Sensitization to nut allergens in children with food allergy and other atopic diseases - just a coexistence?

Introduction: Allergy to nuts, the most common food allergy in childhood, is considered as a significant health problem. Aim: To investigate sensitization to selected nuts in children with or without atopic allergy. Material and methods: Retrospective analysis involved records of 598 children, diagnosed with food allergy. Laboratory data concerned screening for sensitization to major allergens of hazelnut, peanut and walnut. Results: Approximately 77.8% of children with food allergy presented at least one concomitant atopic disease: allergic rhinitis (52.9%), atopic dermatitis (48%) or asthma (31.4%). Nearly one-third experienced at least one episode of anaphylaxis. The nut-specific antibodies were found in 67% of children. Among them, 56% were sensitized to hazelnut, and 54% to peanut. Sensitization to other nuts was less frequent (< 30%). Only 27% of patients were mono-sensitized, the remaining 73% were co-sensitized to two or three of tested nuts. Noteworthy, the occurrence of sensitization varied among age-related groups, and also depended on clinical diagnosis. In patients with sole food allergy the frequency of sensitization was highest in youngest children, whereas, when accompanied by other atopic disease, it was highest in schoolchildren. In children without food allergy, but with another atopic disease, the prevalence of sensitization was relatively low, without any specific pattern. Conclusions: The analysis of sensitization patterns may help to identify patients with an increased risk, and gives the opportunity to introduce more effective prophylaxis. However, since even the first exposure to nuts may be sufficient to trigger the anaphylaxis, this risk should be considered as a serious issue at any age.

Prevalence of nut allergen sensitization among children in central Poland.

Introduction: The diagnostics of plant-derived food allergy may be challenging. However, the recognition of sensitization patterns in defined populations, especially in children, is clinically relevant as it enables the use of secondary prophylaxis to prevent life-threatening complications. Aim: To investigate the rates and sensitization patterns to nut allergens in children from central Poland. Material and methods: The retrospective assessment concerned data of 598 children diagnosed in a single centre due to suspected food allergy. The analysis included the results of component-based multiparametric assay Allergy Explorer2 (ALEX2). Results: The sensitization to particular nut allergens varied among patients, depending on their age and nut type. The sensitization to any nut was found in 67% of children, whereas sensitization to hazelnut and peanut was the most common (56% and 55% of all children, respectively). Hazelnut sensitization was predominant in every age, and its prevalence increased with age, while peanut sensitization was detected in more than half of individuals from all groups, except for teenagers (44% of cases). Among hazelnut molecules sensitization to Cor a 1.04 was the most prevalent (74% of sensitized children), and for peanut allergens - Ara h 1 (65% of sensitized patients). The simultaneous sensitization to hazelnut, peanut and walnut (two or all of them) was found in almost half of the entire group. Conclusions: Component-based diagnostics enables differentiation between primary and cross-reactive sensitization to nut allergens and detects co-sensitization. The clinical relevance of the latter observation is remarkable as co-sensitization increases the risk of life-threatening reactions even in trace nut contamination.

Identification of Lysine Misincorporation at Asparagine Position in Recombinant Insulin Analogs Produced in E. coli.

PURPOSE: Identification of human insulin analogs' impurity with a mass shift +14 Da in comparison to a parent protein. METHODS: The protein sequence variant was detected and identified with the application of peptide mapping, liquid chromatography, tandem mass spectrometric analysis, nuclear magnetic resonance spectroscopy (NMR) and Edman sequencing. RESULTS: The misincorporated lysine (Lys) at asparagine (Asn) position A21 was detected in recombinant human insulin and its analogs. CONCLUSIONS: Although there are three asparagine residues in the insulin derivative, the misincorporation of lysine occurred only at position A21. The process involves G/U or A/U wobble base pairing.

Atom-resonant squeezed light from a tunable monolithic ppRKTP parametric amplifier.

We demonstrate vacuum squeezing at the D1 line of atomic rubidium (795 nm) with a tunable, doubly-resonant, monolithic subthreshold optical parametric oscillator in periodically-poled Rb-doped potassium titanyl phosphate (ppRKTP). The squeezing appears to be undiminished by a strong dispersive optical nonlinearity recently observed in this material.

Mechanism of Binding of Antifungal Antibiotic Amphotericin B to Lipid Membranes: An Insight from Combined Single-Membrane Imaging, Microspectroscopy, and Molecular Dynamics.

