Functional analysis of 3'-UTR hairpins supports a two-tiered model for posttranscriptional regulation of MAT2A by METTL16.

S-adenosylmethionine (SAM) is the methyl donor for nearly all cellular methylation events, so cells need to carefully control SAM levels. MAT2A encodes the only SAM synthetase expressed in the majority of human cells, and its 3'-UTR has six conserved regulatory hairpins (hp1-6) that can be methylated by the N6-methyladenosine methyltransferase METTL16. Hp1 begins 8 nt from the stop codon, whereas hp2-6 are clustered further downstream (∼800 nt). These hairpins have been proposed to regulate MAT2A mRNA levels in response to intracellular SAM levels by regulating intron detention of the last intron of MAT2A and by modulating the stability of the fully spliced mRNA. However, a dissection of these two posttranscriptional mechanisms has not been previously reported. Using a modular reporter system, we show that hp1 functions primarily when the detained intron is included in the reporter and when that intron has a suboptimal polypyrimidine tract. In contrast, the hp2-6 cluster modulates mRNA stability independent of the detained intron, although hp1 may make a minor contribution to the regulation of decay as well. Taken with previously published reports, these data support a two-tiered model for MAT2A posttranscriptional regulation by METTL16 through its interactions with hp1 and hp2-6. In the upstream tier, hp1 and METTL16 control MAT2A intron detention, whereas the second tier involves METTL16-dependent methylation of hp2-6 to control MAT2A mRNA stability. Thus, cells use a similar set of molecular factors to achieve considerable complexity in the posttranscriptional regulation of SAM homeostasis.

Analysis of Heart Rate Variability in Individuals Affected by Amyotrophic Lateral Sclerosis.

INTRODUCTION: Amyotrophic lateral sclerosis (ALS) produces alterations in the autonomic nervous system (ANS), which explains the cardiac manifestations observed in patients. The assessment of heart rate variability (HRV) is what best reflects the activity of the ANS on heart rate. The Polar H7 Bluetooth® device proves to be a non-invasive and much faster technology than existing alternatives for this purpose. OBJECTIVE: The goal of this study is to determine HRV using Polar H7 Bluetooth technology in ALS patients, comparing the obtained measurements with values from healthy individuals. METHOD: The sample consisted of 124 participants: 68 diagnosed with ALS and 56 healthy individuals. Using Polar H7 Bluetooth technology and the ELITE HRV application, various HRV measurements were determined for all participants, specifically the HRV index, RMSSD, RMSSD LN, SDNN index, PNN50, LF, HF, LF/HF ratio, HR average, and HF peak frequency. RESULTS: Statistically significant differences were observed between ALS patients and healthy individuals in the HRV index, RMSSD, RMSSD LN, SDNN index, PNN50, HF, and LF, where healthy individuals exhibited higher scores. For the HR average, the ALS group showed a higher value. Values were similar when comparing men and women with ALS, with only a higher HF peak frequency observed in women. CONCLUSION: The Polar H7 Bluetooth® device is effective in determining heart rate variability alterations in ALS, being a promising prognostic tool for the disease.

Thermal Water Prospection with UAV, Low-Cost Sensors and GIS. Application to the Case of La Hermida.

The geothermal resource is one of the great sources of energy on the planet. The conventional prospecting of this type of energy is a slow process that requires a great amount of time and significant investments. Nowadays, geophysical techniques have experienced an important evolution due to the irruption of UAVs, which combined with infrared sensors can provide great contributions in this field. The novelty of this technology involves the lack of tested methodologies for their implementation in this type of activities. The research developed is focused on the proposal of a methodology for the exploration of hydrothermal resources in an easy, economic, and rapid way. The combination of photogrammetry techniques with visual and thermal images taken with UAVs allows the generation of temperature maps or thermal orthomosaics, which analyzed with GIS tools permit the quasi-automatic identification of zones of potential geothermal interest along rivers or lakes. The proposed methodology has been applied to a case study in La Hermida (Cantabria, Spain), where it has allowed the identification of an effluent with temperatures close to 40 °C, according to the verification measurements performed on the geothermal interest area. These results allow validation of the potential of the method, which is strongly influenced by the particular characteristics of the study area.

