Unveiling Novel Kunitz- and Waprin-Type Toxins in the Micrurus mipartitus Coral Snake Venom Gland: An In Silico Transcriptome Analysis.

Kunitz-type peptide expression has been described in the venom of snakes of the Viperidae, Elapidae and Colubridae families. This work aimed to identify these peptides in the venom gland transcriptome of the coral snake Micrurus mipartitus. Transcriptomic analysis revealed a high diversity of venom-associated Kunitz serine protease inhibitor proteins (KSPIs). A total of eight copies of KSPIs were predicted and grouped into four distinctive types, including short KSPI, long KSPI, Kunitz-Waprin (Ku-WAP) proteins, and a multi-domain Kunitz-type protein. From these, one short KSPI showed high identity with Micrurus tener and Austrelaps superbus. The long KSPI group exhibited similarity within the Micrurus genus and showed homology with various elapid snakes and even with the colubrid Pantherophis guttatus. A third group suggested the presence of Kunitz domains in addition to a whey-acidic-protein-type four-disulfide core domain. Finally, the fourth group corresponded to a transcript copy with a putative 511 amino acid protein, formerly annotated as KSPI, which UniProt classified as SPINT1. In conclusion, this study showed the diversity of Kunitz-type proteins expressed in the venom gland transcriptome of M. mipartitus.

In-depth immunorecognition and neutralization analyses of Micrurus mipartitus and M. dumerilii venoms and toxins by a commercial antivenom.

In Colombia, the Micrurus genus comprises 30 species, including M. mipartitus and M. dumerilii, which are of major clinical relevance due to their wide geographical distribution and the number of snakebites inflicted by them. These neurotoxic envenomations are characterized by neuromuscular paralysis attributed to venom components such as three-finger toxins (3FTx) and phospholipases (PLA2). Additionally, there is limited information available on the neutralizing coverage of commercially available antivenoms, underscoring the need to perform studies to assess the cross-neutralizing ability of these life-saving products. Therefore, we present an in-depth immunorecognition analysis by the anticoral-INS antivenom from Colombia on the M. mipartitus and M. dumerilii venoms. The antivenom cross-recognized the whole venoms and their components with different intensities. For instance, the antivenom showed better recognition on PLA2s than on 3FTxs in both venoms. Moreover, at doses tested, the antivenom totally neutralized the lethal effect of M. dumerilii venom; however, it did not neutralize this effect induced by M. mipartitus venom and its main toxic components from the southwestern region of the department of Antioquia. Furthermore, the anticoral-INS antivenom displayed better cross-immunorecognition of PLA2-predominant Micrurus venoms than of 3FTx-predominant Micrurus venoms. This highlights the need to include venoms from both types of venom patterns in the immunization mixture to produce antivenoms against coral snakes. Finally, our results suggest the need for further research to optimize the composition of immunizing mixtures for antivenom production and improve their efficacy against coral snake envenomation in Colombia and the Americas.

Knowledge about Snake Venoms and Toxins from Colombia: A Systematic Review.

Colombia encompasses three mountain ranges that divide the country into five natural regions: Andes, Pacific, Caribbean, Amazon, and Orinoquia. These regions offer an impressive range of climates, altitudes, and landscapes, which lead to a high snake biodiversity. Of the almost 300 snake species reported in Colombia, nearly 50 are categorized as venomous. This high diversity of species contrasts with the small number of studies to characterize their venom compositions and natural history in the different ecoregions. This work reviews the available information about the venom composition, isolated toxins, and potential applications of snake species found in Colombia. Data compilation was conducted according to the PRISMA guidelines, and the systematic literature search was carried out in Pubmed/MEDLINE. Venom proteomes from nine Viperidae and three Elapidae species have been described using quantitative analytical strategies. In addition, venoms of three Colubridae species have been studied. Bioactivities reported for some of the venoms or isolated components-such as antibacterial, cytotoxicity on tumoral cell lines, and antiplasmodial properties-may be of interest to develop potential applications. Overall, this review indicates that, despite recent progress in the characterization of venoms from several Colombian snakes, it is necessary to perform further studies on the many species whose venoms remain essentially unexplored, especially those of the poorly known genus Micrurus.

Influence of the number of hosts and the risk of predation on the foraging behavior of the parasitoid Eretmocerus eremicus.