Amphotericin B is a lifesaving polyene antibiotic used in the treatment of systemic mycoses. Unfortunately, the pharmacological applicability of this drug is limited because of its severe toxic side effects. At the same time, the lack of a well-defined mechanism of selectivity hampers the efforts to rationally design safer derivatives. As the drug primarily targets the biomembranes of both fungi and humans, new insights into the binding of amphotericin B to lipid membranes can be helpful in unveiling the molecular mechanisms underlying both its pharmacological activity and toxicity. We use fluorescence-lifetime-imaging microscopy combined with fluorescence-emission spectroscopy in the microscale to study the interaction of amphotericin B with single lipid bilayers, using model systems based on giant unilamellar liposomes formed with three lipids: dipalmitoylphosphatidylcholine (DPPC), dimirystoylphosphatidylcholine (DMPC), and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC). The results show that amphotericin B introduced into the water phase as a DMSO solution binds to the membrane as dimers and small-molecular aggregates that we identify as tetramers and trimers. Fluorescence-detected linear-dichroism measurements revealed high orientational freedom of all the molecular-organization forms with respect to the membrane plane, which suggests that the drug partially binds to the membrane surface. The presence of sterols in the lipid phase (cholesterol but particularly ergosterol at 30 mol %) promotes the penetration of drug molecules into the lipid membrane, as concluded on the basis of the decreased orientation angle of amphotericin B molecules with respect to the axis normal to the membrane plane. Moreover, ergosterol facilitates the association of amphotericin B dimers into aggregated structures that can play a role in membrane destabilization or permeabilization. The presence of cholesterol inhibits the formation of small aggregates in the lipid phase of liposomes, making this system a promising candidate for a low-toxicity antibiotic-delivery system. Our conclusions are supported with molecular simulations that reveal the conformational properties of AmB oligomers in both aqueous solution and lipid bilayers of different compositions.

Identification of N-Terminally Truncated Derivatives of Insulin Analogs Formed in Pharmaceutical Formulations.

PURPOSE: Isolation and identification of unknown impurities of recombinant insulin lispro (produced at IBA) formed during accelerated stability testing of pharmaceutical solutions. For comparative purposes also commercially available formulations of recombinant human insulin (Humulin S®; Lilly), recombinant insulin lispro (Humalog®; Lilly), recombinant insulin aspart (NovoRapid® Penfill®; Novo Nordisk), recombinant insulin detemir (Levemir®; Novo Nordisk) and recombinant insulin glargine (Lantus®; Sanofi-Aventis) were analyzed. METHODS: The impurities of insulin analogs were isolated by RP-HPLC and identified with peptide mass fingerprinting using MALDI-TOF/TOF mass spectrometry. RESULTS: The identified derivatives were N-terminally truncated insulin analog impurities of decreased molecular mass of 119, 147 and 377 Da related to the original protein. The modifications resulting in a mass decrease were detected at the N-terminus of B chains of insulin lispro, insulin aspart, human insulin, insulin glargine, insulin detemir in all tested formulations. To our knowledge it is the first time that these impurities are reported. CONCLUSIONS: The following derivatives formed by truncation of the B chain in insulin analogs were identified in pharmaceutical formulations: desPheB1-N-formyl-ValB2 derivative, desPheB1 derivative, pyroGluB4 derivative.

Synthesis, Molecular Structure, Anticancer Activity, and QSAR Study of N-(aryl/heteroaryl)-4-(1H-pyrrol-1-yl)Benzenesulfonamide Derivatives.

A series of N-(aryl/heteroaryl)-4-(1H-pyrrol-1-yl)benzenesulfonamides were synthesized from 4-amino-N-(aryl/heteroaryl)benzenesulfonamides and 2,5-dimethoxytetrahydrofuran. All the synthesized compounds were evaluated for their anticancer activity on HeLa, HCT-116, and MCF-7 human tumor cell lines. Compound 28, bearing 8-quinolinyl moiety, exhibited the most potent anticancer activity against the HCT-116, MCF-7, and HeLa cell lines, with IC50 values of 3, 5, and 7 µM, respectively. The apoptotic potential of the most active compound (28) was analyzed through various assays: phosphatidylserine translocation, cell cycle distribution, and caspase activation. Compound 28 promoted cell cycle arrest in G2/M phase in cancer cells, induced caspase activity, and increased the population of apoptotic cells. Relationships between structure and biological activity were determined by the QSAR (quantitative structure activity relationships) method. Analysis of quantitative structure activity relationships allowed us to generate OPLS (Orthogonal Projections to Latent Structure) models with verified predictive ability that point out key molecular descriptors influencing benzenosulfonamide's activity.

