Immunotherapy in recurrent coccidioidomycosis

A 47-yr-old white woman developed several reactivations of pulmonary foci progressing to cavitation due to Coccidioides immitis. This sequence occurred in the presence of unreactivity to intradermal coccidioidin and unresponsiveness of the patient's lymphocytes in vitro to this antigen. This immunological defect was specific for C. immitis, as the patient was otherwise immunologically normal by several criteria including intradermal testing with other antigens and a normal response of her lymphocytes in vitro to phytohemagglutinin. Immunologic reconstitution was attempted several times with whole leukocytes and with leukocyte extracts (transfer factor). Conversion to intradermal reactivity to coccidioidal antigens was achieved with transfer factor, though the persistence of intradermal reactivity could only be demonstrated with spherulin, a new C, immitis skin-test antigen, and specific lymphocyte reactivity in vitro could not be shown. The patient's disease stabilized for several months, but the overall therapeutic effect of these immunological interventions is not yet certain.     

Peter Brian Medawar

Patients with coccidioidomycosis manifest a wide variety of severity of infection. Patients with unifocal and particularly multifocal dissemination manifest a defect in host defense mechanisms. This study focuses on the in vitro response to PHA as an indicator of immunoregulatory mechanisms. Study subjects are divided into four groups: Group I—17, patients with active disseminated coccidioidomycosis and lesions in multiple organs; Group II—15, patients with active disseminated coccidioidomycosis and a single lesion; Group III—16, patients with active coccidioidomycosis confined to the lungs; and Group IV—18, patients in whom coccidioidomycosis is inactive. The response to PHA was heterogeneous, even within groups. When cells were cultured in the presence of indomethacin or R020-5720, Group I cells demonstrated highly significant augmentation of the response to PHA (P < 0.002), whereas cells from Groups II to IV were unaffected. When adherent cells were removed by passage through a nylon wool column, the effect of indomethacin could no longer be demonstrated. Direct measurement of prostaglandin production (PGE₂ and PGF_2α) and thromboxane suggest that there is no difference in the quantity of these compounds produced by unstimulated or PHA-stimulated cells by Group I versus Group IV cells (P = 0.2-0.4). Indomethacin or R020-5720 markedly inhibit the production of these compounds. Studies of the effect of exogenous prostaglandin on PHA-stimulated cells suggest that Group I cells may be more inhibited by PGF_1α and PGF_2α compared to Group IV cells. The data demonstrate that PHA cultures of cells from Group I patients are suppressed. The suppressor cell can be removed by adherence to nylon wool and is inhibited by indomethacin or R020-5720.     

PLOIDAL CHANGES IN CLONAL CULTURES OF SPIROGYRA COMMUNIS AND IMPLICATIONS FOR SPECIES DEFINITION

A clonal culture of Spirogyra filaments of initially uniform width produced filaments of three additional significantly different widths. Group I filaments of the original clone were 30.9 ± 0.7 μm wide (mean ± SD, N = 50). Group I filaments produced Group II filaments (22.0 ± 1.1 μm) through vegetative growth and sexual reproduction. Zygospores from homothallic Group I filaments produced germlings representative of Groups I and II; zygospores from homothallic Group II filaments produced germlings representative of Group II only. Germlings of Groups III (27.7 ± 1.0 μm) and IV (44.9 ± 0.8 μm) were produced in the cross of I × II. Viable zygospores from homothallic Group III filaments were obtained. Cells of Group IV filaments were initially binucleate and did not conjugate. Of the six intergroup crosses possible, four resulted in conjugation-tube formation only; two crosses yielded zygospores (I × II and III × IV). Germlings from the successful cross of Groups III and IV produced filaments of all four groups. Chromosome counts were: Group I (24), Group II (12), Group III (18), and Group IV (24, one nucleus). Relative nuclear fluorescence values of mithramycin-stained DNA were (mean ± SD, N ≥ 30): Group I (11.1 ± 1.4), Group II (5.7 ± 0.7), Group III (8.8 ± 1.3), and Group IV (10.0 ± 0.9, one nucleus). Cytologically, Group II appears to be a diploid (2x), Group I a tetraploid (4x), and Group III a triploid (3x). Systematically, Groups I, II, and III key out to Spirogyra singularis, S. communis, and S. fragilis, respectively, using Transeau's mongraph of the family Zygnemataceae. These species are interpreted to represent a species complex of S. communis (whose name has priority) with the ancestral haploid (x = 6) missing.     

