Imaging African trypanosomes.

Trypanosoma brucei are extracellular kinetoplastid parasites transmitted by the blood-sucking tsetse fly. They are responsible for the fatal disease human African trypanosomiasis (HAT), also known as sleeping sickness. In late-stage infection, trypanosomes cross the blood-brain barrier (BBB) and invade the central nervous system (CNS) invariably leading to coma and death if untreated. There is no available vaccine and current late-stage HAT chemotherapy consists of either melarsoprol, which is highly toxic causing up to 8% of deaths, or nifurtimox-eflornithine combination therapy (NECT), which is costly and difficult to administer. There is therefore an urgent need to identify new late-stage HAT drug candidates. Here, we review how current imaging tools, ranging from fluorescent confocal microscopy of live immobilized cells in culture to whole-animal imaging, are providing insight into T. brucei biology, parasite-host interplay, trypanosome CNS invasion and disease progression. We also consider how imaging tools can be used for candidate drug screening purposes that could lead to new chemotherapies.

The effect of pyridoxal-5-phosphate on serum alanine aminotransferase activity in dogs suffering from canine babesiosis.

Accurate measurements of serum aminotransferase (ALT) activity in dogs relies on the endogenous pro-enzyme pyridoxal 5-phosphate (P5P). The purpose of this study was to determine whether the exclusion of P5P from the analytical method causes an underestimation of serum ALT activity in dogs suffering from babesiosis and in those manifesting evidence of hepatocellular damage, and to determine if anorexia causes sufficient P5P depletion to affect in vitro serum ALT activity. One-hundred-and-twenty healthy control dogs and 105 Babesia-infected dogs were included in the study. Two methods for ALT measurement were used: Method 1 included P5P, and Method 2 excluded P5P from the reaction mixture. Higher serum ALT activity was measured with Method 1 in the Babesia-infected dogs (P < 0.001), as well as in 14 dogs with suspected hepatocellular damage (P = 0.03). Duration of anorexia had no effect, irrespective of the method used. Although inclusion of P5P to the reaction mixture consistently resulted in higher measured serum ALT activity, the differences were too small to have led to incorrect diagnoses in the Babesia-infected dogs suspected of liver disease.

Estrogen receptor beta dominates in baboon carotid after endothelial denudation injury.

Increasing evidence, mainly from rodents, suggests that the predominant estrogen receptor (ER) in arteries is the newly-described ERbeta. We have investigated the expression of the two ERs in baboon carotid artery before and after denudation injury. Prior to denudation, both full length receptors were detected in semiquantitative RT-PCR; in addition two ERalpha but no ERbeta splicing variants were found. After denudation, ERbeta mRNA increased five-fold and declined, whereas ERalpha mRNA expression remained low. Prior to and after denudation, two ERalpha-specific antibodies showed no reaction with the vessel wall. Instead, two affinity purified antisera to ERbeta demonstrated a weak but distinct reaction over vascular smooth muscle cells with predenudation specimens, escalating post-denudation and declining thereafter. The results suggest that selective targeting to ERbeta should be attempted when designing estrogen-based vasculoprotective drug therapies devoid of uterotrophic side effects.

Review of blackfly (Diptera: Simuliidae) control in South Africa.

The medical, veterinary and economic importance of blackflies in South Africa, and the historical development of blackfly control programmes in various South African rivers, are reviewed in this paper. In 1996 it was estimated that blackflies can cause more than R 88 million damages per annum along the middle and lower Orange River where Simulium chutteri is considered the main pest species. A clear link between the construction of dams and the spread of the blackfly problem was shown. Four phases characterize the development of blackfly control in South Africa: (1) during the 1960s blackflies in the Vaal River were controlled with DDT; (2), during the 1970s and into the 1980s blackflies were controlled using water-flow manipulation; (3) when used at strategic times, water-flow manipulation could be used to enhance the effect of natural predator populations; and (4) during the 1990s the organophosphate temephos and toxins produced by the bacterium Bacillus thuringiensis var. israelensis were tested for their efficacy against blackflies. The larvicides temephos and B. thuringiensis proved to be effective and are still used in several control programmes. The latest research focuses on the factors that influence adult blackfly survival and annoyance, as well as the development of methods that can be used to protect sheep from blackfly attacks.

Changes in the blood-gas status of sheep with experimentally induced heartwater.

The blood-gas status of seven sheep with experimentally induced heartwater during the acute and terminal stages was investigated. Changes in blood gas included a decline in arterial oxygen tension (pO2) combined with a respiratory alkalosis. Although the sheep became hypoxaemic, blood-gas changes associated with respiratory failure were not observed.