BACKGROUND: Parasitoids are natural enemies that can modify their behaviors as they search for hosts based on the characteristics of the sites in which they forage. Theoretical models predict that a parasitoid will stay for longer periods in high-quality sites or patches than in those of low quality. In turn, patch quality may be linked to factors such as the number of hosts and the risk of predation. In the present study, we sought to determine if the factors of the number of hosts, risk of predation, and their interaction influence the foraging behavior of the parasitoid Eretmocerus eremicus (Hymenoptera: Aphelinidae) as theory predicts. To do this, we evaluated different parameters of parasitoid foraging behavior, such as residence time, number of oviposition events, and attacks, in sites of different patch quality. RESULTS: Our results show that when the factors, number of hosts, and, risk of predation, were evaluated separately, E. eremicus resided for longer periods and oviposited more frequently in patches with a high number of hosts and low risk of predation than in other patches. However, when both of these factors were combined, only the number of hosts influenced some aspects of the foraging behavior of this parasitoid, such as the number of oviposition events and attacks. CONCLUSION: For some parasitoids like E. eremicus, theoretical predictions may be fulfilled when patch quality is linked to the number of hosts but will not be fully satisfied when patch quality is related to the risk of predation. Furthermore, at sites with different combinations of host numbers and risk of predation, host number appears to be more critical than predation risk. These results suggest that the performance of the parasitoid E. eremicus to control whiteflies will be mainly mediated by the levels of whitefly infestation and, to a small extent, by the risk of predation to which the parasitoid is subjected. © 2023 Society of Chemical Industry.

Identification of genetic and biochemical mechanisms associated with heat shock and heat stress adaptation in grain amaranths.

Heat stress is poised to become a major factor negatively affecting plant performance worldwide. In terms of world food security, increased ambient temperatures are poised to reduce yields in cereals and other economically important crops. Grain amaranths are known to be productive under poor and/or unfavorable growing conditions that significantly affect cereals and other crops. Several physiological and biochemical attributes have been recognized to contribute to this favorable property, including a high water-use efficiency and the activation of a carbon starvation response. This study reports the behavior of the three grain amaranth species to two different stress conditions: short-term exposure to heat shock (HS) conditions using young plants kept in a conditioned growth chamber or long-term cultivation under severe heat stress in greenhouse conditions. The latter involved exposing grain amaranth plants to daylight temperatures that hovered around 50°C, or above, for at least 4 h during the day and to higher than normal nocturnal temperatures for a complete growth cycle in the summer of 2022 in central Mexico. All grain amaranth species showed a high tolerance to HS, demonstrated by a high percentage of recovery after their return to optimal growing conditions. The tolerance observed coincided with increased expression levels of unknown function genes previously shown to be induced by other (a)biotic stress conditions. Included among them were genes coding for RNA-binding and RNA-editing proteins, respectively. HS tolerance was also in accordance with favorable changes in several biochemical parameters usually induced in plants in response to abiotic stresses. Conversely, exposure to a prolonged severe heat stress seriously affected the vegetative and reproductive development of all three grain amaranth species, which yielded little or no seed. The latter data suggested that the usually stress-tolerant grain amaranths are unable to overcome severe heat stress-related damage leading to reproductive failure.

Physiological Responses of Agave maximiliana to Inoculation with Autochthonous and Allochthonous Arbuscular Mycorrhizal Fungi.

The benefits of mycorrhizal interactions are only known in 8 of 210 recognized Agave taxa. We evaluated the effects of autochthonous and allochthonous arbuscular mycorrhizal fungi (AMF) on growth and nutrient assimilation in Agave maximiliana. The autochthonous consortium (Cn) of eight species was propagated from the rhizospheric soil of A. maximiliana, while Claroideoglomus claroideum (Cc) and Claroideoglomus etunicatum (Ce) were employed as allochthonous AMF. Six treatments were included in the study: Cn, Ce, Cc, Ce + Cc, Tf (fertilized control), and Tn (non-fertilized control, not inoculated). Mycorrhizal colonization increased over time, and the colonization percentages produced by Cn and the allochthonous AMF, both alone and mixed together, were equal at 6, 12, and 18 months. Height increased steadily and was higher in AMF-treated plants from seven months onward. Growth indicators of AMF-treated and AMF-free plants were equal at 6 months, but the beneficial effects of allochthonous and autochthonous AMF were evident in all growth indicators at 18 months and in sugar and mineral (P, K, Ca, Mg, and Fe) content. Arbuscular mycorrhizal fungi significantly improved all growth parameters of A. maximiliana regardless of the origin of the inoculums. This is the first study to report the positive effects of AMF colonization in A. maximiliana.

