Diagnosis, genetic characterization and clinical follow up of mitochondrial fatty acid oxidation disorders in the new era of expanded newborn screening: A single centre experience.

INTRODUCTION: Mitochondrial fatty acid oxidation disorders (FAODs) are a heterogeneous group of hereditary autosomal recessive diseases included in newborn screening (NBS) program in Italy. The aim of this study was to analyse FAODs cases, identified either clinically or by NBS,for clinical and genetic characterization and to evaluate a five years' experience of NBS, in the attempt to figure out the complexity of genotype-phenotype correlation and to confirm the clinical impact of NBS in our centre experience. MATERIALS AND METHODS: We analysed FAODs patients diagnosed either by NBS or clinically, followed since February 2014 to April 2019 at the Regional Screening Centre and Inherited Metabolic Diseases Unit of Verona. Diagnosis was confirmed by plasma acylcarnitines, urinary organic acids, enzymatic and genetic testing. For not clear genotypes due to the presence of variants of uncertain significance, in silico predictive tools have been used as well as enzymatic activity assays. Patients underwent clinical, nutritional and biochemical follow up. RESULTS: We diagnosed 30 patients with FAODs. 20 by NBS: 3 CUD, 6 SCADD, 5 MCADD, 4 VLCADD, 2 MADD. Overall incidence of FAODs diagnosed by NBS was 1:4316 newborns. No one reported complications during the follow up period. 10 patients were diagnosed clinically: 2 CUD, 2 CPT2D, 1 VLCADD, 5 MADD. Mean age at diagnosis was 29.3 years. Within this group, complications or symptoms were reported at diagnosis, but not during follow-up. 12 mutations not previously reported in literature were found, all predicted as pathogenic or likely pathogenic. DISCUSSION AND CONCLUSIONS: Our study highlighted the great phenotypic variability and molecular heterogeneity of FAODs and confirmed the importance of a tailored follow up and treatment. Despite the short duration of follow up, early identification by NBS prevented diseases related complications and resulted in normal growth and psycho-motor development as well.

The role of edible oils in low molecular weight organogels rheology and structure.

In this paper, the role of solvent characteristics on the rheological and physicochemical properties of organogels was investigated using different techniques. Vegetable oils, such as rice, sunflower and castor oil were used as solvents, for producing organogels with monoglycerides of fatty acids or a mixture of fatty alcohols (policosanol) as gelators. Moreover, two non-edible oils (silicon and paraffin oil) were also used for analysing the properties of solvents completely different in nature with respect to the edible ones, for a better interpretation of the given results. Organogels were investigated from a rheological point of view and through a microscopic analysis, given by polarised light (POM) and atomic force (AFM) microscopy, and X-rays to study the crystallinity of the system. The IR technique was used to analyse the intermolecular interactions, resulting in interesting information about the effect of oil polarity on the driving forces promoting structuration. This investigation showed that when solvents of a similar chemical nature are used, their physical properties, mainly oil polarity, are strictly related to the properties of the organogel, such as the onset of crystallisation temperature, the stiffness of the final material and its crystallinity. Anyway, these physical parameters seem insufficient to describe properly the role of solvents when oils of a different chemical nature are compared.

Effect of the monostearate/monopalmitate ratio on the oral release of active agents from monoacylglycerol organogels.

The delivery of active agents from organogels is becoming an important topic owing to the possibility of releasing, in a controlled way, lipophilic agents. Controlled release from foods is a topic with increasing relevance owing to the growing industrial interest towards functional or medical foods, i.e. foods containing nutraceutical agents or drugs. Anyway, release properties are related to the rheological properties of organogels, and, therefore, a deep knowledge of their microstructure and physical characteristics is necessary to design carriers with expected release properties. In this work, two low molecular weight gelators (i.e. glycerol monopalmitate, GMP, and glycerol monostearate, GMS) have been investigated using rheology, microscopy and infrared spectroscopy, IR, aiming at understanding the effects of different gelator ratios on organogel properties. It was observed that GMP, within the range of investigated compositions, seems to be more effective in yielding consistent organogels and this effect was related to differences in microstructure with respect to GMS. Their ability to control the oral release of active agents was investigated, in vitro, using a chemotherapeutic drug for adenocarcinoma of the gastrointestinal tract, 5 fluorouracil (5-FU). A physical model based on carrier erosion was used to describe the release data, evidencing a good agreement with experimental values. Among the tested samples it seems that the use of 90% of GMS (over total organogelator content) yields promising results allowing a good partition of the released drug between the gastric and intestinal tracts with the largest value (although lower than 40% of loaded amount) of the total released drug.

