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Probiotic fermented food mixtures: possible applications in clinical anti-diarrhoea usage.

A probiotic fermented PCMT food mixture was developed by fermentation of an autoclaved and cooled slurry of pearl millet flour, chickpea flour, skim milk powder and fresh tomato pulp (PCMT 2:1:1:1, w/w) with Lactobacillus acidophilus (10(5) cells/ml), a probiotic organism at 37 degrees C for 24 h. Such a fermented mixture inhibited the growth of pathogenic organisms, namely Shigella dysenteriae, Salmonella typhosa and E. coli. A significant decline in pH with a corresponding increase in titratable acidity due to probiotic fermentation occurred in the developed food mixture. Feeding of the freshly developed fermented mixture to mice suffering from E. coli induced diarrhoea, could help to arrest diarrhoea, reduce moisture, protein and ash contents in their faeces. The counts of lactobacilli increased whereas those of E. coli decreased remarkably in the faeces of mice from the 3rd day of the feeding trial till the end of experimental period. The beneficial effect of probiotic feeding may be due to antimicrobial substances produced by L. acidophilus, which might have neutralized the enterotoxins from E. coli. The cost of one 200 ml glass full of this probiotic drink is no more than one rupee.

Animals↗

Alteration of histamine, serotonin and primary prostaglandin in case of diarrhea induced by endotoxin and gastrointestinal absorption of drug.

Effects of endotoxin on the motility, the permeability of mesenteric blood vessel and endogenous substances (electrolyte (Na, K), histamine, serotonin and primary prostaglandin) in the small intestine of mice were investigated. Diarrhea was induced by intraperitoneal injection of Salmonella typhosa endotoxin in mice. It is suggested that the diarrheal action of endotoxin is due to the acceleration of intestinal motility and/or the facilitation of mesenteric blood vessel permeability. On the other hand, from the observation of blood concentration of acetaminophen and sulfisoxazole, inhibition of drug absorption was suggested in cases of diarrhea induced by endotoxin. This inhibition of gastrointestinal absorption is due to the delay of the gastric emptying rate, the increase of intestinal transit time of drug in the absorption site and the acceleration of mesenteric blood vessel permeability. Then it is conceivable that these actions may be mediated by biogenic amines (serotonin and histamine) and primary prostaglandins.

Animals↗

Cocaethylene affects human microvascular endothelial cell p38 mitogen-activated protein kinase activation and nuclear factor-kappaB DNA-binding activity.

BACKGROUND: Cocaethylene (CE) is known to increase the permeability of human microvascular endothelial cell monolayers. The molecular mechanism underlying this increase may involve calcium-modulated signaling pathways such as the p38 mitogen-activated protein kinase (p38 MAPK) and the nuclear factor-kappaB (NF-kappaB) family of transcription factors. The hypothesis of this study was that CE-mediated endothelial permeability change may be mediated by the p38 MAPK and consequently NF-kappaB dimers. METHODS: We used sandwich ELISA to detect phosphorylated p38 MAPK in the cell line human microvascular endothelial cell 1 (HMEC-1) after treatment with 1 mmol/L CE. We used electrophoretic mobility shift assay to detect changes in NF-kappaB dimers present in HMEC-1 and their DNA-binding activity after treatment with CE. Lipopolysaccharide (LPS) from Salmonella typhosa was used as a positive control for all experiments. RESULTS: Treatment with CE and LPS had similar effects on HMEC-1 p38 MAPK phosphorylation and NF-kappaB DNA-binding activity. Both treatments increased the phosphorylation of p38 MAPK, consistent with activation of proinflammatory cell signaling. Treatment of HMEC-1 with CE decreased DNA binding of both the RelA/p50 and p50/p50 dimers of the NF-kappaB transcription factor family, whereas treatment with LPS decreased and then increased the DNA binding of these dimers. CONCLUSION: In addition to increasing HMEC-1 monolayer permeability, CE also alters transcription factor and kinase activity related to inflammation. Thus, CE causes endothelial activation that can elicit a prolonged and organized cellular response, rather than being directly toxic to endothelial cells.

Capillary Permeability↗

Amitriptyline attenuates the febrile response to a pyrogen in rabbits.

