Controversies in point-of-care 3D printing for oncological and reconstructive surgery with free software in oral and maxillofacial surgery: European regulations, costs, and timeframe.

The aim of this paper is to discuss the controversies surrounding the most recent European regulations, as well as the cost, for a 3D printing workflow using free-source software in the context of a tertiary level university hospital in the Spanish public health system. Computer-aided design and manufacturing (CAD/CAM) for head and neck oncological surgery with the printing of biomodels, cutting guides, and patient-specific implants has made it possible to simplify and make this type of highly complex surgery more predictable. This technology is not without drawbacks, such as increased costs and the lead times when planning with the biomedical industry. A review of the current European legislation and the literature on this subject was performed, and comparisons made with the authors' in-house 3D printing setup using free software and different 3D printers. The cost analysis revealed that for the cheapest setup with free software, it would be possible to amortize the investment from case 2, and in all cases the initial investment would be amortized before case 9. The timeframe ranged from 2 weeks with the biomedical industry to 72 h with point-of-care 3D printing. It is now possible to develop point-of-care 3D printing in any hospital with almost any budget.

Supramolecular Assembly of Human Pulmonary Surfactant Protein SP-D.

Pulmonary surfactant protein D (SP-D) is a glycoprotein from the collectin family that is a component of the lung surfactant system. It exhibits host defense and immune regulatory functions in addition to contributing to the homeostasis of the surfactant pool within the alveolar airspaces. It is known that the SP-D monomer forms trimers, which further associate into higher-order oligomers. However, the pathway and the interactions involved in the assembly of SP-D oligomers are not clearly understood. In the current study, a recombinant form of full-length human SP-D (rhSP-D) has been qualitatively and quantitatively studied by atomic force microscopy (AFM) and electrophoresis, with the aim to understand the conformational diversity and the determinants defining the oligomerization of the protein. The rhSP-D preparation studied is a mixture of trimers, hexamers, dodecamers and higher-order oligomeric species, with dodecamers accounting for more than 50% of the protein by mass. Similar structures were also found in hSP-D obtained from proteinosis patients, with the largest fuzzy-ball-like oligomers being more abundant in these samples. The proportion of dodecamer is increased under acidic conditions, accompanied by a conformational change into more compact configurations. Two hexamers appear to be the minimal necessary unit for dodecamer formation, with stabilization of the dodecamer occurring via non-covalent, ionic, and hydrophobic interactions between the individual N-terminal domains and the proximal area of the SP-D collagen stems.

[Electronic prescription for Primary Care: a reliable tool to conceal medication at hospital admission?]. / Prescripción electrónica de Atención Primaria, ¿una herramienta fiable para conciliar medicación al ingreso hospitalario?

OBJECTIVE: To analyze the reliability of an electronic prescription software at primary care (Medoracyl®) to conceal the medication at hospital admission. METHOD: Prospective, comparative study of the home-based validated treatment of patients admitted to the hospital as compared to the one prescribed through Medoracyl®. The medi cation at admission was gathered by medical interview, medi cal records, and revision of the medication brought by the patients; the medication active in Medoracyl® by consulting the application the admission day. The discrepancies between both therapies were analyzed. RESULTS: 47 patients were included with 273 lines of home validated treatments and 274 lines of Medoracyl® treatment. 48 out of 273 lines were in discrepancy (17.6% [95% CI: 13.1- 21.6]). 27 out of 48 were justified, whereas 7.8% represented a discrepancy risk [95% CI: 4.6-11.0]. CONCLUSIONS: Medoracyl® is a useful tool and easy to access that allows knowing more than 90% of the home medication of the patients.

Objetivo: Analizar la fiabilidad de un programa de prescripción electrónica en receta de atención primaria (Medoracyl®) para conciliar la medicación al ingreso hospitalario. Método: Estudio prospectivo comparativo del tratamiento domiciliario validado de pacientes al ingreso, con el prescrito en Medoracyl®. La medicación al ingreso se obtuvo mediante entrevista clínica, informes médicos y revisión de la medicación aportada; la medicación activa en Medoracyl®, mediante consulta a la aplicación el día del ingreso. Se analizaron las discrepancias entre ambos tratamientos. Resultados Se incluyeron 47 pacientes con 273 líneas de tratamiento domiciliario validado y 274 líneas de tratamiento Medoracyl ®. Resultaron 48/273 líneas discrepantes (17,6% [IC95%: 13,1-21,6]). Se consideraron justificadas 27/48, resultando un riesgo final de no concordancia del 7,8% [IC95%: 4,6-11,0]. Conclusiones: Medoracyl® es una herramienta útil y de fácil acceso que permite conocer más del 90% de la medicación domiciliaria de los pacientes.

