Digital technique to analyze the wear of screw-retained implant supported metal-ceramic dental prostheses and natural tooth as antagonist: a pilot study.

The aim of this study was to describe a novel digital technique to analyze the wear of screw-retained implant-supported metal-ceramic dental prostheses and natural tooth as antagonist.Materials and methods Ten patients were consecutively included to rehabilitate partial edentulism by dental implants. Both the screw-retained implant-supported metal-ceramic dental prostheses and the natural tooth as antagonist were submitted to a digital impression through an intraoral scan to generate a Standard Tessellation Language digital file preoperatively (STL1), at 3 months (STL2), and 6 months (STL3) follow-up. Afterwards, an alignment procedure of the digital files (STL1-STL3) was performed on a reverse engineering morphometric software (3D Geomagic Capture Wrap) and volume changes at the screw-retained implant-supported metal-ceramic dental prostheses and the natural tooth as antagonist were analyzed using Student's t-test. Moreover, Gage R&R statistical analysis was conducted to analyze the repeatability and reproducibility of the digital technique.Results Gage R&R showed a variability attributable to the digital technique of 3.8% (among the measures of each operator) and 4.5% (among operators) of the total variability; resulting repeatable and reproducible, since the variabilities were under 10%. In addition, statistically significant differences were shown at the wear volume (µm3) of both the natural tooth as antagonist (p < 0.0001) and the screw-retained implant-supported metal-ceramic dental prostheses between 3- and 6-months follow-up (p = 0.0002).Conclusion The novel digital measurement technique results repeatable and reproducible to analyze the wear of screw-retained implant-supported metal-ceramic dental prostheses and natural tooth as antagonist.

Influence of parallel pins on the angle deviation for placement of dental implants: an in vitro study.

The aim of the present study was to analyze and compare the angle deviation of two, four and six adjacent dental implants placed with and without straight parallel pins. MATERIALS AND METHODS: Two hundred and forty (240) dental implants were selected and randomly allocated into the following study groups: Two dental implants placed with straight parallel pins (Ref.: 144-100, BioHorizons, Birmingham, AL, USA) (n = 10) (2PP); Two dental implants placed without parallel pins (n = 10) (2withoutPP); Four dental implants placed with straight parallel pins hT(n = 10) (4PP); Four dental implants placed without parallel pins (n = 10) (4withoutPP); Six dental implants placed with straight parallel pins (n = 10) (6PP) and Six dental implants placed without parallel pins (n = 10) (6withoutPP). The dental implants randomly assigned to groups 2PP and 2withoutPP were placed into standardized polyurethane models of partially edentulous upper jaws in tooth positions 2.4 and 2.6, the dental implants randomly assigned to groups 4PP and 4withoutPP were placed into standardized polyurethane models of fully edentulous upper jaws in tooth positions 1.6, 1.4, 2.4 and 2.6, and the dental implants randomly assigned to groups 6PP and 6withoutPP were placed into standardized polyurethane models of fully edentulous upper jaws in tooth positions 1.6, 1.4, 1.2, 2.2, 2.4 and 2.6. Afterwards, postoperative CBCT scans and digital impressions were aligned in a 3D implant-planning software to compare the angle deviation (°) of two, four and six adjacent dental implants placed with and without straight parallel pins using the General Linear Model statistical analysis. RESULTS: Statistically significant differences were found between the angle deviation of 2 dental implants placed with straight parallel pins (p < 0.0001) and between the angle deviation of 4 dental implants placed with straight parallel pins (p = 0.0024); however, no statistically significant differences were found in the angle deviation of 6 dental implants placed with straight parallel pins (p = 0.9967). CONCLUSION: The use of a straight parallelization pin results in lower angle deviation between two and four adjacent dental implants; however, it is not effective for a larger number of dental implants.

Natural pentacyclic triterpenoid as allosteric modulators of human 5-lipoxygenase with potential anti-inflammatory activity.

