During nuclear maturation of most eukaryotic pre-messenger RNAs and long non-coding RNAs, introns are removed through the process of RNA splicing. Different classes of introns are excised by the U2-type or the U12-type spliceosomes, large complexes of small nuclear ribonucleoprotein particles and associated proteins. We created intronIC, a program for assigning intron class to all introns in a given genome, and used it on 24 eukaryotic genomes to create the Intron Annotation and Orthology Database (IAOD). We then used the data in the IAOD to revisit several hypotheses concerning the evolution of the two classes of spliceosomal introns, finding support for the class conversion model explaining the low abundance of U12-type introns in modern genomes. The process of RNA splicing is a necessary step in the maturation of nearly all eukaryotic pre-messenger RNAs and many long non-coding RNAs. During this process, introns are excised from primary RNA transcripts, and the flanking exonic sequences are joined together to form functional, mature messenger RNAs ,2. In moThe large-scale and well-organized online databases of genomic data, like Ensembl , UCSC 99 and RefMany features of eukaryotic introns have been examined for clues about their evolutionary history. Introns can be assigned to one of three phases based on their position relative to the codons of the flanking exonic sequence: phase 0 introns fall directly between two codons, phase 1 introns fall between the first and second nucleotides of a single codon, and phase 2 introns fall between the second and third nucleotides of a single codon. It has long been noted that introns are not evenly distributed between the three phases ,28. In cIn an effort to address some of the many open questions about intron evolution, we created the Intron Annotation and Orthology Database (IAOD), a database of intron information for all annotated introns in 24 genomes, including plant, fungal, mammalian, and insect genomes. It also uniquely annotates orthologous introns, and assigns intron class using the intronIC algorithm described herein . The website is publicly accessible at introndb.lerner.ccf.org.To begin, intronIC identifies all intron sequences in an annotation file by interpolating between coding features (CDS or exon) within the longest isoform of each annotated gene. For each intron, sequences corresponding to the 5′ splice site  and branch point sequence (BPS) region (from −55 to −5 relative to the last base of the intron) are scored using a set of position weight matrices (PWMs) representing canonical sequence motifs for both U2-type and U12-type human introns. A small ‘pseudo-count’ frequency value of 0.001 is added to all matrix positions to avoid zero division errors while still providing a significant penalty for low-frequency bases. For all scored motifs, the binary logarithm of the U12/U2 score ratio (the log ratio) is calculated, resulting in negative scores for introns with U2-like motifs, and positive scores for introns with U12-like motifs. Because U2 introns are not known to contain an extended BPS motif, the PWM for the U2-type BPS is derived empirically using the best-scoring U12 BPS motifs from all introns in the final dataset whose 5′SS U12 scores are below the 95th percentile. To identify the most likely BPS for each intron, all 12-mer sequences within the BPS region are scored and the one with the highest U12 log ratio score is chosen. This initial scoring procedure follows the same general approach used by a variety of different groups for bioinformatic identification of U12-type introns ,33–36.et al., the 5′SS and BPS scores together are sufficient to produce good binary clustering of introns into putative types, due to strong correspondence between the 5′SS and BPS scores in U12-type introns  and negative (U2-type) training sequences for the SVM  and U12-type (GT-AG/AT-AC) introns, and sets of high-confidence U2- and U12-type intron sequences with which to train the SVM classifier Figure . Due to ata from  to incluM Figure . In ordeM Figure ).C, which is (roughly) the degree to which misclassification of data in the training set is penalized during the creation of an optimized (i.e. wide) margin separating the positive and negative classes. To our knowledge there is no single, standard approach for establishing the best value of C; we chose to optimize C using an iterative cross-validation method which starts with a wide range of logarithmically-distributed values and narrows that range based upon the best-performing  value of C in each validation round. After several such iterations, the mean of the resulting range is taken as the final value of C to be used to train the classifier. Balanced accuracy is used as a performance metric due to the highly imbalanced nature of the training data, where the negative class (U2-type) greatly outnumbers the positive class (U12-type). Because the human training data is very well-separated, when applied to intron sequences in the human genome values of C ≥10 perform equally well during cross-validation  than taking the default ‘best’ parameter value via the scikit-learn API, which simply returns the first rank-1 parameter value found. For the human genome, this approach results in a classifier which performs perfectly on the training sets, with both F1 and precision-recall AUC scores of 1.0 on held-out training data  and the modified version used for this application is available at (https://github.com/Devlin-Moyer/IAOD).For the purpose of populating the IAOD, intronIC was slightly modified to produce a single output file containing all of the annotation information recorded in the IAOD for each intron—the default version is available for download from , release 39 of Ensembl Metazoa, or release 40 of Ensembl Plants , and al.1; ref. ). LastlyA custom Python script created a PostgreSQL database using the output of intronIC, the lists of gene symbols from BioMart, and the list of orthologous groups of introns. All of the orthologous groups were inserted in a table with two columns: a unique numeric ID for each group and a list of all intron labels belonging to that group. One table for each genome contains, for each intron: the abbreviated sequence (see above), taxonomic and common names of the organism, name of the genome assembly, intronIC score, intron class (determined from the intronIC score), intronIC label, chromosome, start coordinate, stop coordinate, length, strand, rank in transcript, phase, terminal dinucleotides, upstream exonic sequence (50 nt), 3′ terminus with the branch point region enclosed with brackets (40 nt), downstream exonic sequence (50 nt), full intron sequence, Ensembl gene ID, Ensembl transcript ID, and gene symbol. Another table with identical fields contains all U12-type introns from all genomes., an open-source Python web development framework, and Bootstrap 4.0.0, an open-source framework for front-end web development. The search engines use the Django ORM to interact with the PostgreSQL database.The website was constructed using Django 2.0There are four search engines on the website: the main, advanced, U12, and orthologous searches. The main and advanced search interfaces have input fields corresponding to individual columns in the database, so the text input in each field can easily be matched with the appropriate column using the Django ORM. The U12 search engine uses PostgreSQL search vectors to allow users to make full text queries against the database. I.e. users can input a string containing one term corresponding to as many fields as they like and get a result. However, if the search query contains, e.g., the names of two different species or genes, no results will be returned, since no single record (intron) in the database corresponds to multiple species or genes. This limits the number of possible queries, but allows for a simple user interface for simple queries concerning U12-type introns. Since the main and advanced search engines require users to specify which field of the database each term of their query corresponds to, it does not need to use full text search vectors, and can consequently accept multiple search terms for each field. The homolog search engine also makes use of PostgreSQL search vectors to find the row of the homolog table containing the intron ID input by the user.p = 1 − (1 − x)m−1, where x is the proportion of U12-type introns in the genome and m is the average number of introns in the genome. Data from the IAOD was used to obtain n, x, and m for each genome in the database that contained at least one U12-type intron. The dbinom function in R was used to compute the probability of observing the observed number of genes with multiple U12-type introns in each genome. If U12-type introns were randomly inserted a genome, we would expect the distribution of U12-type introns per gene to be binomial with parameters n = number of genes with at least one U12-type intron and To ensure that the observed clustering of U12-type introns in the same genes was not an artefact of U12-type introns with alternative splice sites being recorded as distinct U12-type introns, all intron coordinates listed by intronIC were used to create a graph where each node corresponded to a position within each genome (e.g. GRCh38 + chr1 + 492045 corresponds to base pair 492045 on chromosome 1 in assembly GRCh38 of the human genome) and two nodes are joined with an edge if they appear in the same row of the list of intron coordinates. In this graph, alternatively spliced introns are evident as clusters of >2 nodes, so each cluster represents a single intron, regardless of how many alternative splice sites it possesses. A single edge from each cluster was selected and the corresponding coordinates were matched to the original intronIC output to get accurate counts of the total number of unique introns in each class in all genomes annotated in the IAOD.Homo sapiens using a machine-learning (SVM) classifier to assign each intron a probability of being U12-type  in human. In Arabidopsis thaliana, our method matches the calls in the U12DB 94% of the time (223/238), with similar results  for U12-type introns from the plant-specific database ERISdb  our method categorized only 1/116241 introns as U12-type, suggesting a false-positive rate of less than 0.001%. A total of 8967 U12-type introns were identified using this technique. Groups of analyzed introns in conserved regions of homologous genes were also annotated; collectively, these data constitute the Intron Annotation and Orthology Database (IAOD).We developed the method intronIC , implemented in Python, and used it to perform genome-wide identification of U2- and U12-type spliceosomal introns in 24 eukaryotic species including 14 vertebrate animals, 5 invertebrate animals, 4 plants and two yeasts. For each species, type-specific position-weight matrices (PWMs) for the 5′ splice site and branch point sequences were used to create score vectors for every intron in each genome. These score vectors were then compared against corresponding vectors in high-confidence training sets from Homo sapiens, D. melanogaster, A. thaliana and Ciona intestinalis to the whole genomes of every other organism in the database  replicated their findings in all 21 genomes  are in conserved positions, while out of the 9527 U2-type introns in such regions of the human genome, 2098 (22%) are in conserved positions in A. thaliana. Thus, our comparative analysis is consistent with U12-type intron enrichment in the ancestral eukaryotic genome relative to the most U12-intron-rich extant lineages.Basu et al.  argued tGallus gallus and Tetraodon nigroviridis (Table The majority of introns annotated in the IAOD in both classes begin with GT and end with AG Table , in agreis Table . Inspectis Table  largely is Table ,36,63.Interestingly, We have created a database of intron annotation and homology information and used it to investigate several evolutionary hypotheses regarding the two classes of spliceosomal introns in eukaryotes. We have also created a web-based interface for querying this database to facilitate further investigations. The relationships between intron class, phase, terminal dinucleotides and -1 nucleotides at the 5′ splice site and the nonrandom distribution of U12-type introns annotated in the IAOD do not support many previous models that explain these patterns ,30,50,51introndb.lerner.ccf.org and all code used to create the database and run the website is available at the following GitHub repository https://github.com/Devlin-Moyer/IAOD. The standalone intronIC algorithm is available at https://github.com/glarue/intronIC.The IAOD is publically accessible at gkaa464_Supplemental_FileClick here for additional data file.