Andean crops: kañiwa and tarwi flours used for the development of vegan gluten-free muffins.

BACKGROUND: The flours of two Andean crops, tarwi (Lupinus mutabilis) and kañiwa (Chenopodium pallidicaule Aellen), present an excellent nutritional profile for inclusion in vegan gluten-free muffin formulations. In this study, the proximal composition and techno-functional properties of tarwi and kañiwa flours, and the technological quality of batters and muffins (potato starch-based) formulated with 50% of these flours were evaluated. RESULTS: Andean flours have high protein, fiber, and fat content, and display high water and oil absorption. In premixes formulated with potato starch and Andean flours, a reduction in paste viscosity was observed due to starch dilution and lower water availability. Depending on their ability to interact with water, the batters formulated with these flours had a higher consistency. Confocal laser scanning micrographs showed that batters with Andean flours presented a complex matrix with dispersed starch granules surrounded by proteins and fiber fragments. Muffins made with Andean flours had a slightly lower specific volume than the control, but crumb hardness was not modified by tarwi flour (50%) and a mixture of tarwi (25%) and kañiwa (25%) flours. The intrinsic color of these flours modified crumb color, and their reducing sugar content also favored Maillard reactions in the crust. These color changes are desirable in gluten-free products because such products are frequently pale due to their high starch content. CONCLUSIONS: This study showed that tarwi and kañiwa flours are suitable for developing vegan, gluten-free muffins of good technological quality and improved nutritional profile, adding value to these underutilized ancestral flours. © 2022 Society of Chemical Industry.

Kaposi's Sarcoma-Associated Herpesvirus Fine-Tunes the Temporal Expression of Late Genes by Manipulating a Host RNA Quality Control Pathway.

Kaposi's sarcoma-associated herpesvirus (KSHV) is a human oncogenic nuclear DNA virus that expresses its genes using the host cell transcription and RNA processing machinery. As a result, KSHV transcripts are subject to degradation by at least two host-mediated nuclear RNA decay pathways, the PABPN1- and poly(A) polymerase α/γ (PAPα/γ)-mediated RNA decay (PPD) pathway and an ARS2-dependent decay pathway. Here, we present global analyses of viral transcript levels to further understand the roles of these decay pathways in KSHV gene expression. Consistent with our recent report that the KSHV ORF57 protein increases viral transcript stability by impeding ARS2-dependent decay, ARS2 knockdown has only modest effects on viral gene expression 24 h after lytic reactivation of wild-type virus. In contrast, inactivation of PPD has more widespread effects, including premature accumulation of late transcripts. The upregulation of late transcripts does not require the primary late-gene-specific viral transactivation factor, suggesting that cryptic transcription produces the transcripts that then succumb to PPD. Remarkably, PPD inactivation has no effect on late transcripts at their proper time of expression. We show that this time-dependent PPD evasion by late transcripts requires the host factor nuclear RNAi-defective 2 (NRDE2), which has previously been reported to protect cellular RNAs by sequestering decay factors. From these studies, we conclude that KSHV uses PPD to fine-tune the temporal expression of its genes by preventing their premature accumulation.IMPORTANCE Kaposi's sarcoma-associated herpesvirus (KSHV) is an oncogenic gammaherpesvirus that causes Kaposi's sarcoma and other lymphoproliferative disorders. Nuclear expression of KSHV genes results in exposure to at least two host-mediated nuclear RNA decay pathways, the PABPN1- and PAPα/γ-mediated RNA decay (PPD) pathway and an ARS2-mediated decay pathway. Perhaps unsurprisingly, we previously found that KSHV uses specific mechanisms to protect its transcripts from ARS2-mediated decay. In contrast, here we show that PPD is required to dampen the expression of viral late transcripts that are prematurely transcribed, presumably due to cryptic transcription early in infection. At the proper time for their expression, KSHV late transcripts evade PPD through the activity of the host factor NRDE2. We conclude that KSHV fine-tunes the temporal expression of its genes by modulating PPD activity. Thus, the virus both protects from and exploits the host nuclear RNA decay machinery for proper expression of its genes.

Kaposi's sarcoma-associated herpesvirus ORF57 protein protects viral transcripts from specific nuclear RNA decay pathways by preventing hMTR4 recruitment.

