2,005 research outputs found
Mycobacteriumsmegmatis bioÂąlm formationand slidingmotility are aÂĄected by the serine/threonine protein kinase PknF
Eighteen âeukaryotic-likeâ serine/threonine kinases are present in the Mycobacterium
smegmatis genome. One of them encoded by the ORF 3677 demonstrates high
similarity to the Mycobacterium tuberculosis protein kinase PknF. A merodiploid
strain was generated, which showed reduced growth associated with irregular cell
structure. The merodiploid strain displayed altered colony morphology, defective
slidingmotility and biofilm formation. These data indicate a role for PknF in biofilm
formation, possibly associated with alterations in glycopeptidolipid composition
Mycobacterium tuberculosis Virulence Is Mediated by PtpA Dephosphorylation of Human Vacuolar Protein Sorting 33B
SummaryEntry into host macrophages and evasion of intracellular destruction mechanisms, including phagosome-lysosome fusion, are critical elements of Mycobacterium tuberculosis (Mtb) pathogenesis. To achieve this, the Mtb genome encodes several proteins that modify host signaling pathways. PtpA, a low-molecular weight tyrosine phosphatase, is a secreted Mtb protein of unknown function. The lack of tyrosine kinases in the Mtb genome suggests that PtpA may modulate host tyrosine phosphorylated protein(s). We report that a genetic deletion of ptpA attenuates Mtb growth in human macrophages, and expression of PtpA-neutralizing antibodies simulated this effect. We identify VPS33B, a regulator of membrane fusion, as a PtpA substrate. VPS33B and PtpA colocalize in Mtb-infected human macrophages. PtpA secretion combined with active-phosphorylated VPS33B inhibited phagosome-lysosome fusion, a process arrested in Mtb infections. These results demonstrate that PtpA is essential for Mtb intracellular persistence and identify a key host regulatory pathway that is inactivated by Mtb
A phase I clinical study of inhaled nitric oxide in healthy adults
AbstractBackgroundNitric oxide (NO) is an approved pulmonary vasodilator for neonates and full term infants up to a dose of 80ppm. At 100ppm to 200ppm, NO has potent antimicrobial activities in vitro and in animal studies which suggest its therapeutic use for infectious diseases in humans. However, whether inhaled NO is safe at 160ppm in healthy human adults is unknown. The aim of the phase I study was to assess the safety of delivery and the physiologic effects of intermittent 160ppm NO in healthy human adults.MethodsTen healthy adult volunteers (5 males, 5 females; 20â62years) were recruited and inhaled 163.3ppm (SD: 4.0) NO for 30min, 5 times daily, for 5 consecutive days. Lung function and blood levels of methemoglobin, nitrites/nitrates, prothrombin, pro-inflammatory cytokines and chemokines were determined before and during treatment.ResultsAll individuals tolerated the NO treatment courses well. No significant adverse events occurred and three minor adverse events, not attributable to NO, were reported. Forced expiratory volume in 1sec % predicted and other lung function parameters, serum nitrites/nitrates, prothrombin, pro-inflammatory cytokine and chemokine levels did not differ between baseline and day 5, while methemoglobin increased significantly during the study period to a level of 0.9% (SD: 0.08) (p<0.001).ConclusionThese data suggest that inhalation of 160ppm NO for 30min, 5 times daily, for 5 consecutive days, is safe and well tolerated in healthy individuals
Antimicrobial, Anti-Inflammatory, Antiparasitic, and Cytotoxic Activities of Laennecia confusa
The current paper investigated the potential benefit of the traditional Mexican medicinal plant Laennecia confusa (Cronquist) G. L. Nesom (Asteraceae). Fractions from the hexane, chloroform, methanol, and aqueous extracts were analyzed for antibacterial, antifungal, anti-inflammatory, and antiparasitic activities. The antimicrobial activity of the extracts and fractions was assessed on bacterial and fungal strains, in addition to the protozoa Leishmania donovani, using a microdilution assay. The propensity of the plant's compounds to produce adverse effects on human health was also evaluated using propidium iodine to identify damage to human macrophages. The anti-inflammatory activity of the extracts and fractions was investigated by measuring the secretion of interleukin-6. Chemical analyses demonstrated the presence of flavonoids, cyanogenic