 2) The number of cells were 2.1 x 10(7) CFU/ml in medium which decreased to 5.4 x 10(4) CFU/ml within 5 min by existing 0.1% gentiana violet in the medium as the final concentration, and also decreased under 10(3) CFU/ml 15 min later. 3) Minimum inhibitory concentration (MIC) of gentiana violet to MRSA was between 0.00015% and 0.00063%, and the inhibitory activity was not influenced even if gentiana violet was incubated with the bacteria in the medium for 72 hr at 37 degrees C. 4) By using an ointment containing 0.1% gentiana violet to 12 cases of patients with the MRSA infected skin lesions, MRSA was eliminated completely from the infected areas of the skin within 4 weeks. 5) The side effects of gentiana violet were not observed in all cases during the use of the ointment containing gentiana violet. It is suggested that gentiana violet may be one of the useful drugs for the treatment of the skin lesions infected with MRSA.


[Clinical studies on the treatment of Campylobacter enteritis--emergence of quinolone-resistant Campylobacter jejuni after treatment with new quinolones]. In recent years, new quinolones such as ofloxacin (OFLX) and tosufloxacin (TFLX) have been frequently used in the treatment of bacterial enteritis caused by unknown organisms. The agent of first choice for the treatment of Campylobacter enteritis is one of the macrolides, but new quinolones are often administered accidentally to adult patients with Campylobacter enteritis. We have detected quinolone-resistant strains of Campylobacter jejuni (C. jejuni) after the treatment of some patients with new quinolones, and accordingly we reviewed the treatment of Campylobacter enteritis. We experienced 178 adult patients with Campylobacter enteritis from January 1989 to November 1991 at our hospital. From them, we selected 52 patients (32 males and 20 females) in whom stool culture were performed both before and after treatment. The initially administered antimicrobial agent was a macrolide (rokitamycin, RKM) in 6 cases, a new quinolone in 22 cases, and kanamycin (KM) in 24 cases. The new quinolone used was OFLX in 17 cases, TFLX in 3 cases, and norfloxacin (NFLX) in 2 cases. Fifty-one of the 52 C. jejuni strains isolated before treatment were susceptible to OFLX and erythromycin (EM) according to antimicrobial disc susceptibility tests. C. jejuni was eradicated in all patients treated with RKM or KM. However, treatment failed to achieve bacteriological cure in 8/22 (36.4%) patients given new quinolones. In these patients, the strains of C. jejuni isolated before treatment were susceptible to OFLX, but the strains isolated after treatment were all resistant to OFLX according to disc susceptibility tests.(ABSTRACT TRUNCATED AT 250 WORDS)


[Shigella dysenteriae strains having a provisional serovar isolated from imported diarrheal cases in Tokyo]. Two bacterial strains (ME448 and ME474) isolated from stool cultures of imported cases in Tokyo in 1987 had typical biochemical characteristics of Shigella dysenteriae. The results of antigenic analyses showed that they were serologically identical to each other, but did not belong to any of the established Shigella serovars. These strains were positive for Serény test in guinea pig eye and cell-invasion test in HeLa cells. The strains also had virulence-plasmid encoding outer membrane proteins, indicating that they were pathogenic. We then contacted the Centers for Disease Control in the United States and the Central Public Health Laboratory in the United Kingdom and arranged serological examinations of the strain ME448. From the results, the strain was confirmed to have provisional S. dysenteriae serovar E23507. Although the serovar had been isolated from a Swedish patient who developed diarrhea while in India, this is thought to be the first report of its isolation in Japan.


[Clinical study of eradicated and resistant patients to treatment with ivermectin for strongyloidiasis]. We treated 70 patients with strongyloidiasis (41 males and 29 females) with ivermectin (IVM), and obtained the following results: 1. The eradication rates at 1-2 months, 3-4 months and 5-6 months after treatment were 90.7% (49 of 54 patients), 100.0% (47 of 47 patients) and 95.7% (45 of 47 patients), responsively. Twelve patients were resistant (non-responsive) to treatment. 2. When compared to patients whose parasites were completely eradicated, the resistant patients showed the following results: 1) Incidence of symptoms observed before treatment was significantly lower (50.0% vs. 84.5%). 2) Positive rate of anti-HTLV-I antibody was significantly higher (66.7% vs. 20.7%). 3) Blood eosinophil counts before treatment were significantly lower (266.6 +/- 117.2/mm3 vs. 533.2 +/- 429.7/mm3). 4) Serum IgE levels before treatment were significantly lower (217.2 +/- 442.9 IU/ml vs. 1,076.8 +/- 2,108.0 IU/ml). 5) There were no significant differences in age, sex and dosage of ivermectin. 3. Comparing anti-HTLV-I antibody positive and negative patients, the following results were obtained: 1) Eradicated patients; a) Eosinophils and IgE levels before and after the first administration of medicine in anti-HTLV-I antibody positive patients were significantly lower than those of negative patients. b) Gammaglobulin levels before treatment and after both administrations of the drug, IgG before therapy and OKT4/OKT8 after therapy were significantly higher than in anti-HTLV-I antibody positive patients. 2) Resistant patients; Eosinophils after treatment were significantly lower in anti-HTLV-I antibody negative patients than in positive patients.(ABSTRACT TRUNCATED AT 250 WORDS)


