Endocytosis in plant–microbe interactions

Plants encounter throughout their life all kinds of microorganisms, such as bacteria, fungi, or oomycetes, with either friendly or unfriendly intentions. During evolution, plants have developed a wide range of defense mechanisms against attackers. In return, adapted microbes have developed strategies to overcome the plant lines of defense, some of these microbes engaging in mutualistic or parasitic endosymbioses. By sensing microbe presence and activating signaling cascades, the plasma membrane through its dynamics plays a crucial role in the ongoing molecular dialogue between plants and microbes. This review describes the contribution of endocytosis to different aspects of plant–microbe interactions, microbe recognition and development of a basal immune response, and colonization of plant cells by endosymbionts. The putative endocytic routes for the entry of microbe molecules or microbes themselves are explored with a special emphasis on clathrin-mediated endocytosis. Finally, we evaluate recent findings that suggest a link between the compartmentalization of plant plasma membrane into microdomains and endocytosis.     

Effects of cement dust on volatile oil constituents and antioxidative metabolism of Aleppo pine (Pinus halepensis) needles

The effects of cement dust on the chemical composition of essential oil, lipid peroxidation and antioxidant enzyme activities of Aleppo pine (P. halepensis) needles were studied. Cement dust resulted in a significant decrease in the yield of essential oil with the effect being more pronounced in the close vicinity of the cement factory. A concomitant decrease in all components of the oil was observed and ??-2-carene, trans-carveol, trans-carvyl acetate, ??-terpinyl acetate, ??-copaene, (E,E)-??-farnesene, ??-calacorene, ??-cadinene, spathulenol, humulene oxide II, 8-epi-??-eudesmol, ?-muurolol, cubenol and ethyl hexadecanoate have been proposed as biological indicators of cement dust. Moreover, a redirection of the secondary metabolism toward the biosynthesis of monoterpenes has been evidenced. Malondialdehydes (MDA), a decomposition product of polyunsaturated fatty acids, often considered as a suitable biomarker for lipid peroxidation was induced in the needles exposed to cement dust. Similarly, a remarkable induction of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities was noticed. The positive relationships were observed among activities of antioxidant enzymes, and between MDA content and activities of antioxidant enzymes, indicating the cooperative action of these antioxidant enzymes to cope with the oxidative stress induced by cement dust. The results obtained indicate that P. halepensis needles are useful bio-monitors of cement dust pollution.     

A radioimmunoassay of 15 (S) 15 methyl prostaglandin F2α

A sensitive and relative specific radioimmunoassay for 15 (S) 15 methyl prostaglandin F_2α has been developed to enable the measurements of the concentrations of the drug in biological fluids after its administration for therapeutic abortion. The precision, accuracy and specificity of the assay are described.     

Survival of human rhinovirus type 14 dried onto nonporous inanimate surfaces: effect of relative humidity and suspending medium

To study the survival of human rhinovirus 14 on environmental surfaces, each stainless steel disk (1 cm in diameter) was contaminated with 10 microL (about 10(5) plaque-forming units) of the virus suspended in either 1 chi tryptose phosphate broth (TPB), 5 mg/mL of bovine mucin in normal saline, or undiluted human nasal discharge. The inoculum was dried in a laminar flow cabinet for 1 h under ambient conditions. The disks were then placed in a glass chamber (20 +/- 1 degree C) with the relative humidity at either low (20 +/- 5%), medium (50 +/- 5%), or high (80 +/- 5%) level. At appropriate intervals, the disk to be tested was placed in 1 mL of tryptose phosphate broth and the eluate titrated in A-5 HeLa cells. When the virus was suspended in either tryptose phosphate broth, mucin, or the nasal discharge and subjected to initial drying, there was a 3.0 +/- 1.0, 82.0 +/- 6.7, and 89.0 +/- 3.0% loss in virus infectivity, respectively. The half-life of the TPB-suspended virus was about 14 h at the high relative humidity as compared with less than 2 h at the other two relative humidity levels. The half-lives for the mucin-suspended virus at the high, medium, and low relative humidity were 1.42, 0.55, and 0.24 h, respectively; the corresponding values for the nasal discharge suspended virus being 0.17, 0.25, and 0.09 h.     

