Effect of bacterial lipopolysaccharide on gastric emptying of liquids in rats

The objectives of the present investigation were 1) to study the effect of bacterial lipopolysaccharide (LPS) on rat gastric emptying (GE) and 2) to investigate a possible involvement of the vagus nerve in the gastric action of LPS. Endotoxin from E. coli (strain 055:B5) was administered sc, ip or iv to male Wistar rats (220-280 g body weight) at a maximum dose of 50 µg/kg animal weight. Control animals received an equivalent volume of sterile saline solution. At a given time period after LPS administration, GE was evaluated by measuring gastric retention 10 min after the orogastric infusion of a test meal (2 ml/100 g animal weight), which consisted of 0.9% NaCl plus the marker phenol red (6 mg/dl). One group of animals was subjected to bilateral subdiaphragmatic vagotomy or sham operation 15 days before the test. A significant delay in GE of the test meal was observed 5 h after iv administration of the endotoxin at the dose of 50 µg/kg animal weight. The LPS-induced delay of GE was detected as early as 30 min and up to 8 h after endotoxin administration. The use of different doses of LPS ranging from 5 to 50 µg/kg animal weight showed that the alteration of GE was dose dependent. In addition, vagotomized animals receiving LPS displayed a GE that was not significantly different from that of the sham control group. However, a participation of the vagus nerve in LPS-induced delay in GE could not be clearly demonstrated by these experiments since vagotomy itself induced changes in this gastric parameter. The present study provides a suitable model for identifying the mechanisms underlying the effects of LPS on gastric emptying

Fibronectin in the ascitic fluid of cirrhotic patients: correlation with biochemical risk factors for the development of spontaneous bacterial peritonitis

Cirrhotic patients (23 with alcoholic cirrhosis, 5 with posthepatitic cirrhosis and 2 with cryptogenic cirrhosis) with ascites and portal hypertension were studied and divided into two groups corresponding to high or low risk to develop spontaneous bacterial peritonitis (SBP) related to the concentration of total protein in the ascitic fluid (A-TP): group I (high risk): A-TP£1.5 g/dl and group II (low risk): A-TP>1.5 g/dl. Fibronectin (FN), C3 and C4 concentrations were measured by radial immunodiffusion while total protein was measured by the biuret method. The mean values (group I vs group II) of C3 (12.59 ± 4.72 vs 24.53 ± 15.58 mg/dl), C4 (4.26 ± 3.87 vs 7.26 ± 4.14 mg/dl) and FN (50.47 ± 12.49 vs 75.89 ± 24.70 mg/dl) in the ascitic fluid were significantly lower (P<0.05) in the group considered to be at high risk for SBP. No significant difference was observed in the plasma/ascites fibronectin ratio (3.91 ± 1.21 vs 3.80 ± 1.26) or gradient (131.46 ± 64.01 vs 196.96 ± 57.38) between groups. Fibronectin in ascites was significantly correlated to C3 (r = 0.76), C4 (r = 0.58), total protein (r = 0.73) and plasma FN (r = 0.58) (P<0.05). The data suggest that the FN concentration in ascites is related to the opsonic capacity of this fluid, and that its concentration in the ascitic fluid may be a biochemical risk factor indicator for the development of spontaneous bacterial peritonitis

The role of T cell subsets and cytokines in the regulation of intracellular bacterial infection

Cellular immune responses are a critical part of the host's defense against intracellular bacterial infections. Immunity to Brucella abortus crucially depends on antigen-specific T cell-mediated activation of macrophages, which are the major effectors of cell-mediated killing of this organism. T lymphocytes that proliferate in response to B. abortus were characterized for phenotype and cytokine activity. Human, murine, and bovine T lymphocytes exhibited a type 1 cytokine profile, suggesting an analogous immune response in these different hosts. In vivo protection afforded by a particular cell type is dependent on the antigen presented and the mechanism of antigen presentation. Studies using MHC class I and class II knockout mice infected with B. abortus have demonstrated that protective immunity to brucellosis is especially dependent on CD8+ T cells. To target MHC class I presentation we transfected ex vivo a murine macrophage cell line with B. abortus genes and adoptively transferred them to BALB/c mice. These transgenic macrophage clones induced partial protection in mice against experimental brucellosis. Knowing the cells required for protection, vaccines can be designed to activate the protective T cell subset. Lastly, as a new strategy for priming a specific class I-restricted T cell response in vivo, we used genetic immunization by particle bombardment-mediated gene transfer