Nuclear RNAs are subject to a number of RNA decay pathways that serve quality control and regulatory functions. As a result, any virus that expresses its genes in the nucleus must have evolved mechanisms that avoid these pathways, but the how viruses evade nuclear RNA decay remains largely unknown. The multifunctional Kaposi's sarcoma-associated herpesvirus (KSHV) ORF57 (Mta) protein is required for the nuclear stability of viral transcripts. In the absence of ORF57, we show that viral transcripts are subject to degradation by two specific nuclear RNA decay pathways, PABPN1 and PAPα/γ-mediated RNA decay (PPD) in which decay factors are recruited through poly(A) tails, and an ARS2-mediated RNA decay pathway dependent on the 5' RNA cap. In transcription pulse chase assays, ORF57 appears to act primarily by inhibiting the ARS2-mediated RNA decay pathway. In the context of viral infection in cultured cells, inactivation of both decay pathways by RNAi is necessary for the restoration of ORF57-dependent viral genes produced from an ORF57-null bacmid. Mechanistically, we demonstrate that ORF57 protects viral transcripts by preventing the recruitment of the exosome co-factor hMTR4. In addition, our data suggest that ORF57 recruitment of ALYREF inhibits hMTR4 association with some viral RNAs, whereas other KSHV transcripts are stabilized by ORF57 in an ALYREF-independent fashion. In conclusion, our studies show that KSHV RNAs are subject to nuclear degradation by two specific host pathways, PPD and ARS2-mediated decay, and ORF57 protects viral transcripts from decay by inhibiting hMTR4 recruitment.

Methotrexate reduces keratinocyte proliferation, migration and induces apoptosis in HaCaT keratinocytes in vitro and reduces wound closure in Skh1 mice in vivo.

The aim of the present work was to evaluate MTX treatment (0.1, 1 and 10 µg mL-1) in vitro in order to characterize its effects on cell proliferation alterations in cell cycle of HaCaT keratinocytes and wound healing in a Skh1 mice treated with MTX (low doses 30 mg kg-1, high doses 200 mg kg-1 and repeated doses at 1.5 mg kg-1). We analyzed the cytotoxic effect of methotrexate by a resazurin assay. The effects in the proliferation, cell cycle and apoptosis of HaCaT cells were analyzed by flow cytometry. The effects of MTX on wound healing in vivo were also analyzed. A trend toward reduction in the resazurin assay was found (p > 0.05). Reduced proliferation was also identified in a clonogenic assay and a CFSE assay (p < 0.05) due to the MTX treatment. A reduction in the G2/M and S phases was observed accompanied by apoptosis induction with increased sub G0 phase and annexin V FITC staining. Effect of MTX was evidenced in vivo on the wound closure process after day 10 (p < 0.05) with alterations in tissue architecture and remodeling. There is a marked effect of MTX on wound healing in vivo in Skh1 mice with implications for long-term therapy and surgical interventions.

Niche adaptation and viral transmission of human papillomaviruses from archaic hominins to modern humans.

Recent discoveries on the origins of modern humans from multiple archaic hominin populations and the diversity of human papillomaviruses (HPVs) suggest a complex scenario of virus-host evolution. To evaluate the origin of HPV pathogenesis, we estimated the phylogeny, timing, and dispersal of HPV16 variants using a Bayesian Markov Chain Monte Carlo framework. To increase precision, we identified and characterized non-human primate papillomaviruses from New and Old World monkeys to set molecular clock models. We demonstrate specific host niche adaptation of primate papillomaviruses with subsequent coevolution with their primate hosts for at least 40 million years. Analyses of 212 HPV16 complete genomes and 3582 partial sequences estimated ancient divergence of HPV16 variants (between A and BCD lineages) from their most recent common ancestors around half a million years ago, roughly coinciding with the timing of the split between archaic Neanderthals and modern Homo sapiens, and nearly three times longer than divergence times of modern Homo sapiens. HPV16 A lineage variants were significantly underrepresented in present African populations, whereas the A sublineages were highly prevalent in European (A1-3) and Asian (A4) populations, indicative of viral sexual transmission from Neanderthals to modern non-African humans through multiple interbreeding events in the past 80 thousand years. Remarkably, the human leukocyte antigen B*07:02 and C*07:02 alleles associated with increased risk in cervix cancer represent introgressed regions from Neanderthals in present-day Eurasians. The archaic hominin-host-switch model was also supported by other HPV variants. Niche adaptation and virus-host codivergence appear to influence the pathogenesis of papillomaviruses.