Magnetization switching in high-density magnetic nanodots by a fine-tune sputtering process on a large-area diblock copolymer mask.

Ordered magnetic nanodot arrays with extremely high density provide unique properties to the growing field of nanotechnology. To overcome the size limitations of conventional lithography, a fine-tuned sputtering deposition process on mesoporous polymeric template fabricated by diblock copolymer self-assembly is herein proposed to fabricate uniform and densely spaced nanometer-scale magnetic dot arrays. This process was successfully exploited to pattern, over a large area, sputtered Ni80Fe20 and Co thin films with thicknesses of 10 and 13 nm, respectively. Carefully tuned sputter-etching at a suitable glancing angle was performed to selectively remove the magnetic material deposited on top of the polymeric template, producing nanodot arrays (dot diameter about 17 nm). A detailed study of magnetization reversal at room temperature as a function of sputter-etching time, together with morphology investigations, was performed to confirm the synthesis of long-range ordered arrays displaying functional magnetic properties. Magnetic hysteresis loops of the obtained nanodot arrays were measured at different temperatures and interpreted via micromagnetic simulations to explore the role of dipole-dipole magnetostatic interactions between dots and the effect of magnetocrystalline anisotropy. The agreement between measurements and numerical modelling results indicates the use of the proposed synthesis technique as an innovative process in the design of large-area nanoscale arrays of functional magnetic elements.

Influence of block copolymer feature size on reactive ion etching pattern transfer into silicon.

A successful realisation of sub-20 nm features on silicon (Si) is becoming the focus of many technological studies, strongly influencing the future performance of modern integrated circuits. Although reactive ion etching (RIE), at both micrometric and nanometric scale has already been the target of many studies, a better understanding of the different mechanisms involved at sub-20 nm size etching is still required. In this work, we investigated the influence of the feature size on the etch rate of Si, performed by a cryogenic RIE process through cylinder-forming polystyrene-block-polymethylmethacrylate (PS-b-PMMA) diblock copolymer (DBC) masks with diameter ranging between 19-13 nm. A sensible decrease of the etch depth and etch rate was observed in the mask with the smallest feature size. For all the DBCs under investigation, we determined the process window useful for the correct transfer of the nanometric cylindrical pattern into a Si substrate. A structural and physicochemical investigation of the resulting nanostructured Si is reported in order to delineate the influence of various RIE pattern effects. Feature-size-dependent etch, or RIE-lag, is proved to significantly affect the obtained results.

The relationship between hyperthyrotropinemia and metabolic and cardiovascular risk factors in a large group of overweight and obese children and adolescents.