This study explored new methods to inhibit human 5-lipoxygenase (5-hLOX) by analyzing natural terpenes that share structural similarities with acetoxyboswellic acid (AKBA). Enzymatic assays were used to evaluate the terpene's ability to inhibit the enzyme, potentially providing anti-inflammatory benefits. Our research focused on how certain types of triterpenes can inhibit 5-hLOX allosterically via a newly discovered allosteric site identified by enzyme crystallization. To determine whether natural boswellic acid analogs mimicked the allosteric known inhibitor AKBA, we combined 5-hLOX inhibition with in silico modeling. Our research has discovered that certain amino acids, specifically Arg 138, Arg 101, Arg 68, and Gln129, located in the allosteric 5-hLOX pocket, play a critical role in stabilizing glycyrrhetinic isomers. These amino acids form hydrogen bonds and hydrophobic interactions that contribute to the inhibitory potency of boswellic acid derivatives. We have found that α and ß glycyrrhetinic acid isomers, carbenoxolone, and to a minor extent, prednisolone, have a potent inhibitory effect against 5-hLOX with IC50 values of 8.64, 3.94, 52.98, and 291.20 µM, respectively. These values are in line with our calculated in silico allosteric site binding energy estimations. In contrast, other steroidal or non-steroidal anti-inflammatory agents exhibited inhibitory potencies larger than 500 µM. However, the specific pharmacodynamic mechanisms are currently unknown. We propose that AKBA analogs may lead to the future development of novel anti-inflammatory agents.Communicated by Ramaswamy H. Sarma.

The Expression of Amino Acid and Fatty Acid Receptors Show an Age-Dependent Pattern Involving Oral Cavity, Jejunum and Lower Gut Sensing in Broiler Chickens.

This work aimed to evaluate the gene expression of amino acids (AA) and fatty acids (FA) sensors in the gastrointestinal tract (GIT) of chickens at two different ages (7 and 26 days post-hatch). Sixteen broilers (Ross 308) were selected, and ten sections of the GIT, including upper (tongue base, upper palate, crop, proventriculus), middle (gizzard, duodenum, jejunum, ileum), and lower GIT section (cecum, colon) were collected for analysis. Relative gene expression of AA (T1R1, T1R3, mGluR1, mGluR4, CaSR, GPR139, GPRC6A, GPR92) and FA (FFAR2, FFAR3, FFAR4) sensors were assessed using qPCR. The statistical model included age, GIT section, and gene. In addition, the correlations between gene expressions were calculated. At day 7, a significantly (p = 0.004) higher expression of AA sensors in the oral cavity and FA sensors in the lower GIT section (i.e., cecum and colon) compared to the middle section was recorded. A higher expression of AA compared to FA sensors was detected at the upper GIT section in 7 (p < 0.001) and 26-day-old chickens (p = 0.026). Thus, at day 7, AA sensors were predominantly (p < 0.05) expressed in the upper GIT section (mainly oral cavity), while FA sensors were mainly expressed in the lower GIT section, at cecum (FFR2 and 4) or colon (FFAR3). These results may indicate that in early life, both ends of the GIT are fundamental for feed intake (oral cavity) and development of the microbiota (cecum and colon). In contrast, at 26 days of age, the results showed the emergence of both AA and FA sensors in the jejunum, presumably indicating the essential role of the jejunum in the digestion absorption of nutrients and the signaling to the brain (gut-brain axis) through the enteroendocrine system. Significant positive correlations were observed between T1R1 and T1R3 (r = 0.85, p < 0.001), CaSR and T1R1 (r = 0.78, p < 0.001), CaSR and T1R3 (r = 0.45, p < 0.050), and mGluR1 and FFAR3 (r = 0.46, p < 0.050). It is concluded that the gene expression is greater in the oral cavity for AA sensors and the lower gut for FA sensors. On day 26, the role of jejunum regarding nutrient sensing is highlighted.

Effect of the computer-aided static navigation technique on the accuracy of bicortical mini-implants placement site for maxillary skeletal expansion appliances: an in vitro study.