Electromyographic Activity Evolution of Local Twitch Responses During Dry Needling of Latent Trigger Points in the Gastrocnemius Muscle: A Cross-Sectional Study.

OBJECTIVE: Trigger points (TrPs) are hypersensitive spots within taut bands of skeletal muscles that elicit referred pain and motor changes. Among the variety of techniques used for treating TrPs, dry needling is one of the most commonly applied interventions. The question of eliciting local twitch responses (LTRs) during TrP dry needling is unclear. Our main aim was to investigate the evolution of the electromyographic (EMG) peak activity of each LTR elicited during dry needling into latent TrPs of the gastrocnemius medialis muscle. METHODS: Twenty asymptomatic subjects with latent TrPs in the gastrocnemius medialis muscle participated in this cross-sectional study. Changes in EMG signal amplitude (root mean square [RMS]) with superficial EMG were assessed five minutes before, during, and five minutes after dry needling. The peak RMS score of each LTR was calculated (every 0.5 sec). RESULTS: Analysis of variance revealed a significant effect (F = 29.069, P <0.001) showing a significant decrease of RMS peak amplitude after each subsequent LTR. Differences were significant (P <0.001) during the first three LTRs, and stable until the end of the procedure. No changes (P =0.958) were found for mean RMS data at rest before (mean = 65.2 mv, 95% confidence interval [CI] = 47.3-83.1) and after (61.0 mv, 95% CI = 42.3-79.7) dry needling. CONCLUSIONS: We found that, in a series of LTRs elicited during the application of dry needling over latent TrPs in the medial gastrocnemius muscle, the RMS peak amplitude of each subsequent LTR decreased as compared with the initial RMS peak amplitude of previous LTRs. No changes in superficial EMG activity at rest were observed after dry needling of latent TrPs of the gastrocnemius medialis muscle.

Pain Perception in Clarinetists with Playing-Related Pain After Implementing a Specific Exercise Program.

BACKGROUND: Wind musicians suffer injuries resulting from muscle overuse and poor postural habits, often due to the lack of required physical fitness. For this reason, it is important to study and analyze the characteristics of their activity in order to select appropriate preventive exercises. METHODS: 10 clarinetists, who were experiencing joint pain due to instrumental practice, followed a specific program of physical activity, 3 times a week for 2 months. To assess postural changes after its implementation, the Langlade test and muscle pain scale were used at the beginning and the end of the program. RESULTS: The results show a significant decrease in perceived pain (p<0.001) and changes in the dorsal spine (p=0.001). Given the relationship between the improvement in the Langlade item, which refers to the correct position of the shoulder blades, and muscle pain, with a correlation level of 0.582, it can be said that a change in the shoulder girdle position leads to a decrease of pain in that area. CONCLUSIONS: In this pilot study of 10 clarinetists, a regular program of physical activity for 9 weeks led to an observable change in posture and a reduction in pain using self-report measures.

Randomized trial of cesarean vs vaginal delivery for effects on the pelvic floor in squirrel monkeys.

