Inactivated influenza vaccine adjuvanted with bacterium-like particles induce systemic and mucosal influenza A virus specific T-cell and B-cell responses after nasal administration in a TLR2 dependent fashion.

BACKGROUND: Nasal vaccination is considered to be a promising alternative for parenteral vaccination against influenza virus as it is non-invasive and offers the opportunity to elicit strong antigen-specific responses both systemic and locally at the port of entry of the pathogen. Previous studies showed that non-living bacterium-like particles (BLPs) from the food-grade bacterium Lactococcus lactis are effective stimulators of local and systemic immune responses when administered intranasally. Moreover, in vitro, BLPs specifically interact with human Toll-like receptor 2 (TLR2), suggestive of a role for TLR2 dependent immune activation by BLPs. METHODS: In the present study, we examined the role of TLR2 in vivo in immune activation after nasal administration of BLP mixed with split influenza vaccine (BLP-SV) of influenza A virus (IAV) using TLR2 knockout mice. RESULTS: The systemic Th1 cell and subsequent B-cell responses induced after intranasal BLP-SV vaccination depended on the interaction of BLPs with TLR2. Notably, the BLP-SV-induced class switch to IgG2c depended on the interaction of BLP with TLR2. Local induced IAV-specific Th1 cell responses and the mucosal B-cell responses also depended on interaction of BLP with TLR2. Strongly reduced SIgA levels were observed in TLR2 knockout mice both in the nasal and vaginal lavages. In addition, detailed analysis of the T-cell response revealed that nasal BLP-SV vaccination promoted Th1/Th17 immune responses that coincided with increased IAV-specific IgG2c antibody production. DISCUSSION: Altogether these results indicate that nasal BLP-SV vaccination induces IAV-specific T-cell and B-cell responses, both systemically and at the site of virus entry in a TLR2-dependent manner.

Awake fibrecapnic intubation in head and neck cancer patients with difficult airways: new findings and refinements to the technique.

Recently, we described an adaptation of awake fibreoptic intubation that we call awake fibrecapnic intubation. The aim of this study was to evaluate the efficacy and risk of complications with this novel technique in a consecutive case series of head and neck cancer patients known to have difficult airways. We prospectively studied 40 consecutive intubations in head and neck cancer patients prior to a diagnostic or surgical procedure. Following topical anaesthesia, a flexible bronchoscope was introduced into the pharynx; spontaneous respiration was maintained in all patients. A special suction catheter was advanced into the airway through the suction channel of the bronchoscope for carbon dioxide measurements. When four capnograms were obtained, the bronchoscope was railroaded over the catheter and a tracheal tube was placed. All adverse events and complications were recorded. There were no complications associated with the technique. The median (range) time to intubation was 3 min (1.5-15 min). All patients were intubated successfully, 39 (98%) of them using awake fibrecapnic intubation. There was one patient with severe tumour bleeding and acute airway obstruction caused by advancement of the tube over the bronchoscope. This was not considered to be a complication of the fibrecapnic technique. Awake fibrecapnic intubation is a safe and valuable technique in head and neck cancer patients with a difficult airway.

Compound A and carbon monoxide production from sevoflurane and seven different types of carbon dioxide absorbent in a patient model.

