Resting respiratory tract dendritic cells preferentially stimulate T helper cell type 2 (Th2) responses and require obligatory cytokine signals for induction of Th1 immunity

Consistent with their role in host defense, mature dendritic cells (DCs) from central lymphoid organs preferentially prime for T helper cell type 1 (Th1)-polarized immunity. However, the "default" T helper response at mucosal surfaces demonstrates Th2 polarity, which is reflected in the cytokine profiles of activated T cells from mucosal lymph nodes. This study on rat respiratory tract DCs (RTDCs) provides an explanation for this paradox. We demonstrate that freshly isolated RTDCs are functionally immature as defined in vitro, being surface major histocompatibility complex (MHC) II lo, endocytosishi, and mixed lymphocyte reactionlo, and these cells produce mRNA encoding interleukin (IL)-10. After ovalbumin (OVA)-pulsing and adoptive transfer, freshly isolated RTDCs preferentially stimulated Th2-dependent OVA-specific immunoglobulin (Ig)G1 responses, and antigen-stimulated splenocytes from recipient animals produced IL-4 in vitro. However, preculture with granulocyte/macrophage colony stimulating factor increased their in vivo IgG priming capacity by 2-3 logs, inducing production of both Th1- and Th2-dependent IgG subclasses and high levels of IFN-gamma by antigen-stimulated splenocytes. Associated phenotypic changes included upregulation of surface MHC II and B7 expression and IL-12 p35 mRNA, and downregulation of endocytosis, MHC II processing- associated genes, and IL-10 mRNA expression. Full expression of IL-12 p40 required additional signals, such as tumor necrosis factor alpha or CD40 ligand. These results suggest that the observed Th2 polarity of the resting mucosal immune system may be an inherent property of the resident DC population, and furthermore that mobilization of Th1 immunity relies absolutely on the provision of appropriate microenvironmental costimuli.     

The primary aortoenteric fistula in The Netherlands--the unpublished cases

OBJECTIVE: Primary aortoenteric fistula is a rare disorder of which only four patients have been reported in the Dutch literature so far. The objective of our study was to obtain more realistic figures on the incidence of this condition, with data on the clinical presentation, diagnostic procedures, treatment and results in a group of patients not previously reported as "case histories". METHODS: A questionnaire was sent to all surgical clinics in The Netherlands. Out of 180 questionnaires, 102 have been returned reporting 27 patients to which data of eight others treated in our own institution were added. RESULTS: In all but one of these 29 patients the fistula was caused by an atherosclerotic aneurysm, the one exception being caused by an ingested cocktail pin. Gastrointestinal haemorrhage was the predominant symptom, being present in 28 of the patients, while the complete triad of haemorrhage, pain and a pulsating mass was found in only eight patients. Twenty-seven patients were treated with an in situ graft of which 14 are doing well at long term follow-up. CONCLUSIONS: Primary aortoenteric fistula is far more common than one would expect from the number of patients reported in literature. A high index of suspicion based on a complete physical examination remains the key to a correct diagnosis. Direct closure of the intestine and in situ grafting of the aorta is the treatment of first choice.     

Iterative optimization of high-affinity proteases inhibitors using phage display. 1. Plasmin

We generated a series of libraries having variants of the first Kunitz domain of human lipoprotein-associated coagulation inhibitor (LACI-D1, also known as tissue-factor pathway inhibitor-I) displayed on bacteriophage M13 as pIII-fusions. We varied LACI-DI iteratively in two regions: the P1 region (positions 10-21) and the "second loop", (positions 31-39), which together form one end of the domain. Display-phage library Lib#1 allows 31 200 amino-acid sequences in P1 region (residues 13, 16-19). Preliminary, we screened Lib#1 against human plasmin (PLA, EC 3.4.21.7) immobolized on agarose to enrich for phage displaying variants with PLA affinity. We introduced a 1600-fold increase in second-loop diversity (residues 31, 32, 34, 39) into the population of selectants from Lib#1, yielding Lib#2. Lib#2 (allowing approximately 50 million amino-acid sequences) was screened against PLA-agarose to isolate highest affinity binders. Protein EPI-P211, derived from the best isolate of Lib#2, inhibits PLA with Ki = 2 nM (at least 500-fold better than LACI-D1) and with high specificity. We used amino-acid sequences of PLA-binding selectants to design a PLA-biased library (Lib#3) which we screened against PLA. The protein EPI-P302 (derived from the best binder obtained from Lib#3) has Ki for PLA inhibition of 87 pM, which is 25-fold better than the first-round best binder and > or = 12 500-fold better than LACI-D1. EPI-P302 also shows very high specificity for PLA vs other human proteases and is resistant to inactivation by oxidants and extremes of temperature or pH. Thus, one can use selectants from one library to design target-tailored combinatorial libraries and obtain quite stable, highly specific, very high-affinity binding molecules while maintaining an essentially human framework.     

