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Abstract Soluble di‐iron monooxygenases (SDIMOs) are multi‐component enzymes catalysing the oxidation of various substrates. These enzymes are characterized by high sequence and functional diversity that is still not well understood despite their key role in biotechnological processes including contaminant biodegradation. In this study, we analysed a mutant of Rhodoccocus aetherivorans BCP1 (BCP1‐2.10) characterized by a transposon insertion in the gene smoA encoding the alpha subunit of the plasmid‐located SDIMO SmoABCD. The mutant BCP1‐2.10 showed a reduced capacity to grow on propane, lost the ability to grow on butane, pentane and n‐hexane and was heavily impaired in the capacity to degrade chloroform and trichloroethane. The expression of the additional SDIMO prmABCD in BCP1‐2.10 probably allowed the mutant to partially grow on propane and to degrade it, to some extent, together with the other short‐chain n‐alkanes. The complementation of the mutant, conducted by introducing smoABCD in the genome as a single copy under a constitutive promoter or within a plasmid under a thiostreptone‐inducible promoter, allowed the recovery of the alkanotrophic phenotype as well as the capacity to degrade chlorinated n‐alkanes. The heterologous expression of smoABCD allowed a non‐alkanotrophic Rhodococcus strain to grow on pentane and n‐hexane when the gene cluster was introduced together with the downstream genes encoding alcohol and aldehyde dehydrogenases and a GroEL chaperon. BCP1 smoA gene was shown to belong to the group 6 SDIMOs, which is a rare group of monooxygenases mostly present in Mycobacterium genus and in a few Rhodococcus strains. SmoABCD originally evolved in Mycobacterium and was then acquired by Rhodococcus through horizontal gene transfer events. This work extends the knowledge of the biotechnologically relevant SDIMOs by providing functional and evolutionary insights into a group 6 SDIMO in Rhodococcus and demonstrating its key role in the metabolism of short‐chain alkanes and degradation of chlorinated n‐alkanes.

BackgroundPhthalates are non-persistent chemicals largely used as plasticizers and considered ubiquitous pollutants with endocrine disrupting activity. The exposure during sensible temporal windows as pregnancy and early childhood, may influence physiological neurodevelopment.Aims and ScopeThe aim of this study is to analyze the relationship between the urinary levels of phthalate metabolites in newborn and infants and the global development measured by the Griffiths Scales of Children Development (GSCD) at six months.MethodsLongitudinal cohort study in healthy Italian term newborn and their mothers from birth to the first 6 months of life. Urine samples were collected at respectively 0 (T0), 3 (T3), 6 (T6) months, and around the delivery for mothers. Urine samples were analyzed for a total of 7 major phthalate metabolites of 5 of the most commonly used phthalates. At six months of age a global child development assessment using the third edition of the Griffith Scales of Child Development (GSCD III) was performed in 104 participants.ResultsIn a total of 387 urine samples, the seven metabolites analyzed appeared widespread and were detected in most of the urine samples collected at any time of sampling (66-100%). At six months most of the Developmental Quotients (DQs) falls in average range, except for the subscale B, which presents a DQ median score of 87 (85-95). Adjusted linear regressions between DQs and urinary phthalate metabolite concentrations in mothers at T0 and in infants at T0, T3 and T6 identified several negative associations both for infants’ and mothers especially for DEHP and MBzP. Moreover, once stratified by children’s sex, negative associations were found in boys while positive in girls.ConclusionsPhthalates exposure is widespread, especially for not regulated compounds. Urinary phthalate metabolites were found to be associated to GSCD III scores, showing inverse association with higher phthalate levels related to lower development scores. Our data suggested differences related to the child’s sex.

