Using a single tablet daily to treat latent tuberculosis infection in Brazil: bioequivalence of two different isoniazid formulations (300 mg and 100 mg) demonstrated by a sensitive and rapid high-performance liquid chromatography-tandem mass spectrometry method in a randomised,crossover study

The recommended treatment for latent tuberculosis (TB) infection in adults is a daily dose of isoniazid (INH) 300 mg for six months. In Brazil, INH was formulated as 100 mg tablets. The treatment duration and the high pill burden compromised patient adherence to the treatment. The Brazilian National Programme for Tuberculosis requested a new 300 mg INH formulation. The aim of our study was to compare the bioavailability of the new INH 300 mg formulation and three 100 mg tablets of the reference formulation. We conducted a randomised, single dose, open label, two-phase crossover bioequivalence study in 28 healthy human volunteers. The 90% confidence interval for the INH maximum concentration of drug observed in plasma and area under the plasma concentration vs. time curve from time zero to the last measurable concentration “time t” was 89.61-115.92 and 94.82-119.44, respectively. The main limitation of our study was that neither adherence nor the safety profile of multiple doses was evaluated. To determine the level of INH in human plasma, we developed and validated a sensitive, simple and rapid high-performance liquid chromatography-tandem mass spectrometry method. Our results showed that the new formulation was bioequivalent to the 100 mg reference product. This finding supports the use of a single 300 mg tablet daily strategy to treat latent TB. This new formulation may increase patients’ adherence to the treatment and quality of life.     

Lipid kinases are essential for apicoplast homeostasis in Toxoplasma gondii

Phosphoinositides regulate numerous cellular processes by recruiting cytosolic effector proteins and acting as membrane signalling entities. The cellular metabolism and localization of phosphoinositides are tightly regulated by distinct lipid kinases and phosphatases. Here, we identify and characterize a unique phosphatidylinositol 3 kinase (PI3K) in Toxoplasma gondii, a protozoan parasite belonging to the phylum Apicomplexa. Conditional depletion of this enzyme and subsequently of its product, PI(3)P, drastically alters the morphology and inheritance of the apicoplast, an endosymbiotic organelle of algal origin that is a unique feature of many Apicomplexa. We searched the T. gondii genome for PI(3)P‐binding proteins and identified in total six PX and FYVE domain‐containing proteins including a PIKfyve lipid kinase, which phosphorylates PI(3)P into PI(3,5)P₂. Although depletion of putative PI(3)P‐binding proteins shows that they are not essential for parasite growth and apicoplast biology, conditional disruption of PIKfyve induces enlarged apicoplasts, as observed upon loss of PI(3)P. A similar defect of apicoplast homeostasis was also observed by knocking down the PIKfyve regulatory protein ArPIKfyve, suggesting that in T. gondii, PI(3)P‐related function for the apicoplast might mainly be to serve as a precursor for the synthesis of PI(3,5)P₂. Accordingly, PI3K is conserved in all apicomplexan parasites whereas PIKfyve and ArPIKfyve are absent in Cryptosporidium species that lack an apicoplast, supporting a direct role of PI(3,5)P₂ in apicoplast homeostasis. This study enriches the already diverse functions attributed to PI(3,5)P₂ in eukaryotic cells and highlights these parasite lipid kinases as potential drug targets.     

Glycolytic and non-glycolytic functions of Mycobacterium tuberculosis fructose-1,6-bisphosphate aldolase, an essential enzyme produced by replicating and non-replicating bacilli

The search for antituberculosis drugs active against persistent bacilli has led to our interest in metallodependent class II fructose-1,6-bisphosphate aldolase (FBA-tb), a key enzyme of gluconeogenesis absent from mammalian cells. Knock-out experiments at the fba-tb locus indicated that this gene is required for the growth of Mycobacterium tuberculosis on gluconeogenetic substrates and in glucose-containing medium. Surface labeling and enzymatic activity measurements revealed that this enzyme was exported to the cell surface of M. tuberculosis and produced under various axenic growth conditions including oxygen depletion and hence by non-replicating bacilli. Importantly, FBA-tb was also produced in vivo in the lungs of infected guinea pigs and mice. FBA-tb bound human plasmin(ogen) and protected FBA-tb-bound plasmin from regulation by α₂-antiplasmin, suggestive of an involvement of this enzyme in host/pathogen interactions. The crystal structures of FBA-tb in the native form and in complex with a hydroxamate substrate analog were determined to 2.35- and 1.9-Å resolution, respectively. Whereas inhibitor attachment had no effect on the plasminogen binding activity of FBA-tb, it competed with the natural substrate of the enzyme, fructose 1,6-bisphosphate, and substantiated a previously unknown reaction mechanism associated with metallodependent aldolases involving recruitment of the catalytic zinc ion by the substrate upon active site binding. Altogether, our results highlight the potential of FBA-tb as a novel therapeutic target against both replicating and non-replicating bacilli.     

Effects of four Bacillus spp. of soil origin on the Colorado potato beetle Leptinotarsa decemlineata (Say)

Four species of Bacillus were isolated from soil in an effort to find safe, effective and alternative biological control agents against plant pests. These bacteria were identified as Bacillus pumilus, Bacillus sphaericus, Bacillus megaterium and Bacillus cereus on the basis of fatty acid methyl ester analysis and carbon utilization profiles by using Microbial Identification and Biolog Microplate Systems. Laboratory experiments carried out to determine the insecticidal activities of these isolates showed that B. pumilus caused 95.7 and 26.7% mortality and B. sphaericus caused 74.5 and 23.3% mortality of Leptinotarsa decemlineata larvae and adults, respectively. B. cereus and B. megaterium showed 51.1 and 29.7%, respectively, of L. decemlineata larvae. This study presents at least two Turkish isolates from the genus Bacillus showing high insecticidal activity against L. decemlineata.     