Characterisation of lactic acid producing <Emphasis Type="Italic">Sporolactobacillus</Emphasis> strains from tree barks in Thailand

Eight spore-forming lactic acid producing bacteria were isolated from tree barks in Thailand. They were identified as Sporolactobacillus nakayamae (Group I, three isolates), S. terrae (Group II, two isolates), S. kofuensis (Group III, one isolate) and S. inulinus (Group IV, two isolates) based on their phenotypic characteristics and 16S rRNA gene sequence analyses. Four isolates in Groups I and II produced DL lactic acid (89.60–114.61 g/L), while three isolates in Groups III and IV produced D-lactic acid (88.01–113.78 g/L). Isolate BK65-3 identified as S. inulinus produced the highest D-lactic acid concentrations (101.42 g/L), productivity (1.41 g/L/h), yields (84.52%) and optical purity of D-lactic acid (100%).     

Immunologic and clinical improvement of progressive coccidioidomycosis following administration of transfer factor

Three patients with progressive coccidioidomycosis were given preparations of transfer factor (TF). Adverse reactions to TF were minimal. Following TF administration two of these patients had prolonged clinical remissions in their coccidioidal disease. Cellular immune responses were sequentially evaluated by coccidioidininduced delayed-type skin tests, lymphocyte blast transformation and macrophage inhibition factor production (MIF). These three patients each exhibited different cellular immune patterns before and after TF administration. Two patients converted their coccidioidin skin tests, and one converted lymphocyte transformation response to coccidioidin. Also, TF apparently favorably affected the MIF response in all three patients.     

Intestinal Translocation of Clinical Isolates of Vancomycin-Resistant Enterococcus faecalis and ESBL-Producing Escherichia coli in a Rat Model of Bacterial Colonization and Liver Ischemia/Reperfusion Injury

The objectives of this study were to develop a rat model of gastrointestinal colonization with vancomycin-resistant Enterococcus faecalis (VRE) and extended-spectrum beta-lactamase (ESBL)-producing E. coli and to evaluate intestinal translocation to blood and tissues after total and partial hepatic ischemia. Methods - We developed a model of rat colonization with VRE and ESBL-E coli. Then we studied four groups of colonized rats: Group I (with hepatic pedicle occlusion causing complete liver ischemia and intestinal stasis); Group II (with partial liver ischemia without intestinal stasis); Group III (surgical manipulation without hepatic ischemia or intestinal stasis); Group IV (anesthetized without surgical manipulation). After sacrifice, portal and systemic blood, large intestine, small intestine, spleen, liver, lungs, and cervical and mesenteric lymph nodes were cultured. Endotoxin concentrations in portal and systemic blood were determined. Results – The best inocula were: VRE: 2.4×10¹⁰ cfu and ESBL-E. coli: 1.12×10¹⁰ cfu. The best results occurred 24 hours after inoculation and antibiotic doses of 750 µg/mL of water for vancomycin and 2.1 mg/mL for ceftriaxone. There was a significantly higher proportion of positive cultures for ESBL-E. coli in the lungs in Groups I, II and III when compared with Group IV (67%; 60%; 75% and 13%, respectively; p:0.04). VRE growth was more frequent in mesenteric lymph nodes for Groups I (67%) and III (38%) than for Groups II (13%) and IV (none) (p:0.002). LPS was significantly higher in systemic blood of Group I (9.761±13.804 EU/mL−p:0.01). No differences for endotoxin occurred in portal blood. Conclusion –We developed a model of rats colonized with resistant bacteria useful to study intestinal translocation. Translocation occurred in surgical procedures with and without hepatic ischemia-reperfusion and probably occurred via the bloodstream. Translocation was probably lymphatic in the ischemia-reperfusion groups. Systemic blood endotoxin levels were higher in the group with complete hepatic ischemia.     

