Combined Blockade of TIGIT and CD39 or A2AR Enhances NK-92 Cell-Mediated Cytotoxicity in AML

This study aimed to characterize different natural killer (NK) cell phenotypes on bone marrow and peripheral blood cells from acute myeloid leukemia (AML) patients and healthy donors (HDs). Our data show that CD56^dimCD16⁻ and CD56^brightCD16⁻ NK cells represent the predominant NK cell subpopulations in AML, while the CD56^dimCD16⁺ NK cells are significantly reduced compared to HDs. Moreover, TIGIT⁺ and PVRIG⁺ cells cluster on the CD56^dimCD16⁺ subset whereas CD39⁺ and CD38⁺ cells do so on CD56^brightCD16⁻ NK cells in AML. Furthermore, functional effects of (co-)blockade of TIGIT and CD39 or A2AR on NK cell functionality were analyzed. These experiments revealed that the single blockade of the TIGIT receptor results in an increased NK-92 cell-mediated killing of AML cells in vitro. Combined targeting of CD39 or A2AR significantly augments the anti-TIGIT-mediated lysis of AML cells. Our data indicate that distinct NK cell subsets in AML exhibit different immunosuppressive patterns (via the TIGIT/PVRIG receptors and the purinergic pathway). In summary, we conclude that TIGIT, CD39, and A2AR constitute relevant inhibitory checkpoints of NK cells in AML patients. A combinatorial blockade synergistically strengthens NK-92 cell-mediated cytotoxicity. As inhibitors of TIGIT, CD39, and A2AR are clinically available, studies on their combined use could be conducted in the near future.     

The spray forming of nanostructured Aluminum Oxide

Research Summary Nanostructured ceramics and their composites possess improved properties such as tensile strength, fatigue strength, hardness, and wear resistance. Freestanding, near-net shape, nanostructured Al₂O₃ components can be synthesized via plasma-spray forming. In this study, plasma-spray parameters were optimized and an innovative substrate cooling technique was developed to retain nanosize Al₂O₃ in the spray deposit. Nanosize Al₂O₃ particles were partially melted and trapped between the fully melted coarser, micrometersize Al₂O₃grains. Densification of the spray-deposited Al₂O₃occurred via solidification and sintering. A similar processing approach can be adopted for fabrication of near-net shapes of a variety of nanostructured materials (metals, ceramics, and intermetallics) and their combinations by selecting suitable powder-treatment and plasmaspray parameters.     

Synthesis and in vitro evaluation of alpha-GalCer epimers

alpha-GalCer (also known as KRN7000) is an immunomodulatory glycolipid that is known to potently activate invariant natural killer T (NKT) cells upon CD1d-mediated stimulation. Because Th1 and Th2 cytokines, which are released after alpha-GalCer presentation, antagonize each other's effects, alpha-GalCer analogues that induce a biased Th1/Th2 response are highly awaited. In this context, we report the synthesis and in vitro evaluation of alpha-Gal-D-xylo-Cer and two alpha-Gal-L-lyxo-Cer analogues, one with the natural acyl chain, the other with a truncated chain.     

Recent studies of particle packing in organic coatings

Organic coatings often contain sufficient pigmentation that the pigment volume concentration (PVC) and the critical PVC (CPVC) must be carefully considered. Past work has shown that the CPVC can be identified with the random densest packing of pigment particles after considerations of the adsorbed polymer layer on the particles is taken into account, and that this packing can be predicted by empirical particle packing algorithms developed for mixtures of particle sizes. Recent studies have shown that local fluctuations of pigment and polymer concentrations in a film can lead to local film volumes that can exceed the CPVC even though the average PVC for the whole film (the global PVC) is below the CPVC. These effects are consistent with observations of void formation in films below the CPVC, and can be analyzed by local film statistics. We have also recently performed simulations of particle packing by the compression of multiple hard spheres that allows further insight into local concentration fluctuations in PVC within a film.     

Modification of the fat composition of the Iberian pig using Bacillus licheniformis and Bacillus subtilis

The aim of this work was to study the influence of a diet with probiotic bacteria on the lipid composition of different tissues from Iberian pigs. Cholesterol and fatty acid profiles were measured in liver, subcutaneous fat, and Serratus ventralis muscle samples. Feeding Iberian pigs a mixture of probiotic microorganisms for 45 days prior to slaughter affected the tissue composition. The feeds of the control and probiotic groups were identical except for the added Bacillus licheniformis and Bacillus subtilis (1.28 × 10⁹ cfu/kg feed) in the feed of the probiotic group. The diet with probiotics significantly affected the fatty acid profile of liver, subcutaneous fat and Serratus ventralis muscle samples, but no significant differences were found for the cholesterol content. The livers of the probiotic group contained lower levels of n‐6 fatty acids and higher levels of n‐3 fatty acids. The subcutaneous fat contained lower levels of n‐6 fatty acids. Serratus ventralis muscles from the probiotic group contained a significantly higher percentage of oleic acid than those of the control group. Practical applications: This study deals with the effect of the addition of probiotic bacteria to the diet of pigs on both the cholesterol and fatty acid content. B. subtilis and B. licheniformis can be used in pig feed (Regulation EC no. 2148/2004), but there are no studies about their effects on fresh meat or other tissues, particularly regarding the fat composition. Thus, it is interesting to study how feeding these microorganisms affects the different tissues. Including these microorganisms in the basic diet could serve as a strategy to modify the fat composition and to obtain healthier products from pigs.     

