Growth, survival, digestive activity and respiratory metabolism of Octopus maya juveniles fed with prepared diets.

The effects of two prepared feeds were tested on growth, survival, enzymatic activity, nutritive reserves in the digestive gland and oxygen consumption of Octopus maya juveniles. For the first time, a semihumid paste (HD, control) and a dry diet, in pelleted form (PD, experimental) with the same formulation were used for this species. The experiment lasted for 42 days. Results indicate that growth rates were similar for both diets (p > 0.05); however, survival (70%) was higher with the PD compared to the HD (48%) (p < 0.05). The performance index was higher for octopuses fed the PD (p < 0.05). No differences in acid proteases activity were observed. However, a higher activity of alkaline proteases in the octopuses fed the PD was observed (p < 0.05). Ingestion rate was higher for octopuses fed the PD. Routine energy inversion was similar in both treatments (p > 0.05). A greater energy inversion was observed in octopuses fed the PD, whose active metabolism was double compared to the octopuses fed the HD. Results showed that the PD promoted similar growth compared to the HD diet but favored survival, and a greater investment in the active metabolism, reflected in the apparent heat increase.

Comparative analysis of periodontal pain and quality of life in patients with fixed multibracket appliances and aligners (Invisalign®): longitudinal clinical study.

BACKGROUNDS: The aim of this longitudinal clinical study is to analyse and compare according to location, degree and type, the pain presented by patients during their first year of treatment, as well as the quality of oral life after the placement of two types of orthodontic appliances: conventional brackets and removable Invisalign ® aligners. METHODS: The sample consisted of 140 patients grouped into 2 study groups of 70 patients each. The first group (brackets group- BG), with fixed multibracket appliances, using the MBT technique and a 0.022" slot. The second group (invisaling group- IG), in treatment with removable aligners (IG), using the Invisalign ® system. They were providen with a questionnare where they had to record the degree (mild, moderate or intense), the type and location of the pain monthly during the first year of treatment. The second form was the Spanish version of the OHIP-14, oral quality of life questionnaire, which was provided the twelfth month after the start of treatment. RESULTS: In both groups, we found that the most frequent location of pain occurred during the first phase: mandibular for the IG group and maxillary in the BG group. Throughout the whole analysis, the intensity was mild-moderate with lower values in the conventional brackets' group. The BG group reported acute pain while the IG group reported sensitive pain during the first month; later both reported sensitive pain. CONCLUSIONS: There are differences in terms of periodontal pain in its degree, location, and type according to the different orthodontic techniques used. TRIAL REGISTRATION: The study was approved by the bioethics committee of the University of Salamanca (USAL_20/516).

Alternative splicing of BCL-x is controlled by RBM25 binding to a G-quadruplex in BCL-x pre-mRNA.

BCL-x is a master regulator of apoptosis whose pre-mRNA is alternatively spliced into either a long (canonical) anti-apoptotic Bcl-xL isoform, or a short (alternative) pro-apoptotic Bcl-xS isoform. The balance between these two antagonistic isoforms is tightly regulated and overexpression of Bcl-xL has been linked to resistance to chemotherapy in several cancers, whereas overexpression of Bcl-xS is associated to some forms of diabetes and cardiac disorders. The splicing factor RBM25 controls alternative splicing of BCL-x: its overexpression favours the production of Bcl-xS, whereas its downregulation has the opposite effect. Here we show that RBM25 directly and specifically binds to GQ-2, an RNA G-quadruplex (rG4) of BCL-x pre-mRNA that forms at the vicinity of the alternative 5' splice site leading to the alternative Bcl-xS isoform. This RBM25/rG4 interaction is crucial for the production of Bcl-xS and depends on the RE (arginine-glutamate-rich) motif of RBM25, thus defining a new type of rG4-interacting domain. PhenDC3, a benchmark G4 ligand, enhances the binding of RBM25 to the GQ-2 rG4 of BCL-x pre-mRNA, thereby promoting the alternative pro-apoptotic Bcl-xS isoform and triggering apoptosis. Furthermore, the screening of a combinatorial library of 90 putative G4 ligands led to the identification of two original compounds, PhenDH8 and PhenDH9, superior to PhenDC3 in promoting the Bcl-xS isoform and apoptosis. Thus, favouring the interaction between RBM25 and the GQ-2 rG4 of BCL-x pre-mRNA represents a relevant intervention point to re-sensitize cancer cells to chemotherapy.

