Effects of opsonization and gamma interferon on growth of Brucella melitensis 16M in mouse peritoneal macrophages in vitro

Entry of opsonized pathogens into phagocytes may benefit or, paradoxically, harm the host. Opsonization may trigger antimicrobial mechanisms such as reactive oxygen or nitric oxide (NO) production but may also provide a safe haven for intracellular replication. Brucellae are natural intramacrophage pathogens of rodents, ruminants, dogs, marine mammals, and humans. We evaluated the role of opsonins in Brucella-macrophage interactions by challenging cultured murine peritoneal macrophages with Brucella melitensis 16M treated with complement- and/or antibody-rich serum. Mouse serum rich in antibody against Brucella lipopolysaccharide (LPS) (aLPS) and human complement-rich serum (HCS) each enhanced the macrophage uptake of brucellae. Combinations of suboptimal levels of aLPS (0. 01%) and HCS (2%) synergistically enhanced uptake. The intracellular fate of ingested bacteria was evaluated with an optimal concentration of gentamicin (2 microg/ml) to control extracellular growth but not kill intracellular bacteria. Bacteria opsonized with aLPS and/or HCS grew equally well inside macrophages in the absence of gamma interferon (IFN-gamma). Macrophage activation with IFN-gamma inhibited replication of both opsonized and nonopsonized brucellae but was less effective in inhibiting replication of nonopsonized bacteria. IFN-gamma treatment of macrophages with opsonized or nonopsonized bacteria enhanced NO production, which was blocked by N(G)-monomethyl L-arginine (MMLA), an NO synthesis inhibitor. MMLA also partially blocked IFN-gamma-mediated bacterial growth inhibition. These studies suggest that primary murine macrophages have limited ability to control infection with B. melitensis, even when activated by IFN-gamma in the presence of highly opsonic concentrations of antibody and complement. Additional cellular immune responses, e.g., those mediated by cytotoxic T cells, may play more important roles in the control of murine brucellosis.     

Determination of serum and organ malondialdehyde (MDA) concentration, a lipid peroxidation index, in Trypanosoma brucei-infected rats

Lipid peroxidation was assessed in the sera and various organs of rats experimentally infected with Trypanosoma brucei. Thirty-six adult albino rats divided into 2 groups of eighteen rats each were used in this study. In experiment one, a group of 18 rats were used and they were divided into three groups (A, B and C) of six rats each. Groups B and C rats were infected with 1.54 × 10⁵ trypanosomes per rat intraperitoneally, whereas group A served as uninfected control. The rats were bled on day 0 and subsequently at 7-day intervals for packed cell volume (PCV), sera peroxidation index and parasitaemia. Also, temperature and weight were taken on day 0 and subsequently at 7-day intervals. In experiment 2, 18 rats were also used. Six rats each were sacrificed on days 0, 14 and 28-postinfection. Five rats each were sacrificed on day 14 and day 28 post-infection (PI) from group B, and their organs were promptly collected and washed with normal saline and used for organ malondialdehyde (MDA) concentration. The infection led to an increase in lipid peroxidation index (MDA concentration) of sera samples. The serum MDA concentration of the infected rat group was significantly (p < 0.01) higher than in the uninfected group on days 21 and 28 PI. The increase was however reversed by diminazene aceturate (Berenil; Hoechst, Ireland) treatment at the dosage of 7 mg/kg body weight administered on day 14 PI. The organ lipid peroxidation index also increased significantly (p < 0.05) in the eye, lung and spleen. However, there was no significant (p > 0.05) increase of lipid peroxidation index in the kidney, heart, liver, testes and brain. Also, the mean weekly MDA concentration increased as the disease progressed, the mean weekly temperature and parasitaemia also increased, but the reverse was the case with the mean weekly body weight and PCV which declined as the disease progressed. The findings are indication that oxidative stress plays an important role in the pathology of trypanosomosis.     

Immediate Effect of Acupuncture on Electromyographic Activity of the Upper Trapezius Muscle and Pain in Patients With Nonspecific Neck Pain: A Randomized,Single-Blinded,Sham-Controlled,Crossover Study

Objective

The objective of this study was to assess changes in upper trapezius myoelectric activity and pain in patients with nonspecific neck pain after a single session of acupuncture (ACP).

Soliciting views of various communities on health research: a prelude to engagement in specific research projects

BackgroundMembers of the public are increasingly engaged in health‐service and biomedical research and provide input into the content of research, design and data sharing. As there is variation among different communities on how research is perceived, to engage all sectors of the general public research institutions need to customize their approach.ObjectiveThis paper explores how research institutions and community leaders can partner to determine the best ways to engage different sectors of the public in research.DesignFollowing a literature review, a research institution engaged with four different sectors of the public through their respective representative community‐based organizations (CBOs) by interviews with leaders, community member focus groups and a joint project.SettingSan Diego and Imperial Counties, California, United States of America (USA).ConclusionBefore embarking on more specific research projects, investigators can gain valuable insights about different communities'' attitudes to, and understanding of, health services and biomedical research by interacting directly with members of the community, collaborating with community leaders, and jointly identifying steps of engagement tailored to the community.     