and cardiotonic glycosides, saponins, sesquiterpene lactones, and triterpenes in the chloroform extract. A number of extracts and fractions show antibacterial activity. Of particular interest is antibacterial activity against Staphylococcus aureus and its relative methicillin-resistant strain, MRSA. Hexanic and chloroformic fractions also exhibit antifungal activity and two extracts and the fraction CE 2 antiparasitic activity against Leishmania donovani. All bioactive extracts and fractions assayed were also found to be cytotoxic to macrophages. In addition, the hexane and methane extracts show anti-inflammatory activity by suppressing the secretion of interleukine-6
On the Deformation of a Hyperelastic Tube Due to Steady Viscous Flow Within
In this chapter, we analyze the steady-state microscale fluid--structure
interaction (FSI) between a generalized Newtonian fluid and a hyperelastic
tube. Physiological flows, especially in hemodynamics, serve as primary
examples of such FSI phenomena. The small scale of the physical system renders
the flow field, under the power-law rheological model, amenable to a
closed-form solution using the lubrication approximation. On the other hand,
negligible shear stresses on the walls of a long vessel allow the structure to
be treated as a pressure vessel. The constitutive equation for the microtube is
prescribed via the strain energy functional for an incompressible, isotropic
Mooney--Rivlin material. We employ both the thin- and thick-walled formulations
of the pressure vessel theory, and derive the static relation between the
pressure load and the deformation of the structure. We harness the latter to
determine the flow rate--pressure drop relationship for non-Newtonian flow in
thin- and thick-walled soft hyperelastic microtubes. Through illustrative
examples, we discuss how a hyperelastic tube supports the same pressure load as
a linearly elastic tube with smaller deformation, thus requiring a higher
pressure drop across itself to maintain a fixed flow rate.Comment: 19 pages, 3 figures, Springer book class; v2: minor revisions, final
form of invited contribution to the Springer volume entitled "Dynamical
Processes in Generalized Continua and Structures" (in honour of Academician
D.I. Indeitsev), eds. H. Altenbach, A. Belyaev, V. A. Eremeyev, A. Krivtsov
and A. V. Porubo
Pair-breaking quantum phase transition in superconducting nanowires
A quantum phase transition (QPT) between distinct ground states of matter is
a wide-spread phenomenon in nature, yet there are only a few experimentally
accessible systems where the microscopic mechanism of the transition can be
tested and understood. These cases are unique and form the experimentally
established foundation for our understanding of quantum critical phenomena.
Here we report the discovery that a magnetic-field-driven QPT in
superconducting nanowires - a prototypical 1d-system - can be fully explained
by the critical theory of pair-breaking transitions characterized by a
correlation length exponent and dynamic critical exponent . We find that in the quantum critical regime, the electrical
conductivity is in agreement with a theoretically predicted scaling function
and, moreover, that the theory quantitatively describes the dependence of
conductivity on the critical temperature, field magnitude and orientation,
nanowire cross sectional area, and microscopic parameters of the nanowire
material. At the critical field, the conductivity follows a
dependence predicted by phenomenological scaling theories and more recently
obtained within a holographic framework. Our work uncovers the microscopic
processes governing the transition: The pair-breaking effect of the magnetic
field on interacting Cooper pairs overdamped by their coupling to electronic
degrees of freedom. It also reveals the universal character of continuous
quantum phase transitions.Comment: 22 pages, 5 figure
S-Nitrosylation of α1-Antitrypsin Triggers Macrophages Toward Inflammatory Phenotype and Enhances Intra-Cellular Bacteria Elimination