Effect of Time of Day and Partial Sleep Deprivation on Short‐Term,High‐Power Output

The purpose of this study was to determine whether delaying bedtime or advancing rising time by 4 h affects anaerobic performance of individuals the following day in the morning and afternoon. Eleven subjects participated in the study, during which we measured the maximal, peak, and mean powers (i.e., P_max [force‐velocity test], P_peak, and P_mean [Wingate test], respectively). Measurements were performed twice daily, at 07∶00 and 18∶00 h, following a reference normal sleep night (RN), a partial sleep deprivation timed at the beginning of the night (SDB), and a partial sleep deprivation timed at the end of the night (SDE), and oral temperature was measured every 4 h. Each of the three experimental conditions was separated by a one‐week period. Our results showed a circadian rhythm in oral temperature, and analysis of variance revealed a significant sleep×test‐time effect on peak power (P_peak), mean power (P_mean), and maximal power (P_max). These variables improved significantly from the morning to the afternoon for all three experimental conditions. Whereas the morning‐afternoon improvement in the measures was similar after the RN and SDB conditions, it was smaller following the SDE condition. There was no significant difference in the effect of the two sleep‐deprivation conditions on anaerobic performances at 07∶00 and at 18∶00 h under the SDB condition in comparison with the post‐reference night. However, the performance variables were significantly lower at 18∶00 h after the SDE condition. In conclusion, a 4 h partial sleep deprivation at the end of the night appears to be more disturbing than partial sleep deprivation at the beginning of the night.     

Effect of Time of Day on Aerobic Contribution to the 30‐s Wingate Test Performance

The purpose of this study was to evaluate the effects of time of day on aerobic contribution during high‐intensity exercise. A group of 11 male physical education students performed a Wingate test against a resistance of 0.087 kg · kg^?1 body mass. Two different times of day were chosen, corresponding to the minimum (06:00 h) and the maximum (18:00 h) levels of power. Oxygen uptake (V˙O₂) was recorded breath by breath during the test (30 sec). Blood lactate concentrations were measured at rest, just after the Wingate test, and again 5 min later. Oral temperature was measured before each test and on six separate occasions at 02:00, 06:00, 10:00, 14:00, 18:00, and 22:00 h. A significant circadian rhythm was found in body temperature with a circadian acrophase at 18:16±00:25 h as determined by cosinor analysis. Peak power (P_peak), mean power (P_mean), total work done, and V˙O₂ increased significantly from morning to afternoon during the Wingate Test. As a consequence, aerobic contribution recorded during the test increased from morning to afternoon. However, no difference in blood lactate concentrations was observed from morning to afternoon. Furthermore, power decrease was greater in the morning than afternoon. Altogether, these results indicate that the time‐of‐day effect on performances during the Wingate test is mainly due to better aerobic participation in energy production during the test in the afternoon than in the morning.     

The prevalence and prognostic significance of KRAS mutation in bladder cancer, chronic myeloid leukemia and colorectal cancer

Mutations in the KRAS gene have been shown to play a key role in the pathogenesis of a variety of human tumours. However the mutational spectrum of KRAS gene differs by organ site. In this study, we have analysed the mutational spectrum of KRAS exon 1 in bladder tumours, colorectal cancer (CRC) and chronic myeloid leukemia (CML). A total of 366 patients were included in the present study (234 bladder tumours, 48 CRC and 84 CML). The KRAS mutations are absent in BCR/ABL1 positive CML. This result suggests that BCR/ABL1 fusion gene and KRAS mutations were mutually exclusive. The frequency of KRAS mutations in bladder cancer was estimated at 4.27 %. All of mutations were found in codon 12 and 90 % of them were detected in advanced bladder tumours. However the correlation between KRAS mutations and tumour stage and grade does not report a statistical significant association. The KRAS mutations occur in 35.41 % of patients with CRC. The most frequent mutations were G12C, G12D and G13D. These mutations were significantly correlated with histological differentiation of CRC (p = 0.024). Although the high frequency of KRAS in CRC in comparison to bladder cancer, these two cancers appear to have the same mutational spectrum (p > 0.05).     