PURPOSE: Mild TSH elevations are frequently observed in obese patients, in the absence of any detectable thyroid disease. Our objective is to evaluate the relationship between the raised TSH levels and the biochemical and clinical consequences of obesity. METHODS: This is a retrospective cross-sectional study of a large population of obese children and adolescents. We evaluated 833 subjects (340 m, 493 f), aged 14.4 ± 2.5 (range 5.2-18.5) years, height SDS 0.27 ± 1.04 (-3.49-4.35), and BMI SDS 2.94 ± 0.59 (1.60-4.68). Body composition, free T4, TSH, anti-TPO antibodies, anti-TG antibodies, inflammation markers (total WBC and the subtypes, ultrasensitive C-reactive protein), and metabolic parameters [AST, ALT, γGT, ALP, glycaemia, insulin, total cholesterol (TC), HDL-cholesterol (HDL-C), and LDL-cholesterol (LDL-C), triglycerides (TG)] were measured, and oral disposition index (ODI) and cardiovascular risk factors (TC/HDL-C and TG/HDL-C) were calculated. After exclusion of the subjects showing anti-thyroid antibodies, the remaining 779 (325 m, 454 f) were then subdivided into two subgroups according to a TSH value below (group A) or above (group B) 4.5 mU/L. RESULTS: Clinical characteristics and hematological markers of patients with and without positive anti-thyroid antibodies were similar, with the exception of higher TSH levels in the latter group. Using analysis of covariance, the subjects of group B had significantly higher values of TC (170.3 ± 28.7 vs 163.3 ± 32.9 mg/dL; p < 0.05), systolic (125.8 ± 13.5 vs 124.5 ± 13.1 mm/Hg), and diastolic blood pressure (79.2 ± 8.0 vs 77.9 ± 8.2 mm/Hg) than subjects of group A. No difference was observed in body composition, ODI, and the cardiovascular risk factors between these two groups. CONCLUSION: TSH elevation in overweight and obese children and adolescents, being associated with a higher TC and blood pressure, might negatively influence the cardiac status. Longitudinal studies are requested, however, to confirm this hypothesis and, therefore, to conclude whether a substitutive treatment with l-thyroxine is really needed in these patients.

The innovative "Bio-Oil Spread" prevents metabolic disorders and mediates preconditioning-like cardioprotection in rats.

BACKGROUND AND AIMS: Obesity is often associated with an increased cardiovascular risk. The food industry and the associated research activities focus on formulating products that are a perfect mix between an adequate fat content and health. We evaluated whether a diet enriched with Bio-Oil Spread (SD), an olive oil-based innovative food, is cardioprotective in the presence of high-fat diet (HFD)-dependent obesity. METHODS AND RESULTS: Rats were fed for 16 weeks with normolipidic diet (ND; fat: 6.2%), HFD (fat: 42%), and ND enriched with SD (6.2% of fat + 35.8% of SD). Metabolic and anthropometric parameters were measured. Heart and liver structures were analyzed by histochemical examination. Ischemic susceptibility was evaluated on isolated and Langendorff-perfused cardiac preparations. Signaling was assessed by Western blotting. Compared to ND rats, HFD rats showed increased body weight and abdominal obesity, dyslipidemia, and impaired glucose tolerance. Morphological analyses showed that HFD is associated with heart and liver modifications (hypertrophy and steatosis, respectively), lesser evident in the SD group, together with metabolic and anthropometric alterations. In particular, IGF-1R immunodetection revealed a reduction of hypertrophy in SD heart sections. Notably, SD diet significantly reduced myocardial susceptibility against ischemia/reperfusion (I/R) with respect to HFD through the activation of survival signals (Akt, ERK1/2, and Bcl2). Systolic and diastolic performance was preserved in the SD group. CONCLUSIONS: We suggest that SD may contribute to the prevention of metabolic disorders and cardiovascular alterations typical of severe obesity induced by an HFD, including the increased ischemic susceptibility of the myocardium. Our results pave the way to evaluate the introduction of SD in human alimentary guidelines as a strategy to reduce saturated fat intake.

Neutral wetting brush layers for block copolymer thin films using homopolymer blends processed at high temperatures.