[Rapid diagnosis of influenza infection by PCR method--detection of influenza virus HA gene in throat swab]. We studied the detection of the HA gene of human influenza viruses in throat swabs obtained from the outbreaks of influenza in school children utilizing the polymerase chain reaction (PCR) method. Sensitivity and specificity of the PCR method was compared to conventional virus isolation using MDCK cells. Three pairs of primers for PCR in detecting the HA genes of AH1, AH3, and B influenza viruses showed both subtype and type specificity. The dilution experiments showed that influenza viruses, as few as 1.1-3.5 plaque-forming units per 50 microliters, were sufficient for the detection of HA genes by PCR method and the detection rate by PCR method was 2-3 fold higher than that by conventional method. Our results showed that the PCR method was a fast, sensitive and reliable method for the diagnosis of influenza infections.


[Isolation of vero-cytotoxin-producing Escherichia coli from cattle and serotyping and toxin-typing of the isolated strains]. Two hundred and sixty-six piglets with diarrhea (from 4 farms), 73 healthy pregnant pig (from 2 farms), 27 calves with diarrhea (from 9 farms) and 47 healthy milk cows (from 1 farm) were examined for Vero-cytotoxin-producing Escherichia coli (VTEC), and 52, 11, 15 and 67 strains of VTEC were isolated from 17 piglets, 11 pregnant pigs, 6 calves and 23 milk cows, respectively. All VTEC strains from the piglets produced only VT2vp, while the strains from the healthy pigs did not produce VT2vp, but did VT1 and/or VT2. Most VTEC strains from calves and cows produced VT2vhb and some produced VT2 and VT1. Serotyping of the isolated strains showed that many strains from the piglets belonged either O139:H1, O141:H4 or O141:HUT, but the strains from the pigs were either R-form or O-untypable. Many strains from the calves and cows were serotyped into O116 or O113, but there were several R-form and O-untypable. From these results, it is suggested that VTEC strains, especially from the pregnant pigs, calves with diarrhea and healthy milk cows, which produced the same type of Verotoxins to that produced by human isolates, may become sources of human infections.


[Dynamic process of colonization by Haemophilus influenzae in children--is H. influenzae normal flora in the throat?]. Eighty-seven oropharyngeal isolates of Haemophilus influenzae were obtained by two time cultures six months apart from a total of 288 children who attend a kindergarten. We analyzed the strains by comparing their serotypes, biotypes, beta-lactamase activity and by performing electrophoresis of outer membrane proteins on polyacrylamide gels. Only nineteen strains were not identical, the rest of the 68 strains were classified into 23 types. During 6 months at least 15 types of strains lost from this group and 21 types of new strains were obtained. There were no children who had identical pairs of H. influenzae in their oropharynx during the 6 months. The classification of strains in oropharynx suggested that person-to-person transmission of nontypable H. influenzae can occur. We concluded that oropharyngeal colonization by nontypable H. influenzae is not a normal flora in children.


[Study on local immune response in Escherichia coli-induced experimental urinary tract infection in mice--infiltration of Ia-positive cells, macrophages, neutrophils, T cells and B cells]. We studied the local immune response in a mouse experiment with acute ascending cystitis and pyelonephritis. The experimental infections were induced in BALB/c female mice by transurethral instillation of Escherichia coli O6. Immune response cells were stained, including Ia-positive cells, macrophages, neutrophils, T cells (CD4+ and CD8+) and B cells (IgA, IgM, IgG-positive B cell). They were stained by the immunohistochemical method (ABC method) using monoclonal antibodies against lineage specific antigens except for neutrophils that were readily identified by the standard hematoxylin-eosin. Even in the control mice having no evidence of the infection, mucosa associated lymphoid tissue (MALT) consisted of macrophages, Ia-positive cells and T cells that were sparingly found in the urinary tract tissue and renal parenchyma. Ia-positive cells, macrophages, neutrophils, T cells (CD4+, CD8+) and IgA positive B cells were significantly infiltrated in the bladder submucosa from 6 hours after bacterial inoculation. The infiltration of similar immune response cells was found in the submucosa of the renal pelvis, except for IgA positive B cells that appeared one day after the induction of the infection. In renal parenchyma, Ia-positive cells appeared at 6 hours after introduction of the infection, followed by an infiltration of neutrophils, macrophages and T cells (CD4+, CD8+) at the first day, and IgA positive B cells at the third day. These results are summarized as follows. When microbes invaded the urinary tract tissue, a significant number of Ia-positive cells infiltrated, which were initially present in normal urinary tract tissue. Subsequently, neutrophils, macrophages and T cells (CD4+, CD8+) appeared in the lesion followed by a delayed occurrence of IgA positive B cells.


