Quantifying terpenes in tomato leaf extracts from different species using gas chromatography-mass spectrometry (GC-MS).

Terpenes play a vital role in plant defense; tomato plants produce a diverse range of terpenes within specialized glandular trichomes, influencing interactions with herbivores, predators, and pollinators. This study employed two distinct methods, namely leaf dip and maceration, to extract trichomes from tomato leaves. Terpene quantification was carried out using Gas Chromatography-Mass Spectrometry (GC-MS). The leaf dip method proved effective in selectively targeting trichome content, revealing unique extraction patterns compared to maceration. The GC-MS method demonstrated high linearity, accuracy, sensitivity, and low limits of detection and quantification. Application of the method to different tomato species (Solanum pennellii, Solanum pimpinellifolium, Solanum galapagense, Solanum habrochaites, and Solanum lycopersicum) identified significant variation in terpene content among these species, highlighting the potential of specific accessions for breeding programs. Notably, the terpene α-zingiberene, known for its repellency against whiteflies, was found in high quantities (211.90-9155.13 µg g-1) in Solanum habrochaites accession PI209978. These findings provide valuable insights into terpenoid diversity for plant defense mechanisms, guiding future research on developing pest-resistant tomato cultivars. Additionally, the study underscores the broader applications of terpenes in agriculture.

Production, characterization, and dietary supplementation effect of rumen-protected fat on ruminal function and blood parameters of sheep.

Rumen-protected fat (RPF) was produced in the 1st experimental stage through melt-emulsification technique using buriti oil (BO) as core, at concentrations of 10% (BO10), 20% (BO20), and 30% (BO30) (w/w), and carnauba wax (CW) as encapsulant material. After obtention and characterization, protected fat microspheres were tested in a 2nd experimental stage on the sheep' diet using six castrated 2-year-old male Santa Ines with initial weight 48.9 ± 5.23 kg, fistulated in rumen and distributed in a double Latin square design with 3 treatments × 3 periods, to evaluate rumen pH, temperature, protozoal count, and blood parameters. There was no difference (P > 0.05) among RPF microspheres for microencapsulation yield. However, microencapsulation efficiency increased (P < 0.05) with BO addition ranging from 36 to 61.3% for BO10 and BO30, respectively. The inclusion of BO10 in the sheep's diet did not affect the ruminal dry matter degradability (DMD) of BO over time (P > 0.05); however, BO20 and BO30 had higher (P < 0.05) DMD values than BO10. No significant differences were observed among RPF for rumen pH and temperature (P > 0.05). There was an increase (P < 0.05) in the protozoal population in the rumen environment due to the microencapsulated BO30 inclusion. There was also increase (P < 0.05) in serum albumin, cholesterol, aspartate aminotransferase (ALT), and gamma-glutamyltransferase (GGT), and a reduction (P < 0.05) in serum triglycerides of the sheep when RPF microspheres increased in the diet. Melt-emulsification proved to be a good technique for microencapsulation of buriti oil into the carnauba wax matrix. RPF from buriti oil protected into carnauba wax is recommended for sheep diet because it increases energy density, without adverse effects on the protozoal populations and blood serum metabolites from the bypass effect in the rumen.

Luffa operculata seed proteins: Identification by LC-ESI-MS/MS and biotechnological potential against Candida albicans and C. krusei.

L: operculata is a plant commonly found in the North and Northeast of Brazil. Although the regional population knows its medicinal potential, there are few scientific studies about its antimicrobial potential. Thus, this study aimed to characterize the proteins from L. operculata seeds extracted using different solutions and evaluate their antimicrobial potentials. The protein extracts obtained with NaCl and sodium acetate buffer presented the best inhibitory activities against Candida albicans and C. krusei. The study of the mechanism of action revealed proteins from L. operculata seeds induced pore formation on the membrane and ROS overaccumulation. Scanning Electron Microscopy images also showed severe morphological changes in Candida albicans and C. krusei. Proteins from L.operculata seeds did not show antibacterial activity. The enzymatic assays revealed the presence of proteolytic enzymes, serine and cysteine protease inhibitors, and chitinases in both protein extracts. Proteomic analysis by LC-ESI-MS/MS identified 57 proteins related to many biological processes, such as defense to (a)biotic stress, energetic metabolism, protein folding, and nucleotide metabolism. In conclusion, the L. operculata seed proteins have biotechnological potential against the human pathogenic yeasts Candida albicans and C. krusei.

Novel potent (dihydro)benzofuranyl piperazines as human histamine receptor ligands - Functional characterization and modeling studies on H3 and H4 receptors.

