The Rab-binding profiles of bacterial virulence factors during infection

Legionella pneumophila, the causative agent of Legionnaire's disease, uses its type IV secretion system to translocate over 300 effector proteins into host cells. These effectors subvert host cell signaling pathways to ensure bacterial proliferation. Despite their importance for pathogenesis, the roles of most of the effectors are yet to be characterized. Key to understanding the function of effectors is the identification of host proteins they bind during infection. We previously developed a novel tandem-affinity purification (TAP) approach using hexahistidine and BirA-specific biotinylation tags for isolating translocated effector complexes from infected cells whose composition were subsequently deciphered by mass spectrometry. Here we further advanced the workflow for the TAP approach and determined the infection-dependent interactomes of the effectors SidM and LidA, which were previously reported to promiscuously bind multiple Rab GTPases in vitro. In this study we defined a stringent subset of Rab GTPases targeted by SidM and LidA during infection, comprising of Rab1A, 1B, 6, and 10; in addition, LidA targets Rab14 and 18. Taken together, this study illustrates the power of this approach to profile the intracellular interactomes of bacterial effectors during infection

Characteristics of HIV-infected children seen in Western Kenya

Objectives: To describe the characteristics and outcomes of children registered for care in a large HIV care programme in Western Kenya.Design: A retrospective descriptive study.Setting: USAID-AMPATH HIV clinics in health centres; district and sub-district hospitals; Moi Teaching and Referral Hospital in Western Kenya.Subjects: HIV-infected children below age of 15 years seen in a network of 18 clinics in Western Kenya.Interventions: Paediatric HIV diagnosis and care including treatment and prevention of opportunistic infections and provision of combination antiretroviral therapy (CART).Main outcome measures: Diagnosis, clinical stage and immune status at enrollment and follow-up; hospitalisation and death. Descriptive statistical analyses and chi square tests were performedResults: Four thousand and seventeen HIV-infected children seen between June 2002 and April 2008. Median age at enrollment was four years (0-14.2 years), 51% girls, 25% paternal orphans, 10% total orphans and 13% maternal orphans. At enrollment, 25% had weight-for-Age Z scores (WAZ)> -1 and 21% had WAZ scores < 3. Orphaned children had worse WAZ scores (p=0.0001). Twenty five per cent of children were classified as WHO clinical stage 3 and 4, 56% were WHO clinical stages 1 and 2 with 19% missing clinical staging at enrollment. Cough (25%), gastroenteritis (21%), fever (15%), pneumonia (10%) were the commonest presenting features. Twenty six per cent had been diagnosed with tuberculosis and only 25% started on cotrimoxazole preventive therapy (CPT). Median CD4% at enrollment was 16% (0-64%); latest recorded values were 22% (0-64). Sixty four per cent were on cART (cART+), median age at start was 5.4 (014.4 years).The median initial CD4% among cART+ was 13 (0-62) compared to 24 (0-64) for those not on ART (cART-). Median CD4% for cART+ improved to 22% (0-59); whereas cART- was 23% (0-64) at last appointment. During the period of follow-up, one fifth (19%) of children on cART were lost to follow-up compared to slightly over one third (37%) for those not on cART. Thirty four percent were hospitalised; 41% diagnosed with pneumonia. Six per cent of 4017 were confirmed dead.Conclusions: HIV -infected children were enrolled in care early in childhood. Orphan-hood was prevalent in these children as were gastroenteritis, fever, pneumonia and advanced immuno-suppression. Orphans were more likely to be severely malnourished. Only a quarter of children were put on cotrimoxazole preventive therapy. Children commenced on cART late but responded well to treatment. Loss to follow-up was less prevalent among those on cART

Anaemia and blood transfusion in African children presenting to hospital with severe febrile illness