Protective effect of omega-3 fatty acid against mercury chloride intoxication in mice

The aim of this study was to investigate the protective effect of omega-3 fatty acid in HgCI₂ toxicity in mice. Levels of malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NO) and total sialic acid (TSA), and histopathological changes in selected organs were evaluated. Twenty-eight mice were equally divided into 4 groups, namely Groups I–IV. Group I animals received intraperitoneal (ip) injection of physiological saline solution; Group II animals received ip injection of 0.4 mg/kg/day HgCI₂; Group III animals received ip injection of 0.4 mg/kg/day HgCI₂ in addition to subcutaneous (sc) injection of 0.5 g/kg/day omega-3 fatty acid; and Group IV animals received sc injection of 0.5 g/kg/day omega-3 fatty acid. All treatments lasted 7 days. The levels of MDA, NO and TSA were significantly higher in Group II and lower in Groups III and IV as compared to the Group I. GSH level was the highest in Group IV. In histopathology, severe degeneration in liver and kidney was observed in Group II animals. These degrading changes were seen to be reduced greatly in Group III animals. The results suggested that omega-3 fatty acid might attenuate HgCI₂-induced toxicity by improving antioxidant status and acute phase response in mice.     

Dirofilaria immitis: Parasite-specific humoral and cellular immune responses in experimentally infected dogs

Humoral and cellular immune responses to adult antigens of Dirofilaria immitis were evaluated in experimentally infected dogs during the chronic phase of infection. All infected dogs had significantly elevated IgG (enzyme-linked immunosorbent assay) and IgE (passive cutaneous anaphylaxis) titers against D. immitis adult antigens. However, there was little difference between infected dogs and uninfected controls in cellular-immune responses to D. immitis adult antigen or phytohemagglutinin as assessed by the lymphocyte transformation assay. Although neither cellular nor humoral responses correlated with worm burdens, cellular responses among infected dogs correlated inversely with IgG titers to D. immitis adult antigen. These results are consistent with observations in other nematode and trematode systems which suggest that in chronic tissue helminth infections there is suppression of cellular immune responses to parasite antigens while humoral responses to the same antigens remain relatively preserved.     

An evaluation of reactivity to Coccidioides immitis skin tests in subjects with diabetes mellitus

Persons with diabetes mellitus have an increased incidence of progression and dissemination of infection with Coccidioides immitis. This increased incidence of infection suggests that diabetic subjects may have a defect in cell-mediated immunity (CMI) to coccidioidomycosis. Skin test response is also determined by CMI. A negative coccidioidin skin test is often seen with serious infections and is used as evidence that therapy should be initiated. To evaluate CMI in diabetic subjects we investigated the reactivity to coccidioidin and spherulin of 54 diabetic and 56 nondiabetic subjects. There was no difference between the two groups in the prevalence or size of positive reactions to coccidioidin. These results support the continued use of coccidioidin to evaluate the need for therapy in diabetic subjects who have coccidioidomycosis. Spherulin antigen, however, produced fewer positive reactions in the diabetic group. This decreased spherulin reactivity may be associated with the increased incidence of severe coccidioidal infection in diabetic subjects.     

Cellular immunity in man: correlation of leukocyte migration inhibition factor formation and delayed hypersensitivity

The formation of leukocyte migration inhibition factor (MIF) by the lymphocytes of 13 normal persons immune to the protein antigen keyhole limpet hemocyanin (KLH) has been investigated. KLH-induced MIF formation expressed as percent migration was compared with delayed hypersensitivity, antibody, and in vitro lymphocyte blastogenic responses to this antigen. Individuals were studied 404–840 days (median 540 days) after their last exposure to KLH. Nine persons had delayed hypersensitivity to KLH and 10 had circulating KLH antibody. The lymphocytes of 11 showed an in vitro blastogenic response to KLH stimulation, while the lymphocytes of nine produced MIF after KLH stimulation. The mean percent migration for the subjects with KLH delayed hypersensitivity was 48.2 (range 20.4–70.4) compared with 133 (range 120–161) for the four persons who did not have KLH delayed hypersensitivity (P < 0.05). The correlation coefficient between the precent migration and delayed hypersensitivity was ?0.78 (P < 0.01). No correlation was demonstrated between migration inhibition and the other parameters of immunity.     