The effect of bacterial lipopolysaccharide on gastric emptying in rats suffering from moderate renal insufficiency

The objective of the present study was to evaluate the response of rats suffering from moderate renal insufficiency to bacterial lipopolysaccharide (LPS, or endotoxin). The study involved 48 eight-week-old male SPF Wistar rats (175-220 g) divided into two groups of 24 animals each. One group underwent 5/6 nephrectomy while the other was sham-operated. Two weeks after surgery, the animals were further divided into two subgroups of 12 animals each and were fasted for 20 h but with access to water ad libitum. One nephrectomized and one sham-treated subgroup received E. coli LPS (25 µg/kg, iv) while the other received a sterile, pyrogen-free saline solution. Gastric retention (GR) was determined 10 min after the orogastric infusion of a standard saline test meal labeled with phenol red (6 mg/dl). The gastric emptying of the saline test meal was studied after 2 h. Renal function was evaluated by measuring the plasma levels of urea and creatinine. The levels of urea and creatinine in 5/6 nephrectomized animals were two-fold higher than those observed in the sham-operated rats. Although renal insufficiency did not change gastric emptying (median %GR = 26.6 for the nephrectomized subgroup and 29.3 for the sham subgroup), LPS significantly retarded the gastric emptying of the sham and nephretomized groups (median %GR = 42.0 and 61.0, respectively), and was significantly greater (P<0.01) in the nephrectomized rats. We conclude that gastric emptying in animals suffering from moderate renal insufficiency is more sensitive to the action of LPS than in sham animals

Structural, functional and immunological studies on a polymeric bacterial protein

The characterization of proteins from Brucella spp, the causative agent of brucellosis, has been the subject of intensive research. We have described an 18-kDa cytoplasmic protein of Brucella abortus and shown the potential usefulness of this protein as an antigen for the serologic diagnosis of brucellosis. The amino acid sequence of the protein showed a low but significant homology with that of lumazine synthases. Lumazine is an intermediate product in bacterial riboflavin biosynthesis. The recombinant form of the 18-kDa protein (expressed in E. coli) folds like the native Brucella protein and has lumazine-synthase enzymatic activity. Three-dimensional analysis by X-ray crystallography of the homolog Bacillus subtilis lumazine synthase has revealed that the enzyme forms an icosahedral capsid. Recombinant lumazine synthase from B. abortus was crystallized, diffracted X rays to 2.7-Å resolution at room temperature, and the structure successfully solved by molecular replacement procedures. The macromolecular assembly of the enzyme differs from that of the enzyme from B. subtilis. The Brucella enzyme remains pentameric (90 kDa) in its crystallographic form. Nonetheless, the active sites of the two enzymes are virtually identical at the structural level, indicating that inhibitors of these enzymes could be viable pharmaceuticals across a broad species range. We describe the structural reasons for the differences in their quaternary arrangement and also discuss the potential use of this protein as a target for the development of acellular vaccines.