Ciliate metallothioneins: unique microbial eukaryotic heavy-metal-binder molecules.

This article represents an updated review of ciliate metallothioneins (Tetrahymena species) including a comparative analysis with regard to well-known metallothioneins (MTs) from other organisms and discussion of their exclusive features. It opens with an introduction to ciliates, summarizing the main characteristics of these eukaryotic microorganisms and their use as cellular models to study metallothioneins and metal-eukaryotic cell interactions. It has been experimentally proved that at least three different metal resistance mechanisms exist in ciliates, of which bioaccumulation is the most studied. Structural comparative analysis reveals that Tetrahymena MTs have unique characteristics, such as longer length, a considerably higher cysteine content, different metal-MT stoichiometry values, the presence of new cysteine clusters, and a strictly conserved modular-submodular structure. Gene expression analysis reveals a multistress and differential response to diverse metals and other environmental stressors, which corroborates the classification of these MTs. An in silico analysis of the promoter sequences of some MT genes reveals the presence of conserved motifs that are probably involved in gene expression regulation. We also discuss the great advantages of the first ciliate whole-cell biosensors based on MT promoters from Tetrahymena thermophila to detect heavy metal ions in environmental samples.

Ciliate cryptobiosis: a microbial strategy against environmental starvation.

This review outlines the main features of ciliate resting-cyst formation or encystment. It represents a strategy against several environmental stresses (such as starvation), which involves a highly gene-regulated cell differentiation process and originates a more resistant, differentiated form or resting cyst. This process is mainly characterized by drastic cytoplasmic dehydration that induces a general metabolic rate decrease, intense autophagic activity, the formation of a permeable cyst wall protecting the cell against the adverse environmental conditions, and a gene-silencing mechanism after opening the specific encystment genes.

DNA methylation in ciliates: implications in differentiation processes.

Much experimental evidence on the role of DNA methylation in gene expression has been reported. Here we review reports on DNA methylation in ciliated protozoa, emphasizing its implications in cell differentiation processes. Both types of methylated bases (adenine and cytosine) can be found in macronuclear DNA. The division cycle and conjugation have been studied with regard to adenine methylation, and several different functions have been assigned to the methylation changes detected in these processes. Cytosine methylation changes were analyzed during stomatogenesis of Paramecium and encystment of Colpoda inflata. A comparative analysis with other similar microbial eukaryotic differentiation processes is carried out.

Resting cyst wall glycoproteins isolated from two colpodid ciliates are glycine-rich proteins.

Two glycoproteins bands isolated from the cyst wall protein pattern of two colpodid ciliates, Colpoda inflata (gp46CI) and Colpoda cucullus (gp46CC) were analysed for their amino acid composition. Both glycoproteins are very rich in glycine and have a relatively high hydrophobicity, containing additionally many leucine and alanine residues. Their high degree of similarity is both quantitative and qualitative. Compared with just two previously published reports, their amino acid compositions are similar to those found in the hydrolysed cyst wall total proteins from the ciliates C. steinii and Paraurostyla spp. The amino acid composition corroborates that they are indeed glycoproteins, because asparagine, an amino acid residue suitable for the attachment to N-acetylglucosamine by its amide group (N-glycan), is abundant in both proteins. We discuss our data in relation to other glycine-rich proteins and a comparison with amino acid composition protein databases is carried out.

A rapid bioassay to detect mycotoxins using a melanin precursor overproducer mutant of the ciliate Tetrahymena thermophila.

Four different mycotoxins (patulin, T-2 toxin, diacetoxyscirpenol and roquefortine) were used to study growth inhibitory effects on a melanin precursor overproducer mutant of the ciliate Tetrahymena thermophila. This strain is especially sensitive to diacetoxyscirpenol and T-2 toxin. The secretion capacity of melanin precursors into the culture medium by this mutant and its biosensor capacity are very useful characteristics to elaborate a rapid bioassay to detect some specific mycotoxins.

Karyotypic Variability in Ribosomal DNA Subchromosome Size among Colpodid Ciliates, a Possible Tool To Differentiate Colpodid Species.

Pulsed-field gel electrophoresis has been applied to analyze the karyotypic variability among colpodid ciliates. The 18S ribosomal gene was found at different locations in the electrophoretic pattern, and these size variations in the ribosomal DNA subchromosomal molecule seem to be species specific. This could potentially be a useful new tool with which to differentiate colpodid ciliates.