The objective of the present study was to evaluate and compare the effect of the computer-aided static navigation technique on the accuracy of the maxillary skeletal expansion (MSE) appliances. MATERIAL AND METHODS: Forty orthodontic self-drilling mini-implants were placed in ten anatomically based standardized polyurethane models of a completely edentulous upper maxilla, manufactured using a 3D impression procedure. The four orthodontic self-drilling mini-implants for anchoring the MSE appliance were digitally planned on 3D planning software, based on preoperative cone-beam computed tomography (CBCT) scan and a 3D extraoral surface scan. Afterwards, the surgical templates were virtually planned and manufactured using stereolithography. Subsequently, the orthodontic self-drilling mini-implants were placed an postoperative CBCT scans were performed. Finally, coronal entry-point, apical end-point and angular deviations were calculated using a t-test for independent samples or a non-parametric Signed Rank test. RESULTS: Statistically significant differences were not shown at coronal entry-point (p = 0.13), apical end-point (p = 0.41) and angular deviations (p = 0.27) between the planned and performed orthodontic self-drilling mini-implants. CONCLUSIONS: Computer-aided static navigation technique enables accurate orthodontic mini-implant placement for the MSE appliances.

Intrafamilial phenotypic variability in autosomal recessive DOCK6-related Adams-Oliver syndrome.

Adams-Oliver syndrome (AOS) is diagnosed in presence of aplasia cutis congenita (ACC) of the scalp and terminal transverse limb defects (TTLD). The autosomal recessive (AR) DOCK6-related form of AOS is most often associated with a severe phenotype including also central nervous system and ocular abnormalities. We report a sister and brother with different expression of the phenotype. Both were compound heterozygous pathogenic variants in the DOCK6 gene, including a heterozygous c.5939+2T > C intronic variant that was maternally inherited, and a heterozygous deletion of exons 10 to 21 that was paternally inherited. The sister had microcephaly, periventricular calcifications, minor retinal vasculopathy, and mild impaired neurodevelopment, but only very subtle limb abnormalities and no ACC. Her brother showed a classical DOCK6-related AOS phenotype, including a severe bilateral peripheral ischemic retinopathy. From a review of 22 molecularly confirmed cases with DOCK6-related AOS with ophthalmic examination, we found that 16 of them had retinal vascular pathology (72.7%), confirming as the major ocular anomaly. Documented intrafamilial variability in our family and the evidence revised from previous reports, confirm that AR DOCK6-related AOS expressivity can produce a "milder" phenotype without ACC or TTLD, which could be underdiagnosed in simplex cases because it is difficult to recognize out of a familial context. Therefore, in order to know its real magnitude is required the future inclusion of DOCK6 gene in NGS panels directed to the study of simplex cases of patients with microcephaly, periventricular calcifications, retinal vasculopathy, and/or cardiovascular defects.

Hyperpolarization induces cytosolic alkalization of mouse sperm flagellum probably through sperm Na+/H+ exchanger.

In brief: Hyperpolarization of the membrane potential is a crucial step for mammalian sperm maturation. This work demonstrates that this membrane potential change likely activates a sperm-specific sodium/proton exchanger to induce alkalization in mouse sperm flagellum. Abstract: The sperm-specific sodium/proton exchanger (sNHE) is an indispensable protein for male fertility in mammals. Nevertheless, it is still unknown how mammalian sNHE is regulated. Evidence obtained from sea urchin sNHE indicates that hyperpolarization of plasma membrane potential (Vm), which is a hallmark of mammalian capacitation, positively regulates the sNHE. Therefore, we explored the activity of sNHE in mouse and human sperm by fluorescence imaging of intracellular pH (pHi) with a ratiometric dye, SNARF-5F. A valinomycin-induced Vm hyperpolarization elevated sperm flagellar pHi of WT mouse but not in sNHE-KO mouse. Moreover, this pHi increase was inhibited in a high K+ (40 mM) medium. These results support the idea that mouse sNHE is activated by Vm hyperpolarization. Interestingly, we observed different types of kinetics derived from valinomycin-induced alkalization, including some (30%) without any pHi changes. Our quantitative pHi determinations revealed that unresponsive cells had a high resting pHi (>7.5), suggesting that the activity of mouse sNHE is regulated by the resting pHi. On the other hand, valinomycin did not increase the pHi of human sperm in the head or the flagellum, regardless of their resting pHi values. Our findings suggest that the regulatory mechanisms of mammalian sNHEs are probably distinct depending on the species.