Antibodies against a single fraction of Micrurus dumerilii venom neutralize the lethal effect of whole venom.

The coralsnake Micrurus dumerilii (Elapidae) is reported to cause envenomings of medical importance. Previous studies characterized the protein composition of its venom, with phospholipase A2 (PLA2) proteins the most abundant. However, it is unknown which venom components are responsible for its lethal toxicity. Fractionation of M. dumerilii venom from Colombia was carried out using RP-HPLC and each fraction was screened for lethal effect in mice at a dose of 20 µg by intraperitoneal route. Results showed that only one fraction, F9, was lethal. This fraction displayed PLA2 activity, induced indirect hemolysis in vitro, as well as edema and myotoxicity in vivo. SDS-PAGE of unreduced F9 evidenced two bands of 8 and 15 kDa, respectively, consistent with the detection of proteins with masses of 13,217.77 Da, 7144.06 Da, and 7665.55 Da. Tryptic digestion of F9 followed by nESI-MS/MS revealed peptide sequences matching proteins of the three-finger toxin (3FTx) and PLA2 families. Immunization of a rabbit with F9 proteins elicited antibody titers up to 1:10,000 by ELISA. After serum fractionation with caprylic acid, the obtained IgG was able to neutralize the lethal effect of the complete venom of M. dumerilii using a challenge of 2 ×LD50 at the IgG/venom ratio of 50:1 (w/w). In conclusion, present results show that the lethal effect of M. dumerilii venom in mice is mainly driven by one fraction which contains 3FTx and PLA2 proteins. The antibodies produced against this fraction cross-recognized other PLA2s and neutralized the lethal effect of whole M. dumerilii venom, pointing out to the potential usefulness of F9 as a relevant antigen for improving current coral snake antivenoms.

Actividad de fosfolipasas y proteasas en aislamientos de especies de Candida colonizadoras y causantes de vulvovaginitis en mujeres gestantes / Phospholipase and proteinase activities of isolatesof colonizing Candida spp. causing vulvovaginitis in pregnant women

Introducción. Las proteasas y las fosfolipasas son factores de virulencia de Candida spp. que cumplen un papel importante en la invasión de los tejidos. Entre los factores relacionados con el huésped, se encuentran algunos asociados con las características ambientales y otros con la colonización. Objetivo. Determinar la actividad de fosfolipasas y proteasas en aislamientos de especies colonizadoras y patógenas de Candida spp., aisladas de mujeres gestantes de Cartagena de Indias. Materiales y métodos. Se determinó la actividad de fosfolipasas y proteasas en 56 aislamientos mediante degradación del sustrato y cálculo del coeficiente de actividad enzimática. Se compararon las actividades de fosfolipasas y proteasas, entre los aislamientos colonizadores y los patógenos. Resultados. La actividad de la fosfolipasa fue "muy alta" (< 0,69) en 34 aislamientos e, igualmente, la de la proteasa en 14. No hubo diferencias significativas al comparar las actividades de las fosfolipasas y de las de las proteasas, entre los aislamientos colonizadores y los patógenos. Conclusiones. La actividad de las fosfolipasas predominó como factor de virulencia en los aislamientos estudiados. No obstante, no se encontró una diferencia significativa entre los grupos de aislamientos colonizadores y los patógenos, en cuanto a las actividades de fosfolipasas y proteasas.

Introduction. Proteases and phospholipases are virulence factors of Candida spp. that play an important role in tissue invasion. Among the factors related to the host some are associated with environmental characteristics and others with Candida colonization. Objectives. To determine phospholipase and protease activities in colonizing and pathogenic strains, isolated from pregnant women in Cartagena de Indias. Materials and methods. Phospholipase and protease activity was determined in 56 isolates, evaluating substrate degradation and calculating the enzyme activity coefficient. Phospholipase and protease activities were compared between colonizing and pathogenic strains. Results. "Very high" (<0.69) phospholipase and protease activity was found in 34 and 14 isolates, respectively. There was no significant difference when comparing phospholipase and protease activities between colonizing and pathogenic isolates. Conclusions. Phospholipase activity predominated as a virulence factor in the studied strains, but no significant difference was found between colonizing and pathogenic strains for phospholipase and protease activities.

Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A2 from Coral Snake Micrurus dumerilii to Develop Antivenoms.

Micrurus dumerilii is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases A2 being MdumPLA2 the most abundant protein. Nevertheless, Micrurus species produce a low quantity of venom, which makes it difficult to produce anticoral antivenoms. Therefore, in this work, we present the recombinant expression of MdumPLA2 to evaluate its biological activities and its immunogenic potential to produce antivenoms. For this, a genetic construct rMdumPLA2 was cloned into the pET28a vector and expressed heterologously in bacteria. His-rMdumPLA2 was extracted from inclusion bodies, refolded in vitro, and isolated using affinity and RP-HPLC chromatography. His-rMdumPLA2 was shown to have phospholipase A2 activity, a weak anticoagulant effect, and induced myonecrosis and edema. The anti-His-rMdumPLA2 antibodies produced in rabbits recognized native PLA2, the complete venom of M. dumerilii, and a phospholipase from another species of the Micrurus genus. Antibodies neutralized 100% of the in vitro phospholipase activity of the recombinant toxin and a moderate percentage of the myotoxic activity of M. dumerilii venom in mice. These results indicate that His-rMdumPLA2 could be used as an immunogen to improve anticoral antivenoms development. This work is the first report of an M. dumerilii functional recombinant PLA2.

Habitat-adapted heterologous symbiont Salinispora arenicola promotes growth and alleviates salt stress in tomato crop plants.

To ensure food security given the current scenario of climate change and the accompanying ecological repercussions, it is essential to search for new technologies and tools for agricultural production. Microorganism-based biostimulants are recognized as sustainable alternatives to traditional agrochemicals to enhance and protect agricultural production. Marine actinobacteria are a well-known source of novel compounds for biotechnological uses. In addition, former studies have suggested that coral symbiont actinobacteria may support co-symbiotic photosynthetic growth and tolerance and increase the probability of corals surviving abiotic stress. We have previously shown that this activity may also hold in terrestrial plants, at least for the actinobacteria Salinispora arenicola during induced heterologous symbiosis with a wild Solanaceae plant Nicotiana attenuata under in vitro conditions. Here, we further explore the heterologous symbiotic association, germination, growth promotion, and stress relieving activity of S. arenicola in tomato plants under agricultural conditions and dig into the possible associated mechanisms. Tomato plants were grown under normal and saline conditions, and germination, bacteria-root system interactions, plant growth, photosynthetic performance, and the expression of salt stress response genes were analyzed. We found an endophytic interaction between S. arenicola and tomato plants, which promotes germination and shoot and root growth under saline or non-saline conditions. Accordingly, photosynthetic and respective photoprotective performance was enhanced in line with the induced increase in photosynthetic pigments. This was further supported by the overexpression of thermal energy dissipation, which fine-tunes energy use efficiency and may prevent the formation of reactive oxygen species in the chloroplast. Furthermore, gene expression analyses suggested that a selective transport channel gene, SlHKT1,2, induced by S. arenicola may assist in relieving salt stress in tomato plants. The fine regulation of photosynthetic and photoprotective responses, as well as the inhibition of the formation of ROS molecules, seems to be related to the induced down-regulation of other salt stress response genes, such as SlDR1A-related genes or SlAOX1b. Our results demonstrate that the marine microbial symbiont S. arenicola establishes heterologous symbiosis in crop plants, promotes growth, and confers saline stress tolerance. Thus, these results open opportunities to further explore the vast array of marine microbes to enhance crop tolerance and food production under the current climate change scenario.

Semiactive Knee Orthotic Using a MR Damper and a Smart Insole to Control the Damping Force Sensing the Plantar Pressure.