[Tragedy in the Valley Jaworowa: incredible coincidence or perfect murder?]

The paper presents the case of an unexplained and the most mysterious death in the history of Polish Tatra tourism. It concerns three people of different ages and occurred on August 3, 1925 in the Valley of Jaworowa. Kazimierz Kasznica, his son Waclaw Kasznica and a newly discovered mountaineer Ryszard Wasserberger died suddenly for unknown reasons in 15 minutes during a mountain trek. This story is interesting due to the mysterious, simultaneous death of three people of different ages and due to the fact that Waleria Kasznica - the wife of Kazimierz and the mother of Waclaw Kasznica survived the journey. KEY WORDS.

Fully-resonant, tunable, monolithic frequency conversion as a coherent UVA source.

We demonstrate a monolithic frequency converter incorporating up to four tuning degrees of freedom, three temperature and one strain, allowing resonance of pump and generated wavelengths simultaneous with optimal phase-matching. With a Rb-doped periodically-poled potassium titanyl phosphate (KTP) implementation, we demonstrate efficient continuous-wave second harmonic generation from 795 to 397, with low-power efficiency of 72% and high-power slope efficiency of 4.5%. The measured performance shows good agreement with theoretical modeling of the device. We measure optical bistability effects, and show how they can be used to improve the stability of the output against pump frequency and amplitude variations.

Self-tuning optical resonator.

We demonstrate a nonlinear optical resonator that tunes itself onto resonance with an input beam. In a monolithic Fabry-Perot cavity implemented in rubidium-doped periodically poled potassium titanyl phosphate, an intensity-dependent refractive index produces line pulling by multiple free-spectral ranges (FSRs). In this condition, the cavity passively maintains optical resonance in the face of FSR-scale excursions of the drive laser frequency: when one resonant operating point becomes unstable, the resonator rapidly transitions to another resonant operating point. We demonstrate stable second-harmonic generation with no active feedback to the laser or cavity. The self-tuning effect appears to be supported by a very strong, previously unreported optical nonlinearity.

Membrane Sterols Modulate the Binding Mode of Amphotericin B without Affecting Its Affinity for a Lipid Bilayer.

Membrane-active antibiotics are known to selectively target certain pathogens based on cell membrane properties, such as fluidity, lipid ordering, and phase behavior. These are in turn modulated by the composition of a lipid bilayer and in particular by the presence and type of membrane sterols. Amphotericin B (AmB), the golden standard of antifungal treatment, exhibits higher activity toward ergosterol-rich fungal membranes, which permits its use against systemic mycoses; however, the selectivity for fungal membranes is far from satisfactory leading to severe side effects. Despite decades of research, no consensus has emerged on the origin of AmB specificity for fungal cells and its actual mode of action at the molecular level. Previously, it has been proposed that the specific action of AmB is related to differences in its affinity for membranes of different composition. In this work, we investigate this relationship by employing molecular dynamics simulations to compare the free energy of insertion of AmB into three types of membranes: a pure DMPC bilayer and DMPC bilayers containing 30% of cholesterol or ergosterol. We analyze the orientation of AmB molecules within the bilayer in order to unambiguously establish their membrane binding mode and relate the orientational freedom to the sterol-dependent tightness of lipid packing. Our results strongly indicate that the membrane insertion of AmB proceeds virtually to completion independent of membrane type, and hence the higher toxicity against fungal membranes may rather result from differences in subsequent oligomerization in the membrane and assembly of monomers into functional transmembrane pores. In particular, the latter could be facilitated by sterol-induced ordering of AmB molecules along the membrane normal, revealed by our free energy profiles. Moreover--in contrast to certain claims--we find no stable binding mode corresponding to the horizontal adsorption of AmB on the membrane surface.

Macroscopic quantum state analyzed particle by particle.

Macroscopic quantum phenomena, e.g., superconductivity and squeezing, are believed to result from entanglement of macroscopic numbers of particles. We report the first direct study of this kind of entanglement: we use discrete quantum tomography to reconstruct the joint quantum state of photon pairs extracted from polarization-squeezed light. Our observations confirm several predictions from spin-squeezing theory [Beduini et al., Phys. Rev. Lett. 111, 143601 (2013)], including strong entanglement and entanglement of all photon pairs within the squeezing coherence time. This photon-by-photon analysis may give insight into other macroscopic many-body systems, e.g., photon Bose-Einstein condensates.