A New Digital Method to Quantify the Effects Produced by Carriere Motion Appliance.

The aim of this study was to analyze a novel digital technique to quantify the distal tooth displacement and derotation angle produced by the Carriere Motion Appliance (CMA). Twenty-one patients with a class II molar and canine relationship underwent orthodontic treatment with CMA. All patients were exposed before (STL1) and after the CMA placement (STL2), submitted to a digital impression, and afterwards, data were uploaded to specific cephalometric software to allow automatic mesh network alignment of the STL digital files. Subsequently, the distal tooth displacement of the upper canines and first upper molars, as well as the derotation angle of the first upper molars were analyzed using the Pearson correlation coefficient (ρ). Repeatability and reproducibility were analyzed using Gage R&R statistical analysis. An increase in canine displacement was correlated with an increase in contralateral canine displacement (ρ = 0.759; p < 0.000). An increase in canine displacement was correlated with an increase in molar displacement (ρ = 0.715; p < 0.001). An increase in upper first molar displacement was correlated with an increase in the contralateral upper first molar displacement (ρ = 0.609; p < 0.003) and the canine displacement (ρ = 0.728; p < 0.001). The distal tooth displacement showed a repeatability of 0.62% and reproducibility of 7.49%, and the derotation angle showed a repeatability of 0.30% and reproducibility of 0.12%. The novel digital measurement technique is a reproducible, repeatable, and accurate method for quantifying the distal tooth displacement of the upper canine and first upper molar, as well as the derotation angle of the first upper molars after using CMA.

Programming Positive Mechanofluorescence in Liquid Crystalline Elastomers.

Liquid single crystal elastomers (LSCEs) containing organic fluorophores within their polymeric network are attractive materials to detect forces with simple spectroscopic measurements. Hitherto, all mechanoluminescent LSCEs decrease their emission intensity upon mechanical stimulation; that is, they display negative mechanofluorescence. Such behavior is governed by the mechanically induced approximation of the quenching mesogenic units and the fluorophores. In this work, we propose the integration of fluorescent molecular rotors (FMRs), whose luminescence is not quenched by the mesogens, in LSCEs as a valuable strategy to conceive elastomeric materials programmed with exactly the opposite behavior, i.e., their fluorescence increases upon deformation (positive mechanofluorescence). Specifically, carbazole-indolenine dyes are interesting candidates for this purpose since their luminescence depends mainly on the degree of intramolecular rotation allowed by the local environment. On this basis, the uniaxial deformation of an LSCE, along its anisotropic direction, incorporating such FMRs will place the fluorophores in a more restricted medium, leading to the desired enhanced emission at the macroscale.

Changes in the Color and Brightness of White Spots Associated with Orthodontic Treatment 6 Months after the Application of Infiltrative Resins: Systematic Review and Meta-Analysis.

One of the risks that we find after orthodontic treatment is the secondary appearance of white spot lesions (WLS) after the removal of fixed multi-bracket appliances. Today, there are several treatment methods, resin infiltration being the most used in the most serious cases. The objective of this study is to carry out a systematic review and meta-analysis to determine the efficacy and stability in the variables of color and gloss, six months after resin infiltration. A comprehensive search was performed in the following databases: PubMed, Embase, Google Scholar, Scopus, Medline, and Web of Science. Articles published in the last 10 years were selected, including in vivo studies with a six-month follow-up. PRISMA guidelines were followed to carry out this systematic review. All studies where the application of resin was performed on carious lesions were discarded. Once the inclusion and exclusion criteria were applied, a final sample of four articles was obtained, on which the review and meta-analysis were carried out. Once examined, all authors considered that there was an immediate improvement in both variables. However, statistically significant differences were obtained in the color change outcome, but not in the brightness outcome in the subgroup analysis after six months of icon resin infiltration.