BACKGROUND: The degradation of sevoflurane can lead to the production of compound A (CA) and carbon monoxide (CO) and an increase in temperature of the absorbent. CA is known to be nephrotoxic in rats. These reactions depend on the strong base and water contents of the carbon dioxide absorbent used. The purpose of this study was to measure the maximum amounts of CA and CO produced, and the temperature increase, for seven different carbon dioxide absorbents for sevoflurane containing different contents of strong bases. METHODS: Seven absorbents [some free of strong bases (f)] were employed in hydrated (h) and completely desiccated (d) conditions in a patient model, using a circle anesthesia system connected to an artificial lung. Low-flow anesthesia with an oxygen-nitrous oxide mixture was maintained using 0.8% sevoflurane. For the quantification of CA and CO, a portable gas chromatograph was used. The temperature was measured inside the absorbent. RESULTS: In consecutive order of CA-producing potency, Amsorb(f)(d), Drägersorb(h), Medisorb(h), lithium hydroxide(f)(d), Drägersorb(d), Medisorb(d), Spherasorb(h) and Spherasorb(d) produced small amounts of CA. Loflosorb and Superia, which are free of strong bases, did not produce any CA or CO in hydrated or desiccated conditions. Only desiccated Drägersorb, Medisorb and Spherasorb demonstrated small amounts of CO accompanied by a significant temperature increase. CONCLUSION: In this patient model, we demonstrated that different types of absorbent produced small amounts of CA and CO or none at all. No relationship could be established between temperature and CA concentration.

Carbon monoxide production from desflurane and six types of carbon dioxide absorbents in a patient model.

BACKGROUND: Desflurane is known to produce high concentrations of carbon monoxide (CO) in desiccated sodalime or Baralyme (Allied Healthcare Products, St. Louis, MO). Desiccated absorbents without strong bases like potassium hydroxide or sodium hydroxide are reported to produce less or no CO at all. The purpose of this study is to compare the concentration of CO in an anesthesia circuit for desflurane with six different types of completely desiccated CO(2) absorbents with less strong bases than sodalime. METHODS: A patient model was simulated using a circle anesthesia system connected to an artificial lung. Completely desiccated CO(2) absorbent (950 g) was used in this system. A low flow anesthesia (500 ml min(-1)) was maintained using desflurane. For immediate quantification of CO production a portable gas chromatograph was used. RESULTS: Peak concentrations of CO were very high in Medisorb (Datex-Ohmeda, Hoevelaken, The Netherlands) and Spherasorb (Intersurgical, Uden, The Netherlands) (13317 and 9045 p.p.m., respectively). It was lower with Loflosorb (Intersurgical, Uden, The Netherlands) and Superia (Datex-Ohmeda, Hoevelaken, The Netherlands) (524 and 31 p.p.m., respectively). Amsorb (Armstrong, Coleraine, N. Ireland) and lithium hydroxide produced no CO at all. CONCLUSION: Medisorb and Spherasorb are capable of producing large concentrations of CO when desiccated. Loflosorb and Superia produce far less CO under the same conditions. Amsorb and lithium hydroxide should be considered safe when desiccated.

Predictive value of symptom level measurements for complex regional pain syndrome type I.

The validity with respect to presence or absence of CRPS I according to Veldman's criteria was assessed for measured pain, temperature, volume differences and limitations in range of motion. Evaluated were 155 assessments of 66 outpatients, initially diagnosed with CRPS I, but many of them not so on follow up visits. Pain was measured with VAS and McGill, temperature by infrared thermometry, volume differences by water displacement volumeters and limitations in range of motion by universal goniometers. Sensitivity, specificity, positive and negative predictive value of the measurement instruments at different cut-off points was calculated. Combined symptom scores were evaluated in a similar fashion. High sensitivity was found for the VAS, McGill, and range of motion. The specificity was overall lower, but highest values were obtained for volume differences. The positive predictive value was good for all measurement instruments. Negative predictive value was lower, especially for measurement of temperature and volume asymmetries. If sensitivity and specificity are equally important, VAS>3 cm, McGill>6 words, temperature difference>or=0.4 degrees C, volume difference>6.5% and ROM limitation>15% provide the best results. Using these cut off values, the highest value of sensitivity and of sensitivity and specificity combined, was found for a combination of VAS, McGill and ROM. The highest value of specificity was found for different combinations of 3, 4 and 5 instruments, all containing the VAS. We conclude that the measured pain, temperature, volume and range of motion can be used as diagnostic indicators for establishing presence or absence of CRPS I.

Induction of air embolism in xylem conduits of pre-defined diameter.