These CISe NHJs exhibit surprising non-linear emissions, quantized transitions, large carrier mobility, low trap-state-density, long carrier lifetime and possible up-conversion. They facilitate efficient separation of minority carriers, reduce recombination and essentially function like quantum materials. This approach mitigates the material issues and complex fabrication of incumbent nanoscale heterojunctions; it also overcomes the flexibility and scale-up challenges of conventional planar pn junctions. The self-stabilized CISe NHJ film can be roll-to-roll processed in ambient atmosphere, thus providing a promising platform for a range of optoelectronic technologies. This concept exemplified by CISe compounds can be adapted to create nano-scale pn junctions with other inorganic semiconductors.Pairing semiconductors with electrochemical processing offers an untapped opportunity to create novel nanostructures for practical devices. Here we report the results of one such pairing: the in-situ formation of highly-doped, interface-matched, sharp nanocrystalline homojunctions (NHJs) with single step electrodeposition of two copper-indium-selenide (CISe) compounds on flexible foil. It produces a homogenous film, comprising inherently ordered, 3-dimensional interconnected network of  High performance planar pn junctions often require complex, expensive processing techniques with limited scalability. Fabricating materials systems with low-cost processes that intrinsically host nano-scale pn junctions, such as bulk heterojunctions (BHJs) offers an attractive alternative platform for optoelectronic technologies. Solar cell efficiencies for perovskite quantum dot, organic BHJ and polycrystalline perovskite devices have reached 18–25% range1. But significant challenges remain to improve their stability, toxicity, upscaling and reproducibility6. Here we show how pairing certain inorganic semiconductors, such as copper indium selenide (CISe) compounds with electrochemical processing produces high quality nanostructures that could potentially overcome such issues to enable realistic electro-optic device applications.Semiconductor 2-alloys for over 20% efficiency. The CISe ODCs offer further choice of direct bandgaps (Eg) in the 1.04–1.34 eV range and intrinsic doping with shallow donor and acceptor defects that imparts n- or p-type conductivity9. The intrinsic doping of CISe ODCs with shallow donor  and acceptor (VCu) defects has been theoretically established8. The defects impart n-type or p-type conductivity to the CISe nano grains. These attributes remain largely unexploited for device fabrication. For instance, the dual-type conductivity of CISe-ODCs could realize distinct nano-scale pn junctions10. Additionally, the CISe-ODC compositions are self-stabilized and thus allow single step electrodeposition (SSE) of stoichiometric compounds from a single aqueous electrolyte, which greatly simplifies upscaling to roll-to-roll manufacturing in ambient atmosphere12. This work exploits both ODC attributes to form nanocrystalline homojunction (NHJ) films that could overcome the above issues of the state-of-the-art BHJs and similar nanostructure.The CISe family of ordered defect chalcopyrites (ODC) compounds features a unique combination of desirable functional properties, including: high optical-absorption, high defect tolerance, low toxicity, exceptional longterm stability and established performance of CuInSe2, CuIn3Se5, or mixed CuInSe2-CuIn3Se5 compositions on stainless steel (SS) foil13. A brief low temperature air anneal stabilizes the intrinsic defect distribution and crystallizes the CISe into 4–35 nm grains. The nanocrystalline CISe films comprise strongly bound, spatially resolved, interlinked, space-filling grains, with excellent adhesion10. Initial electro-optical studies indicated that the SSE-made CuInSe2-CuIn3Se5 mixture offered surprising potential for efficient and stable NHJ devices. It revealed unexpected optoelectronic attributes related to the morphology, conductivity and composition of SSE-made CISe films. For instance, electrochemical electroreflectance (EER) spectra for the CISe film indicated high structural order and well crystallized chalcopyrite structured grains of CuInSe2 and CuIn3Se5; and CuInSe2-CuIn3Se5 (≡ CuIn2Se3.5) complexes10. An unexpected trend was discerned, showing lower photocurrent transients for films with CuIn2Se3.5 composition than for the films with excess non-complexed CuInSe2 or CuIn3Se513. These first results highlight the unique and intriguing aspects of specific CISe compositions. They warrant in-depth probing into their electro-optical properties and their relationship to film composition and morphology to fill in the knowledge gaps. A profound understanding will enable exploiting the attributes of the SSE-made CISe films for efficient and easily manufactured pn NHJ devices. Here we combine photoluminescence (PL), time-resolved microwave conductivity (TRMC), capacitance/impedance (C/I) and atomic force microscopy (AFM) methods to uncover more remarkable aspects, concerning the local morphology and carrier dynamics for the CISe NHJ films.Unravelling the reaction mechanism for CISe formation enabled the SSE of stoichiometric CuInSe2+/In3+/Se4+ solution at a fixed potential, Fig. 2 strips with graded composition for the (T) top, (M) middle and (B) bottom sections along the strip length, Fig. 2.SSE produces CISe ODC films from a CuT, M and B sections, CISe films S1 and S2, Table 3Se5 at 4.8°K (inset), which includes emissions from both CuInSe2 and CuIn3Se514, but without the superimposed fine structure that is evident for the new sets of spectra. The 830 nm filter blocks emissions from energies > 1.4 eV and selectively transmits the PL from the shallow states near the edge of Eg for CuIn3Se5, Fig. 2 (0.95 eV), large peak (1.3 eV) for higher order ODCs and a smaller peak (1.73 eV). The 3 sections of film S1 essentially show the same PL intensity and spectral features in both sets of spectra, which likely reflects the nonexistence of significant composition gradient for film S1.PL was measured with a 532 nm laser for the T, M, B sections but with varying compositions for film S2, Table T), S2(M), S2(B), sections while the spectral features remain the same. With the 830 nm filter, S2(T) and S2(M), having Cu:In:Se ratio closer to CuInSe2 show similar PL intensity, while the In-rich S2(B) with composition closer to CuIn2Se3.5 exhibits lower PL response. With the 540 nm filter, S2(T) still shows higher intensity but the peak PL intensity of the S2(M) slightly lower than for S2(B). The presence of varied bulk content of these different stoichiometric ratios is consistent with a respectively weighted intensity of those two regions of PL. The trend for reversal of PL peak intensity could have resulted from local inhomogeneity wherein the complexed and non-complexed [CuIn3Se5] or [CuInSe2] grains respond somewhat differently to the light transmitted by 540 and 830 filters. The intensity variation trend for S2(B) or S2(M) with different filters may not be significant, the overall PL emission intensity could be associated with the extent of the [CuInSe2–CuIn3Se5] complexation in the film as previously observed with other techniques13. The previously trend of lower photocurrent transients was seen for CISe films comprising CuInSe2-CuIn3Se5 complex (≡ CuIn2Se3.5). Conversely, films comprising excess [CuIn3Se5] or [CuInSe2] show higher PL intensity (and faster charge transport) than the films comprising [CuIn3Se5] ≈ [CuInSe2].Figure g fine structure for the 3 regions of film S2 appear identical despite significant composition variation that merely reduces the PL emission intensity from S2(T) to S2(B), Fig. 2 PL peak; enhanced PL for CuIn3Se5; similar PL feature at 1.4 eV; much larger PL at 1.73 eV; 3.5 times higher PL intensity; and more intricate fine structure. The fine structure likely arises from specific sub-Eg emissions, originating from shallow native defect states near the conduction band (CB) or valence band (VB) edge of the nanostructured S1 and S2 films. They are located near the Egs of CuInSe2, CuIn3Se5 and CuIn5Se8 and an unidentified feature at 1.73 eV. The predominantly near-Eg emissions correspond to free-carrier-to-shallow defect transitions and band-to-band transitions from specific states. For example, the transition peaks at: 1.20 eV ≡ shallow donor state in CuIn3Se5; 1.26 eV ≡ CB; 1.31 eV and 1.37 eV ≡ energy levels inside CB or ≡ shallow donor states from CuIn5Se8. Such shallow defect transitions enable efficient charge separation within the p-CuInSe2/n-CuIn3Se5 complex, as evidenced by the EER data10.Surprisingly, the overall PL spectral features and the superimposed sub-E2 Eg, such as: 1.00 eV ≡ (CB → VCu) or ≡ (InCu → VB); 0.93 eV ≡ (InCu → VCu); 0.88 eV ≡ (VSe → VCu) or (InCu → VB). The PL emissions from 1.2 − 1.5 eV occur near the Egs  of CuIn3Se5 and CuIn5Se8. Interestingly, the CLS fitting shows that the superimposed fine structure is typically spaced at 30 mV. It is intriguing that the composition variation does not change the fine structure.The PL spectrum in Fig. g emission peak, centered around 1.73 eV is seen for both samples S1 and S2, but is more pronounced for film S2. It appears to encompass the Egs of In2Se3 (1.8 eV) and Se (1.7 eV). The CISe NHJ band structure could contain defect states associated with these compounds, which were incidentally not discerned by other methods13. Alternatively, the blue shift may ensue from grain size effects. Intra-grain and band-to-band transitions were detected by EER for the p-CuInSe2/n-CuIn3Se5 complex10. The peak also shows ~ 30 mV spaced fine structure, e.g. 1.69, 1.73, 1.76, 1.79 eV. The apparent quantization of states likely results in quantum confined nano-regions in the CISe film that possibly contribute to this large PL emission. The emission of multiple and narrow emission lines is a hallmark of photon upconversion in nanoparticles. It entails stepwise low energy photon absorption, energy transfer and the emission of high energy photons, i.e., quantized electronic transistions can transmute sub-Eg photons into higher energy (above-Eg) photons. Such upconversion phenomena are often observed in core–shell nanoparticles and transition metals18. In the present scenario, it apparently results from the near quantum dot sized CISe nanocrystals that fortuitously lead to the above-Eg quantized states. Significantly this approach allows quantum confinement without introducing external rare earth dopants into the host material as in state-of-the-art upconversion18. Linking this self induced nonlinear optics phenomenon to the CISe NHJ electronic structure could reveal a new avenue for electro-optical applications.The PL data implies that the optical and electronic properties of the CISe film are intimately related via a quantum system, comprising quantized energy levels. The unexpected broadening in PL spectral range resulting from the above-E2 film, S3. The In-rich sections, S1(B) and S2(B) were chosen for these measurements, Table 20. The CISe films: S1(B), S2(B), and S3, were delaminated from the SS foils and transfered to a