BACKGROUND: Severe anaemia in children is a leading cause of hospital admission and a major cause of mortality in sub-Saharan Africa, yet there are limited published data on blood transfusion in this vulnerable group. METHODS: We present data from a large controlled trial of fluid resuscitation (Fluid Expansion As Supportive Therapy (FEAST) trial) on the prevalence, clinical features, and transfusion management of anaemia in children presenting to hospitals in three East African countries with serious febrile illness (predominantly malaria and/or sepsis) and impaired peripheral perfusion. RESULTS: Of 3,170 children in the FEAST trial, 3,082 (97%) had baseline haemoglobin (Hb) measurement, 2,346/3,082 (76%) were anaemic (Hb <10 g/dL), and 33% severely anaemic (Hb <5 g/dL). Prevalence of severe anaemia varied from 12% in Kenya to 41% in eastern Uganda. 1,387/3,082 (45%) children were transfused (81% within 8 hours). Adherence to WHO transfusion guidelines was poor. Among severely anaemic children who were not transfused, 52% (54/103) died within 8 hours, and 90% of these deaths occurred within 2.5 hours of randomisation. By 24 hours, 128/1,002 (13%) severely anaemic children had died, compared to 36/501 (7%) and 71/843 (8%) of those with moderate and mild anaemia, respectively. Among children without severe hypotension who were randomised to receive fluid boluses of 0.9% saline or albumin, mortality was increased (10.6% and 10.5%, respectively) compared to controls (7.2%), regardless of admission Hb level. Repeat transfusion varied from ≤2% in Kenya/Tanzania to 6 to 13% at the four Ugandan centres. Adverse reactions to blood were rare (0.4%). CONCLUSIONS: Severe anaemia complicates one third of childhood admissions with serious febrile illness to hospitals in East Africa, and is associated with increased mortality. A high proportion of deaths occurred within 2.5 hours of admission, emphasizing the need for rapid recognition and prompt blood transfusion. Adherence to current WHO transfusion guidelines was poor. The high rates of re-transfusion suggest that 20 mL/kg whole blood or 10 mL/kg packed cells may undertreat a significant proportion of anaemic children. Future evaluation of the impact of a larger volume of transfused blood and optimum transfusion management of children with Hb of <6 g/dL is warranted. Please see related article: http://dx.doi.org/10.1186/s12916-014-0248-5. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12916-014-0246-7) contains supplementary material, which is available to authorized users

Cordycepin enhances cisplatin apoptotic effect through caspase/MAPK pathways in human head and neck tumor cells

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The iron-chelating drug M30 down-regulates carbon tetrachloride (CCI4)-induced hepatic oxidative stress, inflammation and apoptosis in vitro

Topic: 2 Acute Liver FailureThis journal suppl. entitled: APASL Liver Week 2013BACKGROUND/AIMS: The novel multifunctional brain permeable ironchelator M30 possesses neuroprotective activities against several insults applicable to various neurodegenerative diseases. However, the effect of M30 on CCl4 induced acute liver damage is still unknown. The aim of this study is to investigate whether the multifunctional drug M30 could ameliorate CCl4 induced hepatic injury in human HepG2 cell line. METHODS: HepG2 cells were grown in DMEM supplemented with ...postprin

Zeaxanthin Dipalmitate Therapeutically Improves Hepatic Functions in an Alcoholic Fatty Liver Disease Model through Modulating MAPK Pathway

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WHO guidelines on fluid resuscitation in children: missing the FEAST data.

The World Health Organization recommendations on management of common childhood illnesses affect the lives of millions of children admitted to hospital worldwide. Its latest guidelines,1 released in May 2013, continue to recommend rapid fluid resuscitation for septic shock, even though the only large controlled trial of this treatment (Fluid Expansion as a Supportive Treatment (FEAST) found that it increased the risk of death in African children.2 A subsequent systematic review of bolus resuscitation in children with shock resulting from severe infection also did not support its use.3 Failure to take this evidence into account is not consistent with WHO’s commitment to systematically and transparently assess evidence using the GRADE (Grading of Recommendations Assessment, Development and Evaluation) process when producing guidelines and could endanger the lives of children

Intra‑cardiac masses in adults: A review of echocardiogram records at two echocardiographic laboratories in Enugu, South‑East Nigeria