Abstract Celiac disease (CD) is a complex immune-mediated chronic disease characterized by a consistent inflammation of the gastrointestinal tract induced by gluten intake in genetically predisposed individuals. Although initiated by the interaction between digestion-derived gliadin, a gluten component, peptides, and the intestinal epithelium, the disorder is highly complex and involving other components of the intestine, such as the immune system. Therefore, conventional model systems, mainly based on two- or three-dimension cell cultures and co-cultures, cannot fully recapitulate such a complex disease. The development of mouse models has facilitated the study of different interacting cell types involved in the disorder, together with the impact of environmental factors. However, such in vivo models are often expensive and time consuming. Here we propose an organ ex vivo culture (gut-ex-vivo system) based on small intestines from gluten-sensitive mice cultivated in a dynamic condition, able to fully recapitulate the biochemical and morphological features of the mouse model exposed to gliadin (4 weeks), in 16 h. Indeed, upon gliadin exposure, we observed: i) a down-regulation of cystic fibrosis transmembrane regulator (CFTR) and an up-regulation of transglutaminase 2 (TG2) at both mRNA and protein levels; ii) increased intestinal permeability associated with deregulated tight junction protein expression; iii) induction and production of pro-inflammatory cytokines such as interleukin (IL)-15, IL-17 and interferon gamma (IFNγ); and iv) consistent alteration of intestinal epithelium/villi morphology. Altogether, these data indicate that the proposed model can be efficiently used to study the pathogenesis of CD, test new or repurposed molecules to accelerate the search for new treatments, and to study the impact of the microbiome and derived metabolites, in a time- and cost- effective manner.

Objectives: The study aimed to assess and compare the global development in six-month-old infants before and during the pandemic restrictive social distancing measures.Methods: This cross-sectional nested study involved infants assessed through the Griffiths Scales of Child Development (GSCD) between September 2019 and April 2021. Infants were classified in a pre-COVID or a COVID group, considering the evaluation date and the restrictive measures in place. GSCD subscales and General Development Scores (GDS) were calculated and compared.Results: One hundred and four healthy term-born infants were evaluated. GDS in the COVID group (n:70; median: 94; IQR: 90–100) appeared significantly lower than in the pre-COVID group (n:34; median: 98; IQR: 97–103; p < 0.001). Language and personal-social-emotional subareas scores appeared the most affected. A decreasing trend of GDS along with the severity of restriction was observed.Conclusion: A reduction in infant development scores was observed during pandemic social distancing. Further studies are needed to systematize these findings and to address effective public health policies for infants and families during long-term forced isolation periods.

This study aims to investigate cholesterol metabolism in a mouse model with cystic fibrosis (CF) by the comparison of affected homozygous versus wild type (WT) mice. In particular, we evaluated the effects of a diet enriched with cholesterol in both mice groups in comparison with the normal diet. To this purpose, beyond serum and liver cholesterol, we analyzed serum phytosterols as indirect markers of intestinal absorption of cholesterol, liver lathosterol as indirect marker of de novo cholesterol synthesis, liver cholestanol (a catabolite of bile salts synthesis) and the liver mRNA levels of LDL receptor (LDLR), 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoAR), acyl CoA:cholesterol acyl transferase 2 (ACAT2), cytochrome P450 7A1 (CYP7A1) and tumor necrosis factor alpha (TNFα). CF mice showed lower intestinal absorption and higher liver synthesis of cholesterol than WT mice. In WT mice, the cholesterol supplementation inhibits the synthesis of liver cholesterol and enhances its catabolism, while in CF mice we did not observe a reduction of LDLR and HMG-CoAR expression (probably due to an altered feed-back), causing an increase of intracellular cholesterol. In addition, we observed a further increase (5-fold) in TNFα mRNA levels. This preliminary study suggests that in CF mice there is a vicious circle in which the altered synthesis/secretion of bile salts may reduce the digestion/absorption of cholesterol. As a result, the liver increases the biosynthesis of cholesterol that accumulates in the cells, triggering inflammation and further compromising the metabolism of bile salts.