Studies on the presence of endotoxin activity and inflammatory products (leukotrienes) in cases of experimentally induced heartwater in sheep.

The presence of endotoxin was examined in seven sheep with experimentally induced heartwater. Elevations in endotoxin levels were recorded in one sampling in three of the seven sheep during the acute stage of the disease. The elevations in endotoxin levels were of short duration and decreased in the 24-h follow-up samples. There was no elevation of leukotrienes (B4, C4 and D4) in the blood, or the thoracic or pericardial fluid of any of the sheep.

The clinical pathology of heartwater. III. Changes in blood clotting, blood calcium, blood protein, haematocrit and white-cell counts in sheep with experimentally induced heartwater.

Studies to evaluate changes in blood clotting, blood calcium and protein, the haematocrit and white-cell counts were undertaken in seven sheep with experimentally induced heartwater. A marked decline in thrombocyte count was recorded during the acute stage of the disease. This was associated with increases in both prothrombin time (PT) and activated partial thromboplastin time (APTT); fibrinogen increased while there was no detectable increase in fibrinogen degradation products (FDP). At the same time total serum protein (TSP), albumin and globulin dropped very sharply; total calcium showed a progressive drop but ionized calcium rose initially and was followed by a terminal decline. The total leucocyte count showed a terminal increase while the haematocrit dropped progressively.

A preliminary study on the serum protein response in canine babesiosis.

Total serum protein, albumin, globulin, globulin fractions (alpha, beta and gamma globulins) and an acute-phase protein (alpha1-acid glycoprotein) were evaluated in dogs with naturally occurring mild (Group 1), severe (Group 2) or complicated babesiosis (Group 3). Results showed that the total serum protein, albumin, A/G ratio and alpha globulins were statistically different between Groups 1 and 2. There was no statistical difference between groups with total, beta and gamma globulins. The findings from this study suggest that dogs with mild and severe babesiosis had low total serum proteins, albumin, A/G ratio and alpha globulins; dogs with complicated babesiosis showed no typical serum protein changes or patterns; and that there was no evidence of an acute-phase response detectable on serum protein electrophoresis in any of the 3 groups. A marked acute-phase response was, however, present, as measured by the alpha1-acid glycoprotein, in all 3 groups. As this was a retrospective study, the possibility that the observed responses were due in part to concurrent disease could not be excluded.

Systemic inflammatory response syndrome and multiple-organ damage/dysfunction in complicated canine babesiosis.

This study was designed to document the systemic inflammatory response syndrome (SIRS) and multiple-organ dysfunction syndrome (MODS) in dogs with complicated babesiosis, and to assess their impact on outcome. Ninety-one cases were evaluated retrospectively for SIRS and 56 for MODS. The liver, kidneys, lungs, central nervous system and musculature were assessed. Eighty-seven percent of cases were SIRS-positive. Fifty-two percent of the cases assessed for organ damage had single-organ damage and 48 % had MODS. Outcome was not significantly affected by either SIRS or MODS, but involvement of specific organs had a profound effect. Central nervous system involvement resulted in a 57 times greater chance of death and renal involvement in a 5-fold increased risk compared to all other complications. Lung involvement could not be statistically evaluated owing to co-linearity with other organs, but was associated with high mortality. Liver and muscle damage were common, but did not significantly affect outcome. There are many similarities between the observations in this study and previous human and animal studies in related fields, lending additional support to the body of evidence for shared underlying pathophysiological mechanisms in systemic inflammatory states.

IL-4Rα-responsive smooth muscle cells contribute to initiation of TH2 immunity and pulmonary pathology in Nippostrongylus brasiliensis infections.

Nippostrongylus brasiliensis infections generate pulmonary pathologies that can be associated with strong T(H)2 polarization of the host's immune response. We present data demonstrating N. brasiliensis-driven airway mucus production to be dependent on smooth muscle cell interleukin 4 receptor-α (IL-4Rα) responsiveness. At days 7 and 10 post infection (PI), significant airway mucus production was found in IL-4Rα(-/lox) control mice, whereas global knockout (IL-4Rα(-/-)) and smooth muscle-specific IL-4Rα-deficient mice (SM-MHC(Cre) IL-4Rα(-/lox)) showed reduced airway mucus responses. Furthermore, interleukin (IL)-13 and IL-5 cytokine production in SM-MHC(Cre) IL-4Rα(-/lox) mice was impaired along with a transient reduction in T-cell numbers in the lung. In vitro treatment of smooth muscle cells with secreted N. brasiliensis excretory-secretory antigen (NES) induced IL-6 production. Decreased protein kinase C (PKC)-dependent smooth muscle cell proliferation associated with cell cycle arrest was found in cells stimulated with NES. Together, these data demonstrate that both IL-4Rα and NES-driven responses by smooth muscle cells make important contributions in initiating T(H)2 responses against N. brasiliensis infections.

