FLAgellum Member 8 modulates extravascular trypanosome distribution in the mammalian host

The African trypanosome flagellum is essential in multiple aspects of the parasite development. In the mammalian infective form of this protist, FLAgellar Member 8 (FLAM8) is a large protein distributed along the entire flagellum that is suspected to be involved in host-parasite interactions. Analyses of knockdown and knockout trypanosomes demonstrated that FLAM8 is not essential in vitro for survival, growth, motility and slender to stumpy differentiation. Functional investigations in experimental infections showed that FLAM8 -deprived trypanosomes are able to establish and maintain the infection in the blood circulation, and to differentiate into transmissible stumpy forms. However, bioluminescence imaging revealed that FLAM8 -null parasites exhibit an impaired dissemination in the extravascular compartment, especially in the skin, that is partially restored by the addition of a single rescue copy of FLAM8 . To our knowledge, FLAM8 is the first example of a flagellar protein that modulates T. brucei parasite distribution in the host tissues, contributing to the maintenance of extravascular parasite populations in mammalian anatomical niches.

Contributions of the glycocalyx, endothelium, and extravascular compartment to the blood–brain barrier

The endothelial cells that form the blood–brain barrier (BBB) are coated with glycocalyx, on the luminal side, and with the basement membrane and astrocyte endfeet, on the abluminal side. However, it is unclear how exactly the glycocalyx and extravascular structures contribute to BBB properties. We used two-photon microscopy in anesthetized mice to record passive transport of four different-sized molecules—sodium fluorescein (376 Da), Alexa Fluor (643 Da), 40-kDa dextran, and 150-kDa dextran—from blood to brain, at the level of single cortical capillaries. Both fluorescein and Alexa penetrated nearly the entire glycocalyx volume, but the dextrans penetrated less than 60% of the volume. This suggested that the glycocalyx was a barrier for large but not small molecules. The estimated permeability of the endothelium was the same for fluorescein and Alexa but several-fold lower for the larger dextrans. In the extravascular compartment, co-localized with astrocyte endfeet, diffusion coefficients of the dyes were an order of magnitude lower than in the brain parenchyma. This suggested that the astrocyte endfeet and basement membrane also contributed to BBB properties. In conclusion, the passive transport of small and large hydrophilic molecules through the BBB was determined by three separate barriers: the glycocalyx, the endothelium, and the extravascular compartment. All three barriers must be taken into account in drug delivery studies and when considering BBB dysfunction in disease states.

CORRELATION BETWEEN THE USE OF ALBUMIN INFUSION AND CONCENTRATIONS OF SERUM ALBUMIN, PROINFLAMMATORY CYTOKINES (TNF-, IL6) AND SOFA SCORE IN SEPTIC PATIENTS

Sepsis is a major problem causing high morbidity and mortality in patients who hospitalized in intensive care. In sepsis, there is a change in the distribution of albumin in the intravascular and extravascular compartment, so hipoalbumin is occured. Using albumin in septic patients from several studies that have been done are still controversial. Analyzing albumin infusion with changes in levels of albumin, cytokines and SOFA score in septic patients in intensive care dr. Soetomo. The design of this study is a prospective longitudinal observational. The study was conducted in the intensive care unit or ICU dr. Soetomo in February 2015. The inclusion and exclusion criteria were determined that obtained 15 research subjects. They were examined with serum albumin and proinflammatory cytokines as well as evaluating the SOFA score. Then, it measure with ANOVA test methods. Infusion of albumin positively correlated with increased levels of serum albumin and a negative correlation with the levels of IL6, TNF-a and SOFA score. It also found a correlation between changes in levels of albumin with changing levels of IL6 (p <0:01) and changes in SOFA score (p <0.05).

Biochemical and pharmacokinetic characterisation of two PEGylated variants of dipetarudin

SummaryDipetarudin was coupled to polyethylene glycol (PEG)-5000 residues in order to improve its pharmacokinetic profile and to enhance its anticoagulant efficacy. The resulting compounds, mono-and di-PEGylated dipetarudin were purified by gel filtration. Mono-PEGylated dipetarudin exhibited similar activity like its non-conjugated equivalent both in vitro and in vivo. However, di-PEGylated dipetarudin showed longer distribution and elimination half-lives and higher area under the time-concentration curve in comparison with the unmodified inhibitor which may be attributed to decreased renal clearance. Futhermore, ratio k 12/k 21 decreased when the number of PEG chains coupled to dipetarudin increased. It means that the intercompartment transfer of dipetarudin, characterised by a fast distribution and a high retention in the peripheral compartment, is reverted by coupling to PEG. Thus, the transfer of mono-PEGylated dipetarudin between these compartments is similar in both senses and the transfer of di-PEGylated dipetarudin is slower from vascular to extravascular compartment than vice versa. Our results show that di-PEGylated dipetarudin produces a better and longer anticoagulant effect than unmodified dipetarudin which is a desirable attribute for future therapeutic application.

JAM-C regulates unidirectional monocyte transendothelial migration in inflammation

Monocyte recruitment from the vasculature involves sequential engagement of multiple receptors, culminating in transendothelial migration and extravasation. Junctional adhesion molecule-C (JAM-C) is localized at endothelial intercellular junctions and plays a role in monocyte transmigration. Here, we show that blockade of JAM-B/-C interaction reduced monocyte numbers in the extravascular compartment through increased reverse transmigration rather than by reduced transmigration. This was confirmed in vivo, showing that an anti–JAM-C antibody reduced the number of monocytes in inflammatory tissue and increased the number of monocytes with a reverse-transmigratory phenotype in the peripheral blood. All together, our results suggest a novel mechanism of reducing accumulation of monocytes at inflammation sites by disruption of JAM-C–mediated monocyte retention.

Changes of interstitial fluid volume in superficial tissues detected by a miniature ultrasound device

We evaluated the changes of tissue layer thickness in circumscribed superficial tissue areas with a 10-MHz A-mode and a 20-MHz B-mode ultrasound device under alterations in body posture and plasma volume to detect fluid shifts between the different compartments. In 20 male volunteers, we measured tissue thickness by A mode and corium and subcutis thickness by B mode at the forehead before and 30 min after three procedures: change from upright to supine position (P1); change from upright to 30° head-down-tilt position (P2); infusion of 10 ml/kg body wt of Ringer solution (P3). We found a significant correlation between baseline tissue thickness and the sum of corium and subcutis thicknesses ( r = 0.75, P < 0.01). The changes of body posture and plasma volume resulted in significant increases of tissue thickness (P1, 2.9%; P2, 11.6%; P3, 5.8%) and corium thickness (P1, 4.7%; P2, 8.1%; P3, 9.1%) but not of the sum of chorium and subcutis thicknesses. We conclude that fluid shifts from the intravascular to the extravascular compartment are detectible by evaluating corium thickness with a B-mode, or more easily tissue thickness with an A-mode, ultrasound device.