Binary homopolymer blends of two hydroxyl-terminated polystyrene (PS-OH) and polymethylmethacrylate (PMMA-OH) homopolymers (Mn âˆ¼ 16000 g mol(-1)) were grafted on SiO2 substrates by high-temperature (T > 150 °C), short-time (t < 600 s) thermal treatments. The resulting brush layer was tested to screen preferential interactions of the SiO2 substrate with the different symmetric and asymmetric PS-b-PMMA block copolymers deposited on top of the grafted molecules. By properly adjusting the blend composition and the processing parameters, an efficient surface neutralization path was identified, enabling the formation, in the block copolymer film, of homogeneous textures of lamellae or cylinders perpendicularly oriented with respect to the substrate. A critical interplay between the phase segregation of the homopolymer blends and their grafting process on the SiO2 was observed. In fact, the polar SiO2 is preferential for the PMMA-rich phase that forms a homogeneous layer on the substrate, while the PS-rich phase is located at the polymer-air interface. During the thermal treatment, phase segregation and grafting proceed simultaneously. Complete wetting of the PS rich phase on the PMMA rich phase leads to the formation of a PS/PMMA bilayer. In this case, the progressive diffusion of PS chains toward the polymer-SiO2 interface during the thermal treatment allows tuning of the brush layer composition.

Olive oil and hyperthermal water bigels for cosmetic uses.

Bigels are biphasic systems produced with an organogel (or oleogel) and a hydrogel mixed together at high shear rates. These systems are promising for different uses, among them the formulation of new cosmetic matrices for cosmetic agents delivery is under investigation. In the present paper, a common cosmetic formulation for skin care was enriched with increasing fractions of monoglycerides of fatty acids/olive oil organogels, in order to understand the rheology and the microstructure of these systems. Small amplitude oscillation tests, NMR-self diffusion analysis, contrast phase microscopy and electric conductivity confirmed that the addition of the organogel caused a microstructural change of the starting material, which turned from O/W to a more complex system where, probably, a matrix-in-matrix structure is present at the highest fractions of added organogel.

High aspect ratio PS-b-PMMA block copolymer masks for lithographic applications.

The control of the self-assembly (SA) process and nanostructure orientation in diblock copolymer (DBC) thick films is a crucial technological issue. Perpendicular orientation of the nanostructures in symmetric and asymmetric poly(styrene)-b-poly(methyl methacrylate) (PS-b-PMMA) block copolymer films obtained by means of simple thermal treatments was demonstrated to occur in well-defined thickness windows featuring modest maximum values, thus resulting in low aspect ratio (h/d < 2) of the final lithographic mask. In this manuscript, the thickness window corresponding to the perpendicular orientation of the cylindrical structures in asymmetric DBC is investigated at high temperatures (190 °C ≤ T ≤ 310 °C) using a rapid thermal processing machine. A systematic study of the annealing conditions (temperature and time) of asymmetric PS-b-PMMA (Mn = 67.1, polydispersity index = 1.09) films, with thicknesses ranging from 10 to 400 nm, allowed ordered patterns, with a maximum value of orientational correlation length of 350 nm, to be obtained for film thicknesses up to 200 nm. The complete propagation of the cylindrical structures through the whole film thickness in a high aspect ratio PS template (h/d ≈ 7) is probed by lift-off process. Si nanopillars are obtained having the same lateral ordering and characteristic dimensions of the DBC lithographic mask as further confirmed by grazing-incidence small-angle X-ray scattering experiments.

Rapid thermal processing of self-assembling block copolymer thin films.

Self-assembling block copolymers generate nanostructured patterns which are useful for a wide range of applications. In this paper we demonstrate the capability to control the morphology of the self-assembling process of PS-b-PMMA diblock copolymer thin films on unpatterned surfaces by means of fast thermal treatment performed in a rapid thermal processing machine. The methodology involves the use of radiation sources in order to rapidly drive the polymeric film above the glass transition temperature. Highly ordered patterns were obtained for perpendicular-oriented cylindrical and lamellar PS-b-PMMA block copolymers in less than 60 s. This approach offers the unprecedented opportunity to investigate in detail the kinetics of the block copolymer self-assembly during the early stages of the process, providing a much deeper understanding of the chemical and physical phenomena governing these processes.

Collective behavior of block copolymer thin films within periodic topographical structures.