The different activities of RNA G-quadruplex structures are controlled by flanking sequences.

The role of G-quadruplex (G4) RNA structures is multifaceted and controversial. Here, we have used as a model the EBV-encoded EBNA1 and the Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded LANA1 mRNAs. We have compared the G4s in these two messages in terms of nucleolin binding, nuclear mRNA retention, and mRNA translation inhibition and their effects on immune evasion. The G4s in the EBNA1 message are clustered in one repeat sequence and the G4 ligand PhenDH2 prevents all G4-associated activities. The RNA G4s in the LANA1 message take part in similar multiple mRNA functions but are spread throughout the message. The different G4 activities depend on flanking coding and non-coding sequences and, interestingly, can be separated individually. Together, the results illustrate the multifunctional, dynamic and context-dependent nature of G4 RNAs and highlight the possibility to develop ligands targeting specific RNA G4 functions. The data also suggest a common multifunctional repertoire of viral G4 RNA activities for immune evasion.

Cu4MnGe2S7 and Cu2MnGeS4: two polar thiogermanates exhibiting second harmonic generation in the infrared and structures derived from hexagonal diamond.

The new, quaternary diamond-like semiconductor (DLS) Cu4MnGe2S7 was prepared at high-temperature from a stoichiometric reaction of the elements under vacuum. Single crystal X-ray diffraction data were used to solve and refine the structure in the polar space group Cc. Cu4MnGe2S7 features [Ge2S7]6- units and adopts the Cu5Si2S7 structure type that can be considered a derivative of the hexagonal diamond structure. The DLS Cu2MnGeS4 with the wurtz-stannite structure was similarly prepared at a lower temperature. The achievement of relatively phase-pure samples, confirmed by X-ray powder diffraction data, was nontrival as differential thermal analysis shows an incongruent melting behaviour for both compounds at relatively high temperature. The dark red Cu2MnGeS4 and Cu4MnGe2S7 compounds exhibit direct optical bandgaps of 2.21 and 1.98 eV, respectively. The infrared (IR) spectra indicate potentially wide windows of optical transparency up to 25 µm for both materials. Using the Kurtz-Perry powder method, the second-order nonlinear optical susceptibility, χ(2), values for Cu2MnGeS4 and Cu4MnGe2S7 were estimated to be 16.9 ± 2.0 pm V-1 and 2.33 ± 0.86 pm V-1, respectively, by comparing with an optical-quality standard reference material, AgGaSe2 (AGSe). Cu2MnGeS4 was found to be phase matchable at λ = 3100 nm, whereas Cu4MnGe2S7 was determined to be non-phase matchable at λ = 1600 nm. The weak SHG response of Cu4MnGe2S7 precluded phase-matching studies at longer wavelengths. The laser-induced damage threshold (LIDT) for Cu2MnGeS4 was estimated to be ∼0.1 GW cm-2 at λ = 1064 nm (pulse width: τ = 30 ps), while the LIDT for Cu4MnGe2S7 could not be ascertained due to its weak response. The significant variance in NLO properties can be reasoned using the results from electronic structure calculations.

Nucleotides bearing aminophenyl- or aminonaphthyl-3-methoxychromone solvatochromic fluorophores for the enzymatic construction of DNA probes for the detection of protein-DNA binding.