This work presents the development of semiactive knee orthosis prototype that focus to absorb the forces and impacts in this joint during the human gait. This prototype consists of three subsystems: the first is a wireless and portable system capable of measuring the ground reaction forces in the stance phase of the gait cycle, by means of an instrumented insole with force sensing resistors strategically placed on the sole of the foot, an electronic device allows processing and transmit this information via Bluetooth to the control system. The second is a semiactive actuator, which has inside a magnetorheological fluid, highlighting its ability to modify its damping force depending on the intensity of the magnetic field that circulates through the MR fluid. It is regulated by a Proportional Derivative (PD) controller system according to the values of plantar pressure measured by the insole. The third component is a mechanical structure manufactured by 3D printing, which adapts to the morphology of the human leg. This exoskeleton is designed to support the forces on the knee controlling the action of the magnetorheological actuator by ground reaction forces. The purpose of this assistance system is to reduce the forces applied to the knee during the gait cycle, providing support and stability to this joint. The obtained experimental results indicate that the device fulfills the function by reducing 12 % of the impact forces on the user's knee.

Immunorecognition and Neutralization of Crotalus durissus cumanensis Venom by a Commercial Antivenom Produced in Colombia.

In Colombia, on average 2.9% of the nearly 5600 snakebite events that occur annually involve the rattlesnake Crotalus durissus cumanensis. The envenomation by this snake is mainly characterized by neurotoxicity and the main toxin is crotoxin (~64.7% of the total venom). The Instituto Nacional de Salud (INS) produces a polyvalent antivenom aimed at the treatment of bothropic, crotalid, and lachesic envenomations; nonetheless, its immune reactivity profile and neutralizing capacity over biological activities of the C. d. cumanensis venom has been poorly evaluated. In this sense, the study aims: (1) to describe an in-depth exploration of its immunoreactivity through second-generation antivenomics and HPLC fraction-specific ELISA immunoprofiles; and (2) to evaluate the neutralization pattern of the rattlesnake venom in vitro and in vivo biological activities. The results obtained showed a variable recognition of crotoxin subunits, in addition to a molecular mass-dependent immunoreactivity pattern in which the disintegrins were not recognized, and snake venom metalloproteinases and L-amino acid oxidases were the most recognized. Additionally, a high neutralization of proteolytic and coagulant activities was observed, but not over the PLA2 activity. Further, the median effective dose against C. d. cumanensis venom lethality was 962 µL of antivenom per mg of venom. In conclusion, (1) the antivenom recognition over the crotoxin and the disintegrins of the C. d. cumanensis should be improved, thus aiming upcoming efforts for the exploration of new techniques and approaches in antivenom production in Colombia, and (2) the neutralization activity of the antivenom seems to follow the molecular mass-dependent recognition pattern, although other explanations should be explored.

Anti-Neurotoxins from Micrurus mipartitus in the Development of Coral Snake Antivenoms.

In Colombia, the genus Micrurus includes 30 species, of which M. mipartitus and M. dumerilii are the most widely distributed. Micrurus causes less than 3% of the approximately 5000 cases of snakebite per year. The elapid envenomation caused by the snakes from the Micrurus genus, are characterized by the severity of their clinical manifestations, due to the venom neurotoxic components such as three-finger toxins (3FTx) and phospholipases (PLA2). The treatment for snakebites is the administration of specific antivenoms, however, some of them have limitations in their neutralizing ability. A strategy proposed to improve antivenoms is to produce antibodies against the main components of the venom. The aim of this work was to produce an antivenom, using an immunization protocol including the main 3FTx and PLA2 responsible for M. mipartitus lethality. The antibody titers were determined by ELISA in rabbits' serum. The immunized animals elicited a response against toxins and whole venom. The Immunoglobulin G (IgGs) obtained were able to neutralize the lethal effect of their homologous toxins. A combination of antivenom from M. mipartitus with antitoxins improved their neutralizing ability. In the same way, a mixture of anti 3FTx and PLA2 protected the mice from a 1.5 median lethal dose (LD50) of M. mipartitus venom. The results showed that this might be a way to improve antibody titers specificity against the relevant toxins in M. mipartitus venom and indicated that there is a possibility to develop and use recombinant 3FTx and PLA2 toxins as immunogens to produce antivenoms. Additionally, this represents an alternative to reduce the amount of venom used in anti-coral antivenom production.