We investigated the effect of a tricyclic antidepressant (TCA) agent on fever in rabbits. In the first series of experiments, the TCA agent amitriptyline hydrochloride (AMI) was administered intraperitoneally (i.p.) at a dose of 1 mg/kg or 10 mg/kg daily for three weeks. The rabbit's response to intravenous (i.v.) injection of 0.1 micrograms/kg of the lipopolysaccharide (LPS) of Salmonella typhosa was tested on a weekly basis. In the second series of experiments, rabbits were given simultaneous i.v. injections of either 1 mg/kg or 10 mg/kg of AMI together with 0.1 microgram/kg of LPS. After two weeks of chronic i.p. administration of 10 mg/kg AMI, and for the remainder of the experimental period, the rabbit's response to the LPS was significantly attenuated when compared to control animals. When AMI was administered simultaneously with the LPS, a significant dose-dependent antipyresis was observed from 50 minutes after injection, compared to the response of control animals. AMI had no effect on the body temperature of afebrile animals.

Amitriptyline↗

Submandibular gland peptide-T (SGP-T) modulates ventricular function in response to intravenous endotoxin.

A novel peptide hormone isolated from salivary glands, submandibular gland peptide-T (SGP-T), protects against the enhanced hypotensive response to intravenously administered lipopolysaccharide (LPS; 3.5 mg/kg; Salmonella typhosa) in rats with their submandibular glands removed (sialadenectomy). In this study, we examined the effects of SGP-T on LPS-provoked perturbations of heart function. Sialadenectomy did not alter basal heart function, although the sialadenectomized rats exhibited more pronounced reductions in ventricular peak systolic pressure (VPSP), ventricular pressure at max dP/dt (Pmax dP/dt), and maximum ventricular negative dP/dt (-dP/dt) relative to unoperated controls following LPS challenge. These changes were primarily due to changes in afterload (blood pressure). However, in sialadenectomized rats LPS-induced changes in Pmax -dP/dt (ventricular pressure at maximum -dP/dt) did not correlate with changes in VPSP, which suggests that sialadenectomy may alter the systolic component of the heart beat. The peptide SGP-T, at doses of 1 and 3.5 microg/kg, corrected Pmax -dP/dt and all other endotoxin-induced changes in heart function. Since Pmax -dP/dt is a measure of relaxation during diastole, the submandibular glands appear to play a role in protecting the heart against the adverse effects of acute endotoxin administration on ventricular relaxation. These effects of the submandibular glands may be mediated by the release of the peptide SGP-T.

Animals↗

Weakening of genetic resistance. I. The effect of injection of endotoxin, Freund's complete adjuvant and alloantiserum.

A parent to F1 transplantation combination was used to study the weakening effect of endotoxin, Freund's complete adjuvant and alloantiserum on genetic resistance. The relationship between time of treatment with endotoxin, Salmonella typhosa, and Freund's complete adjuvant, and their weakening effect was assessed by use of the spleen colony technique. CFU growth studies revealed that both endotoxin and alloantiserum were capable of weakening genetic resistance in the spleen but were unable to induce weakening of the resistance in the femoral marrow cavity. These results led us to the conclusion that the agents might not have a direct effect on the effector cells of the resistance. The weakening induced by endotoxin and alloantiserum seemed to be related to a certain immunological phenomenon in the spleen. In this phenomenon macrophages are likely to play a role since a number of agents capable of weakening resistance were known for their capacity to influence the mononuclear phagocytic system.

Animals↗

PREPARATION OF DRIED ACETONE-INACTIVATED AND HEAT-PHENOL-INACTIVATED TYPHOID VACCINES.

Controlled field trials of two dried inactivated typhoid vaccines have been carried out in British Guiana and Yugoslavia under the sponsorship of the World Health Organization. This paper gives details of the methods of preparation of these vaccines in such a manner as to permit of replication by others.Both vaccines were derived from the Ty 2 strain of Salmonella typhosa. For production of the acetone-inactivated vaccine, a portion of the surface agar growth of the Ty 2 strain was treated with acetone by the Landy method modified so that inactivation and handling were done in liquid suspensions rather than on a filter, and the vaccine was dried in the final containers. For the heat-phenol product, another portion of the same growth was processed by heating, the addition of 0.5% phenol and holding at room temperature for 72 hours followed by freeze-drying in the final containers. These vaccines are designed for use within a few hours of the addition of the reconstituting fluid.

Acetone↗

[Bacteremia in febrile children].