Determination of Carbohydrate‐Binding Preferences of Human Galectins with Carbohydrate Microarrays

Galectins are a class of carbohydrate‐binding proteins named for their galactose‐binding preference and are involved in a host of processes ranging from homeostasis of organisms to immune responses. As a first step towards correlating the carbohydrate‐binding preferences of the different galectins with their biological functions, we determined carbohydrate recognition fine‐specificities of galectins with the aid of carbohydrate microarrays. A focused set of oligosaccharides considered relevant to galectins was prepared by chemical synthesis. Structure–activity relationships for galectin–sugar interactions were determined, and these helped in the establishment of redundant and specific galectin actions by comparison of binding preferences. Distinct glycosylations on the basic lactosyl motifs proved to be key to galectin binding regulation—and therefore galectin action—as either high‐affinity ligands are produced or binding is blocked. High‐affinity ligands such as the blood group antigens that presumably mediate particular functions were identified.     

Development, fabrication, and test of a modular eddy current micro sensor on a flexible polymer foil

This paper describes the development and fabrication of a modular eddy current micro sensor on a flexible polymer foil. Due to handling purposes during the micro sensor fabrication process the modular eddy current micro sensors are fabricated on a temporary Si substrate. To enable a release of the micro sensors at the end of the fabrication process, initial investigations concentrated on the proof of principle applying a deep reactive-ion etching (DRIE) process to structure the Si wafer. The DRIE process was used to structure Si frames, which serve as carriers for the modular micro sensors. For the evaluation of the fabricated eddy current micro sensors, electrical resistance measurements were accomplished with the integrated anisotropic magneto-resistance (AMR) sensor. The aim of these investigations was to evaluate the influence of the substrate material on the characteristics of the AMR sensor. The electrical output signal of these micro sensors were subsequently compared to electrical resistance measurements of identical AMR sensors fabricated on a Si substrate and served as reference. To prove the capability of the completed eddy current micro sensors, a defect in form of a scratch was created on the surface of a Cu probe and investigated by eddy current testing. The electrical output signal of the eddy current micro sensor was subsequently compared to an optical measurement of the surface profile of this scratch. There is a high correlation between the optical measurements and the signal of the eddy current micro sensor.     

Inflammation in the Human Periodontium Induces Downregulation of the α1- and β1-Subunits of the sGC in Cementoclasts

Nitric oxide (NO) binds to soluble guanylyl cyclase (sGC), activates it in a reduced oxidized heme iron state, and generates cyclic Guanosine Monophosphate (cGMP), which results in vasodilatation and inhibition of osteoclast activity. In inflammation, sGC is oxidized and becomes insensitive to NO. NO- and heme-independent activation of sGC requires protein expression of the α₁- and β₁-subunits. Inflammation of the periodontium induces the resorption of cementum by cementoclasts and the resorption of the alveolar bone by osteoclasts, which can lead to tooth loss. As the presence of sGC in cementoclasts is unknown, we investigated the α₁- and β₁-subunits of sGC in cementoclasts of healthy and inflamed human periodontium using double immunostaining for CD68 and cathepsin K and compared the findings with those of osteoclasts from the same sections. In comparison to cementoclasts in the healthy periodontium, cementoclasts under inflammatory conditions showed a decreased staining intensity for both α₁- and β₁-subunits of sGC, indicating reduced protein expression of these subunits. Therefore, pharmacological activation of sGC in inflamed periodontal tissues in an NO- and heme-independent manner could be considered as a new treatment strategy to inhibit cementum resorption.     

Identification and Composition of Clasper Scent Gland Components of the Butterfly Heliconius erato and Its Relation to Mimicry

The butterfly Heliconius erato occurs in various mimetic morphs. The male clasper scent gland releases an anti-aphrodisiac pheromone and additionally contains a complex mixture of up to 350 components, varying between individuals. In 114 samples of five different mimicry groups and their hybrids 750 different compounds were detected by gas chromatography/mass spectrometry (GC/MS). Many unknown components occurred, which were identified using their mass spectra, gas chromatography/infrared spectroscopy (GC/IR)-analyses, derivatization, and synthesis. Key compounds proved to be various esters of 3-oxohexan-1-ol and (Z)-3-hexen-1-ol with (S)-2,3-dihydrofarnesoic acid, accompanied by a large variety of other esters with longer terpene acids, fatty acids, and various alcohols. In addition, linear terpenes with up to seven uniformly connected isoprene units occur, e. g. farnesylfarnesol. A large number of the compounds have not been reported before from nature. Discriminant analyses of principal components of the gland contents showed that the iridescent mimicry group differs strongly from the other, mostly also separated, mimicry groups. Comparison with data from other species indicated that Heliconius recruits different biosynthetic pathways in a species-specific manner for semiochemical formation.