Bacterial community structure and petroleum hydrocarbon degradation in the Baltic Sea

The Baltic Sea is unique by its biological, geochemical and physical features. The number of species of larger organisms is small and the species composition is distinctive. On the contrary microbial communities are diverse. Because of the low salinity levels, bacterial communities differ from the ones in the oceans. Knowing the structure of these communities better and how they response to different environmental conditions helps us to estimate how different factors affect the balance and function of the Baltic Sea ecosystem. Bacteria are the key players when it comes to natural biogeochemical processes and human-induced phenomena like eutrophication, oil spills or disposal of other harmful substances to the sea ecosystem. In this thesis, bacterial community structure in the sea surface microlayer and subsurface water of the Archipelago Sea were compared. In addition, the effect of diatom derived polyunsaturated aldehydes on bacterial community structure was studied by a mesocosm experiment. Diesel, crude oil and polycyclic aromatic hydrocarbon degradation capacity of the Baltic Sea bacteria was studied in smaller scale microcosm experiments. In diesel oil experiments bacteria from water phase of the Archipelago Sea was studied. Sediment and iron manganese concretions collected from the Gulf of Finland were used in the crude oil and polycyclic aromatic hydrocarbon experiments. The amount of polycyclic aromatic hydrocarbon degradation genes was measured in all of the oil degradation experiments. The results show how differences in bacterial community structure can be seen in the sea surface when compared to the subsurface waters. The mesocosm experiment demonstrated how diatom-bacteria interactions depend on other factors than diatom derived polyunsaturated aldehydes, which do not seem to have an effect on the bacterial community structure as has been suggested in earlier studies. The dominant bacterial groups in the diesel microcosms differed in samples taken from a pristine site when compared to a site with previous oil exposure in the Archipelago Sea area. Results of the study with sediment and iron-manganese concretions indicate that there are diverse bacterial communities, typical to each bottom type, inhabiting the bottoms of the Gulf of Finland capable to degrade oil and polycyclic aromatic hydrocarbon compounds.  

Identification and in vitro production of Lactobacillus antagonists from women with or without bacterial vaginosis

Lactobacilli isolated from the vaginal tract of women with and without bacterial vaginosis (BV) were identified and characterized for the production of antagonists. Bacterial samples were isolated from healthy women (N = 16), from patients with clinical complaints but without BV (N = 30), and from patients with BV (N = 32). Identification was performed using amplified ribosomal DNA restriction analysis. Production of antagonistic compounds was evaluated by the double-layer diffusion technique using Gram-positive (N = 9) and Gram-negative bacteria (N = 6) as well as yeast (N = 5) as indicator strains. Of a total of 147 isolates, 133 were identified as pertaining to the genus Lactobacillus. Lactobacillus crispatus was the species most frequently recovered, followed by L. johnsonii and L. jensenii. Statistical analysis showed that L. crispatus was more frequent in individuals without BV (P < 0.05). A higher production of antagonists was noted in L. crispatus isolates from healthy women (P < 0.05). More acidic local pH and higher H2O2 production by isolated lactobacilli from healthy women suggest these mechanisms as the possible cause of this antagonism. In conclusion, a significant correlation was detected between the presence and antagonistic properties of certain species of Lactobacillus and the clinical status of the patients.

Randomized clinical trial comparing the efficacy of the vaginal use of metronidazole with a Brazilian pepper tree (Schinus) extract for the treatment of bacterial vaginosis

A 7.4% vaginal extract of the Brazilian pepper tree (Schinus terebinthifolius Raddi) was compared with 0.75% vaginal metronidazole, both manufactured by the Hebron Laboratory, for the treatment of bacterial vaginosis, used at bedtime for 7 nights. The condition was diagnosed using the combined criteria of Amsel and Nugent in two groups of 140 and 137 women, aged between 18 and 40 years. Intention-to-treat analysis was performed. Women were excluded from the study if they presented delayed menstruation, were pregnant, were using or had used any topical or systemic medication, presented any other vaginal infections, presented hymen integrity, or if they reported any history suggestive of acute pelvic inflammatory disease. According to Amsel’s criteria separately, 29 patients (21.2%) treated with the extract and 87 (62.1%) treated with metronidazole were considered to be cured (P < 0.001). According to Nugent’s score separately, 19 women (13.9%) treated with the extract and 79 (56.4%) treated with metronidazole were considered to be cured (P < 0.001). Using the two criteria together, the so-called total cure was observed in 17 women (12.4%) treated with the extract and in 79 women (56.4%) treated with metronidazole (P < 0.001). In conclusion, the cure rate for bacterial vaginosis using a vaginal gel from a pepper tree extract was lower than the rate obtained with metronidazole gel, while side effects were infrequent and non-severe in both groups.