A protective and agonistic function of IL-12p40 in mycobacterial infection.

IL-12p35(-/-)p40(-/-) mice are highly susceptible to Mycobacterium bovis bacillus Calmette-Guérin (BCG) or Mycobacterium tuberculosis infection. In this study IL-12p35(-/-) mice, which are able to produce endogenous IL-12p40, cleared M. bovis BCG and showed reduced susceptibility to pulmonary M. tuberculosis infection, which was in striking contrast to the outcome of mycobacterial infection in IL-12p35(-/-)p40(-/-) mice. Resistance in wild-type and IL-12p35(-/-) mice was accompanied by protective granuloma formation and Ag-specific delayed-type hypersensitivity responses, which were impaired in susceptible IL-12p35(-/- )p40(-/-) mice. Furthermore, IL-12p35(-/-) mice, but not IL-12p35(-/-)p40(-/-) mice, mounted Ag-specific Th1 and cytotoxic T cell responses. In vivo therapy with rIL-12p40 homodimer restored the impaired delayed-type hypersensitivity responses in M. bovis BCG-infected IL-12p35(-/-)p40(-/-) mice and reverted them to a more resistant phenotype. Together, these results show evidence for a protective and agonistic role of endogenous and exogenous IL-12p40 in mycobacterial infection, which is independent of IL-12p70.

Mid-pregnancy genetic terminations of pregnancy--postnatal assessment and management.

STUDY OBJECTIVE: To assess the need for and ability to apply a postnatal assessment protocol (PNAP), consisting of clinical examination, photographs, radiographs, chromosomal analysis and postmortems, of fetuses from mid-pregnancy genetic terminations of pregnancy. DESIGN: Prospective hospital-based study. SETTING: The maternity unit at the Pretoria Academic Hospital. MAIN RESULTS: Fifty consecutively delivered fetuses were assessed by means of the PNAP after genetic termination of pregnancy. A definitive prenatal diagnosis was available in 17 (34%) cases. In 33 (66%) cases the termination was undertaken on the basis of a provisional prenatal diagnosis which was confirmed postnatally in 12 (24%) cases; a definitive postnatal diagnosis could not be confirmed in 5 (10%) cases. In the remaining 16 (32%) cases a totally different postnatal diagnosis was obtained. The definitive postnatal diagnoses in the 28 cases with provisional prenatal diagnoses were confirmed by clinical examination in 13 (26%), by chromosomal analysis in 7 (14%), by postmortem in 5 (10%) and with radiographs in 3 (6%). On retrospective analysis, 22 of the 33 provisional prenatal diagnoses could have been confirmed using available radiographs, chromosome results and photographs only. CONCLUSIONS: Genetic terminations of pregnancy are a subgroup of stillbirths for which a PNAP is essential to ensure that appropriate postnatal genetic counselling can be given to the parent(s).

Structure of carotid artery in baboon and rat and differences in their response to endothelial denudation angioplasty.

BACKGROUND: Most studies dealing with vascular response to injury have been conducted using rodent and rabbit models, although it is expected that the response to injury in these species is dissimilar from man. AIMS: Here we compare the structure of native carotid artery in rat and baboon and the response of these vessels to endothelial denudation angioplasty. METHODS: In both species, the carotid is a musculoelastic artery. Only baboon carotid has a distinct intima, correlating in size with the weight of male baboons. Complete endothelial denudation of left carotid was performed on eight male baboons and 24 male rats by applying an equivalent pull force with a Fogarthy catheter. The animals were sacrificed prior to and 15 min and 2, 3, 4, 7, 14 and 28 days postinjury, one baboon and three rats per time point. RESULTS: Re-endothelialization in the baboon was complete already on day 4, whereas in the rat it was still incomplete on day 28. The proliferative response to injury was far smaller in the baboon than in the rat, the intimal area increased only by 5-fold in baboon compared with 25-fold in rat, and the number of intimal nuclei by 4-fold in baboon compared with 12-fold in rat. Complete compensatory remodelling of the lumen size occurred in the baboon, whereas in the rat remodelling remained incomplete. The cell types participating in the response were, however, similar: deposition of thrombocytes on denuded luminal surface, expression of alpha-actin by intimal cells, and lack of any significant white cell infiltration in the denuded intima. CONCLUSIONS: Baboon carotids are very different from rat carotids both in their native structure and in their response to injury. With the limited amount of information available from human vessels, baboon carotids closely resemble human carotids in both respects.