low-dielectric loss quartz substrate using a double-sided tape transfer method. The samples were excited by 10 Hz 500 nm laser pulses during continuous dry N2 purge.Transient photoconductivity analysis was carried out using TRMC for same films as PL, S1 and S2 along with a third stoichiometric CuInSeB), S2(B), and S3. The TRMC figure of merit is given by φΣμ, corresponding to the peak product of free-carrier generation yield (φ) and sum of free electron and hole mobilities (Σμ). The φΣμ was quite low at (3.9 ± 0.5) × 10–3 cm2/Vs for sample S3, with CuInSe2 composition. Bi-exponential fits of the 500 ns photoconductivity transient decays gave an average lifetime approaching the instrument response of the microwave cavity of less than 30 ns. Sample S1(B) with ~ CuIn2Se3.5 composition showed dramatically improved φΣμ of 1.9 ± 0.4 cm2/Vs, nearly 1000 × larger than S3, yet the average weighted lifetime extracted from the fitted photoconductivity transient was still as short as S3 and nearly instrument response limited; TRMC free carrier lifetimes this short are not promising for useful device applications that rely on bulk charge transport. In contrast, sample S2 (B) with ~ CuIn3Se5 stoichiometry exhibited a φΣμ of ca. 3 cm2/Vs with a remarkable average weighted photoconductivity lifetime of 740 ns (fitted from 500 ns of data). To verify the long lifetime in this sample, the signal collection window was expanded to 2 µs and confirmed the lifetime at 727 ns. Also in contrast to S3 and S1(B) samples, the S2(B) exhibited a strong dependence of photoconductivity lifetime on excitation intensity; above an absorbed photon flux of 1 × 1010 cm−2 the lifetime decreased, but below this absorbed flux there was no intensity dependence. One explanation may be that the sharp pn NHJs in this composition serve as charge recombination centers at higher carrier densities, especially given that for nanocrystals of this size, the microwave photoconductivity signal is dominated by inter-grain carrier dynamics. Even so, the TRMC data confers with the PL data for S1 and S2. It implies that the observed shallow defect transitions may  be responsible for the extraordinary attributes of film S2, and likely contribute to its long carrier lifetime.Figure The following electrical characterization provides further insights into the electronic properties and structure of SSE-made CISe films. AFM topography in Fig. pn junction generates electron–hole pairs that can significantly affect the minority carrier concentration, hence the Fermi energy level and the width of the depletion region. This enables the TUNA photocurrent to distinguish the predominant conductivity type. Figure T) and S4(B) areas, Table 1. The images show that the p-CISe grains (4d) generate higher photocurrent under positive bias, whereas the n-CISe grains (4e) generate more photocurrent under negative bias. The switching of photocurrent polarity with applied bias provides visual evidence that the currents originate from individual p-CISe and n-CISe domains, coexisting within the same film. Both types of grains were present in the entire film, although the S4(T) and S4(B) areas exhibited stronger p-type and n-type response, respectively. Note that the different photocurrent scale bars in Fig. D) ranges in p- and n-grains. These observations are consistent with the Cu-rich and In-rich compositions for the S4(T) and S4(B) sections in Table Shining a laser on a D value was outside the sensitivity range, i.e. < 1014 or > 1021. Electrolyte impedance spectroscopy was performed in the frequency range of 100 kHz to 1 Hz for analogous CISe film S5. Figure V) versus capacitance (C) and 1/C2 for film S5. It shows high capacitance (tens of microns), ND > 1021 cm−3, and flat band potential of − 0.2 V vs. Ag/AgCl. The high values for ND are consistent with the VCu and InCu defect-doped, In-rich n-CISe ODC such as CuIn3Se5. The p-CISe was close to CuInSe2 stoichiometry10. The widely different NDs, outside the SCM range for the two conductivity types is probably the reason for lack of SCM contrast. Based on this rationale and the respective defect structure, we tentatively assign the very high doping ND (> 1021 cm−3) for the n-CISe grains and the very low doping ND (< 1014 cm−3) for the p-CISe grains. Admittance spectroscopy results indicate minimal frequency dependence of defects even at 300°K, Fig. 21. The Mott–Schottky and impedance data, stating the ND and the absence of deep defects corroborates with theoretical treatments that the intrinsic defects in n-CISe are shallow11. The shallow intrinsic defects are uniformly distributed within the nano grains and most of these grains are uniformly and intrinsically doped (1021/cm−3 ≡ one doping site per nm3). The shallow defects allow free carrier flow without trapping, avoid degeneracy and enhance grain-to-grain current flow.Despite the strong photocurrent response, scanning capacitance microscopy (SCM) contrast was not detected for film S4, possibly because its Npn junctions with neutralized electrical charges. Figure pn junctions, Fig. p-grain and n-grain, forming high quality pn NHJ. The sharp pn interface acts as a barrier to recombination of minority carriers; it could enhance current flow through devices, such as solar cells. Besides illustrating the different conductivities and pn junction formation, the combined AFM and EBIC results map out the conductive nature of the distributed grains in addition to the physical distribution of grains.Comparing the topography and AFM micrographs for film S4 on the same 1 µm wide area indicates that the very prominent, bright nanograin agglomerates in Fig. p-CISe and n-CISe nanocrystals that interconnect in a 3-dimensional (3D) network. The electrical potential gradient, ∇U at the pn NHJ interface separates the electron–hole pairs that are generated in each nanograin, Fig. pn junctions enables fast separation of electron and hole carriers. Electrons move through the n-CISe and holes through the p-CISe. The continuous generation of separated electrons and holes contributes to the high carrier density. This creates and maintains a chemical potential gradient, ∇μ to drive the transport of electrons and holes separately along the p-CISe and n-CISe grains, Fig. pn junctions primarily utilize the ∇U while exitonic devices utilize the ∇μ22. Unlike either of such device configurations, the CISe NHJ has the potential to effectively utilize both of the fundamental driving forces, i.e. ∇U  ≈ ∇μ, as shown in Fig. p and n electrodes the CISe NHJ layer forms a device with the configuration of: foil/n-cathode/n-CISe/p-CISe/p-anode, Fig. nF, EpF the valence band (VB) and conduction band (CB) edges for the n-CISe and p-CISe, respectively. The band-to-band transitions in (i) p-CISe and (ii) n-CISe nanograins as well as the intra-grain (i + iii), (ii + iv) transitions nanograins were previously derived from EER spectra10. This combination of intra-grain and band-to-band transitions within the CuInSe2-CuIn3Se5 complex could efficiently separate minority carriers, moving the holes to anode and the electrons to cathode. Suitable carrier selective contacts electrodes are needed to produce highly efficient p-CISe/n-CISe NHJ devices.These results indicate that under specific conditions, SSE spontaneously creates highly-ordered OC of 0.6 V and photocurrent of up to 23 mA/cm2 under 33 mW/cm2 illumination. But the efficiency obtained was ~ 5%, mainly due to the low fill factor that is expected for devices without carrier selective interlayers. Yet, based on the premise of low recombination in CISe NHJ space charge region, the low fill factor likely arises from recombination at the CISe/back contact or the series resistance of the device due to poor CISe/front contact interfaces. Thus, carrier selective electrodes could essentially prevent such interface leakages to maximize device efficiency. These preliminary photovoltaic metrics are especially promising given the completely unoptimized nature of the proof-of-concept devices and the low Eg of the CISe absorber. Thus, if optimized CISe NHJ device can be achieved with fill factor in the range of current peroskite devices of ~ 80%1 the efficiency could reasonably approach the 20% efficiency benchmark.Although solar cell efficiency is not the primary focus of this paper, we have tested several preliminary devices with the architecture of SS/CISe/TCO without appropriate carrier-selective electrodes; they exhibit VD and sharp pn junctions that indicate efficient charge carrier dynamics suggest that the CISe nanocrystals may work as quantum particles. Quantum confinement within CISe nanocrystals evidently generates the unusual opto-electronic properties. Such confinement could produce discrete spaced transitions within quantized states and contribute to the unexpected above-Eg PL emission. The fact that PL spectra are well-resolved even at 300°K attests to the unique quality of SSE-created CISe films. Only very pure and high quality crystals or artificially created quantum dot structures have shown similar transitions and non-linear absorption, and in those cases only under cryogenic conditions unlike here, where it is observed at room temperature. Moreover, the electronic nanostructure of CISe NHJ can tolerate wide composition variation, i.e. CISe films with In/Cu ratios from 1.3 to 2.5 give essentially the same PL spectra. This is in stark contrast to the precise composition and electronic configuration required for standard devices and the associated high fabrication costs. The direct deposition of CISe pn NHJ resolves many issues with the existing approaches, used to fabricate conventional planar pn junctions, perovskite/organic/inorganic bulk heterojunctions or inorganic ordered heterojunctions. Significantly, the CISe grains have highly desirable matching interfaces since they grow into each other as pn junctions. This finding solves a major hurdle for creating inorganic pn junctions. The exceptionally stable, non-toxic CISe NHJ avoids other issues of the nano pn junction devices, in particular the doping difficulties, separate synthesis and physical mixing of two material types. However, many of the carrier selective electrodes deployed in such devices can be utilized to produce the CISe NHJ device of Fig. The electronic properties concur and strongly support the PL and TRMC data. The shallow defects, high Npn NHJ. Its unexpected properties, including PL spectral range broadening, quantized emissions and the long lifetime, imply non-linear behavior similar to that seen in artificially designed quantum dot structures. Quantum confinement within CISe nanocrystals evidently generate discretely spaced quantized states. Specific charge carrier transitions within these states may be responsible for their unusual opto-electronic properties. Notably, the quantum confinement is enabled without external rare earth dopants into the host material. The extraordinary attributes of specific CISe films may be linked to the reaction kinetics during SSE that create special CISe morphologies with CuInSe2-CuIn3Se5 complexes.Coupling electrodeposition with CISe semiconductors creates scalable thin-film of nano