Evaluation of the communication of nursing students in the simulated teleconsultation: A cross-sectional study.

INTRODUCTION: The COVID-19 pandemic presented universities with the challenge of virtualizing the teaching-learning process. Simulated teleconsultation has been used in undergraduate training, which allows nursing students to interact with simulated patients remotely. Studies have identified that distance imposes communication barriers on all elements-sender, receiver and message-and in both forms of transmission: verbal and nonverbal. OBJECTIVE: To describe the communication of nursing students in teleconsultation with simulated patients in the context of primary health care. METHODS: This was a descriptive, cross-sectional study of 92 fifth-year nursing students. The communication variable was measured with the Connect Identify Understand Agree Help scale. In the data analysis, the mean and standard deviation of the scores on the 29 items of the scale were determined, as were the mean values for the total scale and for the 3 domains of the scale. RESULTS: The items that presented an average of less than 1 were primarily those related to the Agree and Help to Act domain. The total mean was 1.15, and the means for the domains Connect, Identify and Understand Problems and Agree and Help to Act were 1.53, 0.90 and 1.28, respectively. A weakness in the exploration of the psychosocial context of the simulated patients was observed. CONCLUSIONS: The results of this study, which evaluated communication in the context of distance care, corroborate the evidence regarding communication in real or simulated face-to-face situations. Studies that compare communication in various teaching-learning contexts, whether real or virtual, face-to-face or at a distance, are recommended.

A Replicable and Reproducible Digital Method for Quantifying Maxillary Sinus Airway Changes after Sinus Lifts Using the Lateral Window Approach Technique-A Retrospective Study.

In the present retrospective study, we aimed to assess the replicability and reproducibility of a novel digital measurement technique for analyzing the volumes of the left and right maxillary sinuses and the nasal and maxillary sinus airway complex after a sinus lift procedure using the lateral window approach, to provide an accurate measurement technique for easily applying in clinical practice and to allow pre-operative assessment of maxillary sinus lift surgery, avoiding complications and making surgery more predictable. MATERIAL AND METHODS: Thirty patients with partially edentulous posterior maxilla were selected and submitted to bilateral sinus lift using the lateral window approach technique, with grafting materials selected and submitted to cone beam computed tomography (CBCT) scans, both pre- and postoperatively. Then, datasets were uploaded to therapeutic digital planning software to measure the volume of the right and left maxillary sinuses and the nasal and maxillary sinus airway complex. Gage R&R statistical analysis was performed to assess the replicability and reproducibility of the digital measurement technique. RESULTS: The variability attributable to the novel digital measurement technique was 3.4% for replicability and 3.4% for reproducibility of the total variability of the samples. CONCLUSION: The novel digital method proposed is a replicable and reproducible technique for analyzing the volume of the right and left maxillary sinuses and the nasal and maxillary sinus airway complex after a sinus lift using the lateral window approach technique, allowing an accurate pre-operative assessment of maxillary sinus lift surgery, avoiding complications and making surgery more predictable.

Influence of the Computer-Aided Static Navigation Technique and Mixed Reality Technology on the Accuracy of the Orthodontic Micro-Screws Placement. An In Vitro Study.