The global economic success of man-made nanoscale materials has led to a higher production rate and diversification of emission sources in the environment. For these reasons, novel nanosafety approaches to assess the environmental impact of engineered nanomaterials are required. While studying the potential toxicity of metal nanoparticles (NPs), we realized that gold nanoparticles (AuNPs) have a growth-promoting rather than a stress-inducing effect. In this study we established stable short- and long-term exposition systems for testing plant responses to NPs. Exposure of plants to moderate concentrations of AuNPs resulted in enhanced growth of the plants with longer primary roots, more and longer lateral roots and increased rosette diameter, and reduced oxidative stress responses elicited by the immune-stimulatory PAMP flg22. Our data did not reveal any detrimental effects of AuNPs on plants but clearly showed positive effects on growth, presumably by their protective influence on oxidative stress responses. Differential transcriptomics and proteomics analyses revealed that oxidative stress responses are downregulated whereas growth-promoting genes/proteins are upregulated. These omics datasets after AuNP exposure can now be exploited to study the underlying molecular mechanisms of AuNP-induced growth-promotion.

Many components of the innate immune system are evolutionarily conserved and shared across many living organisms, from plants and invertebrates to humans. Therefore, these shared features can allow the comparative study of potentially dangerous substances, such as engineered nanoparticles (NPs). However, differences of methodology and procedure between diverse species and models make comparison of innate immune responses to NPs between organisms difficult in many cases. To this aim, this review provides an overview of suitable methods and assays that can be used to measure NP immune interactions across species in a multidisciplinary approach. The first part of this review describes the main innate immune defense characteristics of the selected models that can be associated to NPs exposure. In the second part, the different modes of exposure to NPs across models (considering isolated cells or whole organisms) and the main endpoints measured are discussed. In this synergistic perspective, we provide an overview of the current state of important cross-disciplinary immunological models to study NP-immune interactions and identify future research needs. As such, this paper could be used as a methodological reference point for future nano-immunosafety studies.

Abstract Background In this study, we used targeted next-generation sequencing (NGS) to investigate the genetic basis of congenital hypothyroidism (CH) in a 19-year-old Tunisian man who presented with severe hypothyroidism and goiter. Case presentation The propositus reported the appearance of goiter when he was 18. Importantly, he did not show signs of mental retardation, and his growth was proportionate. A partial organification defect was detected through the perchlorate-induced iodide discharge test. NGS identified a novel homozygous mutation in exon 18 of the SLC26A7 gene (P628Qfs*11), which encodes for a new iodide transporter. This variant is predicted to result in a truncated protein. Notably, the patient's euthyroid brother was heterozygous for the same mutation. No renal acid–base abnormalities were found and the administration of 1 mg of iodine failed to correct hypothyroidism. Conclusions We described the first case of goitrous CH due to a homozygous mutation of the SLC26A7 gene diagnosed during late adolescence.

IntroductionThe single nucleotide polymorphism (SNP) rs4644 at codon 64 of galectin-3 (gal-3, gene name: LGALS3), specifying the variant proline (P64) to histidine (H64), is known to affect the protein’s functions and has been associated with the risk of several types of cancer, including differentiated thyroid carcinoma (DTC).Materials and methodsTo deepen our understanding of the biological effects of this SNP, we analyzed the proteome of two isogenic cell lines (NC-P64 vs. NA-H64) derived from the immortalized non-malignant thyrocyte cell line Nthy-Ori, generated through the CRISPR-Cas9 technique to differ by rs4644 genotype. We compared the proteome of these cells to detect differentially expressed proteins and studied their proteome in relation to their transcriptome.ResultsFirstly, we found, consistently with previous studies, that gal-3-H64 could be detected as a monomer, homodimer, and heterodimer composed of one cleaved and one uncleaved monomer, whereas gal-3-P64 could be found only as a monomer or uncleaved homodimer. Moreover, results indicate that rs4644 influences the expression of several proteins, predominantly upregulated in NA-H64 cells. Overall, the differential protein expression could be attributed to the altered mRNA expression, suggesting that rs4644 shapes the function of gal-3 as a transcriptional co-regulator. However, this SNP also appeared to affect post-transcriptional regulatory mechanisms for proteins whose expression was oppositely regulated compared to mRNA expression. It is conceivable that the rs4644-dependent activities of gal-3 could be ascribed to the different modalities of self-dimerization.ConclusionOur study provided further evidence that rs4644 could affect the gal-3 functions through several routes, which could be at the base of differential susceptibility to diseases, as reported in case-control association studies.