n-CISe and p-CISe grains co-existing in same film. The cell geometry during SSE determines the predominance of Cu-rich p-CISe or In-rich n-CISe grains in the film. The n-CISe exhibits much higher ND and photocurrent than p-CISe. Shallow defects that intrinsically dope the CISe films appear to be uniformly distributed within the nano grains at the rate of one doping site per nm3. SSE naturally creates 3D network of highly-doped, interface-matched, sharp, abrupt p-CISe/n-CISe NHJs in-situ. These CISe pn NHJ potentially utilize both of the fundamental driving forces, i.e. ∇U  ≈ ∇μ, to reduce recombination, enhance carrier separation and transport, and thus boost future device performance. Band-aligned, carrier selective electrodes are sought to transition the CISe NHJ into high performance devices.The SSE-made films comprise pn NHJs that could essentially perform like crystalline 2D planar pn junctions or artificially ordered 3D nanostructures1 without their fabrication complexities. The results also call attention to the rarely explored ODC semiconductors that offer a plethora of relevant properties to produce nanostructures of technological interest. The resulting absorbers/emitters can be directly used in electro-optic devices with minimum manipulation. They offer new perspectives for processing devices with atmospheric roll-to-roll SSE. Such fabrication can impact a variety of the flexible electronics for next-generation optical communications, sensing, and imaging applications27. The resulting device roll can be fashioned into inexpensive flexible solar panels, or directly incorporated in solid-state lighting devices, photoelectrodes and other energy conversion. Overall, the SSE method efficiently balances the critical parameters for inorganic electro-optic devices: performance, stability, cost and scalability, for a variety of flexible devices in simple thin-film form factor.SSE presents a generic, very low-cost processing platform to create high quality 2 SS foil using Technic’s high speed flow cell or on a 3 cm2 SS foil in a beaker. Experimental conditions were varied to obtain samples with different CISe composition, including external dopants. The electrolyte pH was adjusted to ~ 2 during the deposition, with NaOH or HCl solution. Sufficient CuCl solution was added to the electrolyte to deposit roughly CuIn2Se3.5 compositions. Se was added to maintain a [Se2]/[Cu+] ratio between 3 and 6. The formation of In-rich CISe-ODC compounds requires excess of In3+ concentration and slightly higher pH. The films were annealed using an IR lamp (650 W) at 90% power with 2–3 s pulses, repeated 10–15 times in ambient atmosphere. Film composition and thickness were analyzed at multiple spots across the sample area with Spectro Midex XRF spectrometer.The SSE of CISe films was performed in 0.15 M NaCl aqueous electrolyte containing specific Cu:In:Se concentration ratio at 60 °C. The films were deposited on 117 cm2 to the Na2SO3 (pH 3) electrolyte held in a 3-electrode cell. Lock-in ampliflier was used in a frequency range of 100 kHz to 1 Hz to obtain capacitance values and Electrolyte Impedance Spectroscopy.Electrochemical characterization was performed on masked CISe samples, exposing an area of 0.25 cm34. Since the CISe films were deposited onto SS foil, which would short the microwave cavity, the films were transferred to low-dielectric loss quartz substrates (25 × 11 × 1  mm3) using a delamination-and-tape-transfer process. Single wavelength excitation pulses at 10 Hz (ca. 5 ns pulse width) and 500 nm were generated by a 355 nm pumped optical parametric oscillator (Continuum) on the signal arm of the output. A series of absorptive neutral density filters were used to attenuate the excitation power over 3 orders of magnitude, and the beam area was diffused to uniformly illuminate the entire cross-section of the q-band waveguide, providing uniform optical pumping over the entire area of the sample (~ 2.5 cm2). The microwave cavity was purged continuously with dry nitrogen during all measurements.PL emission spectra were obtained at 300°K (room temperature), using Horiba’s Micros setup with a 532 nm laser excitation. Long pass filters were used to block the short wave light below 540 nm or 830 nm; the data were not corrected for the grating spectra. Contactless probing with TRMC provided relevant photoelectric response of CISe films. The technique and its application to photoresponsive materials has been described in explicit detailp-type and n-type grains in the film. CISe sample S1 was cut and mounted on a metal disk with Ag-paste. Topological and nano-electrical AFM-TUNA micrographs were obtained on 1 µm area, using Bruker Dimensional Icon and SCM-PIT or SCM-PTSI probes, under AC bias of 5 V, lift height of 100 nm and scan rate of 0.5 Hz. This system included Kelvin probe force microscopy (KPFM) and scanning capacitance microscopy (SCM). The dark lift mode was used to eliminate the laser effects on the minority carrier concentration, hence the Fermi energy level and the width of the depletion region. The adjunct SCM, KPFM techniques were used to discern surface variations in carrier concentration and work function between the tip/CISe sample.AFM-TUNA measurements used bias polarity effects to distinguish between EBIC micrographs were obtained by Ephemeron, Inc using Mighty EBIC 2.0 on ITO/CISe/SS device at 10 V made from film S6.


Sarcoidosis is a multisystem inflammatory disorder and can affect any organ; however, ureteric involvement is extremely rare with only four cases reported in the literature to date, all of which were diagnosed with surgical ureteral resection including a nephroureterectomy. This study reports the first case of ureteric sarcoidosis controlled with medical therapy where a differential diagnosis was performed based on the diagnostic clue of hypercalcemia. A definitive diagnosis was established without surgical resection of the ureter.A 60-year-old man presented with anorexia and weight loss. Blood tests showed renal dysfunction and hypercalcemia. Computed tomography revealed left hydronephrosis associated with left lower ureteral wall thickening, which showed high signal intensity on diffusion-weighted magnetic resonance imaging. Similarly, we detected a bladder tumor on cystoscopy, and a 2-cm-long stenosis was revealed by retrograde ureterography; therefore, ureteral cancer was suspected. Meanwhile, considering the clinical implication of hypercalcemia, a differential diagnosis of sarcoidosis was established based on elevated levels of sarcoidosis markers. Fluorodeoxyglucose positron emission tomography showed fluorodeoxyglucose accumulation in the left lower ureter, skin, and muscles, suggestive of ureteric sarcoidosis with systemic sarcoid nodules. For a definitive diagnosis, transurethral resection of the bladder tumor and ureteroscopic biopsy were performed. Histopathological examination revealed ureteric sarcoidosis with bladder urothelial carcinoma. Following an oral administration of prednisolone, hypercalcemia instantly resolved, the renal function immediately improved, and the left ureteral lesion showed complete resolution with no recurrence.In this case, the co-occurrence of ureteral lesion with bladder tumor evoked a diagnosis of ureteral cancer. However, considering a case of ureteral lesion complicated with hypercalcemia, assessment for differential diagnosis was performed based on the calcium metabolism and sarcoidosis markers. In cases of suspected ureteric sarcoidosis from the assessment, pathological evaluation with ureteroscopic biopsy should be performed to avoid nephroureterectomy. Sarcoidosis is a multisystem inflammatory disorder characterized by the formation of non-caseating granulomas and can affect any organ . HoweverA 60-year-old man with no medical history or comorbidities presented with anorexia and weight loss (from 59.4 kg before admission to 55.9 kg on admission). No findings were observed on physical examination. Blood tests showed renal dysfunction  and hypercalcemia . A urine test revealed hypercalciuria without proteinuria  revealed left hydronephrosis associated with left lower ureteral wall thickening, which showed high signal intensity on diffusion-weighted magnetic resonance imaging Figs. a, b, 4a.3 2D3, 151.0 ng/L, normal range: 20.0–60.0 ng/L) and sarcoidosis markers: angiotensin-converting enzyme , lysozyme , and soluble interleukin-2 receptor . Neither 25-hydroxyvitamin D (25(OH)D), parathyroid hormone (PTH), nor parathyroid hormone-related peptide (PTHrP) was elevated: levels of 25(OH)D, PTH and PTHrP were 23.3  ng/mL, 15.0  pg/mL and <  1.1  pmol/L, respectively  has been implicated as a potential cause of sarcoidosis . Sarcoid2D3 by non-caseating granulomas has been implicated in calcium metabolism disorder. An elevated level of 1,25-(OH)2D3 enhances intestinal calcium absorption and promotes osteoclastic activity and bone resorption, resulting in hypercalcemia and hypercalciuria [Calcium metabolism disorder, characterized by hypercalcemia and hypercalciuria, which were recognized in this case, is a clinical feature in sarcoidosis. Hypercalcemia and hypercalciuria occur in approximately 5% and 40–62% of sarcoidosis patients, respectively . An incralciuria –10.Calcium metabolism disorder is the most important cause of renal dysfunction. Several different mechanisms, including vasoconstriction of arterioles, acute tubular necrosis, impairment of urinary concentrating ability caused by decreased sensitivity to antidiuretic hormone, and nephrocalcinosis, have been implicated in renal dysfunction . In the The present case met the diagnostic criteria of sarcoidosis: a compatible clinical and/or radiological image, histopathologic evidence of non-caseating granulomas, and exclusion of other diseases with similar findings, such as infections or malignancy . The ind2D3 and sarcoidosis markers . However, it should be noted that urothelial carcinoma is rarely complicated by hypercalcemia as paraneoplastic syndrome [2D3 is similarly reported to be one of the inducers of paraneoplastic hypercalcemia [2D3, would be needed to clarify the cause of hypercalcemia. Finally, ureteroscopy is a useful tool, which can facilitate more precise pathological diagnosis [In the present case, the co-occurrence of ureteral lesion with bladder tumor evoked a diagnosis of ureteral cancer. Meanwhile, the clinical manifestation of hypercalcemia served as a clue to the differential diagnosis of ureteric sarcoidosis through the elevated levels of 1,25-(OH)syndrome –16. In tsyndrome . In uretiagnosis . In the In conclusion, in the cases of ureteral lesions complicated with hypercalcemia, assessments for differential diagnosis based not only on calcium metabolism markers but also on sarcoidosis markers should be performed as a first step. This can help decide whether subsequent examinations for ureteric sarcoidosis are necessary. In the event ureteric sarcoidosis is suspected after the initial assessments, pathological evaluation with ureteroscopic biopsy should be performed, as ureteric sarcoidosis with hypercalcemia can be controlled with medical therapy, thus avoiding nephroureterectomy.