To analyze the effect of a computer-aided static navigation technique and mixed reality technology on the accuracy of orthodontic micro-screw placement. Material and methods: Two hundred and seven orthodontic micro-screws were placed using either a computer-aided static navigation technique (NAV), a mixed reality device (MR), or a conventional freehand technique (FHT). Accuracy across different dental sectors was also analyzed. CBCT and intraoral scans were taken both prior to and following orthodontic micro-screw placement. The deviation angle and horizontal deviation were then analyzed; these measurements were taken at the coronal entry point and apical endpoint between the planned and performed orthodontic micro-screws. In addition, any complications resulting from micro-screw placement, such as spot perforations, were also analyzed across all dental sectors. Results: The statistical analysis showed significant differences between study groups with regard to the coronal entry-point (p < 0.001). The NAV study group showed statistically significant differences from the FHT (p < 0.001) and MR study groups (p < 0.001) at the apical end-point (p < 0.001), and the FHT group found significant differences from the angular deviations of the NAV (p < 0.001) and MR study groups deviations (p = 0.0011). Different dental sectors also differed significantly. (p < 0.001) Additionally, twelve root perforations were observed in the FHT group, while there were no root perforations in the NAV group. Conclusions: Computer-aided static navigation technique enable more accurate orthodontic micro-screw placement and fewer intraoperative complications when compared with the mixed reality technology and conventional freehand techniques.

A Comparative Analysis of the Piezoelectric Ultrasonic Appliance and Trephine Bur for Apical Location: An In Vitro Study.

To compare and contrast the accuracy of piezoelectric ultrasonic insert (PUI) and trephine bur (TB) osteotomy site preparation techniques for apical location. (1) Material and methods: A total of 138 osteotomy site preparations were randomly distributed into one of two study groups. Group A: TB technique (n = 69) and B: PUI technique (n = 69). A preoperative cone-beam computed tomography scan and an intraoral scan were performed and uploaded to implant-planning software to plan the virtual osteotomy site preparations for apical location. Subsequently, the osteotomy site preparations were performed in the experimental models with both osteotomy site preparation techniques and a postoperative CBCT scan was performed and uploaded into the implant-planning software and matched with the virtually planned osteotomy site preparations to measure the deviation angle and horizontal deviation as captured at the coronal entry point and apical end-point between osteotomy site preparations using Student's t-test statistical analysis. (2) Results: The paired t-test found statistically significant differences at the coronal entry-point deviations (p = 0.0104) and apical end-point deviations (p = 0.0104) between the TB and PUI study groups; however, no statistically significant differences were found in the angular deviations (p = 0.309) between the trephine bur and piezoelectric ultrasonic insert study groups. (3) Conclusions: The results showed that the TB is more accurate than the PUI for apical location.

Proteostasis deregulation as a driver of C9ORF72 pathogenesis.

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are two related neurodegenerative disorders that display overlapping features. The hexanucleotide repeat expansion GGGGCC (G4 C2 ) in C9ORF72 gene has been causally linked to both ALS and FTD emergence, thus opening a novel potential therapeutic target for disease intervention. The main driver of C9ORF72 pathology is the disruption of distinct cellular processes involved in the function of the proteostasis network. Here we discuss main findings relating to the induction of neurodegeneration by C9ORF72 mutation and proteostasis deregulation, highlighting the role of the endoplasmic reticulum stress, nuclear transport, and autophagy in the disease process. We further discuss possible points of intervention to target proteostasis mediators to treat C9ORF72-linked ALS/FTD.

Influence of the Computer-Aided Static Navigation Technique on the Accuracy of the Orthodontic Micro-Screws Placement: An In Vitro Study.

To analyze the influence of the computer-aided static navigation technique on the accuracy of placement of orthodontic micro-screws. One hundred and thirty-eight orthodontic micro-screws were randomly assigned to the following study groups: Group A. orthodontic micro-screw placement using a computer-aided static navigation technique (n = 69); B. orthodontic micro-screw placement using the conventional freehand technique (n = 69). In addition, the accuracy in the canine-premolar, premolar and molar sectors was analyzed in each study group. Cone-beam computed tomography and intraoral scans were taken both prior and subsequent to orthodontic micro-screw placement. The images were then uploaded using a 3D implant planning software, where the deviation and horizontal angles were analyzed using a multivariate linear model. These measurements were taken at the coronal entry point and apical endpoint between the planned orthodontic micro-screws. In addition, any complications resulting from micro-screw placement, such as spot perforations, were also analyzed in all dental sectors. The statistical analysis showed significant differences between the two study groups with regard to the coronal entry-point, apical end-point (p < 0.001) and angular deviations (p < 0.001) between the computer-aided static navigation technique and freehand technique study groups. Moreover, statistically significant differences were showed between the different dental sectors (p < 0.001). Additionally, twelve root perforations were observed at the conventional free hand technique study group while there were no root perforations in the computer-aided static navigation technique study group. The results showed that the computer-aided static navigation technique enables a more accurate orthodontic micro-screw placement with less intraoperative complications when compared with the conventional freehand technique.