Antioxidant enzyme activity and lipid peroxidation in the blood of rats co-treated with vanadium (V<Superscript>+5</Superscript>) and chromium (Cr<Superscript>+3</Superscript>)

Selected biochemical parameters were studied in the blood of outbred, male Wistar rats which daily received to drink deionized water (Group I, control) or solutions of: sodium metavanadate (SMV; 0.100 mg V/mL)—Group II; chromium chloride (CC; 0.004 mg Cr/mL)—Group III; and SMV-CC (0.100 mg V and 0.004 mg Cr/mL)—Group IV for a 12-week period. The diet and fluid intake, body weight gain, and food efficiency ratio (FER) diminished significantly in the rats of Groups II and IV, compared with Groups I and III. The plasma total antioxidant status (TAS) as well as the MDA and the l-ascorbic acid level in the erythrocytes (RBCs) remained unchanged in all the groups, whereas the plasma l-ascorbic acid concentration decreased markedly in Group II, compared with Group III. The activities of Cu,Zn-superoxide dismutase (Cu,Zn-SOD), catalase (CAT), cellular glutathione peroxidase (cGSH-Px), and glutathione reductase (GR) in RBCs remained unaltered in all the treated rats. However, the activity of glutathione S-transferase (GST) and the content of reduced glutathione (GSH) in RBCs decreased and increased, respectively, in Groups II, III, and IV, compared with Group I. A vanadium–chromium interaction which affected the GST activity was also found. To summarize, SMV and CC administered separately or in combination in drinking water for 12 weeks did not alter either lipid peroxidation (LPO) or the activities of Cu,Zn-SOD, CAT, cGSH-Px, and GR, which allows a conclusion that both metals in the doses ingested did not reveal their pro-oxidant potential on RBCs.     

Immunology of histoplasmosis: Humoral and cellular activity from a polysacchari-de-protein complex and its deproteinized fraction in experimentally immunized mice

In a previous publication it was reported that a polysaccharide-protein complex (PPC), sensitive to -glucosidase, was isolated from Histoplasma capsulatum. This complex was strongly reactive in an agar gel diffusion assay with sera from patients with histoplasmosis, but was unreactive with sera from patients with coccidioidomycosis. Here, the studies with human sera have been expanded and attempts were made to determine the response of mice immunized with nonviable H. capsulatum or Cocccidioides immitis to PPC or its deproteinized fraction (D-PPC) using more sensitive tests for antibody and including also test for cell-mediated immunity. Histoplasmin and coccidioidin were compared with PPC or its deproteinized fraction (D-PPC) in all assays. In a counterimmunoelectrophoresis (CIE) assay, PPC and D-PPC reacted only with sera from patients with histoplasmosis, whereas cross reactions were noted with histoplasmin and coccidioidin using heterologous sera. Cross-reaction were observed with all four antigen preparations and both types of antisera using a micro complement fixation assay. The assay for macrophage migration inhibitory factor (MIF) was also relatively nonspecific, in that inhibition occurred with cells from animals sensitized with Histoplasma or Coccidioides using both homologous and heterologous antigens. In the footpad assay, histoplasmin and coccidioidin were highly cross-reactive in animals sensitized with the heterologous fungus, but the PPC and D-PPC from H. capsulatum elicited significant reactions only in animals sensitized with Histoplasma.     