Impaired Blood Dendritic Cell Numbers and Functions after Aneurysmal Subarachnoid Hemorrhage

Previous Presentation

Portions of this study were presented at the Annual Congress of Société Française d’Anesthésie et de Réanimation in Paris, September 2012.

Background

Toll-like receptor (TLR) agonists are promising therapy for the prevention of nosocomial infections in critical ill patients. We aimed to analyze the TLR-reactivity of circulating dendritic cells (DC) as assessed by cytokine production after an ex vivo challenge with TLR agonists in aneurysmal subarachnoid hemorrhage (SAH) patients.

Methods and Findings

A single-center prospective observational study took place in one intensive care unit of a teaching hospital. Blood samples were harvested on days 2, 5 and 10 in 21 severe SAH patients requiring mechanical ventilation and 17 healthy controls. DC production of cytokines (Tumour Necrosis Factor, TNF-α; Interleukin, IL-12; and Interferon, IFN-α) was assessed by intracellular immunostaining on TLR-3, 4, 7/8 and 9 stimulations. SAH patients had decreased numbers of blood myeloid (mDCs) and plasmacytoid DCs (pDCs) on days 2, 5 and 10. Compared with the healthy controls, the frequency of mDCs producing TNF-α after TLR-3 stimulation was decreased in the SAH patients. The frequency of myeloid DCs producing IL-12 after TLR-3 and 4 stimulations was also decreased in the SAH patients. In contrast, the mDCs response to TLR-7/8 was not impaired in the SAH patients. The frequency of pDCs producing TNF-α⁺ and IFN-α⁺ on TLR-7/8 stimulation were reduced at all of the tested times in the SAH patients, whereas reactivity to TLR-9 was preserved. On day 2, the pDCs from non-survivor patients (n = 8) had a decreased ability to produce IFN-α on TLR-9 stimulation compared with the survivors.

Conclusions

These data suggest functional abnormalities of circulating pDCs and mDCs that could be important for immunomodulation after SAH.     

Subcutaneous versus Intravenous Administration of Rituximab: Pharmacokinetics,CD20 Target Coverage and B-Cell Depletion in Cynomolgus Monkeys

The CD20-specific monoclonal antibody rituximab (MabThera^®, Rituxan^®) is widely used as the backbone of treatment for patients with hematologic disorders. Intravenous administration of rituximab is associated with infusion times of 4–6 hours, and can be associated with infusion-related reactions. Subcutaneous administration of rituximab may reduce this and facilitate administration without infusion-related reactions. We sought to determine the feasibility of achieving equivalent efficacy (measured by endogenous B-cell depletion) and long-term durability of CD20 target coverage for subcutaneously administered rituximab compared with intravenous dosing. In these preclinical studies, male cynomolgus monkeys were treated with either intravenous rituximab or novel subcutaneous formulation of rituximab containing human recombinant DNA-derived hyaluronidase enzyme. Peripheral blood samples were analyzed for serum rituximab concentrations, peripheral B-cell depletion, and CD20 target coverage, including subset analysis according to CD21+ status. Distal lymph node B-cell depletion and CD20 target coverage were also measured. Initial peak serum concentrations of rituximab were significantly higher following intravenous administration than subcutaneous. However, the mean serum rituximab trough concentrations were comparable at 2 and 7 days post-first dose and 9 and 14 days post-second dose. Efficacy of B-cell depletion in both peripheral blood and distal lymph nodes was comparable for both methods. In lymph nodes, 9 days after the second dose with subcutaneous and intravenous rituximab, B-cell levels were decreased by 57% and 42% respectively. Similarly, levels of peripheral blood B cells were depleted by >94% for both subcutaneous and intravenous dosing at all time points. Long-term recovery of free unbound surface CD20 levels was similar, and the duration of B-cell depletion was equally sustained over 2 months for both methods. These results demonstrate that, despite initial peak serum drug level differences, subcutaneous rituximab has similar durability, pharmacodynamics, and efficacy compared with intravenous rituximab.