We perform a systematic study of the effect of adjacent nanostructures on the confinement of block copolymers (BCP) within pre-patterned trenches in 100 nm thick SiO2 films. Asymmetric PS-b-PMMA BCP with a styrene fraction of 0.71, Mn = 67100 are used. When deposited in the form of thin film, these BCP naturally self-organize upon annealing and form a PS matrix with hexagonally packed PMMA cylinders perpendicularly oriented with respect to the substrate. An accurate study of the confinement of this BCP thin film within isolated trenches is performed as a function of their width (80-260 nm). In this specific configuration the confinement of the BCP thin film within the pre-patterned structures has only been partially achieved. The effect of adjacent trenches on the arrangement of the BCP thin film is investigated using parallel trenches periodically distributed on the surface. The effective confinement of the BCP film is strongly modified by the periodicity of the pre-patterned structures.

Er-doped light emitting slot waveguides monolithically integrated in a silicon photonic chip.

An integrated erbium-based light emitting diode has been realized in a waveguide configuration allowing 1.54 µm light signal routing in silicon photonic circuits. This injection device is based on an asymmetric horizontal slot waveguide where the active slot material is Er(3+) in SiO2 or Er(3+) in Si-rich oxide. The active horizontal slot waveguide allows optical confinement, guiding and lateral extraction of the light for on-chip distribution. Light is then coupled through a taper section to a passive Si waveguide terminated by a grating which extracts (or inserts) the light signal for measuring purposes. We measured an optical power density in the range of tens of µW/cm(2) which follows a super-linear dependence on injected current density. When the device is biased at high current density, upon a voltage pulse (pump signal), free-carrier and space charge absorption losses become large, attenuating a probe signal by more than 60 dB/cm and thus behaving conceptually as an electro-optical modulator. The integrated device reported here is the first example, still to be optimized, of a fundamental block to realize an integrated silicon photonic circuit with monolithic integration of the light emitter.

Electrical pump & probe and injected carrier losses quantification in Er doped Si slot waveguides.

Electrically driven Er(3+) doped Si slot waveguides emitting at 1530 nm are demonstrated. Two different Er(3+) doped active layers were fabricated in the slot region: a pure SiO(2) and a Si-rich oxide. Pulsed polarization driving of the waveguides was used to characterize the time response of the electroluminescence (EL) and of the signal probe transmission in 1 mm long waveguides. Injected carrier absorption losses modulate the EL signal and, since the carrier lifetime is much smaller than that of Er(3+) ions, a sharp EL peak was observed when the polarization was switched off. A time-resolved electrical pump & probe measurement in combination with lock-in amplifier techniques allowed to quantify the injected carrier absorption losses. We found an extinction ratio of 6 dB, passive propagation losses of about 4 dB/mm, and a spectral bandwidth > 25 nm at an effective d.c. power consumption of 120 µW. All these performances suggest the usage of these devices as electro-optical modulators.

Quality of life in patients with pemphigus receiving adjuvant therapy.

BACKGROUND: Pemphigus has a strong effect on patients' quality of life (QOL). AIM: To analyze QOL and psychological well-being within patient groups, subdivided according to their different adjuvant treatments. METHODS: All adult patients with pemphigus enrolled in the study were assessed using the Short Form (SF)-36, the Skindex-29, and the General Health Questionnaire (GHQ)-12 for health status, effect of dermatology-specific aspects, and the presence of psychological comorbidity, respectively. The study population was subdivided into the following treatment groups: (i) those who were untreated or were treated only with corticosteroids (CS) at a dose of ≤ 5 mg/day (no adjuvant treatment, NAT); and patients receiving or not receiving CS ≤ 5 mg/day who also received either (ii) azathioprine (AZ), (iii) cyclophosphamide (CY), (iv) mycophenolate mofetil (MM) or (v) rituximab (RTX). RESULTS: In total, 113 patients were recruited. There were no significant differences between the treatment subgroups in either the SF-36 or Skindex-29 results. However, for the GHQ, there were large differences in QOL scores between patients scoring > 4 points (GHQ+) and those scoring < 4 points (GHQ-), especially for the more 'physical' components of QOL. The overall observed proportion of GHQ+ patients was 33.6%. CONCLUSIONS: We found no significant differences in QOL impairment between the treatment subgroups; however, we observed a strong association between psychiatric morbidity and poorer QOL within each of the treatment groups. This should be of concern for dermatologists, as psychiatric morbidity is associated with poor treatment adherence and dissatisfaction with care.