Histamine acts through four different receptors (H1R-H4R), the H3R and H4R being the most explored in the last years as drug targets. The H3R is a potential target to treat narcolepsy, Parkinson's disease, epilepsy, schizophrenia and several other CNS-related conditions, while H4R blockade leads to anti-inflammatory and immunomodulatory effects. Our group has been exploring the dihydrobenzofuranyl-piperazines (LINS01 series) as human H3R/H4R ligands as potential drug candidates. In the present study, a set of 12 compounds were synthesized from adequate (dihydro)benzofuran synthons through simple reactions with corresponding piperazines, giving moderate to high yields. Four compounds (1b, 1f, 1g and 1h) showed high hH3R affinity (pKi > 7), compound 1h being the most potent (pKi 8.4), and compound 1f showed the best efficiency (pKi 8.2, LE 0.53, LLE 5.85). BRET-based assays monitoring Gαi activity indicated that the compounds are potent antagonists. Only one compound (2c, pKi 7.1) presented high affinity for hH4R. In contrast to what was observed for hH3R, it showed partial agonist activity. Docking experiments indicated that bulky substituents occupy a hydrophobic pocket in hH3R, while the N-allyl group forms favorable interactions with hydrophobic residues in the TM2, 3 and 7, increasing the selectivity towards hH3R. Additionally, the importance of the indole NH in the interaction with Glu5.46 from hH4R was confirmed by the modeling results, explaining the affinity and agonistic activity of compound 2c. The data reported in this work represent important findings for the rational design of future compounds for hH3R and hH4R.

Effectiveness of a reciprocating single file, single cone endodontic treatment approach: a randomized controlled pragmatic clinical trial.

OBJECTIVES: To compare the root filling quality, the sealer extrusion, and the healing rates of apical lesions addressed via two endodontic treatment approaches. The hypothesis tested was that both techniques present similar apical periodontitis healing results. MATERIALS AND METHODS: This study was a parallel-design, pragmatic, and randomized clinical trial. One hundred twenty anterior teeth with necrotic pulps and apical periodontitis were randomly allocated to be either instrumented with hand files and obturated with the lateral compaction technique or instrumented with a single file in a reciprocating movement and obturated with a single-cone technique. The root canal filling quality, the occurrence of sealer extrusion, and apical periodontitis healing were the outcomes of interest. Data were analyzed through chi-square analysis, and the odds ratio for healing was adjusted using a logistic regression model (α = 0.05). RESULTS: No significant differences were observed between the endodontic techniques regarding the root filling quality or sealer extrusion. Six months after treatment, both endodontic techniques presented equivalent results regarding apical periodontitis healing. The healing rate was affected only by the periapical status at baseline. CONCLUSIONS: The obturation of the root canal of the anterior teeth using reciprocating file-matched single cones presented a similar quality to that obtained with manual instrumentation followed by the lateral condensation technique. Similar healing rates of apical lesions were also observed. CLINICAL RELEVANCE: For treating anterior teeth with apical periodontitis, a reciprocating single-file, single-treatment protocol was as effective as a traditional protocol combining hand instrumentation and the lateral compaction obturation technique.

Profile of Educational Technology Use by Medical Students and Evaluation of a New Mobile Application Designed for the Study of Human Physiology.

Nowadays, smartphones represent an invaluable tool to access educational material; however, the available information is not always accurate or evidence-based. Therefore, we aimed to evaluate the use of technology by medical students and assess the effect of a newly developed mobile app for the study of human physiology. We used a standardised questionnaire to assess the profile of educational technology use, from which a mobile app (PhysioQuiz) was developed. The effectiveness and user opinion were assessed in a randomised controlled study (n = 110). Of 1022 students enrolled in medical school, 489 (47.9%) participated in the study. Of the respondents, 96.7% used mobile applications, with the main purpose being entertainment (94.7%) and study (81.9%). Only 6.1% reported use of physiology apps. PhysioQuiz use did not yield higher average grades (p = 0.48); however, user opinion demonstrated that it was useful for assisted learning (82.1%) and identification of non-learned content (78.6%) and considered a tool for self-assessment (89.3%). Mobile app use is widespread among medical students but there is a lack of human physiology education apps. A newly developed app for the study of human physiology was useful for assisted learning and considered a tool for self-assessment.

A systematic review and meta-analysis of the aetiology of acute pancreatitis.

BACKGROUND: Gallstones and alcohol are currently the most frequent aetiologies of acute pancreatitis (AP). The aim of this study is to quantify these aetiologies worldwide, by geographic region and by diagnostic method. METHODS: A systematic review of observational studies published from January 2006 to October 2017 was performed. The studies provided objective criteria for establishing the diagnosis and aetiology of AP for at least biliary and alcoholic causes. A random-effects meta-analysis was used to assess the frequency of biliary (ABP), alcoholic (AAP) and idiopathic AP (IAP) worldwide and to perform 6 subgroup analyses: 2 compared diagnostic methods for AP aetiology and the other 4 compared geographic regions. RESULTS: Forty-six studies representing 2,341,007 patients of AP in 36 countries were included. The global estimate of proportion (95% CI) of aetiologies was 42 (39-44)% for ABP, 21 (17-25)% for AAP and 18 (15-22)% for IAP. In studies that used discharge code diagnoses and in those from the US, IAP was the most frequent aetiology. ABP was more frequent in Latin America than in other regions. CONCLUSION: Gallstones represent the main aetiology of AP globally, and this aetiology is twice as frequent as the second most common aetiology.