Typical and atypical enteropathogenic Escherichia coli bacterial translocation associated with tissue hypoperfusion in rats

Although enteropathogenic Escherichia coli (EPEC) are well-recognized diarrheal agents, their ability to translocate and cause extraintestinal alterations is not known. We investigated whether a typical EPEC (tEPEC) and an atypical EPEC (aEPEC) strain translocate and cause microcirculation injury under conditions of intestinal bacterial overgrowth. Bacterial translocation (BT) was induced in female Wistar-EPM rats (200-250 g) by oroduodenal catheterization and inoculation of 10 mL 10(10) colony forming unit (CFU)/mL, with the bacteria being confined between the duodenum and ileum with ligatures. After 2 h, mesenteric lymph nodes (MLN), liver and spleen were cultured for translocated bacteria and BT-related microcirculation changes were monitored in mesenteric and abdominal organs by intravital microscopy and laser Doppler flow, respectively. tEPEC (N = 11) and aEPEC (N = 11) were recovered from MLN (100%), spleen (36.4 and 45.5%), and liver (45.5 and 72.7%) of the animals, respectively. Recovery of the positive control E. coli R-6 (N = 6) was 100% for all compartments. Bacteria were not recovered from extraintestinal sites of controls inoculated with non-pathogenic E. coli strains HB101 (N = 6) and HS (N = 10), or saline. Mesenteric microcirculation injuries were detected with both EPEC strains, but only aEPEC was similar to E. coli R-6 with regard to systemic tissue hypoperfusion. In conclusion, overgrowth of certain aEPEC strains may lead to BT and impairment of the microcirculation in systemic organs.

Bacterial ecology of tilapia fresh fillets and some factors that can influence their microbial quality

The purpose of research was to investigate the bacterial ecology of tilapia (Oreochromis niloticus) fresh fillets and some factors that can influence its microbial quality. Samples of fish cultivation water (n = 20), tilapia tegument and gut (n = 20) and fresh fillets (n = 20) were collected in an experimental tilapia aquaculture located in the city of Lavras, Minas Gerais, Brazil. Staphylococcus spp., Aeromonas spp., Enterococcus spp. and Enterobacteriaceae were quantified using selective plating. For the enumeration of Pseudomonas spp., the most probable number technique (MPN) was utilized. Bacterial colonies (n = 198) were identified by Gram strain and biochemical tests. Aeromonas spp., Pseudomonas spp., Enterococcus spp. and Enterobacteriaceae were found in the cultivation water (water from a fishpond cultivation), tegument, gut, and fresh fillets. Staphylococcus spp. was not isolated in the cultivation water. Salmonella spp. was not detected. The count variable of 10 to 10³ CFU or MPN.(g or mL)-1. Associated to freshwater tilapia fillet processing, there is a large variety of microorganisms related to foodborne illnesses and fish products deterioration.

Hygienic-sanitary quality of vegetables and evaluation of treatments for the elimination of indigenous E. coli and E. coli O157:H7 from the surface of leaves of lettuce (Lactuca sativa L.)

The purpose of this study is to evaluate the hygienic-sanitary quality of vegetables and irrigation water and assess the effectiveness of lemon juice and vinegar in reducing E. coli strains inoculated on lettuce. One hundred and forty samples of vegetables and 45 samples of irrigation water were investigated for thermotolerant coliforms and Salmonella spp. In order to verify the effectiveness of natural household sanitizers in reducing E. coli in inoculated lettuce, four treatment solutions were tested: fresh lemon juice, alcohol vinegar, lemon juice-vinegar mixture, and lemon juice-vinegar-water mixture. The microbiological analysis revealed high rates of contamination by thermotolerant coliforms and identified the presence of E. coli in 32% of the tested vegetable samples and 56% of the water samples. While no significant statistical difference (p < 0, 05) was identified in the tested solutions, the treatment with a combination of lemon juice and vinegar resulted in the highest Decimal Reductions (DR) of E. coli O157: H7 while the treatment with vinegar alone was the most effective against the indigenous E. coli strain