We designed and synthesized nucleosides bearing aminophenyl- or aminonaphthyl-3-methoxychromone fluorophores attached at position 5 of cytosine or thymine and converted them to nucleoside triphosphates. The fluorophores showed solvatochromic fluorescence with strong fluorescence at 433-457 nm in non-polar solvents and very weak fluorescence at 567 nm in alcohols. The nucleosides and nucleotides also showed only negligible fluorescence in alcohols or water. The triphosphates were substrates for DNA polymerase in the enzymatic synthesis of modified DNA probes that showed only very weak fluorescence in aqueous buffer but a significant light-up and blue shift were observed when they interacted with proteins (histone H3.1 or p53 for double-stranded DNA probes or single-strand binding protein for single-stranded oligonucleotide probes). Hence, nucleotides have good potential in the construction of DNA sensors for studying protein-DNA interactions. The modified dNTPs were also transported into cells using a cyclodextrin-based transporter but they were not incorporated into the genomic DNA.

Novel Digital Measurement Technique to Analyze the Palatine Suture Expansion after Palatine Rapid Maxillary Expansion Technique.

The aim of the study was to validate a novel digital measurement method to quantify the volume of the midpalatal suture after rapid maxillary expansion (RME). MATERIAL AND METHODS: Eight patients with maxillary skeletal transverse deficiency were submitted to palatine suture expansion using the McNamara orthodontic appliance during a period of nine months. After 30 days of treatment, all patients were exposed postoperatively to a cone-beam computed tomography (CBCT) scan. Afterwards, the scans were uploaded into the three-dimensional orthodontic-planning software to allow the volumetric assessment of the palatine suture expansion through palatine rapid maxillary expansion using a McNamara appliance. The repeatability was analyzed by repeating twice the measures by a single operator and reproducibility was analyzed by repeating three times the measures by two operators, and Gage R&R statistical analysis was performed. RESULTS: The expansion of the midpalatal suture by means of the rapid maxillary expansion technique, in terms of digital volume measurement, showed a repeatability value of 0.09% and between the two operators a reproducibility value of 0.00% was shown. CONCLUSIONS: The novel measurement technique demonstrated a high repeatability and reproducibility rate for volume assessment of the palatine suture expansion through palatine rapid maxillary expansion technique.

Acetophenyl-thienyl-aniline-Linked Nucleotide for Construction of Solvatochromic Fluorescence Light-Up DNA Probes Sensing Protein-DNA Interactions.

The synthesis of 2'-deoxycytidine and its 5'-O-triphosphate bearing solvatochromic acetophenyl-thienyl-aniline fluorophore was developed using the Sonogashira cross-coupling reaction as the key step. The triphosphate was used for polymerase synthesis of labelled DNA. The labelled nucleotide or DNA exerted weak red fluorescence when excited at 405 nm, but a significant colour change (to yellow or green) and light-up (up to 20 times) was observed when the DNA probes interacted with proteins or lipids.

Dietary Potassium Downregulates Angiotensin-I Converting Enzyme, Renin, and Angiotensin Converting Enzyme 2.

BACKGROUND: The importance of dietary potassium in health and disease has been underestimated compared with that placed on dietary sodium. Larger effort has been made on reduction of sodium intake and less on the adequate dietary potassium intake, although natural food contains much more potassium than sodium. The benefits of a potassium-rich diet are known, however, the mechanism by which it exerts its preventive action, remains to be elucidated. With the hypothesis that dietary potassium reduces renal vasoconstrictor components of the renin-angiotensin system in the long-term, we studied the effect of high potassium diet on angiotensin-I converting enzyme, renin, and angiotensin converting enzyme 2. METHODS: Sprague Dawley male rats on a normal sodium diet received normal potassium (0.9%, NK) or high potassium diet (3%, HK) for 4 weeks. Urine was collected in metabolic cages for electrolytes and urinary volume measurement. Renal tissue was used to analyze angiotensin-I converting enzyme, renin, and angiotensin converting enzyme 2 expression. Protein abundance analysis was done by Western blot; gene expression by mRNA levels by RT-qPCR. Renal distribution of angiotensin-I converting enzyme and renin was done by immunohistochemistry and morphometric analysis in coded samples. RESULTS: High potassium diet (4 weeks) reduced the levels of renin, angiotensin-I converting enzyme, and angiotensin converting enzyme 2. Angiotensin-I converting enzyme was located in the brush border of proximal tubules and with HK diet decreased the immunostaining intensity (P < 0.05), decreased the mRNA (P < 0.01) and the protein levels (P < 0.01). Renin localization was restricted to granular cells of the afferent arteriole and HK diet decreased the number of renin positive cells (P < 0.01) and renin mRNA levels (P < 0.01). High potassium intake decreased angiotensin converting enzyme 2 gene expression and protein levels (P < 0.01).No morphological abnormalities were observed in renal tissue during high potassium diet.The reduced expression of angiotensin-I converting enzyme, renin, and angiotensin converting enzyme 2 during potassium supplementation suggest that high dietary potassium intake could modulate these vasoactive enzymes and this effects can contribute to the preventive and antihypertensive effect of potassium.