Mini-incubators improve the adventitious rooting performance of Corymbia and Eucalyptus microcuttings according to the environment in which they are conditioned.

We addressed a major challenge in the in vitro clonal propagation of Corymbia citriodora, Eucalyptus urophylla and E. benthamii by using an ex vitro adventitious rooting strategy in a mini-incubator. Mini-incubators were placed in four environments for rooting. A shade house with no fogging system and a greenhouse with no ventilation but with a fogging environment had the best performance in terms of rooting, root growth and survival of microcuttings. Daily recording of the temperature within each mini-incubator in each environment allowed the verification of negative correlations between the maximum average temperature and the survival, adventitious rooting and root growth. The ideal maximum air temperature for the efficient production of clonal plants was 28.4°C (± 5.5°C), and the minimum was 20.3°C (± 6.2°C). E. benthamii was more sensitive to higher temperatures than C. citriodora and E. urophylla. Nevertheless, placing mini-incubators in the shade house with no fogging system resulted in a stable and uniform performance among the three species, with 100.0% survival and 81.4% rooting. Histological sections of the adventitious roots revealed connection with the stem vascular cambium. Therefore, our experimental system demonstrated the potential of mini-incubators coupled with the proper environment to optimize the adventitious rooting performance of microcuttings.

Effectiveness of high-power LEDs to polymerize resin cements through ceramics: An in vitro study.

STATEMENT OF PROBLEM: The cementation of ceramic veneers using light-polymerized resin cement is largely dependent on the proper light activation of the cement. Light activation using high irradiance could shorten the time required to lute multiple restorations. PURPOSE: The purpose of this in vitro study was to evaluate the light transmission of dental light-polymerizing units through ceramic cylinders and its effect on the polymerization kinetics of a resin cement. MATERIAL AND METHODS: Ceramic ingots (IPS Empress Esthetic, shade ET1) were sectioned to produce cylinders 0.5, 1.0, and 2.0 mm thick. Two light-emitting diode units were evaluated: SmartLite Focus and Valo Cordless, the latter used in either Standard or Xtra Power (XP) modes. Light transmission (average of irradiance, total energy, and light-emission profile) through the cylinders was measured (n=3). The polymerization kinetics of a resin cement light polymerized through the ceramic was monitored for 5 minutes (n=3). The degree of conversion was measured again after 72 hours. Data were individually analyzed with 2-way ANOVA and the Tukey HSD test (α=.05). RESULTS: Valo at XP presented the highest values of irradiance and SmartLite the lowest, irrespective of the ceramic thickness. Regarding the total energy, XP showed the lowest values. The total energy and irradiance lessened with the increase in ceramic thickness. In general, except for Valo at XP, the ceramic thickness did not affect the degree of conversion. Valo at XP and interposing 2.0 mm ceramic resulted in the lowest values of Rpmax. CONCLUSIONS: The reduction of total energy and irradiance by ceramic interposition had only a slight effect on polymerization kinetics.

Stearylamine-Containing Cationic Nanoemulsion as a Promising Carrier for Gene Delivery.

The drawbacks related to the use of viral vectors in gene therapy have been stimulated the research in non-viral strategies such as cationic nanoemulsions. The aim of this work was to develop a stearylamine-containing nanoemulsion for gene therapy purpose. The formulation was chosen from a Pseudo-Ternary Phase Diagram and had its long-term stability assessed by Dynamic Light Scattering and Phase Analysis Light Scattering during 180 days at 4 degrees C, 25 degrees C and 40 degrees C. Besides, studies of sterilization and scale up of the product were conducted. It was demonstrated that the proposed system was stable up to 180 days when stored at 4 degrees C and could be sterilized by a 0.22 microm filter pore without changes on its characteristics. The scale up was possible by adjusting the volume to the sonication time. Because the nanoemulsion presented a droplet size smaller than 200 nm and a zeta potential higher than 30 mV, this system was able to correctly complex the plasmid model PIRES2-EGFP, as confirmed by the agarosis gel electrophoresis assay. The nanoemulsion toxicity evaluated over lung fetus human cells (MRC-5) was dose-dependent. However, it does not appear to be a limiting factor for further experiments aiming gene transfection. As a conclusion, stearylamine-containing cationic nanoemulsions can be used for gene therapy, since it presents suitable characteristics, stability and possibility of sterilization.