Abstract Background Three-dimensional (3D) printing is a novel innovation in the field of craniomaxillofacial surgery, however, a lack of evidence exists related to the comparison of the trueness of skull models fabricated using different technology-based printers belonging to different cost segments. Methods A study was performed to investigate the trueness of cone-beam computed tomography-derived skull models fabricated using different technology based on low-, medium-, and high-cost 3D printers. Following the segmentation of a patient’s skull, the model was printed by: (i) a low-cost fused filament fabrication printer; (ii) a medium-cost stereolithography printer; and (iii) a high-cost material jetting printer. The fabricated models were later scanned by industrial computed tomography and superimposed onto the original reference virtual model by applying surface-based registration. A part comparison color-coded analysis was conducted for assessing the difference between the reference and scanned models. A one-way analysis of variance (ANOVA) with Bonferroni correction was applied for statistical analysis. Results The model printed with the low-cost fused filament fabrication printer showed the highest mean absolute error ( $$1.33\pm 0.24 \text{mm}$$ 1.33 ± 0.24 mm ), whereas both medium-cost stereolithography-based and the high-cost material jetting models had an overall similar dimensional error of $$0.07\pm 0.03 \text{mm}$$ 0.07 ± 0.03 mm and $$0.07\pm 0.01 \text{mm}$$ 0.07 ± 0.01 mm , respectively. Overall, the models printed with medium- and high-cost printers showed a significantly ( $$p

This article presents the effects of thermal annealing at elevated temperatures (> glass transition temperature Tg) on the performance of polymer parts via material extrusion 3D printing. Both semi-crystalline and amorphous filaments were used. As-printed parts were designed to be amorphous and then annealed at 60, 110 and 150 °C for different durations ranging from 50 to 6400 s. The flexural strength and Young’s modulus increased by a maximum of approximately 10%. The increase was ascribed to crystallisation development during annealing, as confirmed by thermal and morphology characterisations. Hence, this effect was only observed with semi-crystalline materials. On the other hand, all the annealed parts expanded in the thickness direction and shrank in the perpendicular plane. The maximum linear strain reached 20%, while the volume strain was negligible. These morphology changes after annealing reversed the strain-hardening of the strand and led to inferior strand performance against local tensile deformation. The degradation can outweigh the benefits of crystallinity development. For amorphous parts, the degradation reached approximately 25% in both the flexural strength and the modulus.

Abstract Background Premature ovarian insufficiency (POI) is an ovarian defect characterized by primary or secondary amenorrhea, hypergonadotropism and hypoestrogenism which occurs before the age of 40 years with a major genetic component. In this study we performed clinical evaluation and genetic analysis of a group of 18 patients with POI. The study involved 18 consecutive women with POI. Karyotiping and genetic analysis for research of mutations in GDF9 (Growth Differentation Factor 9) and BMP15 (Bone morphogentic protein 15) genes and FMR1 (Fragile X Mental Retardation 1) premutation were carried out. In vitro functional study of the novel BMP15 mutation was performed using COV434 (Human ovarian granulosa tumour cells 434) cells of ovarian granulosa, which consistently express BMP responsive element, and luciferase reporter assay. Results Three patients (17%) had a family history of POI. Ten patients (56%) had a family history of autoimmune diseases and nine patients (50%) showed a personal history of one or more autoimmune diseases. Of patients for whom morphological assessment was available, almost half (44%) had poor follicle assets or small ovaries’s size at pelvic US. Two patients (13%) showed reduced bone density at DEXA (Dual Energy X-ray Absorptiometry). All the women had normal female kariotype and no mutations in the GDF-9 gene or FMR1 premutations were found. A novel heterozygous mutation c.406G > C (V136L) of BMP15 gene was identified in one patient. After transfection in COV434 cells, BMP15 variant showed a significantly reduced luciferase activity compared to wild type. Conclusions POI is a multifactorial disease with several health implications. Autoimmunity and genetics represent the most common aetiology. We identified and characterized a novel BMP15 mutation, providing an additional elucidation of molecular basis of this complex disorder.