Atomic filtering for hybrid continuous-variable/discrete-variable quantum optics.

We demonstrate atomic filtering of frequency-degenerate photon pairs from a sub-threshold optical parametric oscillator (OPO). The filter, a modified Faraday anomalous dispersion optical filter (FADOF), achieves 70% peak transmission simultaneous with 57 dB out-of-band rejection and a 445 MHz transmission bandwidth. When applied to the OPO output, only the degenerate mode, containing one-mode squeezed vacuum, falls in the filter pass-band; all other modes are strongly suppressed. The high transmission preserves non-classical continuous-variable features, e.g. squeezing or non-gaussianity, while the high spectral purity allows reliable discrete-variable detection and heralding. Correlation and atomic absorption measurements indicate a spectral purity of 96% for the individual photons, and 98% for the photon pairs. These capabilities will enable generation of atom-resonant hybrid states, e.g. "Schrödinger kittens" obtained by photon subtraction from squeezed vacuum, making these exotic states available for quantum networking and atomic quantum metrology applications.

Interferometric measurement of the biphoton wave function.

Interference between an unknown two-photon state (a "biphoton") and the two-photon component of a reference state gives a phase-sensitive arrival-time distribution containing full information about the biphoton temporal wave function. Using a coherent state as a reference, we observe this interference and reconstruct the wave function of single-mode biphotons from a low-intensity narrow band squeezed vacuum state.

Ultranarrow Faraday rotation filter at the Rb D1 line.

We present a theoretical and experimental study of the ultranarrow bandwidth Faraday anomalous dispersion optical filter operating at the rubidium D1 line (795 nm). This atomic line gives better performance than other lines for key figures of merit, e.g., simultaneously 71% transmission, 445 MHz bandwidth, and 1.2 GHz equivalent-noise bandwidth.

Abnormal sperm morphology is associated with sensitization to inhaled allergens.

BACKGROUND: Allergy is associated with the loss of tolerance of environmental antigens, combined with a pathological immune response. There were no studies up to date that would show whether the quality of semen decreases in people with allergic diseases. MATERIAL AND METHODS: The research included men who reported to the Gynecological Outpatient Clinic due to reproductive difficulties, defined as the lack of pregnancy after one year of regular intercourse. Semen quality was assessed according to the World Health Organization (WHO) standard. All patients underwent skin prick tests with the most important inhalation allergens (such as hazel, silver birch, mugwort, rye, dog, cat, Dermatophagoides farinae, Dermatophagoides pteronyssinus, alder, Alternaria alternata, Cladosporium herbarum, and grass mix). The data was statistically analyzed. RESULTS: Results of 52 patients aged 25-52 years (34.62 ± 4.96) were analyzed. The mean BMI (Body mass index) was 28.25 (+ -3.77). It was found that 38 men (73%) had increased body weight, and 14 men (26.9%) were obese (BMI > = 30). 13 patients were smokers (25%), and 24 patients (46%) had skin tests positive for at least one inhaled allergen. Sperm tail defects were statistically more significant in patients allergic to birch, rye, cat, alder, and grass. In patients allergic to Alternaria alternata, head defects were statistically more significant (p < .05). No association was found between allergy to house dust mites, mugwort, hazel, and dogs and the deterioration of semen. CONCLUSION: Allergy due to inhalation allergens had an influence on the quality of male semen. Further research is necessary to establish the immunological bases of this phenomenon.

Self-assembly, stability and conductance of amphotericin B channels: bridging the gap between structure and function.

Amphotericin B (AmB), one of the most powerful but also toxic drugs used to treat systemic mycoses, is believed to selectively permeabilize fungal cell membranes to ions in a sterol-dependent manner. Unfortunately, the structure of the biologically active AmB channels has long eluded researchers, obstructing the design of safer alternatives. Here, we investigate the structural and thermodynamic aspects of channel formation, stability, and selective ion conduction. We combine fluorescence lifetime imaging and molecular simulations to trace the process of channel assembly until the formation of stable, roughly octameric double-length channels (DLCs). This stoichiometry is confirmed by matching the predicted channel conductances with the past results of patch-clamp measurements. We then use free energy calculations to explain the effect of sterols on DLC stability and discuss the observed cation selectivity in structural terms, addressing several long-standing controversies in the context of their physiological relevance. Simulations of ion permeation indicate that only solvated ions pass through DLCs, revealing surprising solvation patterns in the channel lumen. We conclude our investigation by inspecting the role of the tail hydroxyl in the assembly of functional channels, pointing at possible origins of the cholesterol-ergosterol selectivity.