Changes in Biochemical Composition and Energy Reserves Associated With Sexual Maturation of Octopus maya.

Climate conditions are related to changes in the biochemical composition of several tissues and associated to the processes of growth and sexual development in cephalopods. The biochemical composition (protein, glucose, cholesterol, acylglycerides) and hemocytes of the hemolymph, the hepatosomatic and gonadosomatic indices, and the reserves of the gonad, hepatopancreas and muscle (lipids, glycogen, and caloric value of muscle) of Octopus maya were determined and related to sex and season. A total of 154 wild animals were used (≈50 caught per season) and the multivariate analysis of the biochemical indicators of the tissues allowed following the variations during winter, dry and rainy season. The permutational MANOVA showed that both sex and season contributed significantly to variations in metabolites and energy reserves. However, the non-significant interaction term indicated that the biochemical composition changed with the seasons in a similar way and regardless of sex. The pattern observed in metabolites and reserves indicates a variation associated with growth and the reproductive peak, but may also reflect a physiological response to seawater temperature. The present study provides reference values for several physiological indicators in O. maya that may be useful for programs monitoring wild populations, as well as to design diets and management protocols to produce octopus under controlled conditions.

Best compromise nutritional menus for childhood obesity.

Childhood obesity is an undeniable reality that has rapidly increased in many countries. Obesity at an early age not only increases the risks of chronic diseases but also produces a problem for the whole healthcare system. One way to alleviate this problem is to provide each patient with an appropriate menu that is defined by a mathematical model. Existing mathematical models only partially address the objective and constraints of childhood obesity; therefore, the solutions provided are insufficient for health specialists to prepare nutritional menus for individual patients. This manuscript proposes a multiobjective mathematical programming model to aid in healthy nutritional menu planning that may prevent childhood obesity. This model provides a plan for combinations and amounts of food across different schedules and daily meals. This approach minimizes the major risk factors of childhood obesity (i.e., glycemic load and cholesterol intake). In addition, this approach considers the minimization of nutritional mismatch and total cost. The model is solved using a deterministic method and two metaheuristic methods. Test instances associated with children aged 4-18 years were created with the support of health professionals to complete this numerical study. The quality of the solutions generated using the three methods was similar, but the metaheuristic methods provided solutions in a shorter computational time. These results are submitted to statistical hypothesis tests to be validated. The numerical results indicate proper guidelines for personalized plans for individual children.

Thiophene-linked tetramethylbodipy-labeled nucleotide for viscosity-sensitive oligonucleotide probes of hybridization and protein-DNA interactions.