Contextualized genome-scale model unveils high-order metabolic effects of the specific growth rate and oxygenation level in recombinant Pichia pastoris.

Pichia pastoris is recognized as a biotechnological workhorse for recombinant protein expression. The metabolic performance of this microorganism depends on genetic makeup and culture conditions, amongst which the specific growth rate and oxygenation level are critical. Despite their importance, only their individual effects have been assessed so far, and thus their combined effects and metabolic consequences still remain to be elucidated. In this work, we present a comprehensive framework for revealing high-order (i.e., individual and combined) metabolic effects of the above parameters in glucose-limited continuous cultures of P. pastoris, using thaumatin production as a case study. Specifically, we employed a rational experimental design to calculate statistically significant metabolic effects from multiple chemostat data, which were later contextualized using a refined and highly predictive genome-scale metabolic model of this yeast under the simulated conditions. Our results revealed a negative effect of the oxygenation on the specific product formation rate (thaumatin), and a positive effect on the biomass yield. Notably, we identified a novel positive combined effect of both the specific growth rate and oxygenation level on the specific product formation rate. Finally, model predictions indicated an opposite relationship between the oxygenation level and the growth-associated maintenance energy (GAME) requirement, suggesting a linear GAME decrease of 0.56 mmol ATP/gDCW per each 1% increase in oxygenation level, which translated into a 44% higher metabolic cost under low oxygenation compared to high oxygenation. Overall, this work provides a systematic framework for mapping high-order metabolic effects of different culture parameters on the performance of a microbial cell factory. Particularly in this case, it provided valuable insights about optimal operational conditions for protein production in P. pastoris.

Acrosomal alkalization triggers Ca2+ release and acrosome reaction in mammalian spermatozoa.

The sperm acrosome reaction (AR), an essential event for mammalian fertilization, involves Ca2+ permeability changes leading to exocytosis of the acrosomal vesicle. The acrosome, an intracellular Ca2+ store whose luminal pH is acidic, contains hydrolytic enzymes. It is known that acrosomal pH (pHacr ) increases during capacitation and this correlates with spontaneous AR. Some AR inducers increase intracellular Ca2+ concentration ([Ca2+ ]i ) through Ca2+ release from internal stores, mainly the acrosome. Catsper, a sperm specific Ca2+ channel, has been suggested to participate in the AR. Curiously, Mibefradil and NNC55-0396, two CatSper blockers, themselves elevate [Ca2+ ]i by unknown mechanisms. Here we show that these compounds, as other weak bases, can elevate pHacr , trigger Ca2+ release from the acrosome, and induce the AR in both mouse and human sperm. To our surprise, µM concentrations of NNC55-0396 induced AR even in nominally Ca2+ free media. Our findings suggest that alkalization of the acrosome is critical step for Ca2+ release from the acrosome that leads to the acrosome reaction.

Volatile and non-volatile/semi-volatile compounds and in vitro bioactive properties of Chilean Ulmo (Eucryphia cordifolia Cav.) honey.