Evidence for a spinal serotonergic control of the peripheral inflammation in the rat

We investigated the effect of serotonergic agonists and antagonists injected intrathecally by direct punction of the spinal cord at the lumbar level (between L5-L6) on peripheral inflammatory edema. Edema was induced by carrageenan injected subcutaneously in one hindpaw 30 min after spinal treatments. Serotonin (0.1, 1, 10 pmol) caused a graded-inhibition of the inflammatory paw edema. The corticosteroid inhibitor aminoglutethimide (100 mg/kg, p.o. 1.5 h before spinal treatment) did not modify this effect. The 5-HT1A agonist buspirone and the 5-HT1B/1D agonist sumatriptan (0.1, 1.0 and 10 nmol) also inhibited paw edema. The 5-HT1,2 antagonist methysergide (10 and 100 pmol) enhanced edema, but higher doses ( 4 and 8 nmol) diminished edema. NAN-190 (5-HT1 antagonist; 1 and 10 nmol) increased paw edema, while ritanserin (5-HT2 antagonist; 1 nmol) inhibited paw edema. Ondansetron (5-HT3 antagonist; up to 10 nmol) did not affect edema, but metoclopramide (5-HT3 antagonist / 5-HT4 agonist; 5, 10 and 30 pmol) inhibited edema. These data suggest that a tonic release of serotonin in the spinal cord may occurs during ongoing peripheral inflammation, modulating the neurogenic component of edema either by an inhibitory action on 5-HT1 receptors or by a stimulatory action on 5-HT2 receptors. A disfunction in such mechanism may be involved in the pathophysiology of certain types of headaches or migraine, which seem to depend on neurogenic vasodilation, and may also help to explain the therapeuthic effectiveness of some serotonergic agents in these conditions.     

ApoB-100 <Emphasis Type="Italic">R3500Q</Emphasis> mutation in the Lebanese population: Prevalence and historical review of the literature

An interesting mutation affecting the Apo-B gene, R3500Q, is known to display variable geographical distribution in the world and is mostly implicated in the pathogenesis of Familial Hypercholesterolemia (FH). The aim of this study is to determine the prevalence of this mutation in the Lebanese population and compare it to the available international literature. DNA from 160 unrelated healthy donors from our HLA-bank was used and the ApoB genotype was determined using the CardioVascular Disease (CVD) StripAssay (this assay is based on a Polymerase Chain Reaction-Reverse Hybridization technique). The R3500Q mutation was not observed in the general Lebanese population. Since the mutation frequency is elevated in Central Europe and tends to decrease as one moves east and south, it disappears completely in the Mediterranean regions such as Spain, Turkey and Israel; therefore, it is rather expected to be absent in Lebanon as well. Our report adds a valuable piece of information regarding this mutation in an Arab country and paves the way for future research involving patients diagnosed with FH in order to assess the role of the R3500Q mutation in the development of this clinical entity.     

Permanence and Extinction of a Diffusive Predator–Prey Model with Robin Boundary Conditions

The main concern of this paper is to study the dynamic of a predator–prey system with diffusion. It incorporates the Holling-type-II and a modified Leslie–Gower functional responses under Robin boundary conditions. More concretely, we study the dissipativeness of the system by using the comparison principle, and we derive a criteria for permanence and for predator extinction.     

Strong distance-dependent effects for a spatially aggregated tropical species

A fundamental aspect of the Janzen–Connell Hypothesis (JCH) is that distance- and density-dependent mortality reduce the local dominance of species and cause regular rather than random or aggregated spatial patterns. Despite this explicit linkage between process and pattern, very few studies have explored how JCH processes translate into the spatial distributions of adult populations. In field experiments, we assessed germination, mortality and growth of conspecific and heterospecific seedlings beneath and away from Esenbeckia leiocarpa, a highly aggregated tropical tree species. We also investigated the effects of vertebrates using exclosures in the field, and the effects of pathogens in a soil sterilization shadehouse experiment. Germination of conspecifics underneath Esenbeckia was reduced by 64 % and mortality increased 28–123 % when compared to seedlings growing under other tree species; 99–100 % of Esenbeckia seedlings died under conspecifics. Heterospecifics were much less affected by Esenbeckia canopies. However, we found no evidence that either vertebrate herbivores or soil pathogens affected seed germination and seedling performance. Although many tropical tree species are aggregated, our results are the first to demonstrate strong negative distance-dependence for an aggregated species and one of the few to explore germination in the context of the JCH and show differences between conspecifics and heterospecifics, thus suggesting a broader role for Janzen–Connell processes as determinants of the distribution and abundance of tree species in tropical forests.     

Ultra-High-Sensitive Sensor Based on Surface Plasmon Resonance Structure Having Si and Graphene Layers for the Detection of Chikungunya Virus

Chikungunya virus has been discovered in about 60 countries of the world. It leads to joint pain, joint swelling, headache, muscle pain, and fatigue of the human body. In this work, a surface plasmon resonance (SPR)based sensor is developed to detect chikungunya virus through normal and infected platelets and plasma blood cells. The proposed SPR-based sensor uses silicon and graphene layers coated over the base of a glass prism sputtered with a silver layer. The graphene layer has the advantage of enhancing the biomolecules adsorption on the metal layer. The silicon layer between silver and graphene enhances the sensor performance. The number of graphene layers along with the thicknesses of silicon and silver layers is optimized to get the highest sensitivity of the detector. To investigate the effect of the light source wavelength, simulations are performed for four different wavelengths. The highest sensitivities exhibited by the SPR-based sensor are 393 and 160 deg/RIU for the platelets and plasma cells, respectively.