Among conductive polymers, poly(3,4 ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) has been widely used as an electrode material for supercapacitors, solar cells, sensors, etc. Although PEDOT:PSS-based thin films have acceptable properties such as good capacitive and electrical behaviour and biocompatibility, there are still several challenges to be overcome in their use as an electrode material for supercapacitors. For this reason, the aim of this work is to fabricate and characterise ternary nanocomposites based on PEDOT:PSS and graphene oxide (GO), blended with green additives (glucose (G) or ascorbic acid (AA)), which have the benefits of being environmentally friendly, economical, and easy to use. The GO reduction process was first accurately investigated and demonstrated by UV-Vis and XRD measurements. Three-component inks have been developed, and their morphological, rheological, and surface tension properties were evaluated, showing their printability by means of Aerosol Jet® Printing (AJ®P), an innovative direct writing technique belonging to the Additive Manufacturing (AM) for printed electronics applications. Thin films of the ternary nanocomposites were produced by drop casting and spin coating techniques, and their capacitive behaviour and chemical structures were evaluated through Cyclic Voltammetry (CV) tests and FT-IR analyses. CV tests show an increment in the specific capacitance of AAGO-PEDOT up to 31.4 F/g and excellent overtime stability compared with pristine PEDOT:PSS, suggesting that this ink can be used to fabricate supercapacitors in printed (bio)-electronics. The inks were finally printed by AJ®P as thin films (10 layers, 8 × 8 mm) and chemically analysed by FT-IR, demonstrating that all components of the formulation were successfully aerosolised and deposited on the substrate.

Heated build plates are common yet essential components in many additive manufacturing (AM) machines. However, their influence on the air temperature above and the subsequent impact on the AM process are barely discussed. This article proposes a simple model for the air temperature above the plate Ta(z), which exponentially decays from the plate temperature to the far environment temperature by a characteristic dimension lc in the range of 2–9 mm. The model has been empirically validated for three fused filament fabrication (FFF) machines, including open and closed systems, small and large build chambers. Temperature simulations in printed parts suggest that Ta(z) plays a critical role when printing at low layer thicknesses, such as 300 µm. Besides, adding an enclosure to FFF machines can significantly raise the far environment temperature inside the chamber, delaying the part cooling. However, its effect is noticeable for the portion above 10 mm only.

Collagen has grown increasingly present in bioprinting, however collagen bioprinting has mostly been limited to the extrusion printing of collagen type I to form weak collagen hydrogels. While these weak collagen hydrogels have their applications, synthetic polymers are often required to reinforce gel-laden constructs that aim to replicate dense collagenous tissues found in vivo. In this study, aerosol jet printing (AJP) was used to print and process collagen type I and II into dense constructs with a greater capacity to replicate the dense collagenous ECM found in connective tissues. Collagen type I and II was isolated from animal tissues to form solutions for printing. Collagen type I and II constructs were printed with 576 layers and measured to have average effective elastic moduli of 241.3 ± 94.3 and 196.6 ± 86.0 kPa (±SD), respectively, without any chemical modification. Collagen type II solutions were measured to be less viscous than type I and both collagen type I and II exhibited a drop in viscosity due to AJP. Circular dichroism and SDS-PAGE showed collagen type I to be more vulnerable to structural changes due to the stresses of the aerosol formation step of aerosol jet printing while the collagen type II triple helix was largely unaffected. SEM illustrated that distinct layers remained in the aerosol jet print constructs. The results show that aerosol jet printing should be considered an effective way to process collagen type I and II into stiff dense constructs with suitable mechanical properties for the replication of dense collagenous connective tissues.