In this study, the filled natural rubber (NR) was prepared with organic montmorillonite (OMMT) and carbon black (CB). The effects of the amount of OMMT on the properties of CB/NR composites were investigated by measuring the physical and mechanical properties, compression set and compression heat properties, processing properties and damping properties. The formulation was optimized depending on the different conditions of end applications and the damping properties of rubber were maximized without affecting the other properties of the rubber. The results showed that the rubber composite system filled with 2 phr (parts per hundreds of rubber) OMMT had better mechanical properties and excellent damping performance. With the development of rail transit, it has become a major problem to prepare high-performance  rubber shock absorbers to improve NVH  performance. Rubber can transform the mechanical energy to heat energy, and so can reduce or even eliminate the vibration, and has been widely used as a damping material ,2. In reTg), storage modulus, and the damping properties of composite materials. The results showed that the storage modulus increased with the increase of the dosage and the loss factor decreases gradually. Guo et al. [Wang  reportedo et al.  reportedo et al.  grafted o et al.  preparedOrganic montmorillonite (OMMT) has attracted extensive attention due to its good reinforcing effect, low price, abundant reserves, simple preparation and environmental protection. As a result, it is used as a new reinforcing filler in rubber. The influence of OMMT content on swelling behavior was investigated, showing that it can significantly improve the delayed expansion property of rubber . Chen etIt is rarely reported that OMMT was used to improve the vibration damping performance of rubber, while the layered structure of it is worth studying. At the same time, it can be found from the previous research that the damping performance of rubber is mainly affected by the structure of rubber and its filler. In this study, the processing performance, mechanical properties and damping properties of rubber composites were all improved only with little amount of OMMT. To characterize the damping performance of rubber material with OMMT (widely used as nanofillers in polymeric composites), the OMMT/CB/NR composites were obtained by mechanical mixing reaction intercalation method. In addition, studies on the influence of OMMT content and the composition of rubber components on its mechanical, dynamic mechanical properties and damping properties were performed. The characterization of the samples was done by rubber processing analysis (RPA), dynamic mechanical analysis (DMA), scanning electron microscope (SEM) and the mechanical analysis instruments.N-cyclohexyl thio-phthalimide , N-1,3-dimethylbutyl-N’-phenyl-p-phenylenediamine , Poly   and Zinc oxide (ZnO-80) were purchased from Ningbo Actmix Rubber Chemicals Co., Ltd, Ningbo, China The remaining ingredients were provided by Jinchangsheng Co., Guangzhou, Guangdong, China. Natural rubber  was used as basis material. Sulfur (99.5% purity) was purchased from Jinchangsheng Co., Guangzhou, Guangdong, China. Organic montmorillonite  was purchased from Zhejiang Fenghong clay chemical Co., Anji, Zhejiang, China. Cabon Black (N330) was purchased from Cabot Corporation, Tianjin, China. N-cyclohexyl thio-phthalimide, a kind of scorch retarder), 4020 , RD , a kind of antioxidant), mixed for 3 min. The following were carbon black N330 and OMMT, mixed for 3 min. After that, sulfur and ZnO (Zinc oxide) were added into the blended compound. Next, set the distance between the two rollers to 1mm, made the “triangle bag” in the process for 5 times. Lastly, the compound obtained was molded into sheets in a Type Plate Vulcanizer (XLB-400 × 400 × 2 50 T)  at 150 °C for 15 min.The formula of this study was listed in Mooney viscosity test was carried out on a CL-2000G Mooney Viscosity Meter at 100 °C  according to the standard of ASTM-D1646. Vulcanization performance was conducted using a MDR2000 Rotor Rheometer at 150 °C  according to the standard of ASTM-GB/T 16584-1996. Processing performance of each content were tested with three samples.1 according to HG/T 3849-2008, GB/T 529-2008 and HG/T 3321-2012. Hardness was measured by GT-GS-MB Shore A Hardness Tester  according to GB/T 531.2-2009. Compression set was tested according to GB/T 7759.1-2015 . Compression heat was tested by GT-RH-200  according to GB/T1687.3-2016.  at an impact speed of 14 m·s−1 according to GB/T 1681-2009. Five samples of each content were tested for mechanical properties.Tensile properties, tear strength and elasticity modulus were determined using AT-7000M Universal Electronic Tensile Machine  at a tensile speed of 500 mm·min⁻Dynamic rheological measurements of unvulcanized rubber compounds were carried out on a RPA8000 Rubber Processing Analyzer  by a shear mode at a temperature of 60 °C, a frequency of 1 Hz, and a strain amplitude in the range of 0.28–200%, according to ASTM D 6204. Three samples of each content were tested for data of RPA.The DMA was carried out using a DMA 242 Dynamic Mechanical Analyzer  in a double cantilever beam deformation mode within a temperature range of −80 to 80 °C with a heating rate of 3 °C/min and at a fixed frequency of 1 Hz. The experiment process was carried out in the atmosphere of nitrogen. Three samples of each content were tested for data of DMA.The micromorphology of OMMT in rubber matrix was investigated using a JEOL JSM-7500F scanning electron microscope (SEM) . Resolution: 1.0 nm at 15 kV; 1.4 nm at 1 kV. Acceleration voltage: 0.1–30 kV. One sample for each content was tested for SEM.L) increased gradually with an increase in the content of OMMT. It was determined by the viscosity of the rubber compound, which indicates the strength of physical interaction when the rubber is not cross-linked. Therefore, when the content of OMMT was increased, the physical interactions of the rubber gradually increased. The maximum torque (MH) increased appreciably with addition of 2 phr OMMT. However, the increment was not obvious with more amounts of OMMT addition. MH depends on the filler and the vulcanization system, and it reflected the superposition of physical interactions and chemical interactions after cross-linking. When OMMT was used in rubber, the MH-ML of compound increased, and the viscosity became larger thereby producing the agglomeration effect. Therefore, the larger content of OMMT, the stronger the agglomeration effect, and thus it damaged the mechanical properties of the material.Normally, the change in torque during vulcanization is proportional to the density of cross-linking including the physical and chemical cross-linking. The vulcanization properties of each content were tested. It can be seen from H and MH-ML as discussed in the above analysis.Hardness was measured at five different locations of each sample. It can be seen from In general, polymers are not used in their pure form, which means that the CB, silica and OMMT play an important role in the polymer reinforcement. The fillers in polymer strongly modifies the viscoelastic behavior, especially the dynamic mechanical properties of the composite. In dynamic mechanics measurements of shear modulus, the Payne effect is often referred to as the progressive breakdown of packing-packing interactions, where G’ is used commonly ,24,25.RPA8000 was used to conduct three-dimensional strain scanning on CB/NR compounds filled with different contents of OMMT, and the interaction between packing-packing and the interaction between packing-rubber were accurately characterized. Each content was tested with three samples and the experimental results were shown in In order to further study the dispersion of fillers in the CB/NR composites, we marked it in the RPA scans. The difference values of G’(1)-G’(3) at low strain could indicate the strength of the packing network, and the different values of G’(3)-G’(0) without adding filler at low strain could indicate the strength of the filler and the rubber . It can It can be seen from Tg of the OMMT/CB/NR composites slightly shifts toward the high temperature direction for the addition amount of 2, 4, and 6 phr OMMT. The slight increase of Tg is due to the fact that the rubber macromolecular segment is restricted by the sheet structure of OMMT and the exercise capacity is thus reduced. In the case of the 8 phr content, the agglomeration was produced and Tg was found decrease slightly, which is consistent with the analysis results of RPA. The peak value of Tanδ increased after OMMT was added, which was because the density of cross-linking within the vulcanized rubber increased. In the glass transition zone, the rubber was deformed by the external force and the flaky filler OMMT was oriented subsequently, which would generate friction with the rubber. This will transform the mechanical energy generated by the vibration into the heat energy loss, and then improve the damping of the material. As a result, the damping value of the material increased and the damping temperature range (Tanδ ≥ 0.3) was widened (As showed in  widened . FinallyThe CB/NR composite filled with 2 phr OMMT was found to have excellent mechanical properties and was up to the standard of the performance requirements of automobile damping vibration absorbing parts. Compared with the composite without OMMT, the crosslink density of the rubber was increased; the scorch time was extended; and the vulcanization rate was decreased. The OMMT is beneficial to improve the performance of the anti-vibration parts of the automobile and the processing safety. Meanwhile, the addition of OMMT could increase the dispersibility of CB and reduce the final temperature rise of compression heat generation. Both dynamic and static compression permanent deformations were quite small. After filling the OMMT, the system dynamic mechanical and damping properties were enhanced and the effective damping temperature range was broadened.