Ulmo honey originating from Eucryphia cordifolia tree, known locally in the Araucania region as the Ulmo tree is a natural product with valuable nutritional and medicinal qualities. It has been used in the Mapuche culture to treat infections. This study aimed to identify the volatile and non-volatile/semi-volatile compounds of Ulmo honey and elucidate its in vitro biological properties by evaluating its antioxidant, antibacterial, antiproliferative and hemolytic properties and cytotoxicity in Caco-2 cells. Headspace volatiles of Ulmo honey were isolated by solid-phase microextraction (SPME); non-volatiles/semi-volatiles were obtained by removing all saccharides with acidified water and the compounds were identified by GC/MS analysis. Ulmo honey volatiles consisted of 50 compounds predominated by 20 flavor components. Two of the volatile compounds, lyrame and anethol have never been reported before as honey compounds. The non-volatile/semi-volatile components of Ulmo honey comprised 27 compounds including 13 benzene derivatives accounting 75% of the total peak area. Ulmo honey exhibited weak antioxidant activity but strong antibacterial activity particularly against gram-negative bacteria and methicillin-resistant Staphylococcus aureus (MRSA), the main strain involved in wounds and skin infections. At concentrations >0.5%, Ulmo honey reduced Caco-2 cell viability, released lactate dehydrogenase (LDH) and increased reactive oxygen species (ROS) production in a dose dependent manner in the presence of foetal bovine serum (FBS). The wide array of volatile and non-volatile/semi-volatile constituents of Ulmo honey rich in benzene derivatives may partly account for its strong antibacterial and antiproliferative properties important for its therapeutic use. Our results indicate that Ulmo honey can potentially inhibit cancer growth at least partly by modulating oxidative stress.

Dynamic genome-scale metabolic modeling of the yeast Pichia pastoris.

BACKGROUND: Pichia pastoris shows physiological advantages in producing recombinant proteins, compared to other commonly used cell factories. This yeast is mostly grown in dynamic cultivation systems, where the cell's environment is continuously changing and many variables influence process productivity. In this context, a model capable of explaining and predicting cell behavior for the rational design of bioprocesses is highly desirable. Currently, there are five genome-scale metabolic reconstructions of P. pastoris which have been used to predict extracellular cell behavior in stationary conditions. RESULTS: In this work, we assembled a dynamic genome-scale metabolic model for glucose-limited, aerobic cultivations of Pichia pastoris. Starting from an initial model structure for batch and fed-batch cultures, we performed pre/post regression diagnostics to ensure that model parameters were identifiable, significant and sensitive. Once identified, the non-relevant ones were iteratively fixed until a priori robust modeling structures were found for each type of cultivation. Next, the robustness of these reduced structures was confirmed by calibrating the model with new datasets, where no sensitivity, identifiability or significance problems appeared in their parameters. Afterwards, the model was validated for the prediction of batch and fed-batch dynamics in the studied conditions. Lastly, the model was employed as a case study to analyze the metabolic flux distribution of a fed-batch culture and to unravel genetic and process engineering strategies to improve the production of recombinant Human Serum Albumin (HSA). Simulation of single knock-outs indicated that deviation of carbon towards cysteine and tryptophan formation improves HSA production. The deletion of methylene tetrahydrofolate dehydrogenase could increase the HSA volumetric productivity by 630%. Moreover, given specific bioprocess limitations and strain characteristics, the model suggests that implementation of a decreasing specific growth rate during the feed phase of a fed-batch culture results in a 25% increase of the volumetric productivity of the protein. CONCLUSION: In this work, we formulated a dynamic genome scale metabolic model of Pichia pastoris that yields realistic metabolic flux distributions throughout dynamic cultivations. The model can be calibrated with experimental data to rationally propose genetic and process engineering strategies to improve the performance of a P. pastoris strain of interest.

Morphometric evaluation of wound healing in burns treated with Ulmo (Eucryphia cordifolia) honey alone and supplemented with ascorbic acid in guinea pig (Cavia porcellus).