Isolation and characterization of bacterial strains with a hydrolytic profile with potential use in bioconversion of agroindustial by-products and waste

There is a trend towards the use of novel technologies nowadays, mainly focused on biological processes, for recycling and the efficient utilization of organic residues that can be metabolized by different microorganisms as a source of energy. In the present study the isolation of bacterial strains from six different agro-industrial by-products and waste was performed with the objective of evaluating their hydrolytic capacities and suitability for use in bioconversion of specific substrates. The 34 isolated strains were screened in specific culture media for the production of various hydrolytic enzymes (lipase, protease, cellulase, and amylase). It was found that 28 strains exhibited proteolytic activity, 18 had lipolytic activity, 13 had caseinolytic activity, 15 had amylolytic activity, and 11 strains exhibited cellulolytic activity. The strains that showed the highest hydrolytic capacities with biotechnological potential were selected, characterized genotipically, and identified as Bacillus, Serratia, Enterococcus, Klebsiella, Stenotrophomonas, Lactococcus, and Escherichia genera. It was concluded that the strain isolates have a high potential for use in the bioconversion of agro-industrial waste, both as a pure culture and as a microbial consortium.

Green and brown propolis: efficient natural biocides for the control of bacterial contamination of alcoholic fermentation of distilled beverage

This study aimed to evaluate the efficiency of natural biocides, brown and green propolis, for the control of bacterial contamination in the production of sugarcane spirit. The treatments consisted of brown and green propolis extracts, ampicillin, and a control and were assessed at the beginning and end of harvest season in ten fermentation cycles. In the microbiological analyses, the lactic acid bacteria were quantified in the inoculum before and after the treatment with biocides, and the viability of yeast cells during fermentation was evaluated. The levels of acids, glycerol, total residual reducing sugars, and ethanol were analyzed for the wine resulting from each fermentation cycle. A reduction in the number of bacterial contaminants in the inoculum in the treatments with the natural biocides was observed, but it did not affect the viability of yeast cells. The control of the contaminants led to the production of higher levels of ethanol and reduced acidity in the wine produced. The results of the use of brown and green propolis to control the growth microorganisms in the fermentation of sugarcane spirit can be of great importance for using alternative strategies to synthetic antibacterials in fermentation processes including other distilled beverage or spirits.

Individual Crimes or a Sociological Phenomenon: A Critical discourse analysis of the 2014 Royal Canadian Mounted Police National Operational Overview on missing and murdered Indigenous women in Canada