The release study of ACTH was tested using a modified USP apparatus 2 with Enhancer cells. The permeation study was conducted with vertical Franz cells. Rheograms of hydrogels were made with the use of a universal rotational rheometer. The dependence of the amount of released and permeated hormone on the ointment concentration was found. Based on the test of ACTH release from semi-solid formulations and evaluation of rheological parameters, it was found that glycerol ointment is the most favourable base for ACTH. The ACTH release and permeation process depends on both viscosity and ACTH concentration. The higher the hormone concentration, the higher the amount of released ACTH but it reduces the amount of ACTH penetrating through porcine skin.The transdermal route of administration of drug substances allows clinicians to obtain a therapeutic effect bypassing the gastrointestinal tract, where the active substance could be inactivated. The hormonal substance used in the study-corticotropin (ACTH)-shows systemic effects. Therefore, the study of the effect of the type of ointment base and drug concentration on the release rate and also permeation rate in in vivo simulated conditions may be a valuable source of information for clinical trials to effectively optimize corticotropin treatment. This goal was achieved by preparation ointment formulation selecting the appropriate ointment base and determining the effect of ACTH concentration on the release and permeation studies of the ACTH. Semi-solid preparations containing ACTH were prepared using Unguator CITO  Corticotropin (ACTH) is a hormone secreted by the anterior pituitary gland. It is a polypeptide consisting of 39 amino acid residues with a 4.5 kDa molecular weight ,2,3. It ® Gel, contains purified pig corticotropin and is administered by subcutaneous or intramuscular injection. This preparation is used in the treatment of, among others: West’s syndrome, bending attacks and mental retardation, usually diagnosed in the first year of life, an exacerbated form of multiple sclerosis, allergic conjunctivitis, ulcerative colitis, lupus erythematosus and vesicular epithelial separation  within 360 min. The release profiles was made based on obtained results.A modified USP Apparatus 2 equipped with 200 mL flat bottom vessels and mini paddles  was used for drug release studies. Excess amount of the ACTH ointment was loaded inside a special semisolid holder, Enhancer fluoropolymer cells (ERWEKA GmbH) with an exposed area of 3.8 cm2 values were calculated.The release results were fitted with different kinetics models such as zero order (% ACTH release vs. time), first order (log of % ACTH remaining vs. time), Higuchi’s model (% ACTH release vs. square root of time) and Korsmeyer-Peppas model (log of % ACTH release vs. log time). For each model R0-6h)-the area under the concentration-time-change curve was calculated. Area under the curve of concentration changes-time was counted as a numerical integral, using the parabol method. The size of the area under the concentration-time curve (AUC0-6h) was used to calculate the degree of relative availability  of ACTH 20 mg/g and ACTH 25 mg/g of ointment relative to the ACTH 15 mg/g reference, according to Equation (2):The availability of ACTH from the ointment (AUC2) between the ACTH concentration in the ointment and the total amount of released hormone after six hours and AUC were calculated.The obtained results were compared to the F-9 formulation, containing 15 mg/g of ACTH based on glycerol ointment. The existing interdependencies  with inner diameters 15 mm and permeation area of 1.77 cmon study . An accu2. Each of the formulations was applied to the skin surface.Porcine skin was used to study the permeation profile of ACTH due to the limited availability and difficulties associated with the use of ex vivo human skin. Nevertheless, porcine skin is a suitable alternative due to the similarities in thickness and permeability properties to human skin . The mod−1and 30 s−1, with shear stress measurement in relation to the shear rate (step by step) and a flow curve (flow test and hysteresis loops). Five repetitions were conducted for each experiment, and the mean values of the calculated parameters are presented (n = 5).Rheological tests were performed only for ointments releasing ACTH-for all hydrogels with corticotropin (F-7–F-9) and for hydrogel unloaded with ACTH. The analysis was performed using universal Reometry RM 200 Touch rotational rheometer by Lamy Rheology Instruments  for testing the rheological properties of samples. It was equipped with a CP1 Plus thermostat from Lamy Rheology Instruments with Rheomatic Lamy Instruments software. The tests were conducted in a system plate-plate geometry, using the MS CP 2445 Lamy Rheology Instruments measuring system with a diameter of 24 mm (α = 0.45°). Before measurement, the samples were placed in a CLW 53 STD incubator  at a temperature of 25.0 ± 0.5 °C. After 30 min, the sample was placed on the lower plate of the Peltier effect thermostatic system made of stainless steel, and the upper plate was lowered. The height of the gap between the plates was 50 μm. Excess sample was removed with a spatula. All rheological tests were carried out at 25.0 ± 0.5 °C. For the tested ointments, viscosity was measured at two shear rates, 15 s−1. The obtained results are presented in the form of shear rate dependence on shear stress as flow curves and also in the form of hysteresis loop for step-by-step test. The analysis was carried out in the tangential stress range 5.0–200.0 sp < 0.05 was calculated using Student’s t-test.Mean values with standard deviation were calculated and statistically analyzed using the Microsoft Excell package and Statistica  option: Industrial analysis, experimental design (DOE). The Statistica Pharmaceutical Kit: Statistica “Release Profiles” was used to analyze the results of ACTH release from the ointment bases. The Weibull method. Statistically significant significance at Analyzing the results of ACTH release test it can be stated that the degree of ACTH release depends on several factors: the base used, the concentration of ACTH, rheological parameters of ointment. Among the bases used, the most favourable ointment base for preparation of ointment from ACTH was hydrogel ointment (glycerol ointment), whereas the optimal concentration of corticotropin in the ointment was 20 mg/g. The higher the hormone concentration, the greater its amount was released into the acceptor fluid. The process of release at the latest began from ointment at concentration of 15 mg/g and as the concentration increased, more corticotropin was released at specific intervals in relation to ointments with the lowest concentration. The degree and rate of release of ACTH depend on the concentration of the active substance in the ointment and on the rheological properties of semisolid formulations. The addition of hormone causes an increase in viscosity in relation to the unloaded hydrogel itself. At the same time, a further increase in the concentration to 20 mg/g and 25 mg/g results in a increase in viscosity and release but increase in viscosity at 25 mg/g ACTH provide decrease in released amount of ACTH.Based on a study of corticotropin penetration through the natural membrane that is the skin of the pig’s ear, it was found that ACTH penetrates the skin. The inverse dependence of the amount permeated and the rate of hormone permeation on its concentration was obtained. The most advantageous formulation was the ointment with the lowest concentration , from which the corticotropin penetrated in the highest amounts and the fastest. With increased concentration and increased viscosity the amount of penetrating corticotropin decreased. These results do not confirm the relation obtained from the release test with the use of an artificial cellulose membrane. This fact proves different membrane properties and unpredictable character of processes taking place in the skin. Therefore, it is important to check the behavior of the active substance in simulated in vivo conditions. Based on the obtained results, it was found that there was a possibility of using ACTH in the form of ointments in clinical practice . Semi-solid dosage forms are a convenient form of skin application for potential patients, and unlike injections, a safe and non-invasive method of therapy.


The development of electrochemical biosensors is an important challenge in modern biomedicine since they allow detecting femto- and pico-molar concentrations of molecules. During this study, pillared graphene structures supported by vertically aligned carbon nanotubes (VACNT-graphene) are examined as the potential recognition element of DNA biosensors. Using mathematical modeling methods, the atomic supercells of different (VACNT-graphene) configurations and the energy profiles of its growth are found. Regarding the VACNT-graphene doped with DNA nitrogenous bases, calculated band structure and conductivity parameters are used. The obtained results show the presence of adenine, cytosine, thymine, and guanine on the surface of VACNT-graphene significantly changes its conductivity so the considered object could be the prospective element for DNA biosensing. The application of impedance technology has reduced the detection of complementary DNA sequence limits to 5 × 10−12 mol L−1 [−21 mol L−1 [−15 mol L−1 [DNA and microRNA play a key role in storing and transmitting genetic information. Currently, molecular analysis of DNA can diagnose more than 400 diseases . It is k mol L−1 . The dis mol L−1 , and for mol L−1 .−7 mol/L and a sensitivity of 53.89 μA mmol/L−1cm−2 for glucose detection with an excellent stability and reproducibility. An electrochemical chiral sensor via an integrated polysaccharides/3D nitrogen-doped graphene-CNT frame exhibited excellent reproducibility, stability, and selectivity [2-CNTs@GONR nanocomposite that exhibited satisfactory stability and accuracy for quercetin detection in actual samples [The high sensitivity of CNTs coupled with the larger detecting surface area of graphene makes the hybrid material on their base an ideal candidate for the role of a biosensor element. Zou et al., fabricated a novel glucose biosensor by loading a glassy carbon electrode with a 1-methyl imidazole-based ionic liquid-functionalized graphene/CNT composite . The bioectivity . Mousaabectivity . Three-dectivity . Zhao et samples .3 gas significantly increased the conductivity of CNTs [2O-graphene sheet nanostructures can be well used as FET to detect polycyclic aromatic hydrocarbons [Methods of mathematical modeling results serve as a guideline for the design and optimization of biosensors on the base of graphene and CNTs. Khodadadi et al., fabricated an epirubicin biosensor based on the interactions of DNA with polypyrrole and nitrogen-doped reduced graphene during the study of docking theoretical investigations confirming the binding of epirubicin and guanine sides in the ds-DNA structure . The ana of CNTs . This ef of CNTs . Studies of CNTs . To reve of CNTs . The simocarbons .