The immunology of Leishmania/HIV co-infection

Leishmaniases are emerging as an important disease in human immunodeficiency virus (HIV)–infected persons living in several sub-tropical and tropical regions around the world, including the Mediterranean. The HIV/AIDS pandemic is spreading at an alarming rate in Africa and the Indian subcontinent, areas with very high prevalence of leishmaniases. The spread of HIV into rural areas and the concomitant spread of leishmaniases to suburban/urban areas have helped maintain the occurrence of Leishmania/HIV co-infection in many parts of the world. The number of cases of Leishmania/HIV co-infection is expected to rise owing to the overlapping geographical distribution of the two infections. In Southwestern Europe, there is also an increasing incidence of Leishmania/HIV co-infection (particularly visceral leishmaniasis) in such countries as France, Italy, Spain and Portugal. Studies suggest that in humans, very complex mechanisms involving dysregulation of host immune responses contribute to Leishmania-mediated immune activation and pathogenesis of HIV. In addition, both HIV-1 and Leishmania infect and multiply within cells of myeloid or lymphoid origin, thereby presenting a perfect recipe for reciprocal modulation of Leishmania and HIV-1-related disease pathogenesis. Importantly, because recovery from leishmaniases is associated with long-term persistence of parasites at the primary infection sites and their draining lymph nodes, there is very real possibility that HIV-mediated immunosuppression (due to CD4⁺ T cell depletion) could lead to reactivation of latent infections (reactivation leishmaniasis) in immunocompromised patients. Here, we present an overview of the immunopathogenesis of Leishmania/HIV co-infection and the implications of this interaction on Leishmania and HIV disease outcome.     

Detection, purification and characterisation of a secretory alkaline phosphatase from Onchocerca species

An Onchocerca secretory alkaline phosphatase (E.C. 3.1.3.1) of molecular weight 90 kDa when in crude extract, but which dimerises to about 180 kDa upon purification, was detected, purified and characterised. The enzyme was found to be secreted by both O. ochengi and O. volvulus worms. It was shown to be of Onchocerca origin by Western blotting with bovine onchocerciasis sera and by its time-dependent release in cultures. The O. ochengi enzyme was purified to near homogeneity by a combination of polyethylene glycol precipitation, DEAE-cellulose chromatography and preparative electrophoresis. About 0.96 mg of the active enzyme was purified from 48.4 mg of the crude parasite-released products, giving a purification fold of 71.45 and a yield of 8.7%. The purified enzyme exhibited a typical Michaelis-Menten kinetics with optimum activity on p-nitrophenylphosphate (p-NPP) at pH 10.2. Its apparent K(m) for p-NPP was 0.56+/-0.03 mM and it required Mg(2+) and dithiothreitol (DTT) for stability throughout its purification. Sodium dodecyl sulphate at 2% (w/v) did not inhibit the enzyme activity, but apparently stabilised it during freezing. Inorganic phosphate inhibited the enzyme competitively with an apparent inhibition constant (K(i)) of 3.33+/-0.04 mM, whereas l-phenylalanine inhibited it in a mixed way with a K(i) of 3.18+/-0.03 mM. While contributing to the understanding of metabolism in Onchocerca, the present apparently unique enzyme which is likely to serve in the nutrition of the parasite could be further characterised as a macrofilaricide target or diagnostic marker in onchocerciasis.     

Colon Interposition Graft for Corrosive Esophageal Stricture: Midterm Functional Outcome

Background

Corrosive esophageal stricture is a major cause of morbidity among Nigerians. In most cases, this follows accidental or parasuicidal ingestion of caustic sodium hydroxide solution (NaOH) often used in the local production of soaps. Various treatment modalities have been advocated for the treatment of esophageal stricture. In this study, we review the results of our adopted technique in the past 10 years for pedicled colonic interposition.

Methods

This is a retrospective study of 21 patients who underwent substernal isoperistaltic pedicled colonic interposition graft for management of corrosive esophageal stricture. The right colon was pulled up into the neck in all the patients without resecting the strictured esophagus.

Results

Long segment strictures and multiple strictures were the main indications for the procedure. The mean duration of the procedure was 339.6 ± 71.1 min. The average intraoperative blood loss was 673.1 ± 398.1 mL. There were two (9.5 %) hospital mortalities. Graft infarction (9.5 %), cervical fistulae (19.0 %), and reflux neo-esophagitis (14.3 %) were the main non-fatal complications. In the mid-term, dysphagia was completely relieved in a little over 84 % (16/19) of patients, while one patient (4.8 %) still experienced reflux neo-esophagitis requiring treatment. There was no case of gross regurgitation or nocturnal aspiration in the mid-term.

Conclusions

Although the use of pedicled colonic interposition graft offers a good mid-term functional outcome with relief of dysphagia, early postoperative morbidity is high. Graft infarction is the single most important factor for poor functional outcome and every effort must be made to prevent its occurrence.