Effect of zinc and melatonin supplementation on cellular immunity in rats with toxoplasmosis

The effects of zinc (Zn) and/or melatonin supplementation on cellular immunity were investigated in rats infested with Toxoplasma gondii. Fifty Sprague-Dawley male rats were used for this study. All animals were fed a normal diet, ad libitum, containing 97 mg Zn/kg. They were divided into five experimental groups, as follows. Group I (n=10) received intraperitoneal injections of zinc sulfate at a dose of 3 mg/kg/d for 3 wk. Group II (n=10) received intraperitoneal injections of melatonin at a dose of 3 mg/kg/d for 3 wk. Group III (n=10) received intraperitoneal injections of zinc sulfate (3 mg/kg/d) and melatonin (3 mg/kg/d) for 3 wk. Group IV (n=10) was infested controls. Group V (n=10) was healthy controls. There were no differences in the percentage of CD3+ lymphocytes among all groups. For groups I–III, the CD4+ and CD8+ ratios were higher than those of the groups IV and V controls (p<0.01). Similarly, the total lymphocyte ratios in groups I–III were higher than those of infested and healthy controls (p<0.01). The total lymphocyte ratios in group III were significantly higher than those of groups I and II (p<0.01). The plasma Zn levels in the supplemented groups were significantly higher than those of control groups IV and V (p<0.01). These results suggest that melatonin and/or Zn supplementation may activate cellular immunity by stimulating CD4+ and CD8+ production in infected rats with T. gondii.     

A conventional polymerase chain reaction-based method for the diagnosis of human schistosomiasis in stool samples from individuals in a low-endemicity area

The aim of this study was to evaluate the efficacy of a polymerase chain reaction (PCR)-based method to detect Schistosoma mansoni DNA in stool samples from individuals living in a low-endemicity area in Brazil. Of the 125 initial stool samples, 80 were ELISA reactive and eggs were identified in 19 of the samples by parasitological examination. For the PCR evaluations, 56 stool samples were selected and divided into five groups. Groups I-IV were scored negative for S. mansoni eggs by parasitological examination. Groups I and II were ELISA reactive, whereas Groups III and IV were ELISA nonreactive. Groups II and III were positive for other intestinal parasites. PCR testing scored eight samples as positive from these four groups. Group V represented the S. mansoni -positive group and it included ELISA-reactive samples that were scored positive for S. mansoni by one or more parasitological examinations (6/19 were positive by Kato-Katz method, 9/17 by saline gradient and 10/13 by Helmintex®). PCR scored 13 of these 19 samples as positive for S. mansoni . We conclude that while none of these methods yielded 100% sensitivity, a combination of techniques should be effective for improving the detection of S. mansoni infection in low-endemicity areas.     

Immunofluorescent test for simultaneous detection of Edwardsiella ictaluri and Flavobacterium columnare

Enteric septicemia of catfish (ESC) and columnaris disease are 2 bacterial diseases significantly affecting the aquaculture industry, and thus rapid diagnosis of disease is imperative for making judicious management decisions. A rapid indirect fluorescent antibody (IFA) test with antibody conjugated fluorochromes having 2 different spectral properties (Alexa Fluor 488-emitting green fluorescence, and Alexa Fluor 594-emitting red fluorescence) was compared with bacteriological culture (accepted standard) for simultaneous detection of Edwardsiella ictaluri (EI) and Flavobacterium columnare (FC) in 3 groups of experimentally infected channel catfish (Ictalurus punctatus Rafinesque), and a fourth group that acquired an aquarium-infection with F. columnare. A total of 303 samples (derived from kidney, brain and nares) from 101 fish were concurrently examined by both tests. Fish in the 3 experimentally infected groups (I to III) were culture positive for the bacteria with which they were infected, and fish in Group IV, (the spontaneously infected fish) revealed F. columnare only. The IFA test compared favorably in sensitivity (EI= 80.7 %; FC = 87.2%) and specificity (EI = 83.9%; FC = 88.9%) with the standard bacteriological culture. The positive predictive value (EI = 96.2% Group I, 90.8% Group II, 93.7% Groups I and II combined; FC = 95.2% Group II, 95.3% Groups II, III and IV combined) was high, while the negative predictive value (EI = 66.7% Group I, 31.3% Group II, 59.5% Groups I and II combined; FC = 73.7% Group II, 72.7% Groups II, III and IV combined) was relatively low. The IFA test will serve as an efficient tool for rapid simultaneous detection of E. ictaluri and F. columnare in outbreaks of disease.     