Phospholipase and proteinase activities of isolates of colonizing Candida spp. causing vulvovaginitis in pregnant women / Actividad de fosfolipasas y proteasas en aislamientos de especies de Candida colonizadoras y causantes de vulvovaginitis en mujeres gestantes

Introduction: Proteases and phospholipases are virulence factors of Candida spp. that play an important role in tissue invasion. Among the factors related to the host some are associated with environmental characteristics and others with Candida colonization. Objectives: To determine phospholipase and protease activities in colonizing and pathogenic strains, isolated from pregnant women in Cartagena de Indias. Materials and methods: Phospholipase and protease activity was determined in 56 isolates, evaluating substrate degradation and calculating the enzyme activity coefficient. Phospholipase and protease activities were compared between colonizing and pathogenic strains. Results: "Very high" (<0.69) phospholipase and protease activity was found in 34 and 14 isolates, respectively. There was no significant difference when comparing phospholipase and protease activities between colonizing and pathogenic isolates. Conclusions: Phospholipase activity predominated as a virulence factor in the studied strains, but no significant difference found between colonizing and pathogenic strains for phospholipase and protease activities.

Introducción: Las proteasas y las fosfolipasas son factores de virulencia de Candida spp. que cumplen un papel importante en la invasión de los tejidos. Entre los factores relacionados con el huésped, se encuentran algunos asociados con las características ambientales y otros con la colonización. Objetivo: Determinar la actividad de fosfolipasas y proteasas en aislamientos de especies colonizadoras y patógenas de Candida spp., aisladas de mujeres gestantes de Cartagena de Indias. Materiales y métodos. Se determinó la actividad de fosfolipasas y proteasas en 56 aislamientos mediante degradación del sustrato y cálculo del coeficiente de actividad enzimática. Se compararon las actividades de fosfolipasas y proteasas, entre los aislamientos colonizadores y los patógenos. Resultados: La actividad de la fosfolipasa fue "muy alta" (< 0,69) en 34 aislamientos e, igualmente, la de la proteasa en 14. No hubo diferencias significativas al comparar las actividades de las fosfolipasas y de las de las proteasas, entre los aislamientos colonizadores y los patógenos. Conclusiones: La actividad de las fosfolipasas predominó como factor de virulencia en los aislamientos estudiados. No obstante, no se encontró una diferencia significativa entre los grupos de aislamientos colonizadores y los patógenos, en cuanto a las actividades de fosfolipasas y proteasas.

Influence of Educational Background, Childhood Socioeconomic Environment, and Language Use on Cognition among Spanish-Speaking Latinos Living Near the US-Mexico Border.

OBJECTIVES: We investigated the impact of culturally relevant social, educational, and language factors on cognitive test performance among Spanish speakers living near the US-Mexico border. METHODS: Participants included 254 healthy native Spanish speakers from the Neuropsychological Norms for the US-Mexico Border Region in Spanish (NP-NUMBRS) project (Age: M = 37.3, SD = 10.4; Education: M = 10.7, SD = 4.3; 59% Female). A comprehensive neuropsychological battery was administered in Spanish. Individual test scaled scores and T-scores (based on region-specific norms adjusted for age, education, and sex) were averaged to create Global Mean Scaled and T-scores. Measures of culturally relevant factors included a self-reported indicator of educational quality/access (proportion of education in Spanish-speaking country, quality of school/classroom setting, stopped attending school to work), childhood socioeconomic environment (parental education, proportion of time living in Spanish-speaking country, childhood socioeconomic and health status, access to basic resources, work as a child), and Spanish/English language use and fluency. RESULTS: Several culturally relevant variables were significantly associated with unadjusted Global Scaled Scores in univariable analyses. When using demographically adjusted T-scores, fewer culturally relevant characteristics were significant. In multivariable analyses, being bilingual (p = .04) and working as a child for one's own benefit compared to not working as a child (p = .006) were significantly associated with higher Global Mean T-score, accounting for 9% of variance. CONCLUSIONS: Demographically adjusted normative data provide a useful tool for the identification of brain dysfunction, as these account for much of the variance of sociocultural factors on cognitive test performance. Yet, certain culturally relevant variables still contributed to cognitive test performance above and beyond basic demographics, warranting further investigation.

Physicochemical Properties, Antioxidant Capacity, Prebiotic Activity and Anticancer Potential in Human Cells of Jackfruit (Artocarpus heterophyllus) Seed Flour.