BACKGROUND: Vaginal delivery is a risk factor in pelvic floor disorders. We previously described changes in the pelvic floor associated with pregnancy and parturition in the squirrel monkey, a species with a humanlike pattern of spontaneous age- and parity-associated pelvic organ prolapse. OBJECTIVE: The potential to prevent or diminish these changes with scheduled cesarean delivery (CD) has not been evaluated. In a randomized, controlled trial, we compared female squirrel monkeys undergoing spontaneous vaginal delivery with those undergoing scheduled primary CD for pelvic floor muscle volumes, muscle contrast changes, and dynamic effects on bladder neck position. STUDY DESIGN: Levator ani, obturator internus, and coccygeus (COC) muscle volumes and contrast uptake were assessed by magnetic resonance imaging in 20 nulliparous females examined prior to pregnancy, a few days after delivery, and 3 months postpartum. The position of bladder neck relative to bony reference line also was assessed with abdominal pressure using dynamic magnetic resonance imaging. RESULTS: Baseline measurements of 10 females randomly assigned to scheduled primary CD were not different from those of 10 females assigned to spontaneous vaginal delivery. Levator ani and obturator internus muscle volumes did not differ between groups, while volumes were reduced (P < .05) in the observation immediately after pregnancy. The COC muscles increased (P < .05) immediately after delivery for females in the spontaneous vaginal delivery group, but not for females in the scheduled CD group. Position of the bladder neck descended (P < .05) by 3 months postpartum in both groups. CONCLUSION: Scheduled CD diminishes changes in COC muscle volume and contrast reported to be associated with spontaneous vaginal delivery in squirrel monkeys. However, pelvic support of the bladder was not protected by this intervention suggesting that effects of pregnancy and delivery are not uniformly prevented by this procedure.

Functional morphology of the tubular genital organs in the female owl monkey (Aotus spp.).

BACKGROUND: Studies on reproductive morphology are important to understand the reproductive cycle of non-human primates. METHODS: This study describes the functional morphology of the adult female tubular genital organs in 41 Aotus (12.8 ± 6.8 years old, ranging from 3 to 25 years), with respect to reproductive status and number of parturitions. RESULTS: In females with developing embryos, endometrial glands showed a higher secretion than other females, and the embryo implantation occupied this secretive endometrium. Changes in the thickening, number of layers, and keratinization in the vaginal epithelium suggest that vaginal cytology may be an indicator of the estrous cycle. Non-pregnant multiparous females had a larger uterine body than nulliparous females. Number of parturitions and reproductive state had an impact on tubular genital organs in female owl monkeys. CONCLUSIONS: These results can be useful for the development of biotechnologies of reproduction and for improvement of the management of this species.

F-box and leucine-rich repeat protein 5 (FBXL5) is required for maintenance of cellular and systemic iron homeostasis.

Maintenance of cellular iron homeostasis requires post-transcriptional regulation of iron metabolism genes by iron regulatory protein 2 (IRP2). The hemerythrin-like domain of F-box and leucine-rich repeat protein 5 (FBXL5), an E3 ubiquitin ligase subunit, senses iron and oxygen availability and facilitates IRP2 degradation in iron replete cells. Disruption of the ubiquitously expressed murine Fbxl5 gene results in a failure to sense increased cellular iron availability, accompanied by constitutive IRP2 accumulation and misexpression of IRP2 target genes. FBXL5-null mice die during embryogenesis, although viability is restored by simultaneous deletion of the IRP2, but not IRP1, gene. Mice containing a single functional Fbxl5 allele behave like their wild type littermates when fed an iron-sufficient diet. However, unlike wild type mice that manifest decreased hematocrit and hemoglobin levels when fed a low-iron diet, Fbxl5 heterozygotes maintain normal hematologic values due to increased iron absorption. The responsiveness of IRP2 to low iron is specifically enhanced in the duodena of the heterozygotes and is accompanied by increased expression of the divalent metal transporter-1. These results confirm the role of FBXL5 in the in vivo maintenance of cellular and systemic iron homeostasis and reveal a privileged role for the intestine in their regulation by virtue of its unique FBXL5 iron sensitivity.

Lack of association between pelvic outlet diameter and pelvic organ prolapse in squirrel monkeys.

INTRODUCTION AND HYPOTHESIS: The aim was to test the hypothesis that the pelvic outlet diameter (POD) is associated with pelvic organ prolapse (POP) in squirrel monkeys. METHODS: Magnetic resonance images (MRI) were obtained from 55 females with and without POP. Commercial software was used by two observers to measure the POD. POD, age, parity, and weight for the two groups were compared using Student's t test. Associations of age, parity, and body weight with POD were evaluated with linear regression analysis. Multiple regression analysis examined the relationships of age, parity, and POD with POP. Receiver operating curve methods were used to set thresholds. RESULTS: Observers concurred on measurements (Cronbach's alpha = 0.96 with lower 95% confidence limit of 0.94). POD was not related to parity (p = 0.10) or weight (p = 0.053), but was inversely related to age (p = 0.011). Animals with POP did not differ from those without POP with regard to age (p = 0.10), weight (p = 0.17), or POD (p = 0.99). The groups differed with regard to parity (p = 0.007) and multiple regression methods demonstrated that only parity had a significant relationship with POP (p = 0.002). Parity greater than 2 had 64% specificity and 89% sensitivity for POP. CONCLUSIONS: POD size does not contribute to POP in squirrel monkeys. This study confirms that among variables of age, body weight, POD, and parity, only parity appears as a risk factor for POP in squirrel monkeys.