Pro gradu -tutkielma käsittelee Kanadan poliisivoimien, Royal Canadian Mounted Police (RCMP), vuoden 2014 operationaalista raporttia Kanadan kadonneista ja murhatuista alkuperäiskansojen naisista. Heitä katoaa ja murhataan Kanadassa suhteessa huomattavasti enemmän kuin muita naisia. Aihe on hyvin ajankohtainen sekä kriittinen ja siihen on paikallisten organisaatioiden lisäksi kiinnittänyt huomiota mm. Amnesty International ja Yhdistyneet Kansakunnat. Tilannetta pidetään jo ihmisoikeuskriisinä ja sen puolesta puhuminen voidaan nähdä osana laajempaa kansannousua alkuperäiskansojen oikeuksien puolesta. Kanadan hallitusta sekä poliisivoimia on syytetty välinpitämättömyydestä ja rasismista, poliiseja jopa väkivallasta alkuperäiskansojen naisia kohtaan. Kanadan hallitus ei myönnä ongelman olevan sosiologinen ilmiö, vaan pääministerin mukaan naisten katoamiset ja murhat ovat yksittäisiä rikoksia. Tilanteen taustalla on laajempi ongelma, joka juontaa juurensa kolonialismista, alkuperäiskansojen pakkokoulutuksesta ja heidän kulttuurinsa sekä identiteettinsä tuhoamisesta. Ennakkoluulot ja stereotypiat elävät yhä vahvana luoden heikon aseman alkuperäiskansojen edustajille ja tässä tapauksessa erityisesti naisille, joita myös usein syytetään omista ongelmistaan. Tutkielma selvittää, onko tämä operationaalinen raportti eriarvoisuutta, stereotypioita sekä ennakkoluuloja ylläpitävä ja tukeeko se näkemystä sosiologisesta ilmiöstä. Poliisivoimat ovat yhteiskunnassa vaikutusvaltaisessa asemassa ja voivat edesauttaa, jatkaa ja voimistaa syrjivää ajattelua, käytöstä ja viestintää. Vaikka he myös ajavat alkuperäiskansojen etuja, tulee heidän toimintaansa ja täten tätä kyseistä raporttia tarkastella kriittisesti. Tutkielma keskittyy kriittiseen diskurssianalyysiin (Fairclough 1995, Wodak & Meyer 2009). Raportin teksti ei ole avoimesti diskriminoivaa ja keskittyy tilastojen ja lukujen raportointiin sekä vertailuun, luoden samalla kuvan asianomaisista sekä tilanteesta yleisesti. Makroanalyysi (van Dijk 1977, 1980) mahdollistaa korkeamman tason ajatusten ja ideoiden esille tuonnin paljastaen ns. rivien välistä löytyvän viestinnän. Näistä makroanalyysin tuloksista etsitään erilaisia diskurssistrategioita (Reisigl & Wodak 2001), joiden avulla luodaan mm. kuva positiivisesta itsestä ja negatiivisesta toisesta. Näitä tuloksia tuetaan lyhyellä multimodaalisella analyysillä, joka keskittyy lähinnä raportin kahteen kuvaan (Kress & van Leeuwen 2006). Lopuksi tuloksista keskustellaan suhteessa yhteiskunnan valtasuhteisiin (Foucault 1976), rasismiin ja seksismiin. Analyysin perusteella raportti on selkeästi diskriminoiva. Se tuo esiin vanhoja stereotypioita ja ennakkoluuloja, esittää tilanteen alkuperäiskansojen sisäisenä ongelmana, jättää mainitsematta tilanteen taustat ja luo hyvin positiivisen kuvan poliisivoimista (RCMP). Tällä tavoin se ylläpitää yhteiskunnan epätasapainoisia valtasuhteita ja alkuperäiskansojen naisten heikompaa ja marginalisoitua asemaa. Raportti todistaa kuinka syvällä Kanadan yhteiskunnassa jopa institutionaalisella tasolla nämä ajatukset piilevät, sillä diskriminaatio ulottuu tapausten syistä niiden käsittelyyn ja raportointiin. Tilanne on huolestuttava ja voidaan nähdä sosiologisena ilmiönä. Poliisivoimien tulisi lisätä sisäistä koulutusta asian suhteen sekä hallituksen tutkia tätä ilmiönä ja paneutua sen selvittämiseen laaja-alaisesti ja läpinäkyvästi.

Individual Crimes or a Sociological Phenomenon: A Critical discourse analysis of the 2014 Royal Canadian Mounted Police National Operational Overview on missing and murdered Indigenous women in Canada