Macronuclear DNA demethylation is involved in the encystment process of the ciliate Colpoda inflata.

Ciliate encystment is an eukaryotic cell differentiation process which involves a specific gene expression, to form the resting stage. In this study, we investigate, for first time, the DNA methylation pattern changes during encystment in the ciliate Colpoda inflata, and the 5-azacytidine effect on growing cells and encystment. Results indicate that 5-methylcytosine is present in macronuclear DNA of this ciliate and the 5-azacytidine treatment induces encystment in growth conditions. From restriction enzyme digestion and 5-azacytidine experiments, we conclude that a specific DNA demethylation is probably involved in the encystment gene expression of this ciliate.

Age-related excretion of six glycosidases in rat urine.

The activity of beta-N-acetylglucosaminidase (NAG), beta-galactosidase, alpha-L-fucosidase, beta-glucuronidase, beta-glucosidase and alpha-mannosidase was determined in the urine of rats at progressive ages from newborn to old animals. The age-dependence of urinary creatinine, protein and pH values was also studied. Enzyme activity, related to urinary creatinine, was significantly higher in the newborn group than other ages. The excretion of NAG increased significantly in adult rats (3-6 months old) compared to young rats (1 month old). Most of the enzyme activities were diminished in old rats (25 months old). Increased proteinuria and creatinine excretion were observed in rats since 3 months of age. Age-related differences among enzyme activities therefore should be considered when these urinary glycosidases are to be studied in rats.

Detection of antigenic cyst wall elements in Colpoda inflata: an immunoelectron microscopic study and immunoblotting identification of cyst wall polypeptides.

By using an antiserum against isolated cyst walls from resting cysts of the ciliate Colpoda inflata, cyst wall polypeptides have been identified by immunoblotting test. Likewise, an immunoelectron microscopical study on both complete resting cysts and isolated cyst walls to localize the cyst wall proteins recognized by the antiserum, has been carried out. The immunoblotting test showed that three main polypeptide bands were recognized by the antiserum, with tentative molecular weights of 61, 66 and 70 kDa respectively. This methodology provides a better identification of cyst wall proteins after electrophoretic separation of cyst wall samples from ciliate resting cysts.

New methods for the assay of 5-isosorbide mononitrate and its validation.

Differential scanning calorimetry (DSC) is a method which has been applied to obtain thermal information about both raw material and the determination of the content of 5-ISMN in dry mixtures with lactose. The main advantage this offers, compared to other methods, is that it is not necessary to use a standard, it only being necessary to know the data of its fusion heat. HPLC and TLC-densitometry are considered for the determination of 5-ISMN: lactose and pharmaceutical dosage forms. Linearity test, repeatability and accuracy were satisfactory in both methods. Recovery data in pharmaceutical formulations (expressed as the percentage of the label claim) from HPLC and TLC did not give any significant difference (P = 95%). The results show that the chromatographic methods are simple, fast and reliable procedures for the determination of 5-ISMN.

Towards a generalized model of encystment (cryptobiosis) in ciliates: a review and a hypothesis.

In this work we propose a generalized model to explain encystment (cryptobiosis) in ciliates. The model is elaborated from both structural and physiological studies previously reported, and some hypothetical considerations. The main features of this model are the following: (a) starvation is considered as the most important inducer of ciliate encystment, and it constitutes the first trigger of this cell differentiation process; (b) the "switch on" of encystment genes involves the appearance of several new transcripts; (c) starvation involves cell autophagy and protein turnover, which provides material to synthesize the new cystic proteins; (d) cytoplasmic dehydration has an important role during the resting cyst formation; (e) a gradual loss of intracellular water can inactivate cell metabolism, leading to an ametabolic state (cryptobiosis); (f) as a late precystic event, the encysting cell forms a barrier (cyst wall), that isolates the cell from the hostile environment.

Cortical morphogenesis and conjugation process in Caenomorpha medusula (Ciliophora, Heterotrichida).

The new oral infraciliature comes from the posterior longitudinal proliferation of numerous somatic kineties of the parental perizonal zone during the bipartition of Caenomorpha medusula. The new perizonal zone of the opisthe originates from proliferation of the anterior extremes of all the somatic kineties that make up the parental perizonal zone. The proter retains both, the oral infraciliature and the perizonal zone of the parental cell. Conjugation in C. medusula presents three maturation divisions and three postzygotic divisions. During conjugation the oral infraciliature of each conjugant degenerates to be replaced by a new one.