Temperature dependence of macroscopic L-type calcium channel currents in single guinea pig ventricular myocytes

INTRODUCTION: Lowering temperature greatly reduces calcium influx through calcium channels. Studies on a number of tissues demonstrate that the peak inward current, ICa, exhibits Q10 values ranging from 1.8 to 3.5; however, it remains unclear which component(s) of calcium channel gating may give rise to this large temperature sensitivity. Components of gating that may affect channel availability include phosphorylation and changes in [Ca2+]i, processes that vary in pertinence depending on the channel examined. This study addresses this problem by examining the temperature sensitivity (from 34 degrees to 14 degrees C) of cardiac ICa under control conditions, during attenuation or activation of protein kinase A (PKA) activity, and when intracellular [Ca2+] has been elevated. METHODS AND RESULTS: ICa was studied using the whole cell configuration of the patch champ technique. In control, lowering temperature from 34 degrees to 24 degrees C resulted in a shift in the potential for maximum slope (Va) and the peak current (Ymax) toward more positive membrane potentials. The Q10 values for the decrease in Ymax and the macroscopic slope conductance (Gmax), which reflects the number of available channels, were 3.15 +/- 0.19 and 2.57 +/- 0.13, respectively. At 0 mV the Ca2+ current decayed biexponentially, and the two time constants (tau 1 and tau 2) showed Q10 values of 1.79 +/- 0.21 and 2.06 +/- 0.38, while their contribution to the total current (I1 and I2) showed a Q10 of 5.99 +/- 0.83 and 1.61 +/- 0.22. In myocytes loaded with inhibitors of the PKA cycle sufficient to inhibit the increase of ICa to 1 microM isoprenaline, the Q10 values for some of the kinetic parameters were increased with the Q10 for I1 increasing to 17.06 +/- 3.48. Stimulation of ICa by exposing myocytes to 1 microM isoprenaline reduced the temperature sensitivity of Ymax, Gmax and I1, yielding respective values of 2.00 +/- 0.18, 1.85 +/- 0.07, and 2.04 +/- 0.15. Raising [Ca2+]i to enhance Ca2+i-dependent inactivation, while affecting inactivation and activation kinetics, affected temperature sensitivity little compared to control. The Q10 for time to peak changed little under experimental conditions (2.3 to 2.4) CONCLUSIONS: Increasing the phosphorylated states of calcium channels, but not Ca2+i-dependent inactivation, reduces temperature sensitivity of certain gating parameters. The data suggest that the rate of the transitions between the unavailable and also between the various closed states are changed in the opposite direction to that induced by PKA-dependent phosphorylation. Processes, e.g., inhibitory mechanisms, may be involved to maintain channels in unavailable or "unphosphorylated" states, and it may be these that contribute to the high Q10 of macroscopic channel currents.     

Recognition of depressed affect in hospitalized psychiatric patients: staff and patient perceptions

A comparison of self-report vs. observer rating of depressed mood in a heterogenous inpatient population revealed wide variations in concordance among diagnostic groups. Patients diagnosed as having Affective Psychosis and "Other' illnesses showed the highest correlation between four self-report scales and an observer rating scale. Patients with a diagnosis of depressive Neurosis showed only modest correlation, while Schizophrenics revealed no significant correlation, on these instruments, suggesting inconsistent communication of affect from Schizophrenic patients to observers. In contrast, when self-report scales were intercorrelated, patients in all four diagnostic categories showed highly significant correlations, indicating that they were consistently reporting their affective state on these instruments. The implications of these findings for future research as well as for practical clinical management are discussed.     