Background: Transthoracic echocardiography (TTE) is an excellent initial diagnostic technique used to evaluate and diagnose cardiac masses, even though transoesophageal echocardiography (TEE) provides superior image resolution and better visualization of cardiac masses, especially in patients with suboptimal transthoracic echocardiographic studies. TTE is the clinical procedure of choice for identification of left ventricular thrombi. TTE has greater than 90% sensitivity and greater than 85% specificity for detection of left ventricular thrombi and is probably superior to the sensitivity and specificity of TEE, especially for apical thrombi.Aims: The study aimed to identify the common types of cardiac masses and their commonest locations in the heart.Materials and Methods: We did a retrospective review of our echocardiogram reports from May 2003 to July 2012 to identify the frequency of intra‑cardiac masses in adults, as well as the gender distribution and commonest location of these masses.Results: There were 2,814 echo examinations in adults over this period, comprising 1,661 males (59.1%) and 1,153 females (40.9%). Intra‑cardiac masses were found in 20 of these patients representing 0.7% of the study population. Thrombi were the commonest masses noted in our study, and there were more masses in the atria than in the ventricles. The left heart chambers also had more masses than the right heart chambers. There was no sex difference in the frequency of cardiac masses.Conclusion: Intra‑cardiac masses are rare, and transthoracic echocardiography is still valuable in the diagnosis and initial characterization of cardiac masses.Keywords: Adults, echocardiography, intra‑cardiac masses, NigeriaNigerian Journal of Clinical Practice • Oct-Dec 2013 • Vol 16 • Issue

The molecular mechanism of the anti-oxidative effects of catechin on palmitic acid-induced cytotoxicity in astrocytes

Poster PresentationAIMS: Metabolic syndrome (MS) encompasses a group of problems which will put a person at a high risk of developing cardiovascular diseases, including heart attack and stroke. Effective prevention or treatment of MS significantly reduces the risk for developing serious complications. Palmitic acid (PA) is a saturated fatty acid, when being excessive, is a significant risk factor for development of MS or stroke. However, damage by MS to astrocytes is relatively unexplored. Catechin is an effective antioxidant which would be beneficial to neurons subjected to reactive oxygen species (ROS) damage as well ...postprin

In Vivo Comparative Evaluation of Effects of Artemeter-Lumefantrine, Sulphadoxine-Pyrimetamine and Halofantrine on G6PD Activities, Haemoglobin Concentration and Malaria Parasite Clearance Rate in Malaria Infected Adults

Background: The use of antimalaria combination therapy especially regimens containing an artemisin-based compound has been recommended as a good first-line treatment for malaria by WHO. However, limited reports exist on the effects of the ACT and other antimalarial drugs on some vital biological parameters such as G6PD activity and haemoglobin concentration. In this study, we investigated the effects of Artemeter-Lumefantrine, sulphadoxine-pyrimetamine combination therapies and Halofantrine monotherapy on G6PD activity, Haemoglobin level and parasite clearance rate in malaria-infected adults treated with the drugs in Enugu, Nigeria. Methodology: Forty malaria-infected adults aged between 20 and 30 years were used. The subjects were divided into four groups (A, B, C and D). The groups A, B and C were given Artemeter-Lumefantrine, sulphadoxine-pyrimetamine and halofantrine respectively, while group D was a control group (malaria-positive control). Blood samples of the subjects were collected through venepuncture at baseline (Day 0) and after treatment on Day 4, for comparative analysis of G6PD activity, haemoglobin concentration and parasite clearance for each group. Results: The result of this study showed that sulphadoxine-pyrimetamine significantly (p&lt;0.05) lowered haemoglobin concentration as compared with Halofantrine, Artemeter-lumefantrine and control. The haemoglobin concentration of the last three groups did not show any significant difference (p&gt;0.05) between each other. The G6PD activity of the group treated with sulphadoxine-pyrimetamine was significantly (p&lt;0.05) higher as compared with that treated with Artemeter-lumefantrine but non-significant (p&gt;0.05) as compared with halofantrine and control. Parasite clearance rate was significantly (p&lt;0.05) highest with the Halofantrine group (76%, p&lt; 0.05) while Sulphadoxine-pyrimetamine had the lowest (52%) parasite clearance. Conclusion: This study therefore indicated that antimalaria drugs as well as malaria parasite could cause a reduction in haemoglobin concentration with sulphadoxine-pyrimetamine causing significant (p&lt;0.05) increase in G-6-PD activity. Keywords: Antimalarials, Artemether-Lumefantrine, G6PD, sulphadoxine-pyrimetamine, Parasite clearance, Halofantrine, Haemoglobin concentratio