Cytosine 2'-deoxyribonucleoside dCTBdp and its triphosphate (dCTBdpTP) bearing tetramethylated thiophene-bodipy fluorophore attached at position 5 were designed and synthesized. The green fluorescent nucleoside dCTBdp showed a perfect dependence of fluorescence lifetime on the viscosity. The modified triphosphate dCTBdpTP was substrate to several DNA polymerases and was used for in vitro enzymatic synthesis of labeled oligonucleotides (ONs) or DNA by primer extension. The labeled single-stranded ONs showed a significant decrease in mean fluorescence lifetime when hybridized to the complementary strand of DNA or RNA and were also sensitive to mismatches. The labeled dsDNA sensed protein binding (p53), which resulted in the increase of its fluorescence lifetime. The triphosphate dCTBdpTP was transported to live cells where its interactions could be detected by FLIM but it did not show incorporation to genomic DNA in cellulo.

Sea Surface Temperature Modulates Physiological and Immunological Condition of Octopus maya.

Octopus maya is a valuable endemic species of the Yucatán Peninsula (YP). This area can be divided into distinct regions depending on the presence of cold waters associated to upwelling events during spring and summer. This study was designed to determine if the physiological and immunological condition of O. maya show a relationship with variation of the sea surface temperature associated with the seasonal upwelling. A total of 117 organisms were collected from February to July in three fishing zones: Ría Lagartos located in the upwelling zone; Seybaplaya corresponding to the non-upwelling zone, and Sisal, the transitional zone. The organisms were examined in terms of physiological (total weight, the weight of the gonad and digestive gland, osmotic pressure, hemocyanin, protein, glucose, and cholesterol concentrations in plasma), and immunological variables (total hemocyte count, hemagglutination, phenoloxidase system activity, total phenoloxidase plasma activity, and lysozyme activity). Multivariate one-way ANOVA showed overall significant differences between groups of octopus by month/zone of capture, indicating that the physiological-immunological condition of O. maya is related to a temperature gradient. Wild octopuses captured at the upwelling zone and the transitional zone (Ría Lagartos and Sisal) in February, March, and April -with temperatures lower than 27°C- were in better conditions: larger size, high concentrations of hemocyanin, and low activity of the phenoloxidase system. Octopuses captured in the warmer waters (28-30°C) of the non-upwelling and transitional zones (Seybaplaya and Sisal) during June and July, could be reflecting the metabolic stress through immunological compensation mechanisms with higher activity of the phenoloxidase system, despite having a lower concentration of hemocytes, hemocyanin, and proteins. Although the movement of individual O. maya along the YP throughout their life cycle has not yet been determined, direct development and benthic behavior could limit the mobility of the organisms in such a way that their physiological and immunological condition might reflect adaptation to the regional environment. This information could help understand the performance of octopuses in their distribution area, which sustains an important fishery.

Brightly Fluorescent 2'-Deoxyribonucleoside Triphosphates Bearing Methylated Bodipy Fluorophore for in Cellulo Incorporation to DNA, Imaging, and Flow Cytometry.

Synthesis of cytosine, uracil, and 7-deazaadenine 2'-deoxyribonucleosides and triphosphates (dNTPs) bearing hexamethylated phenyl-bodipy fluorophore attached at position 5 of pyrimidines or at position 7 of 7-deazapurine was developed. All the title labeled nucleosides and dNTPs displayed bright green fluorescence with very high quantum yields. The modified dNmBdpTPs were good substrates to diverse DNA polymerases and were used for in vitro enzymatic synthesis of labeled DNA by primer extension or PCR. In combination with cationic cyclodextrin-peptide-based dNTP transporter, the dNmBdpTPs were successfully used for staining of genomic DNA in living cells for applications in confocal microscopy and in flow cytometry. The best performing cytosine nucleotide dCmBdpTP was used to monitor mitosis in live cells.

Klotho Gene and Protein in Human Placentas According to Birth Weight and Gestational Age.

