Caecal ligation and puncture in the rat mimics the pathophysiological changes in human sepsis and causes multi-organ dysfunction.

Sepsis is a major clinical challenge that is associated with encephalopathy and multi-organ dysfunction. Current therapeutic interventions are relatively ineffective and the development of novel treatments is hampered by the lack of a well-characterised animal model. Therefore, the behavioural, metabolic, physiological and histological changes resulting from 'through and through' caecal ligation and puncture (CLP) in the rat were investigated to determine its suitability as an animal model of human sepsis. CLP resulted in bacteraemia, characterised by the presence of multiple enteric species within 18-20 h. Locomotor activity was reduced within 4 h of CLP and this reduction increased with time. Pyrexia was evident 4-5 h after CLP and was followed by hypothermia beginning 17 h after intervention. CLP resulted in reduced white blood cell and platelet counts and an increased neutrophil: lymphocyte ratio within 18-20 h. It also resulted in decreased blood glucose, but not lactate levels. CLP caused histopathological changes in the cerebral cortex, liver, lungs and vascular system indicative of multi-organ dysfunction. Therefore, CLP in the rat mimics the cardinal clinical features of human sepsis and the subsequent development of multi-organ dysfunction. It appears to be the best available animal model currently available, in which to investigate the underlying pathophysiology of sepsis and identify therapeutic targets.

Occludin expression in microvessels of neoplastic and non-neoplastic human brain.

The tight junction protein occludin 'glues' normal, adjacent brain microvessel endothelial cells together. Malignant brain tumours cause cerebral oedema because they have leaky endothelial tight junctions, which allow plasma fluid to enter the brain from the microvessel lumen. In order to identify molecular abnormalities in tumour endothelial tight junctions, we investigated occludin expression in microvessels from adult human non-neoplastic brain tissue using immunohistochemistry and immunoblotting. The proportions of microvessels immunolabelling for occludin were >2/3 in 5/5 non-neoplastic brain tissue samples, >1/3 in 5/5 low grade (Daumas-Duport I or II) astrocytomas and <1/3 in 5/5 high grade (III or IV) astrocytomas and 6/6 metastatic adenocarcinomas. Six non-neoplastic brain tissue immunoblots gave a 55-kDa occludin band, three low-grade astrocytomas gave 55-kDa and 60-kDa bands, 13 high-grade astrocytomas gave 60-kDa or no band and four adenocarcinomas did not give an occludin band. Expression of 55-kDa occludin inversely correlated with the presence of contrast enhancement on computed tomograms (P < 0.001). Electron microscopy showed open endothelial tight junctions in 0/2 non-neoplastic human brain specimens and 2/2 high-grade astrocytomas. We suggest that loss of 55-kDa occludin expression in human brain tumours may contribute to endothelial tight junction opening. Characterizing the molecular pathology of brain endothelial tight junctions may facilitate the design of novel drugs against cerebral oedema.

Pathophysiology of septic encephalopathy: a review.

OBJECTIVES: Encephalopathy is a common complication of sepsis. This review describes the different pathologic mechanisms that may be involved in its etiology. DATA SOURCES: The studies described here were derived from the database PubMed (http:¿¿www.nlm.nih.gov) and from references identified in the bibliographies of pertinent articles and books. The citations are largely confined to English language articles between 1966 and 1998. Older publications were used if they were of historical significance. STUDY SELECTION: All investigations in which any aspect of septic encephalopathy was reported were included. This selection encompasses clinical, animal, and in vitro cell culture work. DATA EXTRACTION: The literature cited was published in peer-reviewed clinical or basic science journals or in books. DATA SYNTHESIS: Contradictions between the results of published studies are discussed. CONCLUSIONS: The most immediate and serious complication of septic encephalopathy is impaired consciousness, for which the patient may require ventilation. The etiology of septic encephalopathy involves reduced cerebral blood flow and oxygen extraction by the brain, cerebral edema, and disruption of the blood-brain barrier that may arise from the action of inflammatory mediators on the cerebrovascular endothelium, abnormal neurotransmitter composition of the reticular activating system, impaired astrocyte function, and neuronal degeneration. Currently, there is no treatment.

Faecal peritonitis causes oedema and neuronal injury in pig cerebral cortex.

Encephalopathy is a common complication of sepsis. However, little is known about the morphological changes that occur in the brain during sepsis. Faecal peritonitis was induced in pigs that were killed 8 h later and frontal cortex samples were taken immediately after death. The tissue was investigated using light and electron microscopy and compared with frontal cortex samples taken from sham-operated controls. Septic pigs had 49.5% more perimicrovessel oedema than sham pigs. However, the tight junctions between cerebral microvessel endothelial cells appeared morphologically intact in both septic and sham pigs. Sepsis also resulted in neuronal injury, disruption of astrocytic end-feet and swollen, rounded erythrocytes. These morphological changes may be sufficient to underlie the clinical features seen in septic encephalopathy.