Pro gradu -tutkielma käsittelee Kanadan poliisivoimien, Royal Canadian Mounted Police (RCMP), vuoden 2014 operationaalista raporttia Kanadan kadonneista ja murhatuista alkuperäiskansojen naisista. Heitä katoaa ja murhataan Kanadassa suhteessa huomattavasti enemmän kuin muita naisia. Aihe on hyvin ajankohtainen sekä kriittinen ja siihen on paikallisten organisaatioiden lisäksi kiinnittänyt huomiota mm. Amnesty International ja Yhdistyneet Kansakunnat. Tilannetta pidetään jo ihmisoikeuskriisinä ja sen puolesta puhuminen voidaan nähdä osana laajempaa kansannousua alkuperäiskansojen oikeuksien puolesta. Kanadan hallitusta sekä poliisivoimia on syytetty välinpitämättömyydestä ja rasismista, poliiseja jopa väkivallasta alkuperäiskansojen naisia kohtaan. Kanadan hallitus ei myönnä ongelman olevan sosiologinen ilmiö, vaan pääministerin mukaan naisten katoamiset ja murhat ovat yksittäisiä rikoksia. Tilanteen taustalla on laajempi ongelma, joka juontaa juurensa kolonialismista, alkuperäiskansojen pakkokoulutuksesta ja heidän kulttuurinsa sekä identiteettinsä tuhoamisesta. Ennakkoluulot ja stereotypiat elävät yhä vahvana luoden heikon aseman alkuperäiskansojen edustajille ja tässä tapauksessa erityisesti naisille, joita myös usein syytetään omista ongelmistaan. Tutkielma selvittää, onko tämä operationaalinen raportti eriarvoisuutta, stereotypioita sekä ennakkoluuloja ylläpitävä ja tukeeko se näkemystä sosiologisesta ilmiöstä. Poliisivoimat ovat yhteiskunnassa vaikutusvaltaisessa asemassa ja voivat edesauttaa, jatkaa ja voimistaa syrjivää ajattelua, käytöstä ja viestintää. Vaikka he myös ajavat alkuperäiskansojen etuja, tulee heidän toimintaansa ja täten tätä kyseistä raporttia tarkastella kriittisesti. Tutkielma keskittyy kriittiseen diskurssianalyysiin (Fairclough 1995, Wodak & Meyer 2009). Raportin teksti ei ole avoimesti diskriminoivaa ja keskittyy tilastojen ja lukujen raportointiin sekä vertailuun, luoden samalla kuvan asianomaisista sekä tilanteesta yleisesti. Makroanalyysi (van Dijk 1977, 1980) mahdollistaa korkeamman tason ajatusten ja ideoiden esille tuonnin paljastaen ns. rivien välistä löytyvän viestinnän. Näistä makroanalyysin tuloksista etsitään erilaisia diskurssistrategioita (Reisigl & Wodak 2001), joiden avulla luodaan mm. kuva positiivisesta itsestä ja negatiivisesta toisesta. Näitä tuloksia tuetaan lyhyellä multimodaalisella analyysillä, joka keskittyy lähinnä raportin kahteen kuvaan (Kress & van Leeuwen 2006). Lopuksi tuloksista keskustellaan suhteessa yhteiskunnan valtasuhteisiin (Foucault 1976), rasismiin ja seksismiin. Analyysin perusteella raportti on selkeästi diskriminoiva. Se tuo esiin vanhoja stereotypioita ja ennakkoluuloja, esittää tilanteen alkuperäiskansojen sisäisenä ongelmana, jättää mainitsematta tilanteen taustat ja luo hyvin positiivisen kuvan poliisivoimista (RCMP). Tällä tavoin se ylläpitää yhteiskunnan epätasapainoisia valtasuhteita ja alkuperäiskansojen naisten heikompaa ja marginalisoitua asemaa. Raportti todistaa kuinka syvällä Kanadan yhteiskunnassa jopa institutionaalisella tasolla nämä ajatukset piilevät, sillä diskriminaatio ulottuu tapausten syistä niiden käsittelyyn ja raportointiin. Tilanne on huolestuttava ja voidaan nähdä sosiologisena ilmiönä. Poliisivoimien tulisi lisätä sisäistä koulutusta asian suhteen sekä hallituksen tutkia tätä ilmiönä ja paneutua sen selvittämiseen laaja-alaisesti ja läpinäkyvästi.