The general aim of this study was to evaluate physicochemical properties, prebiotic activity and anticancer potential of jackfruit (Artocarpus heterophyllus) seed flour. The drying processes of jackfruit seeds were performed at 50, 60 and 70 °C in order to choose the optimal temperature for obtaining the flour based on drying time, polyphenol content and antioxidant capacity. The experimental values of the moisture ratio during jackfruit seed drying at different temperatures were obtained using Page's equation to establish the drying time for the required moisture between 5 and 7% in the flour. The temperature of 60 °C was considered adequate for obtaining good flour and for performing its characterization. The chemical composition, total dietary fiber, functional properties and antioxidant capacity were then examined in the flour. The seed flour contains carbohydrates (73.87 g/100 g), dietary fiber (31 g/100 g), protein (14 g/100 g) and lipids (1 g/100 g). The lipid profile showed that the flour contained monounsaturated (4 g/100 g) and polyunsaturated (46 g/100 g) fatty acids. Sucrose, glucose, and fructose were found to be the predominant soluble sugars, and non-digestible oligosaccharides like 1-kestose were also found. The total polyphenol content was 2.42 mg of gallic acid/g of the sample; furthermore, the antioxidant capacity obtained by ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) was 901.45 µmol Trolox/100 g and 1607.87 µmol Trolox/100 g, respectively. The obtained flour exhibited good functional properties, such as water and oil absorption capacity, swelling power and emulsifier capacity. Additionally, this flour had a protective and preventive effect which is associated with the potential prebiotic activity in Lactobacillus casei and Bifidobacterium longum. These results demonstrate that jackfruit seed flour has good nutritional value and antioxidant and prebiotic activity, as well as potential protective effects and functional properties, making it an attractive food or ingredient in developing innovative functional products.

Demographically-adjusted norms for selected tests of verbal fluency: Results from the Neuropsychological Norms for the US-Mexico Border Region in Spanish (NP-NUMBRS) project.

OBJECTIVE: Verbal fluency tests are sensitive to various disorders affecting the central nervous system and are commonly included in neuropsychological evaluations. We aimed to develop normative data for two verbal fluency tests in a sample of native Spanish-speakers living in the US-Mexico border region. METHOD: Participants included 254 adults from the Neuropsychological Norms for the US-Mexico Border Region in Spanish (NP-NUMBRS) Project (Age: range = 19-60; Education: range = 0-20, 59% female). Participants completed two verbal fluency tests (i.e., letter [PMR] and semantic/category fluency [Animal Naming]) as part of a larger neuropsychological test battery. We examined linear and nonlinear effects of demographic factors (age, education, and gender) on verbal fluency raw scores, and developed T-scores using fractional polynomial equations controlling for demographics. We also calculated the rates of "impairment" (T-scores < 40) that would be obtained by applying the newly developed norms and available norms for non-Hispanic English-speakers on comparable tests. RESULTS: There were positive small effects of age and medium effects of education on verbal fluency raw scores. The normalized distribution of T-scores with the new norms showed expected psychometric properties. However, rates of impairment for both letter and semantic fluency were significantly higher when applying non-Hispanic White norms, and significantly lower when applying non-Hispanic Black norms. CONCLUSIONS: We provide norms for Spanish-speakers living along the US-Mexico border region for two verbal fluency tests that are co-normed with a more extensive neuropsychological battery. These regional norms will improve interpretation of verbal fluency test performance in Spanish-speakers living in the US-Mexico borderland.

Demographically adjusted norms for the Trail Making Test in native Spanish speakers: Results from the neuropsychological norms for the US-Mexico border region in Spanish (NP-NUMBRS) project.

OBJECTIVE: Despite the wide use of the Trail Making Test (TMT), there is a lack of normative data for Spanish speakers living in the USA. Here we describe the development of regional norms for the TMT for native Spanish speakers residing in the Southwest Mexico-Border Region of the USA. METHOD: Participants were 252 healthy native Spanish speakers, 58% women, from ages 19 to 60, and ranging in education from 0 to 20 years, recruited in San Diego, CA and Tucson, AZ. All completed the TMT in Spanish along with a comprehensive neuropsychological test battery as part of their participation in the Neuropsychological Norms for the US-Mexico Border Region in Spanish (NP-NUMBRS) project. Univariable and interactive effects of demographics on test performance were examined. T-scores were calculated using fractional polynomial equations to account for linear and any non-linear effects of age, education, and sex. RESULTS: Older age and lower education were associated with worse scores on both TMT A and B. No sex differences were found. The newly derived T-scores showed no association with demographic variables and displayed the expected 16% rates of impairment using a -1 SD cut point based on a normal distribution. By comparison, published norms for English-speaking non-Hispanic Whites applied to the current data yielded significantly higher impairment for both TMT A and B with more comparable rates using non-Hispanic African Americans norms. CONCLUSIONS: Population-specific, demographically adjusted regional norms improve the utility and diagnostic accuracy of the TMT for use with native Spanish speakers in the US-Mexico Border region.