Starch, gloves and extradural catheters.

We have investigated contamination of extradural catheters during normal handling with starch powdered gloves. In the laboratory, extradural catheters were handled in sterile fields with both powdered and non-powdered gloves, simulating preparation for patient insertion. The catheters together with glove samples were then examined using a Zeiss 940 scanning electron microscope. Microscopy of samples revealed starch contamination of the catheters handled with powdered gloves, especially in the side hole areas. We conclude that extradural catheters may be contaminated easily by starch powder from surgical gloves. This powder may then be deposited into the extradural space. The effect of starch in the extradural space is not known but starch is known to cause inflammatory and granulomatous reactions in other parts of the body as well as being directly allergenic. Powder contamination of catheters may be avoided easily by the use of powder-free gloves and we feel that these should be used whenever possible.

A narrow range, medium molecular weight pentastarch reduces structural organ damage in a hyperdynamic porcine model of sepsis.

OBJECTIVE: to compare diafiltered 6% pentastarch (Pentafraction--PDP, MWn 120,000 and MWw 280,000) and native pentastarch (Pentaspan--PSP, MWn 63,000 and MWw 264,000 dalton) in a porcine model of faecal peritonitis. DESIGN: Randomised prospective study in 12 adolescent pigs. INTERVENTIONS: Prior to infection the study solution was infused to increase Qt by 25%. Thereafter adjustments in infusion rate were made (up to 1 l/h) in an attempt to maintain Qt at 25% above baseline values. MEASUREMENTS AND RESULTS: Animals were sacrificed at 8 h. Tissue was excised from the right lobe of liver and from the right lung and fixed for later electron microscopy and digital morphometric analysis. Patent sinusoidal lumen was significantly greater in group PDP compared to PSP (11.3% +/- 2.3% of liver tissue versus 4.8% +/- 1.1%, p < 0.05) and this was accounted for by a significantly lower proportion of sinusoidal lumen occluded with white cells (2.1% +/- 0.6% versus 6.6% +/- 1.9%, p < 0.05). Similarly, patent capillary represented a significantly higher proportion of lung tissue for group PDP versus PSP (26.2% +/- 1.9% versus 18.5% +/- 2.7%, p < 0.05). The arithmetic mean alveolar capillary barrier thickness was significantly greater in group PSP than in group PDP (4.3 +/- 0.3 microns versus 2.5 +/- 0.3 microns, p < 0.01). CONCLUSIONS: The molecular weight profile of Pentafraction was associated with less structural organ damage including less tissue oedema and less white cell occlusion.

Advantages of a narrow-range, medium molecular weight hydroxyethyl starch for volume maintenance in a porcine model of fecal peritonitis.

OBJECTIVE: To compare the effectiveness of two hydroxyethyl starch solutions of different molecular weight ranges for volume maintenance in a porcine model of fecal peritonitis. DESIGN: Randomized prospective trial. SETTING: Laboratory investigation. SUBJECTS: Adolescent female pigs weighing approximately 30 kg. INTERVENTIONS: We compared diafiltered 6% pentastarch with 6% high molecular weight hetastarch for volume maintenance in a porcine model of fecal peritonitis. The number average molecular weight of pentastarch is higher than hetastarch, although the weight average molecular weight is lower, i.e., a narrow range of medium weight molecules. The infusion rate of each agent was adjusted to maintain baseline arterial Hct for less than or equal to 7 hr after instrumentation and induction of fecal peritonitis. MAIN OUTCOME MEASUREMENTS: The volume of fluid required to maintain arterial Hct was compared along with comparisons of hemodynamic and histologic responses associated with the two agents. RESULTS: Significantly less pentastarch was required to prevent hemoconcentration than hetastarch (109 +/- 22.8 vs. 150 +/- 10.3 mL/kg; p less than .05) while hemodynamics, colloid osmotic pressure, and oxygen transport responses were similar. Capillary patency was greater (21.99 +/- 3.68 vs. 10.09 +/- 1.17%; p less than .05) and mean alveolar capillary barrier thickness was less (2.36 +/- 0.13 vs. 3.06 +/- 0.17 microns; p less than .05) with pentastarch than with hetastarch, as judged by electron microscopy. CONCLUSIONS: These data suggest that pentastarch is better retained in the circulation in capillary leak syndromes compared with hetastarch.