Bacteremia in children under 2 years of age is not uncommon. Several studies indicated that a small proportion of young children with fever who do not appear to be seriously ill may have bacteremia, and a small proportion of this subgroup may go on to develop serious illness such as meningitis. The rationale for this study was to assess both the incidence of bacteremia in febrile children in Taipei area and the predictive value of the white blood cell count and degree of body temperature for bacteremia. Three hundred febrile children (B. T. greater than or equal to 39 degrees C), without underlying disease or readily recognizable viral illness, who visited the emergency service of Chang Gung Memorial Hospital, were enrolled. Blood cultures and complete blood counts were obtained. Clinically important pathogens were identified in 6 of 300 cases (2%). The incidence and pathogens of various age group were: (1) Neonate, 2/15 (13%), one each of Group D streptococcus & streptococcus viridans; (2) 1 month to 1 year old, 3/66 (5%), 2 Escherichia coli and 1 Salmonella Group D; (3) 1 to 2 years old, 0/49; (4) 2 to 5 years old, 0/72; (5) 5 to 10 years old, 0/67; (6) 10 to 15 years old, 1/26 (4%), Salmonella typhosa. Neither the white blood cell count, the absolute segmented or band neutrophil count, nor the degree of body temperature had predictive value for bacteremia.

Adolescent↗

Enhancing effect of phorbol esters on induction of differentiation of mouse myeloid leukemia cells by human urinary protein and lipopolysaccharide.

12-O-Tetradecanoylphorbol-13-acetate, a potent promoter of carcinogenesis in mouse skin, enhanced differentiation of cultured mouse myeloid leukemia cells (M1) induced by human urinary protein or by lipopolysaccharide from Salmonella typhosa. 12-O-Tetradecanoylphorbol-13-acetate enhanced differentiation of all the markers tested, such as phagocytosis, Fc rosette formation, lysozyme activity, and morphological change. Other potent tumor-promoting macrocyclic plant diterpenes also enhanced the induction of differentiation, but no-tumor-promoting diterpenes did not. These findings were in marked contrast with generally accepted findings on the inhibitory effect of 12-O-tetradecanoylphorbol-13-acetate on terminal differentiation observed in other cell culture systems but consistent with the observations with some kinds of leukemia cells.

Animals↗

Immunoradiometric assay of endotoxin in serum.

We describe an immunoradiometric assay for the specific detection and quantitation of endotoxin from Escherichia coli 026, with use of 125I-labeled antibody. The sensitivty of this assay is 1 microgram/L.E. coli 026 endotoxin was detected in serum of rats for as long as 24 h after intraperitoneal injection of 0.1 mg of endotoxin per 100 g body weight. The assay is specific for the O-polysaccharide portion of this endotoxin, as confirmed by results after protein digestion and oxidation of the endotoxin, as well as by failure to detect endotoxin in serum after similar injections of E. coli 0127 and Salmonella typhosa 0901 endotoxins.

Animals↗

Effect of Ascaris suum and other adjuvants on the potentiation of the IgE response in guinea-pigs.

The potentiation effect of various adjuvants on the production of guinea-pig IgE was investigated using Freund's complete and incomplete adjuvant, the lipopolysaccharide of Escherichia coli and Salmonella typhosa, Bordetella pertussis, and the nematodes Nippostrongylus brasiliensis and Ascaris suum. While all the antigens had a variable effect on the potential of the IgG response, only infection with A. suum resulted in an enhanced IgE response to the antigen, egg albumin. Maximum potentiation occurred when primary immunization and nematode infection were accomplished simultaneously.

Adjuvants, Immunologic↗

Constitutive production and release of a lymphokine with macrophage-activating factor activity distinct from gamma-interferon by a human T-cell leukemia virus-positive cell line.

Culture supernatant fluids from the human T-cell leukemia virus-positive cell line C10/MJ-2 were found to contain a soluble factor with macrophage-activating factor (MAF) activity. The MAF activity of this culture supernatant fluid was stable at 100 degrees for 2 min and was unaffected by treatment with human anti-gamma-interferon (IFN-gamma) monoclonal antibody. Both treatments neutralized greater than 97% IFN-gamma activity from the supernatant fluid as measured by virus neutralization. Furthermore, this MAF activity was not due to contamination with endotoxins since the Limulus amebocyte lysate assay was negative (less than 0.125 ng/ml) and preincubation of the C10/MJ-2 supernatant fluid with polymyxin B did not diminish its activating potential. By contrast, human IFN-gamma rendered human monocytes tumoricidal only when combined with Salmonella typhosa lipopolysaccharide (LPS) at a minimum dose of 0.2 ng/ml. The concentrations of both LPS and IFN-gamma were crucial to achieve activation since IFN-gamma at doses less than 10 units/ml did not activate human monocytes even when combined with maximal doses of LPS (0.5 ng/ml). Finally, when human IFN-gamma admixed with LPS was preincubated with polymyxin B, its activating potential was completely abrogated. Collectively, these data suggest that the human T-cell line C10/MJ-2 constitutively produces a diffusable product distinct from IFN-gamma which activates human monocytes to lyse tumor cells. Thus, this cell line could provide a good source of a unique human MAF for future purification procedures.