Biochemical Properties Affecting the Nutritional Quality, Safety, and Aroma of Dry-Cured Products Manufactured from Meat of Rare Native Pig Breeds.

The aim of study was to compare the biochemical properties affecting the nutritional quality, safety, and aroma of dry-cured products manufactured from valuable meat of rare native pig breeds: Pulawska (Pul) and Zlotnicka Spotted (ZS). The count of lactic acid bacteria (4.4 log cfu/g) and the release of palmitic (23.1% and 25.9%), oleic (44.1% and 42.2%), and linoleic acids (8.3% and 7.8%), as well as arginine (30.0 and 44.3 mg/kg), histidine (25.8 and 20.6 mg/kg), and lysine (26.8-22.9 mg/kg), shaped the final pH (5.3 and 5.4) in Pul and ZS products during the 4 week maturing, respectively. Lastly, Pul and ZS meat differed in the proportion of decanoic, lauric, stearic, arachidic, and conjugated linoleic acids. The high content of putrescine (23.7 mg/kg), cadaverine (54.3 mg/kg), and tyramine (57.2 mg/kg), as well as a twofold greater share of histamine (163.2 mg/kg) and tryptamine (9.1 mg/kg), indicated a more advanced decarboxylation of ZS meat. Volatile compounds differentiating Pul and ZS meat were primarily hexanal, 3-hydroxybutan-2-one, phenylacetalaldehyde, 2,3-dimethyl-2-cyclopenten-1-one, 2-cyclopenten-1-one, and 3-methyl- and 2-cyclopenten-1-one. Most marked volatile compounds were obtained as a result of microbial activity (acetic acid, 3-methylbutan-1-ol, ethanol, acetone, and 3-hydroxybutan-2-one), advanced lipid oxidation, and decomposition of secondary oxidation products (hexanal, phenylacetaldehyde, and 2-cyclopenten-1-one).

High-Grade Gliomas in Children-A Multi-Institutional Polish Study.

Due to the rarity of high-grade gliomas (HGG) in children, data on this topic are scarce. The study aimed to investigate the long-term results of treatment of children with HGG and to identify factors related to better survival. We performed a retrospective analysis of patients treated for HGG who had the main tumor located outside the brainstem. The evaluation of factors that correlated with better survival was performed with the Cox proportional-hazard model. Survival was estimated with the Kaplan-Meier method. The study group consisted of 82 consecutive patients. All of them underwent surgery as primary treatment. Chemotherapy was applied in 93% of children with one third treated with temozolomide. After or during the systemic treatment, 79% of them received radiotherapy with a median dose of 54 Gy. Median follow-up was 122 months, and during that time, 59 patients died. One-, 2-, 5-, and 10-year overall survival was 78%, 48%, 30% and 17%, respectively. Patients with radical (R0) resection and temozolomide-based chemotherapy had better overall survival. Progression-free survival was better in patients after R0 resection and radical radiotherapy. The best outcome in HGG patients was observed in patients after R0 resection with immediate postoperative temozolomide-based chemotherapy and radical radiotherapy.

A novel potassium channel in skeletal muscle mitochondria.

In this work we provide evidence for the potential presence of a potassium channel in skeletal muscle mitochondria. In isolated rat skeletal muscle mitochondria, Ca(2+) was able to depolarize the mitochondrial inner membrane and stimulate respiration in a strictly potassium-dependent manner. These potassium-specific effects of Ca(2+) were completely abolished by 200 nM charybdotoxin or 50 nM iberiotoxin, which are well-known inhibitors of large conductance, calcium-activated potassium channels (BK(Ca) channel). Furthermore, NS1619, a BK(Ca)-channel opener, mimicked the potassium-specific effects of calcium on respiration and mitochondrial membrane potential. In agreement with these functional data, light and electron microscopy, planar lipid bilayer reconstruction and immunological studies identified the BK(Ca) channel to be preferentially located in the inner mitochondrial membrane of rat skeletal muscle fibers. We propose that activation of mitochondrial K(+) transport by opening of the BK(Ca) channel may be important for myoprotection since the channel opener NS1619 protected the myoblast cell line C2C12 against oxidative injury.