Pressure sensing is not a new concept and can be applied by using different transduction mechanisms and manufacturing techniques, including printed electronics approaches. However, very limited efforts have been taken to realise pressure sensors fully using additive manufacturing techniques, especially for personalised guide prosthetics in biomedical applications. In this work, we present a novel, fully printed piezoresistive pressure sensor, which was realised by using Aerosol Jet® Printing (AJP) and Screen Printing. AJ®P was specifically chosen to print silver interconnects on a selective laser sintered (SLS) polyamide board as a customised substrate, while piezoresistive electrodes were manually screen-printed on the top of the interconnects as the sensing layer. The sensor was electromechanically tested, and its response was registered upon the application of given signals, in terms of sensitivity, hysteresis, reproducibility, and time drift. When applying a ramping pressure, the sensor showed two different sensitive regions: (i) a highly sensitive region in the range of 0 to 0.12 MPa with an average sensitivity of 106 Ω/MPa and a low sensitive zone within 0.12 to 1.25 MPa with an average sensitivity of 7.6 Ω/MPa with some indeterminate overlapping regions. Hysteresis was negligible and an electrical resistance deviation of about 14% was observed in time drift experiments. Such performances will satisfy the demands of our application in the biomedical field as a smart prosthetics guide.

Research in bioprinting is booming due to its potential in addressing several manufacturing challenges in regenerative medicine. However, there are still many hurdles to overcome to guarantee cell survival and good printability. For the 3D extrusion-based bioprinting, cell viability is amongst one of the lowest of all the bioprinting techniques and is strongly influenced by various factors including the shear stress in the print nozzle. The goal of this study is to quantify, by means of in silico modeling, the mechanical environment experienced by the bioink during the printing process. Two ubiquitous nozzle shapes, conical and blunted, were considered, as well as three common hydrogels with material properties spanning from almost Newtonian to highly shear-thinning materials following the power-law behavior: Alginate-Gelatin, Alginate and PF127. Comprehensive in silico testing of all combinations of nozzle geometry variations and hydrogels was achieved by combining a design of experiments approach (DoE) with a computational fluid dynamics (CFD) of the printing process, analyzed through a machine learning approach named Gaussian Process. Available experimental results were used to validate the CFD model and justify the use of shear stress as a surrogate for cell survival in this study. The lower and middle nozzle radius, lower nozzle length and the material properties, alone and combined, were identified as the major influencing factors affecting shear stress, and therefore cell viability, during printing. These results were successfully compared with those of reported experiments testing viability for different nozzle geometry parameters under constant flow rate or constant pressure. The in silico 3D bioprinting platform developed in this study offers the potential to assist and accelerate further development of 3D bioprinting.

Soy food health effects are due to isoflavone (ISF) components functioning as phytoestrogens similar to natural estrogen. Epidemiological data suggest a role of estrogens in pathogenesis of thyroid diseases explaining their prevalence in women.Aim of this study was to determine in vitro effects of isoflavones, soybean extract and Cimicifuga extract on iodide uptake, a crucial step in thyroid hormone biosynthesis, by using a differentiated TSH-dependent cell line (FRTL5).Estradiol didn’t influence iodide uptake and sodium-iodide symporter (NIS) mRNA expression in FRTL-5 cells. None of analyzed glycoside or aglycone isoflavone determined a significant inhibition of iodide uptake, while both isoflavone rich and triterpene rich extracts determined a significant reduction of iodide uptake at the higher doses.While soybean and Cimicifuga extracts inhibited iodide uptake at high doses, none of analyzed isoflavone exerted this effect. These elevated doses, however, are never reached in the blood following intake of food supplements.