The effects of dobutamine, dopexamine and fluid on hepatic histological responses to porcine faecal peritonitis.

We studied the effects of two catecholamines with differing receptor profiles on hepatic blood flow and hepatic structure in a porcine model of faecal peritonitis. We treated animals with dopexamine (group Dp) or dobutamine (group Db) and fluid, or fluid alone as a control, to achieve a 25% increase in Qt from baseline values. After the induction of faecal peritonitis the increased Qt was maintained throughout the 8 h study period by adjustment of the fluid infusion rate. The dose of catecholamines remained constant. Hepatic blood flow was correspondingly maintained at above baseline values throughout the study. Post-mortem liver biopsy specimens were analysed from experimental animals and 5 sham animals who had not been instrumented or infected. In experimental animals there was a reduction in sinusoidal patency between sham and group Dp (76% of total sinusoid vs 51%, p less than 0.05) and group Dp and control (51% vs 33%, p less than 0.05) or groups Dp and Db (51% vs 34%, p less than 0.05) animals. This was accounted for by an increase in sinusoidal leukocytes and endothelial swelling. In addition to the changes noted above there was marked hepatocellular destruction in group Db. We conclude that maintenance of organ blood flow does not guarantee structural integrity in the sepsis syndrome and hepatocellular damage was greater in group Db than group Dp or control.

Clinical pulmonary function and industrial respirator wear.

This investigation was the initial step in determining a clinical pulmonary test which could be used to evaluate workers as to their suitability to industrial respirator wear. Sixty subjects, 12 superior, 37 normal, and 11 moderately impaired with respect to lung function tests were evaluated with a battery of clinical pulmonary tests while wearing an industrial respirator. The respirator was a full-face mask (MSA-Ultravue) "demand" breathing type equipped with an inspiratory resistance of 85 mm H2O at 85 L/min air flow and an expiratory resistance of 25 mmH2O at 85 L/min air flow. Comparisons of these tests were made between the three groups of subjects both with and without a respirator. It appears that those lung tests which measure the flow characteristics of the lung especially those that are effort dependent are more susceptible to change as a result of respirator wear. Hence, the respirator affects the person with superior lung function to a greater degree than the moderately impaired person. It was suggested that the clinical test of 15 second maximum voluntary ventilations (MVV.25) may be the test of choice for determining worker capability in wearing an industrial respirator.

The physiological responses of mild pulmonary impaired subjects while using a "demand" respirator during rest and work.

This investigation determined the cardiorespiratory responses of subjects with normal lung function and exercise tolerance and compared them with subjects with moderate and severe impairment of lung function and exercise tolerance. Comparisons were made during work while wearing an industrial respirator. The respirator was a full-face mask (MSA-ultravue) "demand" breathing type equipped with an inspiratory resistance of 85 mm H2O at 85 L/min air flow and an expiratory resistance of 25 mm H2O at 85 L/min air flow. Physiologically and subjectively the response of the normal and moderately impaired subjects to respirator wear during rest, 35%, 50% and 63% of their maximal workloads were not different. However, when the moderately impaired worked at 63% of their maximum the workload was equivalent to 50% of maximum of the normal subject. Significant differences in the peak flow/pressure ratio of the severely impaired compared to the normals and moderately impaired were found. By relating work performance to the dyspnea index (VI/MBC) it was suggested that the relationship between maximal lung function and maximal work performance needs to be identified both with and without respirators. This relationship may prove suitable in predicting performance during respirator wear.

Primary atrophic rhinitis: a scanning electron microscopic (SEM) study.

The surface features of atrophic rhinitis are shown and it is suggested that these explain the majority of symptoms. It seems clear that any lesion preventing the formation or maturation of large numbers of motile cilia, or the production of mucus capable of forming confluent sheets suitable for continuous propulsion, may cause atrophic rhinitis. In both familial and idiopathic forms of the disease, both abnormalities are present. It would be most interesting to know the ultrastructure of the cilia in transverse section of this condition. Transverse electron microscopic studies in Retinitis Pigmentosa, Usher's syndrome and Kartagener's syndrome now becoming known as the low cilia motility diseases show clearly the primary lesions in the micro-tubules and dynein arms of the cilia. A similar transmission electron microscopic study in atrophic rhinitis may show the fundamental cilial lesions. Surgical closure of the nasal passages has much to offer the patient with severe symptoms as the clinical features of the disease improve with the increasing normal micro-anatomical features demonstrated in this paper as a result of closure. This improvement in structure is not in all cases sufficient to fulfil the above criteria so a complete cure is improbable in the majority of cases.