Spark plasma sintering of B4C and B4C-TiB2 composites: Deformation and failure mechanisms under quasistatic and dynamic loading

Spark plasma sintering (SPS) was employed to consolidate powder specimens consisting of B₄C and various B₄C-TiB₂ compositions. SPS allowed for consolidation of pure B₄C, B₄C-13 vol.%TiB₂, and B₄C-23 vol.%TiB₂ composites achieving ≥99 % theoretical density without sintering additives, residual phases (e.g., graphite), and excessive grain growth due to long sintering times. Electron and x-ray microscopies determined homogeneous microstructures along with excellent distribution of TiB₂ phase in both small and larger-scaled composites. An optimized B₄C-23 vol.%TiB₂ composite with a targeted low density of ～3.0 g/cm³ exhibited 30–35 % increased hardness, fracture toughness, and flexural bend strength compared to several commercial armor-grade ceramics, with the flexural strength being strain rate insensitive under quasistatic and dynamic loading. Mechanistic studies determined that the improvements are a result of a) no residual graphitic carbon in the composites, b) interfacial microcrack toughening due to thermal expansion coefficient differences placing the B₄C matrix in compression and TiB₂ phase in tension, and c) TiB₂ phase aids in crack deflection thereby increasing the amount of intergranular fracture. Collectively, the addition of TiB₂ serves as a toughening and strengthening phase, and scaling of SPS samples show promise for the manufacture of ceramic composites for body armor.     

Maize and pigeon pea sole crop and intercrop nutrient response functions for Tanzania

The development of ecological circular agriculture has been highly encouraged by the Chinese government to recycle agricultural wastes, reduce mineral fertilizer input, and protect the environment. Biogas slurry, a byproduct of biogas engineering developed in rural areas of China, could be used as N fertilizer for crop growth. The field experiments were conducted in 2014 and 2015 to study the plant growth responses and environmental impacts of applying biogas slurry to two-season Zizania aquatica Turcz. growth. The potential factors that restrict the rational use of biogas slurry were also clarified. Mineral N fertilizer can be completely or partly substituted by N fertilizer from biogas slurry to satisfy Z. aquatica plant growth. It was not at the cost of sacrificing yield, dry matter accumulation, N accumulation and physiological N use efficiency in the above-ground parts. However, the growth inhibition occurs when the N quantity in biogas slurry was 2 or 2.7 times higher than that of mineral N fertilizer. Vitamin C in non-shell swollen culms (as edible part) of Z. aquatica significantly increased after biogas slurry application. Biogas slurry application substantially increased the N concentrations, i.e., total N, NH₄ ⁺–N, and NO₃ ^?–N in floodwater and delayed the time to reach national discharge standards. However, biogas slurry application did not affect the N concentrations in percolating water compared with the treatment with mineral N fertilizer only. Applying biogass slurry did not generate potential pollution risks by trace elements (Cu, Zn, Pb, Cr, Cd, As, and Hg) in the non-shell swollen culm and soil, and did not increase the nitrate content in non-shell swollen culm. We found the NH₄ ⁺–N concentration in biogas slurry can account for 77–93% of total N and reflects the N level in biogas slurry to a great degree. Semi-quantitative color-based colorimetric methods possessing simple and fast characteristics should be developed to determine the NH₄ ⁺–N concentration with the purpose of promoting reasonable use of biogas slurry in area of crop cultivation. Otherwise, excessive use of biogas slurry can adversely affect crops and increase environmental risks.     

Boundary scan: the Internet of test

Boundary scan enables us to reuse tests developed at different design levels and in different life-cycle phases. By facilitating communication between previously isolated areas, built-in boundary scan becomes what the authors call the “Internet of Test”     

Hairline lowering during foreheadplasty

Many patients seeking rejuvenation of their foreheads have high hairlines and are troubled by the prospect that surgery will worsen the deformity. Hairline elevation occurs in both coronal and endoscopic foreheadplasty techniques and is at least part of the reason that many surgeons do not always recommend these procedures when otherwise indicated. Although a pretrichial "hairline" incision prevents hairline retro-displacement, it results in forehead shortening only and not true hairline lowering. When a pretrichial incision is used in combination with a posterior scalp advancement flap, however, true hairline lowering is possible. Experience with this technique encompasses 27 procedures performed over a 5-year period. Patients ranged in age from 35 to 71 years. A significant improvement was demonstrable in all cases and corresponded with a high degree of patient satisfaction. No serious complications were seen. A high hairline must be recognized as the source of both a disproportionate and aged appearance. The ability to lower the hairline and place it in a more proportionate, youthful relationship with the rest of the face adds a new dimension to foreheadplasty equal to or greater in importance to an overall improved appearance as other maneuvers typically performed during the procedure. For many patients, the benefits of hairline lowering far outweigh the trade-off of a more anteriorly situated, and possibly more visible, scar.