The fused filament fabrication (FFF) process is characterized by fast local temperature decrease. Such characteristic usually leads to lower crystallinity degree in final printed parts when semi-crystalline polymers are used. By melt-compounding poly(lactic acid) (PLA) with its nucleating agent Talc, filaments of PLA/Talc composite were produced and used in FFF. Through investigations in static/dynamic mechanical and thermal properties, and morphology, the influence of Talc on the PLA based composite in FFF are revealed: after melt compounding with Talc, PLA matrix can crystallize considerably during fast cooling in FFF; as a consequence, final parts exhibited higher degree of crystallinity than the case of neat PLA; furthermore, melt crystallization process releases heat, promoting coalescence between adjacent layers, and ultimately contributes to higher parts stiffness and better mechanical integrity. This paper presents a first example in which in-situ melt crystallization is recovered for semi-crystalline polymers during FFF via composition modification. With this example, an increase in crystallinity from 3.6% to 12% can lead to 15% increase in flexural modulus for FFF printed parts. Research results also provide insights in material property reinforcement, heat treatment on finished parts, and explorations in other semi-crystalline materials in FFF. Keywords: Crystallinity, Fused filament fabrication (FFF), Melt crystallization, PLA, Talc

In this study, an original and green procedure to produce water-based solutions containing nanometric recycled carbon particles is proposed. The nanometric particles are obtained starting from carbon waste ashes, produced by the wooden biomass pyro-gasification plant CMD (Costruzioni motori diesel) ECO20. The latter is an integrated system combining a downdraft gasifier, a spark-ignition internal combustion engine, an electric generator and syngas cleaning devices, and it can produce electric and thermal power up to 20 kWe and 40 kWth. The carbon-based ashes (CA) produced by the CMD ECO20 plant were, first, characterized by using differential scanning calorimetry (DSC) and microcomputed tomography (microCT). Afterward, they were reduced in powder by using a milling mortar and analyzed by scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) spectrometry, thermogravimetric analysis (TGA), X-ray diffraction (WAXD) and Fourier-transform infrared (FTIR) spectroscopy. The optimization of an original procedure to reduce the dimensions of the ashes in an aqueous solution was then developed by using ball milling and sonication techniques, and the nanometric dimensions of the particles dispersed in water were estimated by dynamic light scattering (DLS) measurements in the order of 300 nm. Finally, possible industrial applications for the nanomaterials obtained from the waste ashes are suggested, including, for example, inks for Aerosol Jet® Printing (AJ® P).

ZrO2 ceramics are currently used in a broad range of industrial applications. However, the machining of post-sintered ZrO2 ceramic is a difficult task, due to its high hardness and brittleness. In this study, micro-milling of ZrO2 with two kinds of diamond-coated end mills has been conducted on a Kern MMP 2522 micro-milling center (Kern Microtechnik GmbH, Eschenlohe, Germany). To achieve a ductile mode machining of ZrO2, the feed per tooth and depth of cut was set in the range of a few micrometers. Cutting force and machined surface roughness have been measured by a Kistler MiniDynamometer (Kistler Group, Winterthur, Switzerland) and a Talysurf 120 L profilometer (Taylor Hobson Ltd., Leicester, UK), respectively. Machined surface topography and tool wear have been examined under SEM. Experiment results show that the material can be removed in ductile mode, and mirror quality surface with Ra low as 0.02 μm can be achieved. Curled and smooth chips have been collected and observed. The axial cutting force Fz is always bigger than Fx and Fy, and presents a rising trend with increasing of milling length. Tool wear includes delamination of diamond coating and wear of tungsten carbide substrate. Without the protection of diamond coating, the tungsten carbide substrate was worn out quickly, resulting a change of tool tip geometry.