Background: Human α1-antitrypsin (hAAT) is a circulating anti-inflammatory serine-protease inhibitor that rises during acute phase responses. in vivo, hAAT reduces bacterial load, without directly inhibiting bacterial growth. In conditions of excess nitric-oxide (NO), hAAT undergoes S-nitrosylation (S-NO-hAAT) and gains antibacterial capacity. The impact of S-NO-hAAT on immune cells has yet to be explored.Aim: Study the effects of S-NO-hAAT on immune cells during bacterial infection.Methods: Clinical-grade hAAT was S-nitrosylated and then compared to unmodified hAAT, functionally, and structurally. Intracellular bacterial clearance by THP-1 macrophages was assessed using live Salmonella typhi. Murine peritoneal macrophages were examined, and signaling pathways were evaluated. S-NO-hAAT was also investigated after blocking free mambranal cysteine residues on cells.Results: S-NO-hAAT (27.5 uM) enhances intracellular bacteria elimination by immunocytes (up to 1-log reduction). S-NO-hAAT causes resting macrophages to exhibit a pro-inflammatory and antibacterial phenotype, including release of inflammatory cytokines and induction of inducible nitric oxide synthase (iNOS) and TLR2. These pro-inflammatory effects are dependent upon cell surface thiols and activation of MAPK pathways.Conclusions: hAAT duality appears to be context-specific, involving S-nitrosylation in a nitric oxide rich environment. Our results suggest that S-nitrosylation facilitates the antibacterial activity of hAAT by promoting its ability to activate innate immune cells. This pro-inflammatory effect may involve transferring of nitric oxide from S-NO-hAAT to a free cysteine residue on cellular targets
Disruption of the serine/threonine protein kinase H affects phthiocerol dimycocerosates synthesis in Mycobacterium tuberculosis
Mycobacterium tuberculosis possesses a complex cell wall that is unique and essential for interaction of the pathogen with its human host. Emerging evidence suggests that the biosynthesis of complex cell-wall lipids is mediated by serine/threonine protein kinases (STPKs). Herein, we show, using in vivo radiolabelling, MS and immunostaining analyses, that targeted deletion of one of the STPKs, pknH, attenuates the production of phthiocerol dimycocerosates (PDIMs), a major M. tuberculosis virulence lipid. Comparative protein expression analysis revealed that proteins in the PDIM biosynthetic pathway are differentially expressed in a deleted pknH strain. Furthermore, we analysed the composition of the major lipoglycans, lipoarabinomannan (LAM) and lipomannan (LM), and found a twofold higher LAM/LM ratio in the mutant strain. Thus, we provide experimental evidence that PknH contributes to the production and synthesis of M. tuberculosis cell-wall components
Strain-Transcendent Immune Response to Recombinant Var2CSA DBL5-Δ Domain Block P. falciparum Adhesion to Placenta-Derived BeWo Cells under Flow Conditions
BACKGROUND: Pregnancy-associated malaria (PAM) is a serious consequence of the adhesion to the placental receptor chondroitin sulfate A (CSA) of Plasmodium falciparum-infected erythrocytes (PE) expressing the large cysteine-rich multi-domain protein var2CSA. Women become resistant to PAM, and develop strain-transcending immunity against CSA-binding parasites. The identification of var2CSA regions that could elicit broadly neutralizing and adhesion-blocking antibodies is a key step for the design of prophylactic vaccine strategies. METHODOLOGY: Escherichia coli expressed var2CSA DBL domains were refolded and purified prior to immunization of mice and a goat. Protein-G-purified antibodies were tested for their ability to block FCR3(CSA)-infected erythrocytes binding to placental (BeWo) and monkey brain endothelial (ScC2) cell lines using a flow cytoadhesion inhibition assay mimicking closely the physiological conditions present in the placenta at shear stress of 0.05 Pa. DBL5-Δ, DBL6-Δ and DBL5-6-Δ induced cross-reactive antibodies using Alum and Freund as adjuvants, which blocked cytoadhesion at values ranging between 40 to 96% at 0.5 mg IgG per ml. Importantly, antibodies raised against recombinant DBL5-Δ from 3 distinct parasites genotypes (HB3, Dd2 and 7G8) showed strain-transcending inhibition ranging from 38 to 64% for the heterologuous FCR3(CSA). CONCLUSIONS: Using single and double DBL domains from var2CSA and Alum as adjuvant, we identified recombinant subunits inducing an immune response in experimental animals which is able to block efficiently parasite adhesion in a flow cytoadhesion assay that mimics closely the erythrocyte flow in the placenta. These subunits show promising features for inclusion into a vaccine aiming to protect against PAM
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