A new method is presented that enables the induction of embolisms in a fraction of all xylem vessels, based on diameter, at one cut end of a stem segment. The method is based on the different capillary characteristic of xylem vessels of different cross-sectional size. To verify the method, air embolisms were induced in cut xylem vessels of chrysanthemum (Dendranthemaxgrandiflorum Tzvelev cv. Cassa) stem segments at different xylem tensions and compared with the distribution of gas-filled vessels as visualized by cryo-scanning electron microscopy (Cryo-SEM). At -6 kPa xylem pressure, air-entrance was only induced in large diameter vessels (>30 microm), while at -24 kPa embolisms were induced in almost all xylem vessels (>10 microm). Although the principle of the embolization method worked well, smaller diameter vessels were observed to be embolized than was expected according to the calculations. The role of cross-sectional shape and contact angle between xylem sap and vessel wall at the menisci are discussed. After correction for the observed (diameter independent) deviation from circularity of the cross-sectional vessel shape the contact angle was calculated to be approximately 55 degrees. Hydraulic resistance (Rh) measurements before and after embolization showed that the effect of embolizing only large diameter cut xylem vessels had only a small influence on overall Rh of a stem segment. Embolizing all cut xylem vessels at one cut end almost trebled overall Rh. The difference was discussed in the light of the networking capacity of the xylem system.

Xylem hydraulic conductivity related to conduit dimensions along chrysanthemum stems.

The stem xylem conduit dimensions and hydraulic conductivity of chrysanthemum plants (Dendranthema x grandiflorum Tzvelev cv. Cassa) were analysed and quantified. Simple exponential relations describe conduit length distribution, height dependency of conduit length distribution, and height dependency of stem hydraulic conductivity. These mathematical descriptions can be used to model the xylem water transport system. Within a chrysanthemum stem of 1.0 m, the conduit half-length (the length within which 50% of the conduits have their end) was 0.029 m at soil surface and decreased by half at a height of 0.6 m. With each 0.34 m increase in height up the stem, the hydraulic conductivity decreased by 50%. The resistance calculated from conduit lumen characteristics was 70% of the measured resistance. The remaining unexplained part of the hydraulic resistance is at least partly caused by inter-conduit connections.

Molecular characterization of the CER1 gene of arabidopsis involved in epicuticular wax biosynthesis and pollen fertility.

The aerial parts of plants are coated with an epicuticular wax layer, which is important as a first line of defense against external influences. In Arabidopsis, the ECERIFERUM (CER) genes effect different steps of the wax biosynthesis pathway. In this article, we describe the isolation of the CER1 gene, which encodes a novel protein involved in the conversion of long chain aldehydes to alkanes, a key step in was biosynthesis. CER1 was cloned after gene tagging with the heterologous maize transposable element system Enhancer-Inhibitor, also known as Suppressor-mutator. cer1 mutants display glossy green stems and fruits and are conditionally male sterile. The similarity of the CER1 protein with a group of integral membrane enzymes, which process highly hydrophobic molecules, points to a function of the CER1 protein as a decarbonylase.

A Seed Shape Mutant of Arabidopsis That Is Affected in Integument Development.

A seed shape mutant of Arabidopsis was isolated from an ethyl methanesulfonate-treated population. Genetic analysis revealed that the heart-shaped phenotype was maternally inherited, showing that this is a testa mutant. This indicated the importance of the testa for the determination of the seed shape. This recessive aberrant testa shape (ats) gene was located at position 59.0 on chromosome 5. A comparison was made between ovules and developing and mature seeds of the wild type and of the mutant using light and scanning electron microscopy. We showed that the mutant seed shape is determined during the first few days after fertilization, when the embryo occupies only a very small part of the seed. The integuments of ats ovules consisted of only three rather than five cell layers. In double mutants, the effect of ats was additive to other testa mutations, such as transparent testa, glabra (ttg), glabrous2 (gl2), and apetala2 (ap2). The ats mutation resulted in a reduced dormancy, which was maternally inherited. This effect of a testa mutation on germination was also seen in ttg seeds, in which the outer layer of the testa was disturbed. This indicated the importance of the testa as a factor in determining dormancy in Arabidopsis.