[A case of pneumonia due to Mycoplasma pneumoniae accompanying high adenosine deaminase activity in pleural effusion]. We experienced a 5-year-old male case of Mycoplasma pneumoniae pneumonia accompanying Adenosine Deaminase (ADA) activity in pleural effusion. Chest roentgenograms revealed the infiltration in the left upper lung field and the left pleural effusion. In serum, the M. pneumoniae CF titer increased to 1:512. The pleural effusion was yellowish in color, with a specific gravity of 1.030, protein 3.7 g/dl, glucose 101 g/dl, and ADA 50 IU/l. Pleural effusion accompanying M. pneumoniae pneumonia is rare, and the high ADA activity in this case has been reported only in one other case. This is a report of a high activity of ADA in the pleural fluid by M. pneumoniae pneumonia.


[A case of eosinophilic meningitis due to Angiostrongylus cantonensis]. We report a case of eosinophilic meningitis caused by Angiostrongylus cantonensis. This patient, a 50-year-old male, had been eating uncooked slugs for 40 years. His chief complaints on admission were headache, fever and general fatigue. Neurological examination and CT findings were normal, but the CSF contained increased cells, most of which were eosinophilic cells. The presence of eosinophilic cells in the CSF is by itself abnormal. We therefore suspected eosinophilic meningitis and performed immunological tests. Since the gelatin particle method and immunological antigen antibody reaction were positive. We diagnosed the patient as having eosinophilic meningitis caused by Angiostrongylus cantonensis. There are only 27 reported cases of this disease in Japan, and most of them have been reported in Okinawa-Prefecture.


[Detection of Neisseria gonorrhoeae in first-voided urine sediments from male urethritis patients by polymerase chain reaction]. Neisseria gonorrhoeae was detected from first-voided urine sediments of male patients with urethritis by polymerase chain reaction (PCR). Urine and urinary sediment were treated with proteinase K, and DNA was further purified by phenol extraction. Two oligonucleotides based on sequences within a ribosomal RNA gene from N. gonorrhoeae were used as primers for the PCR. A DNA fragment of 206 bp specific for N. gonorrhoeae was amplified by PCR and detected by agarose gel electrophoresis. In 19 specimens of urine sediments collected from 21 patients in whom N. gonorrhoeae was isolated from urethral swab by culture, 206 bp DNA fragment was amplified by PCR. In all specimens of urine sediments from 24 patients in whom cultures for N. gonorrhoeae were negative, no DNA was amplified by the PCR. The overall coincidence rate between the PCR for detecting N. gonorrhoeae in first-voided urine sediments and culture in urethral swab was 95.6% (43/45). PCR procedure for detection of pathogens from first-voided urine sediments would be noninvasive and would be applied for the diagnosis of gonococcal urethritis and chlamydial urethritis.


A clinical study on patients with urinary tract infection due to coagulase-negative staphylococci. To assess the clinical significance of coagulase-negative staphylococci (CNS) in patients with urinary tract infection (UTI), the clinical characteristics of a total of 117 patients (106 complicated UTI patients, 11 uncomplicated UTI patients) from whom CNS were isolated at urinary colony counts of 10(5) or more per ml were studied. Of the complicated UTI patients, 95 patients (89.6%) suffered from no symptoms while 11 (10.4%) had fever of 38 degrees C or greater, which was strongly suspected to be due to genitourinary tract infections. Six of these patients were managed by indwelling urinary catheters. On the other hand, all of the patients with uncomplicated UTI were young women and had typical symptoms of acute cystitis. These results suggest that CNS, which hitherto have been considered mere contaminants or benign colonization rather than true pathogens, can also cause complicated UTI requiring chemotherapy under certain conditions such as indwelling urinary catheterization and acute cystitis in sexually active women.


[Vectors of Japanese spotted fever].