BACKGROUND: In the context of the search for cost-efficient treatments, Ulmo (Eurcyphia cordifolia) honey is an excellent alternative for treating burn wounds and could have a profound medical, social, and economic impact. Ascorbic acid is an enzymatic co-factor necessary for the synthesis of collagen and the proliferation of fibroblasts and has been proposed as a coadjuvant to strengthen the healing effects of honey. The aim of this work was to evaluate by morphometric studies the healing wounds caused by burns treated with Ulmo honey alone and supplemented with ascorbic acid in guinea pig (Cavia porcellus). METHODS: Fifteen guinea pigs were used and divided randomly into three groups: positive control (C+), experimental with unsupplemented honey (H), and experimental with supplemented honey (SH). A uniform deep burn covering 1 cm2 of the back skin was performed. The following indices were calculated for the morphometric study: superficial contraction index of the wound, deep contraction index of the wound, wound severity index, global healing index, global contraction index, and dermal proliferation area. RESULTS: The superficial contraction index of the wound, global healing index, global contraction, and dermal proliferation area values of the experimental with supplemented honey group were higher than the other groups (P < 0.05). CONCLUSIONS: According to these results, the combination of honey with an antioxidant (ascorbic acid) promotes an appropriate action to support the healing effect. This study showed that by supplementing the Ulmo honey with ascorbic acid, the healing and contraction effects can be strengthened in burn wounds compared to unsupplemented honey. These results were proof of the synergy between honey and ascorbic acid in healing burn wounds.

Derivatives of 3H-spiro1-benzofuran-2, 1’-cyclohexanes: immunostimulants for veterinary use / Derivados de 3H-espiro1-benzofurano-2, 1’-ciclohexanos: inmunoestimulantes de uso veterinario

Four 3H-spiro1-benzofuran-2, 1’-cyclohexanes were synthesized from filifolinol, two of which are reported for the first time. Docking molecular studies were carried out to determine in silico whether these derivatives have similar immunostimulant activity to that reported for filifolinol, and its oxidation product, filifolinone. Through of the study of interactions of these compounds with the heterodimer of the protein present in teleost TLR1-TLR2, filifolinol, 3’-filifolinchloride and filifolinyl acetate shows similar interactions between them, allowing to predict that they would have similar immunostimulant activity, but different to filifolinone and filifolinane or that they would act by a different mechanisms.

Cuatro 3H-spiro1-benzofuran-2, 1'-ciclohexanos se sintetizaron a partir de filifolinol, dos de los cuales son reportados por primera vez. Se llevaron a cabo estudios de docking molecular para determinar in silico si estos derivados tienen actividad inmunoestimulante similar a la reportada para filifolinol y su producto de oxidación, filifolinona. A través del estudio de las interacciones de estos compuestos con el heterodímero de la proteína presente en teleósteos TLR1-TLR2 se estableció que el filifolinol, 3'-cloruro de filifolinilo y acetato de filifolinilo tienen interacciones similares con el heterodímero, lo que permite predecir que entre ellos tendrían una actividad simi- lar, pero diferente a la de la filifolinona y filifolinano o que estos últimos actuarían por diferentes mecanismos.

Chemical, technological and in vitro antioxidant properties of mango, guava, pineapple and passion fruit dietary fibre concentrate.

The aim of this work was to determine the chemical, technological and in vitro antioxidant properties of co-products from the industrialisation of some tropical exotic fruits, such as mango, pineapple, guava and passion fruit, and to evaluate their potential use as dietary fibre sources for food enrichment. Proximate compositions were determined, as well as the total, insoluble and soluble fibre contents. The water holding, oil holding and swelling capacities were also determined. For the antioxidant activity, three different test systems were used (ABTS, DPPH and FRAP). The dietary fibre content of the co-products varied in a range between 69.1 and 81.5g/100g on a dry matter basis with a good balanced ratio between insoluble dietary fibre and soluble dietary fibre. Phenol recovery was dependent on the fruit type and the solvent system used. Methanol:acetone was a more efficient solvent for extracting phenols than ethanol. There was a good correlation between total phenol content and antioxidant capacity of the fruit extracts. All samples analysed had good antioxidant capacity. The results of this study indicate that exotic fruit fibres obtained as co-products in the process to obtain juice may be considered a good source of natural compounds with significant antioxidant activity.