THE PROCESSES OF ANTHER DEHISCENCE AND POLLEN DISPERSAL: I. THE OPENING MECHANISM OF LONGITUDINALLY DEHISCING ANTHERS.

The process of anther opening was followed in different species by means of light and electron microscopical techniques, stereo microscopical observation inside living tissues and micromanipulation. From these data and a review of the mainly very old literature on this subject, the opening process is outlined. It consists of the following stages: (1) Expansion of the epidermis and endothecium cells and deposition of U-shaped wall thickenings inside the latter. (2) Enzymatic opening of the septum between two locules. (3) Mechanical rupture of the tapetum. (4) Mechanical opening of the stomium by the centripetal force of the highly turgescent epidermis and endothecium. (5) Outward bending of the locule walls by the centrifugal force of the dehydrating epidermis and endothecium. Steps (4) and (5) clearly reflect the mechanical functions of the two latter tissues. The roles of the other anther tissues and the filament during the earlier stages are discussed.

THE PROCESSES OF ANTHER DEHISCENCE AND POLLEN DISPERSAL: II. THE FORMATION AND THE TRANSFER MECHANISM OF POLLENKITT, CELL-WALL DEVELOPMENT OF THE LOCULUS TISSUES AND A FUNCTION OF ORBICULES IN POLLEN DISPERSAL.

The development of the locule of Gasteria verrucosa (Mill.) H. Duval and Lilium hybrida cv. Enchantment, especially the border between the sporophyte and the gametophyte, is investigated by means of light and electron microscopy, histochemistry and micromanipulation and related to pollen dispersal. The pecto-cellulosic cell walls of both the middle layer and the tapetum and a part of the endothecium disappear, and the tapetum cells are covered with sporopollenin-containing tapetal membranes and orbicules. The cell contents of the tapetum turn into the hydrophobic pollenkitt. In the locule, similar changes are observed as the pecto-cellulosic and callose walls of the meiocytes disappear and the future pollen grains are covered with the sporopollenin-containing exine. Due to their non-wettability, these hydrophobic substances on both sides of the sporophyte-gametophyte surface are important in pollen dispersal. The transfer of pollenkitt from the tapetum to the locule appears to be due to capillary forces in the locule after the continuous expansion of the pollen, rather than to active flow or to direct attraction by the pollen grains.

Structure and function of the microtubular cytoskeleton during pollen development in Gasteria verrucosa (Mill.) H. Duval.

In a study of pollen development in Gasteria verrucosa, the changes in the spatial organization of microtubules were related to the processes of cell division, nuclear movement and cytomorphogenesis. Sections of polyethylene-glycol-embedded anthers of G. verrucosa were processed immunocytochemically to record the structure and succession of fluorescently labeled microtubular configurations. Using microspectrophotometric measurements the relative quantity of tubulin in microtubules per unit of cytoplasm was determined. Cell dimensions and nuclear positions were measured to relate changes in cell shape and nuclear movements to microtubular configurations. Microtubules were detected in the different cells during microsporogenesis and microgametogenesis. In microspore mother cells which are approximately isodiametric at interphase, microtubules were predominantly arranged in a criss-cross pattern. The microtubules probably function as a flexible cytoskeleton which sustains the integrity of the cytoplasm. Bundles of microtubules were observed in the microspores, in the generative cells and during nuclear division, where they functioned in establishing and maintaining cell and spindle shapes. Microtubules radiating from nuclear membranes appeared to fix the nucleus in position. In prophase of meiosis and after microspore mitosis, periods a high fluorescence intensity were distinguished indicating a variation in the quantity of microtubules.