[Vectors of Japanese spotted fever]. In the southeast coast of Shikoku, most of the Japanese spotted fever (JSF) patients were found to be infected with the rickettsial pathogen through bamboo plantation, and also the eschar was frequently noticed in skin inspection, so that it was clinically speculated to be caused by ticks. According to out field research, the tick fauna was very rich throughout that area, and it was permissible enough to determine ticks as the vectors, based on arising of anti-SF group rickettsiae (SFGR) antibody in mice inoculated with some tick emulsions, findings of rickettsiae reactive to patient sera or a species-specific monoclonal antibody to JSFR in the hemolymph cells of some ticks, and electron microscopical observations of SFGR in various internal organs including the salivary gland of ticks. Also the transovarial and transstadial transmission of SFGR in ticks were supposed. Our results suggest that common species (adult/nymph) of the genus Haemaphysalis may be the most related vectors to man, based on their dominances, SFGR-prevalences, behavior and incidences of infestation but also indicate that various genera and species should be kept as the potential vectors, in spite of JSFR isolation from a restricted species of ticks in the future.


[Detection of hepatitis delta virus (HDV) markers in HBV carriers]. To investigate the prevalence and clinical features of hepatitis delta virus (HDV) superinfection in hepatitis B virus (HBV) carriers, antibody to hepatitis delta antigen (anti-HD) was determined in the sera of 328 HBV carriers in Japan. 1) Of the 328 HBV carriers, six (1.8%) were seropositive for anti-HD by enzyme linked immunosorbent assay. IgM-antibody to hepatitis delta antigen was detected in 2/6 patients with a high anti-HDV titer. None of the patients was positive for hepatitis dealt antigen. 2) HBV carriers with chronic liver disease had a greater frequency of seropositivity of anti-HD than asymptomatic HBV carriers. These data indicate that HDV superinfection may be an etiologic agent of chronic liver disease in HBV carriers.


[Clinical study of albendazole therapy for strongyloidiasis]. We reported the efficacy of albendazole (ABZ) for the treatment of 27 patients with strongyloidiasis. Twenty-seven patients, 23 males and 4 females, received 200 mg of ABZ one hour before breakfast and supper for 3 days and this treatment was repeated 2 weeks later. The following results were obtained: 1) The eradication rate at 2 weeks after the initial treatment was 70.4% (19 of 27 patients) and 2 weeks after the second course was 66.7% (16 of 24 patients). 2) One patients (3.7%) complained of abdominal pain after the first treatment. Four patients (14.8%) complained of headache (n = 2), nausea (n = 1) and exanthema (n = 1) after the second treatment. But all symptoms were mild and required no treatment and subsided in a few days. 3) Positive rate of HTLV-1 antibody was 45.8% in the patients. As described above, side effects occurred in some cases, although they were mild and transient. From these results, it can be concluded that on increased dose of ABZ could be much more favorable for the treatment of strongyloidiasis.


Drug resistant plasmid of Actinobacillus pleuropneumoniae isolated from swine pleuropneumonia in Thailand. Actinobacillus pleuropneumoniae, isolated from the pneumonic lung tissue of pigs, was resistant to streptomycin, sulfadiazine and tetracycline. This isolate, TA5, possessed 2 plasmids (pTA51 = 3.7 Kb, pTA52 = 6.2 Kb); one of the plasmids (pTA52) was coded for resistance to SM and SA. Another isolate of A. pleuropneumoniae, TA8, also possessed 2 plasmids (pTA81 = 3.7 Kb, pTA82 = 4.4 Kb), with a plasmid (pTA82) coded for resistance to SM and SA. All 4 plasmids were not transmissible and small.


[An epidemiological study on Bordetella pertussis infection in Fukui Prefecture from 1986 to 1991--especially observation on the patients of culture-confirmed pertussis]. In order to clarify the epidemiological situation of whooping cough in Fukui prefecture, 478 nasopharyngeal swabs from patients with pertussis-like symptoms were submitted to bacterial isolation. Laboratory data of these patients with clinical informations at the departments of pediatrics of 6 hospitals in Fukui city from June 1986 to May 1991 were also examined in relation to the above bacterial isolation. The results observed in culture positive patients were as follows: 1) B. pertussis were isolated from 83 patients (41 men and 42 women). 2) Isolates were classified into 3 serotypes. Most dominant type was 1.3.6 (80 strains, 96.4%), and followed by 1 (2 strains) and 1.4.5 (1 strain). 3) Most of the patients were non-vaccinees less than 3 years of age (69 of 83 (83.1%). This evidence suggested strongly that pertussis vaccine was highly effective to prevent pertussis. 4) The patients under 3 years of age whose leucocyte counts were > or = 15000/mm3 and lymphocyte rate to total leucocyte were > or = 70% in their peripheral blood at the time of nasopharyngeal swabs sampling were only 49.3% (33 of 67). Therefore, diagnosis of pertussis by leucocyte findings alone were considered to be inadequate. 5) The area of the patient's residence covered 7 regions of 8 health center districts in Fukui prefecture. The areal distribution of the number of patients from June 1986 to May 1991 was relatively proportional to the population of each district.(ABSTRACT TRUNCATED AT 250 WORDS)