Erbium emission in MOS light emitting devices: from energy transfer to direct impact excitation.

The electroluminescence (EL) at 1.54 µm of metal­oxide­semiconductor (MOS) devices withEr3C ions embedded in the silicon-rich silicon oxide (SRSO) layer has been investigated under different polarization conditions and compared with that of erbium doped SiO2 layers. EL time-resolved measurements allowed us to distinguish between two different excitation mechanisms responsible for the Er3C emission under an alternate pulsed voltage signal (APV). Energy transfer from silicon nanoclusters (Si-ncs) to Er3C is clearly observed at low-field APV excitation. We demonstrate that sequential electron and hole injection at the edges of the pulses creates excited states in Si-ncs which upon recombination transfer their energy to Er3C ions. On the contrary, direct impact excitation of Er3C by hot injected carriers starts at the Fowler­Nordheim injection threshold (above 5 MV cm(-1)) and dominates for high-field APV excitation.

Measuring health-related quality of life in clinical trials in cystic fibrosis.

The inclusion of health-related quality of life (HRQoL) as an outcome measure in cystic fibrosis (CF) clinical trials can supply important patient-reported information not captured by other endpoints. Both an appropriate HRQoL measure and sound methodology are required in order to draw valid inferences about treatments and HRQoL. This paper provides the current consensus of the HRQoL Outcomes Group. Particular consideration has been given to the appropriateness of measurement scales, the rationale for including specific domains as endpoints, the importance of considering baseline ceiling effects and the difficulties of data interpretation. Guidance is provided on HRQoL measurement in National and European CF clinical trials.

Modulation of Toll-like receptors in psoriatic patients during therapy with adalimumab.

Toll-like receptors (TLRs) are a key part of the innate immune system that detect pathogen-associated molecular patterns (PAMPs) of microorganisms and their stimulation results in the activation of signaling pathways leading to the modulation of inflammatory and immune responses. Since psoriasis is a complex, inflammatory and immune skin disease, characterized by an abnormal immune response and increased proliferation of keratinocytes, with an increased production of proinflammatory cytokines, TLRs could play an important role in the pathogenesis of the disease. We propose to assess the modulation of TLR expression on psoriatic skin of patients treated with Adalimumab and systemic conventional therapies. We therefore recruited fifteen patients: ten were treated with adalimumab and five with systemic conventional therapies; their clinical conditions were analyzed by PASI index and skin biopsies were evaluated for TLR1 and TLR2 expression by immunohistochemistry assays. Our data suggest adalimumab is not only able to improve the clinical condition of psoriatic patients, but also to modulate TLR1 and TLR2 expression involved in psoriasis, as in healthy skin. Adalimumab is a most promising biological drug able to orchestrate immune and inflammatory responses in psoriatic lesions, recovering TLR expression on basal keratinocytes and improving clinical conditions of psoriatic patients, with no evident side effects.

Morpheaform extra-ocular sebaceous carcinoma.

BACKGROUND: Sebaceous carcinoma is an unusual, aggressive, malignant tumor derived from the epithelium of sebaceous glands. It may arise in ocular or extra-ocular sites and is often evident as an ulcerated or non-ulcerated cystic nodule measuring up to 8 cm in diameter. A significant number of extra-ocular sebaceous carcinomas have been associated with metastases and high mortality rate. METHODS AND RESULTS: We reviewed the literature in the context of our report of a 68-year-old woman who had a morpheaform plaque of the upper lip of 15 years duration, extending into the left naso-genal groove, which histologically also showed marked stromal fibrosis. Three years after excision, there were no signs of recurrence or metastases. CONCLUSIONS: This patient is unusual in clinical and histologic pattern, as a morpheaform appearance has only been described previously in only one sebaceous carcinoma. In addition, no metastatic disease 15 years after tumor onset is a fortunate and remarkable feature.