The Neuropsychological Norms for the U.S.-Mexico Border Region in Spanish (NP-NUMBRS) Project: Overview and considerations for life span research and evidence-based practice.

OBJECTIVE: This paper summarizes the findings of the Neuropsychological Norms for the U.S.-Mexico Border Region in Spanish (NP-NUMBRS) Project and offers a roadmap for future research. METHODS: The NP-NUMBRS project represents the largest and most comprehensive co-normed neuropsychological battery to date for native Spanish-speaking healthy adults from the U.S. (California/Arizona)-Mexico borderland region (N = 254; ages 19-60 years). These norms provide demographic adjustments for tests across numerous domains (i.e., verbal fluency, processing speed, attention/working memory, executive function, episodic memory [learning and delayed recall], visuospatial, and fine motor skills). CONCLUSIONS: This project: 1) shows that the NP-NUMBRS norms consistently outperformed previously published norms for English-speaking non-Hispanic (White and African-American) adults in identifying impairment; 2) explores the role of Spanish-English bilingualism in test performance; and 3) provides support for the diagnostic validity of these norms in detecting HIV-associated neurocognitive impairment. Study limitations include the limited assessment of sociocultural variables and generalizability (e.g., other Latina/o populations, age limit [19 - 60 years]). Future research is needed to: 1) investigate these norms with U.S.-dwelling Spanish-speakers of non-Mexican heritage and other clinical subpopulations; 2) expand coverage of cognitive domains (e.g. language, visuospatial); 3) develop large normative datasets for children and older Latina/o populations; 4) examine how sociocultural factors impact performance (e.g., bilingualism, acculturation); 5) investigate these norms' diagnostic and ecological validity; and 6) develop norms for neurocognitive change across time. It is hoped that the NP-NUMBRS norms will aid researchers and clinicians working with U.S.-dwelling Spanish-speakers from the U.S.-Mexico borderland to conduct research and evidence-based neuropsychological evaluations in a more culturally responsive and ethical manner.

Demographically-adjusted norms for the processing speed subtests of the WAIS-III in a Spanish-speaking adult population: Results from the Neuropsychological Norms for the U.S.-Mexico Border Region in Spanish (NP-NUMBRS) project.

OBJECTIVE: The Wechsler Adult Intelligence Scale (WAIS) processing speed subtests are among the most ubiquitous indices of processing speed in the field. The aim of this study was to develop and examine demographically-adjusted normative data for Spanish language versions of the WAIS-III Digit Symbol Coding (DSC) and Symbol Search (SS) subtests for US-dwelling Spanish-speakers living in the US/Mexico border region. METHODS: The sample included 203 healthy participants who were part of the larger Neuropsychological Norms for the US-Mexico Border Region in Spanish (NP-NUMBRS) project (DSC: n = 201; SS: n = 200). RESULTS: Older age and higher education were both related to lower scores on the DSC and SS subtests (all ps < .0001). There were no significant effects for gender (all ps > .05). Raw-to-scaled score conversions were calculated for both subtests, and fractional polynomial equations were derived to compute demographically-adjusted T-scores accounting for age, education, and gender for each subtest and the Processing Speed Index. Published norms for English-speaking non-Hispanic white adults slightly overestimated impairment rates (T-scores <40) on both the DSC and SS subtests, while the norms for English-speaking non-Hispanic Black/African Americans and the new NP-NUMBRS norms Spanish-speakers both yielded impairment rates that fell within expected limits for healthy controls (i.e. 13%-14%). CONCLUSIONS: This study suggests that population-specific normative data can improve the diagnostic validity of these measures for U.S.-dwelling Spanish-speakers living in the US/Mexico border region. Future research is needed to investigate the utility of these norms for other U.S.-dwelling Spanish-speaking subpopulations (e.g. Caribbean, Central American, South American).