Cell Line↗

[Immunization with a complex preparation: a new method of influenza prevention. The basis for using a complex preparation for preventing influenza].

A combined preparation for influenza prevention (CPIP) consisting of an interferon inducer stimulating immunogenesis and killed influenza vaccine is proposed. Twenty five inducers-stimulators have been tested: polynucleotides, polysaccharide and lipopolysaccharide extracted from Salmonella typhosa. Intranasal administration of CPIP to laboratory animals markedly stimulates interferon, secretory, and circulating antibody synthesis. Resistance to fatal influenza infection develops within 18 hours after administration of CPIP and its intensity increases in the following 14 days (the observation period.

Animals↗

Effects of low-dose total-body irradiation on canine bone marrow function and canine lymphoma.

Low-dose total-body irradiation, 150 rad given in 10 fractions over 5 weeks, is a useful treatment modality for favorable-prognosis lymphomas. Little is known, however, about the effects of this regimen on normal bone marrow. Six healthy beagle dogs and 5 dogs of various breeds with lymphoma were treated with total-body irradiation. Three of the 5 lymphomatous dogs achieved remissions of limited duration. No changes in hemograms or in bone marrow cellularity (as assessed by needle marrow biopsies) could be detected during or after treatment. Bone marrow progenitor cells were studied weekly during treatment and for 4 weeks thereafter using in vitro growth assays for GM-CFC and M-CFC. These studies demonstrated significant reductions (P less than 0.001) of granulocyte and macrophage progenitor cells with subsequent recovery toward normal pre-irradiation and sham irradiation values. Two additional dogs were injected with sublethal doses of Salmonella typhosa endotoxin 2 weeks after completion of the irradiation regimen. Their bone marrow GM-CFC responses were dramatically blunted compared to nonirradiated controls whereas their peripheral leukocyte responses and serum CSF levels were comparable to nonirradiated controls. These studies suggest that total-body irradiation may induce bone marrow injury that may be clinically significant if patients so treated are further stressed by infections or myelosuppressive drugs.

Animals↗

[Electrocardiographic study of sinus bradycardia associated with enterohemorrhagic Escherichia coli O157: H7 infection].

A severe outbreak of hemorrhagic colitis occurred at a kindergarten in Saitama, Japan in October, 1990. Children who were affected by enterohemorrhagic E. coli O157: H7 infection showed apparent bradycardia as well as severe bloody diarrhea, generalized convulsion, or hemolytic uremic syndrome. Cardiac involvement such as bradycardia observed in the patients of this outbreak has not been described in previous reports about EHEC infection, while bradycardia has been well known in typhoid fever due to salmonella typhosa infection. Electrocardiographic examination was performed to evaluate bardicardia, utilizing electrocardiography at rest and Holter's twenty-four hour electrocardiography. In the report, we demonstrate that the bradicardia was due to reduced frequency of sinus node. Both average heart rate and average minimum heart rate of the patients at night (74.0 +/- 5.6 BPM and 57.0 +/- 5.1 BPM, respectively) decreased significantly, as compared with controls (84.6 +/- 9.3 BPM and 66.3 +/- 8.0 BPM respectively) (p < 0.01). CVRR of the patients (0.120 +/- 0.019, respectively) increased significantly as compared with controls (0.090 +/- 0.010, respectively). These results indicate that an activated parasympathetic nervous system, that is, activation of the vagal nerve, might have induced the sinus bradycardia observed in the patients with EHEC infection.

Bradycardia↗

Removal of the submandibular glands increases the acute hypotensive response to endotoxin.

We investigated the roles of the submandibular glands and of their sympathetic innervation on the acute hypotensive response induced by the i.v. injection of endotoxin (10 mg/kg; Salmonella typhosa) into rats anesthetized with sodium pentobarbital. Endotoxin given to unoperated and sham-operated rats induced a transient 30-40 mm Hg drop in blood pressure. Rats that had their submandibular glands removed (sialadenectomy) or bilateral resection of the superior cervical ganglia (ganglionectomy), exhibited significantly larger drops in blood pressure (approximately 65 mm Hg) that were maintained for the 60 min of the experiment. This increased responsiveness to endotoxin in sialadenectomized and ganglionectomized animals was not related to changes in haematocrit, white blood cell counts, or neutrophil activity. Pretreatment of rats with pentoxifylline (PTX) produced substantial protection against the hypotensive effects of endotoxin in ganglionectomized and sialadenectomized rats, although this xanthine did not affect the responses of sham-operated rats. These results suggest that the submandibular gland elaborates a factor that protects against acute hypotension induced by endotoxin and that this factor is under the control of the cervical sympathetic nervous system.

Animals↗