Introduction: Fetal growth restriction may be the consequence of maternal, fetal, or placental factors. The insulin-like growth factors (IGFs) are major determinants of fetal growth, and are expressed in the mother, fetus and placenta in most species. Previously we reported higher placental protein content of IGF-I, IGF-IR, and AKT in small (SGA) compared with those from appropriate for gestational age (AGA) placentas. The protein Klotho, has been reported in placenta and may regulate IGF-I activity. In this study we determined Klotho gene expression and protein immunostaining in term (T-SGA y T-AGA) and preterm (PT-SGA y PT-AGA) human placentas. In addition, we assessed the effect of Klotho on the IGF-IR and AKT activation induced by IGF-I. Methods: Placentas (n = 1 17) from 32 T-SGA (birth weight (BW) = -1.74 ± 0.08 SDS), 37 T-AGA (BW = 0.12 ± 0.12 SDS), 20 PT-SGA (BW = -2.08 ± 0.14 SDS), and 28 PT-AGA (BW = -0.43 ± 0.13 SDS) newborns were collected. mRNA expression by RT-PCR in the chorionic (CP) and basal (BP) plates of the placentas, and the presence of Klotho was evaluated by immunohistochemistry (integral optical density, IOD). In addition, we developed placental explants that were incubated with IGF-I in the presence or absence of Klotho. Results: We found a lower mRNA expression and protein immunoreactivity of Klotho in the CP of SGA (term and preterm) compared with AGA placentas. We also observed a significant reduction in IGF-IR tyrosine activation induced by IGF-I 10 nM when preincubated with 2.0 nM of Klotho (2.4 ± 0.5 arbitrary units vs. 1.3 ± 0.3 AU), and similar results we observed on AKT and ERK42/44 activation. Conclusion: We describe for the first time that Klotho mRNA and protein varies according to fetal growth and gestational age. In addition, Klotho appears to down-regulate the activation induced by IGF-I on IGF-IR and AKT, suggesting that Klotho may be regulating IGF-I activity in human placentas according to intrauterine fetal growth.

Digestive Physiology of Octopus maya and O. mimus: Temporality of Digestion and Assimilation Processes.

Digestive physiology is one of the bottlenecks of octopus aquaculture. Although, there are successful experimentally formulated feeds, knowledge of the digestive physiology of cephalopods is fragmented, and focused mainly on Octopus vulgaris. Considering that the digestive physiology could vary in tropical and sub-tropical species through temperature modulations of the digestive dynamics and nutritional requirements of different organisms, the present review was focused on the digestive physiology timing of Octopus maya and Octopus mimus, two promising aquaculture species living in tropical (22-30°C) and sub-tropical (15-24°C) ecosystems, respectively. We provide a detailed description of how soluble and complex nutrients are digested, absorbed, and assimilated in these species, describing the digestive process and providing insight into how the environment can modulate the digestion and final use of nutrients for these and presumably other octopus species. To date, research on these octopus species has demonstrated that soluble protein and other nutrients flow through the digestive tract to the digestive gland in a similar manner in both species. However, differences in the use of nutrients were noted: in O. mimus, lipids were mobilized faster than protein, while in O. maya, the inverse process was observed, suggesting that lipid mobilization in species that live in relatively colder environments occurs differently to those in tropical ecosystems. Those differences are related to the particular adaptations of animals to their habitat, and indicate that this knowledge is important when formulating feed for octopus species.

Inhibition of non-templated nucleotide addition by DNA polymerases in primer extension using twisted intercalating nucleic acid modified templates.

A simple and elegant method for inhibition of non-templated nucleotide addition by DNA polymerases and for following DNA 3'-heterogeneity in enzymatic DNA synthesis by primer extension (PEX) is described. When template bearing ortho-twisted intercalating nucleic acid (ortho-TINA) at the 5'-end is used, non-templated nucleotide addition is reduced in both the A- and B-family DNA polymerases (KOD XL, KOD (exo-), Bst 2.0, Therminator, Deep Vent (exo-) and Taq). Formation of a single oligonucleotide product was observed with ortho-TINA modified template and KOD XL, KOD (exo-), Bst 2.0, Deep Vent (exo-) and Taq DNA polymerases. This approach can be applied to the synthesis of both unmodified and base-modified oligonucleotides.