TAV-in-TAV in patients with prosthesis embolization: Impact of commissural alignment and global outcomes.

BACKGROUND: Optimal strategies to manage embolization of transcatheter aortic valve implantation (TAVI) devices are unclear; valve-in-valve (ViV) is often used. We aimed to describe through one-single center experience its rate, causes, consequences, and management as well as the rate and relevance of commissural alignment (CA) in this context. METHODS: We identified across 1038 TAVI cases, those cases requiring ViV for the management of first device embolization. CA (absence or mild misalignment) after first and second device was assessed by CT or fluoroscopy. RESULTS: A total of 23 cases (2.2%) were identified, 52.3% embolized towards the aorta and 47.7% towards the ventricle. Suboptimal implant height (38%) and embolization at the time of post-dilation (23%) were the most frequent mechanisms together with greater rate of bicuspid valve (p < 0.001) and a trend to greater annular eccentricity. Procedural and 1-year death occurred in 13% and 34%, respectively (vs. 1.1% and 7.8% in the global cohort, p < 0.001). CA was present in 76.9% of the prostheses initially implanted but was only spontaneously achieved in 30.8% of the second ViV device. Adequate CA of both prostheses was identified in only two cases (8.7%). There were no cases of coronary obstruction. CONCLUSIONS: TAVI device embolization mechanisms can often be predicted and prevented. Mortality following bail-out ViV is higher than in regular TAVI procedures but 2/3 of these patients survived beyond 1-year follow-up. In them, valve degeneration or coronary re-access might be particularly challenging since CA was rarely achieved with both devices suggesting that greater efforts should be made in this regard.

Hemerythrin-like domain within F-box and leucine-rich repeat protein 5 (FBXL5) communicates cellular iron and oxygen availability by distinct mechanisms.

Iron regulatory proteins play a principal role in maintaining cellular iron homeostasis by post-transcriptionally regulating factors responsible for iron uptake, utilization, and storage. An E3 ubiquitin ligase complex containing FBXL5 targets IRP2 for proteasomal degradation under iron- and oxygen-replete conditions, whereas FBXL5 itself is degraded when iron and oxygen availability decreases. FBXL5 contains a hemerythrin-like (Hr) domain at its N terminus that mediates its own differential stability. Here, we investigated the iron- and oxygen-dependent conformational changes within FBXL5-Hr that underlie its role as a cellular sensor. As predicted, FBXL5-Hr undergoes substantive structural changes when iron becomes limiting, accounting for its switch-like behavior. However, these same changes are not observed in response to oxygen depletion, indicating that this domain accommodates two distinct sensing mechanisms. Moreover, FBXL5-Hr does not behave as a dynamic sensor that continuously samples the cellular environment, assuming conformations in equilibrium with ever-changing cellular iron levels. Instead, the isolated domain appears competent to incorporate iron only at or near the time of its own synthesis. These observations have important implications for mechanisms by which these metabolites are sensed within mammalian cells.

Structural and molecular characterization of iron-sensing hemerythrin-like domain within F-box and leucine-rich repeat protein 5 (FBXL5).

Mammalian cells maintain iron homeostasis by sensing changes in bioavailable iron levels and promoting adaptive responses. FBXL5 is a subunit of an E3 ubiquitin ligase complex that mediates the stability of iron regulatory protein 2, an important posttranscriptional regulator of several genes involved in iron metabolism. The stability of FBXL5 is regulated in an iron- and oxygen-responsive manner, contingent upon the presence of its N-terminal domain. Here we present the atomic structure of the FBXL5 N terminus, a hemerythrin-like α-helical bundle fold not previously observed in mammalian proteins. The core of this domain employs an unusual assortment of amino acids necessary for the assembly and sensing properties of its diiron center. These regulatory features govern the accessibility of a mapped sequence required for proteasomal degradation of FBXL5. Detailed molecular and structural characterization of the ligand-responsive hemerythrin domain provides insights into the mechanisms by which FBXL5 serves as a unique mammalian metabolic sensor.