−24 g), which is even better than the carbon nanotube-based nanosensors [Recently, such hybrid 3D materials as pillared graphene structures supported by vertically aligned CNTs (VACNT-graphene) have become increasingly popular. This material is successfully used for hydrogen storing and thermal transport and is used in nanoelectronic devices and supercapacitors ,37,38,39osensors . Here, mTo build atomic structures of VACNT-graphene, graphene nanoparticles with sizes 34.44 Å × 34.08 Å  and SWCNTs with chirality indexes , , , , ,  were used as the most often synthesized ,42,43. Ri − E0 was calculated, where E0 was the energy per atom of the initial GNM and Ei was the energy per atom of the current structure. Occurring at the next stage, the energy profile of the considered junction’s growth was constructed. To this aim, the author-developed technique called “virtual growing” was used, which was previously successfully tested for seamless junctions on the base of GNMs and SWCNTs of the armchair-type . RegardiThe SWCNT is the only tube with conductive properties among those considered, so the junction on its base was considered as the potential biosensor. Probably, the electronic structure of this particular compound will react more than others to the presence of other molecules near its structure. Since surface area and intertube bridging are key parameters for different properties of pillared graphene ,49,50, tSince the DNA molecule is too small for quantum chemical calculation, we performed a study of the interaction between the VACNT-graphene and nitrogenous bases: guanine, adenine, thymine, and cytosine often abbreviated by G, A, T and C, respectively. The search for the optimal location of nitrogenous bases on the surface of the VACNT-graphene supercell was performed using the SCC DFTB 2 method using the 16 × 16 × 16 Monkhorst–Pack sampling and 3ob-3-1 parameterization . As a ref = −0.4 eV, and for the composite decorated with adenine, thymine, or cytosine this value shifted to −0.24 eV. The exception was the addition of guanine—in this case the noticeable peak at −0.24 eV is observed. It also draws attention to the fact that the DOS of VACNT-graphene+A and VACNT-graphene+T are similar. This may be caused by the fact that adenine and thymine form a base pair in DNA [Band structure analysis showed that addition of adenine and guanine to the surface of the VACNT-graphene decreases its Fermi level from −4.44 to −4.87 eV, while the addition of just adenine lowers to −4.88 eV, and of just thymine further reduces to −4.89 eV. r in DNA .2/h). Similar to the case of seamless junctions between SWCNT and graphene of the armchair-type, the anisotropy of transmission is not observed [The electron transmission function T(E), electric resistance R, and electrical conductivity G were calculated at 300 K on the basis of the Keldysh nonequilibrium Green function method  and Landobserved . When thTo explain the presence of peaks on the graph of the integral characteristic T(E) , we calcAb initio calculations confirmed that band structures, as well as the conductive properties of this junction, were sensitive to the presence of the DNA nitrogenous bases near its surface. Absorption of guanine, adenine, cytosine, and thymine led to the appearance of the noticeable peak at the Fermi level where they significantly affected the composite’s conductivity. When the current flew along the zigzag directions, the presence of the DNA nitrogenous bases increased the conductivity of the VACNT-graphene by 5–10%, while, when the current flew along in the armchair direction, its conductivity dramatically decreased by 38–45%. The obtained results indicate that the VACNT-graphene composite may be a prospective candidate for the role of a recognition element in electrochemical biosensor of DNA. The future task is to provide studies of interaction between VACNT-graphene and large DNA molecules. Since the typical DNA molecules are large, the sizes of VACNT-graphene should be enlarged too. Our preliminary calculations, for example, showed adsorption of B-DNA dodecamer [The atomic supercells of VACNT-graphene with SWCNTs of the most often synthesized chirality indexes , , , ,  were obtained using the original molecular modeling methods. Our calculations showed that the growth of all considered structures was energetically favorable, which opens a prospect for its potential manufacturing. Containing semiconductive tubes, the VACNT-graphene structure is a prospective candidate for the role of a biosensor element. Its geometrical structure facilitates adsorption of different molecules on its surface. The first place for such absorption is located in the area of defects formed at the graphene and CNT junction during growth. Another place was found in the pores of CNTs. The presence of conductive CNT and superconductive graphene allows the assumption that conductive properties of the composite can be changed by adsorption of different molecules. odecamer  on the s


A suitable way to modify the electronic properties of graphene—while maintaining the exceptional properties associated with its two-dimensional (2D) nature—is its functionalisation. In particular, the incorporation of hydrogen isotopes in graphene is expected to modify its electronic properties leading to an energy gap opening, thereby rendering graphene promising for a widespread of applications. Hence, deuterium (D) adsorption on free-standing graphene was obtained by high-energy electron ionisation of D One of the most pursued key goals in graphene functionalisation is the achievement of a two-dimensional 2D) material with unvaried mechanical strength and stiffness but with different and tailored electronic and transport properties. In fact, while the extremely high mobility of graphene renders it appealing for applications, its semi-metallic nature represents a hindrance. Graphane is the forecasted semiconducting ,2,3 formD materiagraphane . Graphangraphane ,5,6, cougraphane ,8,9,10,  project ) has bee project . The sta project . Since TAll these potential outcomes of graphane share the need to incorporate significant hydrogen (or its isotopes) quantities in the carbon mesh. Several experiments have been carried out till now, mainly using hydrogen as the element to functionalise graphene, which also presents very similar chemical reactivity to that of deuterium. Graphene hydrogenation can be achieved by different methods, ranging from hot ,14,15 ancracking ,18; the cracking , to chemcracking ,20,21, tcracking ,23,24. TThe upper limit for hydrogen (deuterium) uploading can be due both to the presence of defects/edges (in 2D graphene flakes) or to the influence of the substrate . To overcome these drawbacks we propose to expose to deuterium high-quality, three-dimensional (3D), warped, free-standing graphene. Here, we study the effect of deuterium incorporation in nanoporous graphene samples: NPG is constituted by a compact, bi-continuous interconnected 3D arrangement of high-quality graphene veils, with 1000 m fermion ,26, comp fermion ,28. The  fermion  associat [3 bond . Thus, i [3 bond , by measIn this paper, after characterising and describing how to prepare a properly clean NPG sample, we describe its direct exposure to a deuterium ion beam into a deuterium atmosphere, with the ions generated by the irradiation of the gas with 1.8 MeV electrons; the Drgy ions . Furtherrgy ions .Nanoporous graphene was synthesised by using a nano-porous Ni-based chemical vapour deposition (CVD) method ,33,34,35Deuteration of NPG was achieved at the CIEMAT laboratory in Madrid . This inThis chamber was mounted in the beam line of a Van de Graaff electron accelerator. A 0.05 mm aluminium foil separated the gas chamber and the accelerator beam line in vacuum. In this way, 1.8 MeV electrons passed through the aluminium foil into the gas chamber. Once in the gas chamber, the 1.8 MeV electron beam was collimated in such a way that it passed between the two copper electrodes without touching the electrodes or the sample, just ionising the deuterium gas between the two electrodes without irradiating the sample. Once the deuterium gas was ionised by the 1.8 MeV electron beam, due to the action of an electric field perpendicular to the beam, free electrons generated by the ionising beam drifted to the grounded electrode and deuterium positive ions were directed to the negative (−500 V) electrode, where the NPG sample was placed. The ion density current reaching the graphene sample was measured during the experiment, being 1.4 mA/cmAfter deuteration, the D-NPG sample was eventually transferred in dry ambient conditions to the XPS chamber.The XPS and ultraviolet photoelectron spectroscopy (UPS) experiments were performed at Sapienza University in Rome, in the Nanostructures at Surfaces laboratory. The UHV chambers had a base pressure in the high 10tails in ,41,42,43The Raman experiments were carried out at the Optical Spectroscopy of Nanostructured Materials laboratory, at Sapienza University of Rome. As the excitation source, a Nd:YVO4 laser at 532 nm (DPSS series by Lasos) was employed. The measurements were acquired in a backscattering configuration, by focusing the laser and collecting the signal with an Olympus 50× objective (NA = 0.50). The laser power was kept below 100 W measured at the entrance of the objective to avoid sample damage, having checked that no changes were induced in the spectra with such a power, and that powers higher by about one order of magnitude were needed to induce possible damages. A 200-mm focal length monochromator (Isoplane 160) equipped with a 150 or 300 grooves/mm grating was employed to spectrally analyse the Raman signal, which was detected by a back-illuminated Si CCD Camera (model 100BRX by Princeton Instruments). Rejection for elastically scattered laser light has been achieved by a sharp high-pass Razor edge filter at 535 nm (Semrock).Scanning electron microscopy (SEM) images of NPG at the In order to overcome the limitations in hydrogen/deuterium incorporation, a cleaning protocol should be adopted, so to minimise defects and contamination of NPG samples. In fact, both the synthesis and the deuterium upload procedures, despite being carried out in controlled and the cleanest possible conditions, led to slight yet unavoidable oxygen contamination. A proper preparation procedure of the pristine NPG sample before exposure to deuterium can be achieved by annealing at 600 The NPG sample microscopically appears with an apparent non-homogeneous texture by optical microscopy a, while As we will discuss in the following, the Raman spectra acquired in our NPG samples show a very slight presence of defect/disorder related peaks, typical of a very high-quality and low-defect one-to-few layer continuous graphene sheet with a turbostratic stacking, thereby having only weak bonding between the sheets, as discussed in previous papers ,28.NPG was measured by XPS on pristine samples and after an annealing procedure in UHV, aiming at reducing possible residual contamination. The C 1s and the O 1s core levels as functions of annealing temperature and time are displayed in Ox bonds ,45,46, dResidual oxygen is the only intense contaminant present, with a concentration of ≥20 at.% estimated by taking into account the relative excitation cross sections of the O 1s and C 1s core levels , as showhisation . This clhisation . We remaRaman spectroscopy represents a non-invasive, all-optical technique particularly effective to probe the formation of new bonds with other atomic species, to achieve information on the number of layers, or to verify the presence of defects, doping, strain and lattice deformation in graphene ,51,52 aned domes ,58,59. Ied domes . In the To have a comprehensive characterisation of the deuterated sample, Raman measurements were performed both before and after the D-exposure. Raman spectroscopy has been by far among the most effective and well used techniques for estimating graphene lattice quality . The Ram K point . Further K point . However K point ,16,60,61In Both the G peak at 1578 cmh higher ,16,60,61The C 1s core level spectra of the D-NPG sample as-adsorbed and after subsequent temperature annealing steps, along with the data of the UHV-clean one for comparison, are plotted in The experimental data were fitted with components representing the different chemically shifted environments, after subtracting a Shirley background. In particular, we used the asymmetric Doniach–Sunjic (DS) lineshape for the main sper ≤ 0.1 ,63,64), The most prominent component of the C 1s spectra is associated with the speriments ,28. Theritations ,65,66,67urations ,45,46. Wurations ,69,70,71Deuterium uptake induces the following changes in the C 1s lineshape: (i) increase of the spcase see . We can position . Anothersorption . After t0 ∘C see . FurtherThe adsorption of H on graphene is known to turn graphene from a semimetal to a semiconductor ,2,21,72.In the UHV-clean NPG we observe the typical electronic spectral density of graphene, characterised by (i) the linear behaviour towards the Fermi level (Eσ−π band , evidencgraphene .The observed gap opening is further proof of the achievement of semiconducting graphane after chemisorption of D, in unprecedented, highly D-uploaded graphene with very high quality. We underline that the high percentage of spNanoporous graphene has been efficiently functionalised with deuterium atoms by Ddetector .