Embryo production and progesterone profiles in ewes superovulated with different hormonal treatments

The superovulatory response and embryo yield following hormonal treatments of Merino ewes during late spring and their estrous cycle were evaluated. Ewes (n=17) were treated with progestagen-impregnated sponges and assigned to Group I (800 IU PMSG plus 11.5 mg FSH-p); Group II (1200 IU PMSG); Group III (1600 IU PMSG). Ewes were naturally mated and followed by laparotomy 6 days later. After laparotomy, ewes were injected with a prostaglandin analogue (PGF) and serum samples were obtained prior to surgery and then for 25 days to measure progesterone (P4) by radioimmunoassay. There were no differences among groups neither for estrous incidence (Group I: 83.3%; Group II: 83.3%; Group III: 100%), nor for the time interval to estrous onset (Group I: 26.4±2.4 h; Group II: 28.8±2.9 h; Group III: 24.0±3.8 h). Group I had more corpora lutea than Group II (14.2±1.2 and 6.2±0.8; P<0.05), and Group III was intermediate (11.0±3.0). There was a low incidence of persistent follicles in all treatments (Group I: 0.5±0.5; Group II: 0.6±0.4; Group III: 1.8±1.2). Number of collected ova were 9.0±2.6, 3.8±0.6 and 6.5±0.9 for Groups I, II and III, respectively. Significant differences in number of ova were detected between Groups I and II. Unfertilized ova did not differ among groups (Group I: 3.5±1.0; Group II: 2.8±0.8; Group III: 5.2±1.4; P>0.05). Embryos and high viability embryos were higher (P<0.05) in Group I (5.2±1.9 and 4.8±2.0) than in Group II (1.0±0.5 and 1.0±0.5) or Group III (1.2±0.6 and 1.0±0.5). Total plasma progesterone (P4) and P4 per corpus luteum before PGF administration did not vary (P>0.05) among groups (Group I: 71.0±14.7 and 4.9±0.7 nmol/l; Group II: 50.6±13.3 and 7.9±1.6 nmol/l; Group III: 90.4±42.6 and 6.8±1.8 nmol/l). There was a significant and positive correlation between P4 before PGF administration and number of corpora lutea (r=0.76). No significant differences were detected among groups for: interval PGF to P4 <3.18 nmol/l (Group I: 2.7±0.3 days; Group II: 1.8±0.6 days; Group III: 2.2±0.5 days), cycle length (Group I: 18.3±1.4 days; Group II: 17.9±0.5 days; Group III: 16.8±0.9 days), duration of P4 levels <3.18 nmol/l (Group I: 11.3±1.9 days; Group II: 7.1±1.0 days; Group III: 7.2±2.4 days), duration of P4 levels ≥3.18 nmol/l (Group I: 7.0±1.3 days; Group II: 10.8±0.8 days; Group III: 9.5±1.7 days) and peak of P4 (Group I: 7.4±0.4 nmol/l; Group II: 10.8±1.6 nmol/l; Group III: 9.2±1.9 nmol/l). It was concluded that PMSG–FSH-p treatment was more efficient than PMSG alone for superovulation and embryo production in ewes while P4 profiles were similar among groups.     