Effect of partial substitution of wheat flour by quinoa (Chenopodium quinoa Willd.) and tarwi (Lupinus mutabilis Sweet) flours on dough and bread quality.

Bread is the main important food product worldwide. In this study, eleven bread formulations were developed by partial substitution of wheat flour with quinoa and tarwi flours, to evaluate the effect on the rheological and pasting properties of mixtures, as well as on the physicochemical and textural properties of the final product. Partial substitution with quinoa flour generated similar thermomechanical and textural properties in the dough, and similar bread technological characteristics related to the control bread (100% wheat). In the case of tarwi, the increase in the concentration of this legume showed a negative effect on the bread quality parameters (specific volume, crumb porosity, textural properties, etc.). A negative technological impact of high percentages of wheat flour substitution by the mixture of both Andean flours was found, but it was contrasted with a positive effect on nutritional quality, particularly evidenced by a high content of proteins and dietary fiber. An optimal formulation considering technological and nutritional quality was obtained, presenting the maximum analyzed substitution level (13.35% quinoa flour and 6.65% tarwi flour). This study showed that these Andean grains are suitable for developing bread of good technological quality and improved nutritional profile, adding value to these underused ancestral flours.

Immunodetection of hemocytes, peneidins and α2-macroglobulin in the lymphoid organ of white spot syndrome virus infected shrimp.

Viral diseases restrict the development of the world shrimp industry and there are few studies on cell response to the presence of viral infections. We performed immunohistochemistry assays to characterize hemocytes subpopulations involved in the immune process occurring in the LO of Litopenaeus vannamei shrimp. Tissue sections of animals that increased their LO spheroids and hemocytes infiltration after WSSV induced infection, were used. Three MABs namely, 40E10 (recognizing small granule hemocytes), 40E2 (recognizing large granule hemocytes), and 41B12, which recognize α(2)-macroglobulin were used. Additionally one polyclonal antibody was used against the penaeidins antimicrobial peptides, and to detect WSSV a commercial immunohistochemistry kit (DiagXotics) was used. Numerous small granule hemocytes were detected in the stromal matrix of LO tubules, whereas large granule hemocytes were less numerous and located mainly in hemal sinuses. The exocytosis of two molecules, which have been related to the phagocytosis process, i.e. penaeidins, and α(2)-macroglobulin, was detected in the external stromal matrix and the outer tubule walls. α(2) -macroglobulin inhibits phenoloxidase activity and its strong release in LO tissue may explain the absence of melanization in the immune processes occurring in it. The immunolabeling of vesicles within the LO spheroids with MABs 41B12 40E10 and antipenaedin antibody suggests that LOS are formed by phagocytic cells derived from small granule and hyaline hemocytes, with a possible role of peneidins and α(2)-macroglobulin acting as opsonines.

Muscular Response in ALS Patients during Maximal Bilateral Isometric Work of the Biceps Brachii until Fatigue.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative and fatal disease, characterized by the loss of motor neurons and progressive musculoskeletal deterioration. The clinical onset is mainly bulbar or spinal. Considering that there is no effective medical treatment, there is a need to understand the muscle activation patterns to design better physical exercise routines. The objective of this study was to determine muscle strength and fatigue in patients with ALS performing a unilateral exercise, and according to sex and type of ALS. A cross-sectional, analytical study was conducted with 23 patients. Five maximal unilateral isometric contractions were performed with the right and left biceps brachii. Muscle activation was calculated by surface electromyography bilaterally in the biceps brachii, triceps brachii, rectus femoris anterior, and tibialis anterior. The results showed more accentuated fatigue in men than in women, between the first and last contractions performed and especially on the dominant side (p = 0.016). In addition, there was evidence of a coactivation effect on the muscles around the work joint, which reflects a growing activation of synergists, regardless of sex or type of ALS. These findings support the use of systematic and extensive resistance exercise as a non-invasive option for maintaining the functional capacity of patients with ALS.