After the EFA; the varimax rotation method was employed for a factor through commonality analysis; reducing the 14 initial factors to 10. Only question 30 presented commonality lower than 0.5; and the other questions returned values higher than 0.5 in the commonality analysis. Regarding the reliability of the instrument; all of the questions presented reliability as the values varied between 0.953 and 0.956. Thus; the instrument demonstrated construct validity and reliabilityThe purpose of this study was to validate the construct and reliability of an instrument to assess the work environment as a single tool based on quality of life (QL), quality of work life (QWL), and organizational climate (OC). The methodology tested the construct validity through Exploratory Factor Analysis (EFA) and reliability through Cronbach’s alpha. The EFA returned a Kaiser–Meyer–Olkin (KMO) value of 0.917; which demonstrated that the data were adequate for the factor analysis; and a significant Bartlett’s test of sphericity ( A dynamic work environment marked by fast and continuous changes, both in social relations and work transformations, demands that managers and companies rethink their positions regarding the workplace, work methods, and the elements that compromise the realization of work activities by the worker adequately . The worUnderstanding the complex work environment is quite valuable for organizations, as it enables them to elucidate situations that generate health problems and consequently entail high absence rates and turnover, and low productivity at work. In this context, managers’ responsibility for the health and welfare of the workers has increased ,7. EnvirHealth-focused environmental assessment is conducted through research instruments or tools, investigating situations that might compromise workers’ health and consequently affect productivity at the workplace. Assessing this environment is a complex task, seeing that it comprises both micro-environmental  and macro-environmental  factors, which influence an organization’s progress .These factors in the work environment include quality of life (QL), quality of work life (QWL), and organizational climate (OC). Health condition is a fundamental aspect of QL and work capability , and a pSimilarly, QWL is seen as the degree in which an individual satisfies their personal and work needs through participation at the workplace . It is rFinally, OC is linked to motivation, loyalty, and the identity of each company . As collIt is noteworthy that the human being is complex and holistic, and therefore seeks in an individualized way to satisfy his needs. The theoretical concept of the three focus areas of this study, namely, QL, QWL, and OC, have the same strand and these focus areas are directly related to welfare, satisfaction in the workplace, and productivity, and when used in a unique way they become useful tools for managerial practice.In the current research scenario, work environment assessment tools or instruments focus on three areas: QL, QWL, and OC . They feIn this context, the applicability of these individual instruments becomes costly and challenging, as the investigation takes longer and the results cannot be compared and linked to other tools, thus returning unreliable responses. Therefore, the scientific gap for the development of this study reveals itself, with the realization that companies need practical and all-encompassing tools that provide more complete responses about the work environment.Taking that into account, this study’s general purpose was to validate the construct and reliability of an instrument capable of assessing the work environment as a single tool based on QL, QWL, and OC.This study is justified because, despite the existence of QL and QWL assessment instruments, their structures are based on classic theoretical models developed over two decades ago, specifically for the American labor culture . SimilarThe proposal of this new instrument model intends to fill this gap in the academic and organizational context. It will allow the researcher or manager to identify, with a single tool, the problems and/or situations in the work environment that might potentially compromise workers’ health and affect productivity, be it related to QL, QWL, or OC.The first element in the development of a research instrument is the construction of a conceptual structure  containing the questions and factors related to the theme . ObserviThe theoretical construct initially contained 23 factors and 83 questions. Afterward, a board of examiners composed of five experts  in the field assessed the instrument and suggested the removal of 9 factors and 23 questions, resulting in a final theoretical construct comprising 14 factors and 50 questions. The factors represented the areas encompassed by the instruments . Within them, factors represented by questions were created, aiming to assess the collaborator’s satisfaction.After establishing the instrument’s theoretical construct, its validation was started. The validity of an instrument is directly related to how it measures what it intends to measure . Thus, aThe validation of assessment instruments commonly employs the techniques of content validity, face validity, criterion validity, and construct validity . The valThe first step to validate the instrument was testing content validity, seeking to identify the clarity of language, practical pertinence, and theoretical relevance. This verification was conducted by evaluator-judges selected according to their field, following the procedures described by the authors : the insAfter content validation, a pilot test was applied to a group of 100 workers of the company Beta, the focus of this study, aiming to test the feasibility of the study’s design and the clarity of the instrument. In this moment all workers signed an Informed Consent, of ethics committee project CAAE 35553720.0.0000.5547. The test revealed that the workers understood the instrument, eliminating the need for changes.Subsequently, the construct validity was tested through exploratory and confirmatory factor analysis. To allow this analysis, the aforementioned company Beta, which employs 565 workers, was contacted and authorized the survey. For a reliability level of 95% and a margin of error of 5%, the necessary sample would be 229 workers. For its validation, the instrument was applied to 231 workers, belonging to the same group as the pilot study, at this moment, all workers signed the Free and Informed Consent Term, according to the Ethics Committee project CAAE 35553720.0.0000.5547. These company employees over 18 years old were selected, and those who would be excluded would be given incomplete or blank questionnaires; however, no questionnaire was given this way.r values are distributed as follows: r = 0–0.25, very low correlation; r = 0.26–0.49, low correlation; r = 0.5–0.69, moderate correlation; r = 0.7–0.89, high or strong correlation; r = 0.9–1.0, very high or very strong correlation [In order to assess construct validity, Pearson’s correlation was employed. Pearson’s correlation is commonly used to verify the intensity of the existing linear association between variables and it measures the linear association between quantitative variables . This corelation . PearsonThe next step was reliability analysis through test–retest of the instrument and analyzing its internal consistency. Reliability or trustworthiness seeks to identify how consistent are the scores obtained by the same subjects surveyed when they are re-examined, which occurs through the application of the same instrument on another occasion .The test–retest procedure was employed to verify the instrument’s reliability. The instrument was applied by the researcher in the same company Beta and the workers answered the questions (test). After seven days, it was reapplied to the same group (retest).The instrument’s reliability was verified through Cronbach’s alpha, calculating the existing correlation between each item of the test and the remaining items or the total . Cronbach’s alpha has particular characteristics that seek solutions in a population surveyed, not simply a measure by itself, and these alpha values change according to the group in which the measure is adapted .Having verified the instrument’s reliability, the next step would be the criterion validation, aiming to examine whether the measuring scale was adequate to what was expected in relation to other variables, which are selected through expressive criteria ,39. For Finally, a sensitivity analysis sought to ascertain whether the instrument was identifying its actual goal. The sensitivity analysis aimed to identify whether this study actually measured its intended objective among the population assessed and to detect significant changes in the construct studied .The instrument’s sensitivity was assessed through indicators of central tendency and dispersion  of both sexes, only men, and only women so as to compare the results. These data were obtained in the application of the instrument during the test stage, and they revealed that the instrument measured its intended target.The stages that required statistical assessment were performed through SPSS 22.0  for Windows.In order to validate this instrument, a survey was conducted with 231 collaborators of the aforementioned company, the focus of this study. The anthropometric data showed that the respondents were, on average, 32.3 years old (SD ± 10.91). The sample contained 139 male respondents and 92 female respondents. In terms of educational background, 9 had basic education, 70 had a high school degree, 42 were undergraduate students, 53 had a college degree, and 57 had post-graduate specializations. These 231 collaborators were distributed in the three hierarchical levels of the company . Within the questionnaires, information about the organization’s hierarchical levels was collected, but it did not include this as a demographic variable in order for this instrument to meet all hierarchical levels. Also pointed out that the hierarchical pattern was implicit in the educational pattern and that this varied from company to company.Thus, the anthropometric data revealed that the respondents were 32 years old on average, mostly male, had a high school degree, and were mostly single.χ² = 7465.349, Df = 1225, p ≤ 0.000), allowing the EFA to be conducted.The Kaiser–Meyer–Olkin (KMO) index, used to verify the suitability of the application of Exploratory Factor Analysis (EFA) for this study’s data set, returned a value of KMO = 0.917, which demonstrated that the data were suitable for factor analysis. Bartlett’s sphericity test was significant .The number of factors estimated was determined by assessing the scree plot, employing question retention through the Kaiser–Guttman criterion of the components with eigenvalues higher than 1 . The anaSeeking to test the unidimensionality of the instrument, the EFA was conducted for one factor through commonality, employing the principal component method. Only question 30 presented commonality lower than 0.5. The others presented values higher than 0.5, demonstrating the existence power of each question. According to , the higAfter the commonality analysis employing the principal component method, the component matrix was verified. Employing the principal component analysis as the extraction method and the varimax rotation method with Kaiser normalization, the instrument went from 14 factors initially to 10 factors after the statistical analysis.The 50 questions were distributed into 10 factors, each allocated in one or more domains. However, the extraction and rotation method revealed the highest score in the factors, determining in which one the question must remain. It was possible to observe that factor 1 was composed by questions 22–25, 28–30, 39, and 43; factor 2 by questions 26, 27, 31–33, 45–47, and 50; factor 3 by questions 4, 8–11, and 18; factor 4 by questions 34–37, 41, and 42; factor 5 by questions 1–3, 5, and 6; factor 6 by questions 15–17; factor 7 by questions 19–21; factor 8 by questions 38, 40, 44, 48, and 49; factor 9 by questions 12–14; and factor 10 was composed by question 7.The factors were named according to the area which they encompassed, as pointed by the instrument’s theoretical construct and displayed by Factor 1—Health comprised five questions . Factor 2—Emotional and Psychological comprised six questions . Factor 3—Spiritual comprised three questions (12–14). Factor 4—Sleep and Rest comprised only one question (7). Factor 5—Work and Life comprised three questions (15–17). Factor 6—Work Conditions comprised three questions (19–21). Factor 7—Leadership Management comprised nine questions . Factor 8—Remuneration and Functional Assistance also comprised nine questions . Finally, Factor 10—Personal Relations and Company Image comprised five questions .Seeking to analyze the construct validity of the instrument, the correlation matrix demonstrated the existing relationships between the pairs of variables.In the EFA application, Pearson’s correlation was conducted in the test–retest stage of the instrument, adopting a significance level of 0.01. The analysis of the correlation matrix revealed that all of the 50 questions presented a positive correlation.r values from 0 to 0.25 indicate a very low correlation, from 0.26 to 0.49 indicate a low correlation, from 0.5 to 0.69 indicate a moderate correlation, from 0.7 to 0.89 indicate a high or strong correlation, and from 0.90 to 1.0 indicate a very high or very strong correlation. The results showed that none of the questions presented very low correlations. Eight questions presented low correlations, thirty-eight presented moderate correlations, and four presented high or strong correlations, as displayed in According to , the r vThus, the 50 questions of the instrument were correlated. Even though eight questions presented a low correlation, they were significantly correlated.In order to determine the instrument’s internal consistency in terms of reliability, Cronbach’s alpha was calculated for the existing correlation between each item of the test and the remaining items or their total . The Cronbach’s alpha of the 50 questions is exhibited in Although determinant values higher than 0.7 are considered ideal , values The Cronbach’s alpha values presented in the instrument ranged 0.953–0.956. The only item that obtained 0.956 was question 5, while the others stayed within the interval mentioned.The results showed that all of the factors presented values higher than 0.7, which indicated the reliability of the proposed instrument.The full instrument is available in the χ² = 7465,349, Df = 1225, p ≤ 0.000). The EFA composed of the total of variables in the instrument revealed that nine questions should be removed from the initial fourteen. For that purpose, an EFA was conducted for one question through the commonality analysis, which presented only one question under the necessary parameter. However, this question remained in the instrument because it was the only one that addressed the theme of autonomy granted by the company to make decisions.The construct validation was conducted through Exploratory Factor Analysis, obtaining the values for KMO (0.917) and Bartlett’s test (After the commonality analysis, the component matrix was observed. The instrument, which at first featured fourteen factors, contained ten factors after the statistical analysis.Seeking to analyze the construct validity of the instrument, the correlation matrix was examined in the application of the instrument’s test–retest. All of the questions of the matrix presented positive correlations. Eight questions presented low correlation, thirty-eight presented moderate correlation, and four presented high or strong correlation.Regarding reliability, Cronbach’s alpha was calculated in order to determine the instrument’s internal consistency, pointing to values ranging from 0.953 and 0.956, which indicated that all of the questions presented reliability.Thus, it is possible to conclude that the proposed instrument is valid and applicable, seeing that it met all of the parameters necessary for its validation. It assesses elements of QL, QWL, and OC as a single tool that allows managers to comprehend the situation of the work environment.