Vegetation and soils of the Meikle Kilrannoch ultramafic sites

Summary

The vegetation of two ultramafic sites (MK1 and MK2) at Meikle Kilrannoch are described. MK1 is dome shaped and has much weathering bedrock whilst MK2 is flatter, lacks weathering bedrock and has probably been entirely peat covered. Six vegetation Groups were recognized: I, high-level blanket bog; II and III, grass heath (with II more sedge-rich); IV, dwarf shrub heath; V and VI, debris (an open vegetation on stony skeletal soils). Vegetation maps were produced which had five mapping units: Group I, Groups II and IH combined, Group IV, Groups V and VI combined, and non-vegetated eroding peat. Three main soil types occur: peat, which underlies Group I; a complex of freely draining magnesian brown soils and imperfectly drained magnesian gleys which bear Groups II–IV; and skeletal soils, derived either from weathering bedrock (MK1 only) or ultramafic drift, which bear vegetation in Groups V or VI. The rare plants are commonest in, but not restricted to, debris vegetation on skeletal soils. The likely plant toxicity of soil magnesium at the sites is reaffirmed but it is suggested that the low plant cover in the debris is more likely to result from low nutrients or intensive frost action or both. Floristic differences between MK1 and MK2 are discussed and quantified for Lychnis alpina (which had 68000individuals on the former site and 46 on the latter) but the causes of the differences remain unexplained.     

Production of macrophage migration inhibitory factor and lymphotoxin by leukocytes from normal and Wiskott-Aldrich syndrome patients

The production of macrophage migration inhibitory factor (MIF) and lymphotoxin (LT) by cultured leukocytes from patients with Wiskott-Aldrich syndrome (WAS) and normal controls was studied. The presence of these lymphokines in leukocyte culture supernatants usually correlated directly with the dose of stimulant used. Doses of nonspecific mitogens and specific antigens, which produced maximal in vitro lymphocyte transformation, stimulated maximal production of these mediators. When the incorporation of tritiated thymidine by stimulated leukocyte cultures from patients with Wiskott-Aldrich syndrome (WAS) was deficient, they usually produced less MIF and lymphotoxin than normal. However, when their in vitro lymphoproliferative responses were normal, the lymphotoxin activity in supernatants of WAS leukocyte cultures was normal.     

Insecticidal bacterium isolated from an ant lion larva from Munakata,Japan

Twenty bacteria were isolated from four ant lion larvae. The isolates were classified into three groups by biological characteristics. Since Group I, Group II and Group III were isolated from individual larvae Kuo1, Kuo3, 4 and Kuo2, respectively, with exception of one isolate Kuo2‐1, each ant lion tested had its own dominant bacterial flora. Groups I and II were closer to Serratia liquefaciens and Enterobacter cloacae, respectively, whereas Group III could not be identified by the test used. The phylogenetic analysis of GroEL amino acid sequences revealed that Group I, II and III were related to those of Serratia spp., E. cloacae and Salmonella spp. –Escherichia/Shigella spp., respectively. Among these groups, Group I was highly virulent against Bombyx mori and Periplaneta americana, and caused 100% mortality within 24 h. The other two groups (Group II and III) were avirulent to these insect species. The culture filtrate of Group I caused killing activity to B. mori larvae and the insecticidal substance was purified from culture filtrate of Group I bacterium. Since the insecticidal activity highly correlated with proteolytic activity in the chromatographies, Group I bacterium may secret insecticidal proteinase in vitro.     

The Impact of Magnesium on Isometric Twitch Parameters and Resting Membrane Potential of the Skeletal Muscle in Diabetic Rats

To present the relationship between oral magnesium supplementation, blood glucose, and changes in isometric twitch parameters, resting membrane potential (RMP), in the gastrocnemius muscle in diabetic rats. Sixty rats were used in this study. The rats were divided into four groups: control (drinking tap water, Group I, n = 15), control with treated with magnesium sulfate (10 g/L) (Group II, n = 15), diabetic (Group III, n = 15), and diabetic with treated with magnesium sulfate (10 g/L) (Group IV, n = 15). In Group II and IV, the level of plasma magnesium was increased comparing to those of the control group (p < 0.05). Isometric twitch tensions were decreased significantly in the Group III, but Group IV isometric twitch tensions were increased significantly. Group IV RMP values were close to the Group I. Hyperglycemia decreases gastrocnemius muscle isometric twitch tension and increases RMP in diabetic rats. Magnesium treatment can prevent these diabetic complications.