Transcriptome and small RNAome profiling uncovers how a recombinant begomovirus evades RDRγ-mediated silencing of viral genes and outcompetes its parental virus in mixed infection.

Tomato yellow leaf curl virus (TYLCV, genus Begomovirus, family Geminiviridae) causes severe disease of cultivated tomatoes. Geminiviruses replicate circular single-stranded genomic DNA via rolling-circle and recombination-dependent mechanisms, frequently generating recombinants in mixed infections. Circular double-stranded intermediates of replication also serve as templates for Pol II bidirectional transcription. IS76, a recombinant derivative of TYLCV with a short sequence in the bidirectional promoter/origin-of-replication region acquired from a related begomovirus, outcompetes TYLCV in mixed infection and breaks disease resistance in tomato Ty-1 cultivars. Ty-1 encodes a γ-clade RNA-dependent RNA polymerase (RDRγ) implicated in Dicer-like (DCL)-mediated biogenesis of small interfering (si)RNAs directing gene silencing. Here, we profiled transcriptome and small RNAome of Ty-1 resistant and control susceptible plants infected with TYLCV, IS76 or their combination at early and late infection stages. We found that RDRγ boosts production rates of 21, 22 and 24 nt siRNAs from entire genomes of both viruses and modulates DCL activities in favour of 22 and 24 nt siRNAs. Compared to parental TYLCV, IS76 undergoes faster transition to the infection stage favouring rightward transcription of silencing suppressor and coat protein genes, thereby evading RDRγ activity and facilitating its DNA accumulation in both single and mixed infections. In coinfected Ty-1 plants, IS76 efficiently competes for host replication and transcription machineries, thereby impairing TYLCV replication and transcription and forcing its elimination associated with further increased siRNA production. RDRγ is constitutively overexpressed in Ty-1 plants, which correlates with begomovirus resistance, while siRNA-generating DCLs (DCL2b/d, DCL3, DCL4) and genes implicated in siRNA amplification (α-clade RDR1) and function (Argonaute2) are upregulated to similar levels in TYLCV- and IS76-infected susceptible plants. Collectively, IS76 recombination facilitates replication and promotes expression of silencing suppressor and coat proteins, which allows the recombinant virus to evade the negative impact of RDRγ-boosted production of viral siRNAs directing transcriptional and posttranscriptional silencing.

Leadership and the finder's advantage in mantled howler monkeys.

Leading collective movements and arriving first at feeding sites may improve food acquisition. Specifically, the first individual to discover and exploit a feeding site may gain a feeding advantage known as the "finder's advantage." The aim of this research was to verify if the probability of leading group movements to feeding sites in mantled howler monkeys (Alouatta palliata) varied by sex and reproductive status, and whether finders had higher foraging success than followers. We studied 18 adult individuals from two groups in La Flor de Catemaco over a year (978 h), and sampled group movements (n = 211) and foraging behavior (n = 215 feeding episodes). Gestating females were leaders and finders of group movements to feeding sites more often than expected but were also replaced in the leading position more frequently than individuals of other sex/reproductive states. Feeding behavior was not influenced by the order of arrival at feeding sites per se, but gestating females had higher food intake rate, bite rate, and feeding time when arriving earlier (i.e., occupying front group positions) than later. Therefore, leadership and the finder's advantage occur in this species and are probably employed by gestating females to maintain their energetic condition.

Reference Values in the 6-Minute Walk Test in Chilean Children Aged 3-10 Years and Relationship With Cardiometabolic Risk.

PURPOSE: The aim of this study was to evaluate the performance of healthy Chilean children aged 3-10 years in the 6-minute walking test (6MWT) and cardiometabolic risk variables and to determine sex- and age-specific reference values. METHODS: This study involved 1165 healthy children (age = 6.36 [1.70] y old). The 6MWT was used to evaluate exercise performance. Furthermore, anthropometric measures were collected, like weight, height, body mass index, waist circumference, and skin folds. Resting heart rate and blood pressure (BP) were also evaluated. RESULTS: The prevalence of overweight and obesity was 35.0% and 25.4% in preschoolers and 29.0% and 36.2% in school-age children, respectively, showing significant differences (P < .05) between age groups. The distance walked (6-min walk distance) increased significantly year on year at ages from 3 to 10 years. According to the regression analysis, 6MWT performance was positively related to age, systolic BP, and height, whereas it was negatively related to ∑4 skinfold fat, resting heart rate, diastolic BP, and waist circumference. CONCLUSIONS: This study provides a reference equation and an age- and sex-adjusted percentile curve to assess the predicted 6MWT performance in a cohort of prepubertal Chilean children. The 6-minute walk distance depends mainly on age; however, other variables, such as resting heart rate, BP, skinfold fat, and waist circumference, add significant information and should be taken into account.

The Biology and Genomics of Human Hair Follicles: A Focus on Androgenetic Alopecia.

Androgenetic alopecia is a highly prevalent condition mainly affecting men. This complex trait is related to aging and genetics; however, multiple other factors, for example, lifestyle, are also involved. Despite its prevalence, the underlying biology of androgenetic alopecia remains elusive, and thus advances in its treatment have been hindered. Herein, we review the functional anatomy of hair follicles and the cell signaling events that play a role in follicle cycling. We also discuss the pathology of androgenetic alopecia and the known molecular mechanisms underlying this condition. Additionally, we describe studies comparing the transcriptional differences in hair follicles between balding and non-balding scalp regions. Given the genetic contribution, we also discuss the most significant risk variants found to be associated with androgenetic alopecia. A more comprehensive understanding of this pathology may be generated through using multi-omics approaches.

Hybrid Therapies for Supporting Critically Ill Patients with Acute Kidney Injury: When, how, and for Whom?

UNASSIGNED: Acute kidney injury (AKI) is common in critically ill patients. There is no specific pharmacological treatment for established severe AKI. Therefore, the conventional therapeutic strategy is limited to the use of kidney replacement therapy (KRT) to maintain homeostasis. Hybrid therapies optimize the advantages of intermittent and continuous modalities of KRT, combining lower hourly efficiency, longer application time, at lesser cost, but also adding different physicochemical principles of extracorporeal clearance. The sum of convection and diffusion, with or without adsorption or apheresis, and in different time combinations gives hybrid techniques great flexibility in prescribing a personalized treatment adapted to the needs of each patient at any given time. Hybrid therapies are increasingly being used due to their flexibility, which is determined by the combination of equipment, membranes, and available resources (machines and health-care personnel experience). The required technology is widely available in most intensive care units and uses low-cost consumables compared to other types of AKI treatment modalities, favoring its widespread use. Hybrid therapies are feasible and provide a viable form of KRT, either alone or as a transition therapy from continuous kidney replacement therapy to intermittent hemodialysis. (Rev Invest Clin. 2023;75(6):337-47).

Comprehensive cardiac evaluation to maximal exercise in a contemporary population of prepubertal children.

BACKGROUND: Heart rate (HR) is a biomarker used to measure physiological function, health status and cardiovascular autonomic function. The purpose of this study was to determine sex- and age-specific reference values for cardiac autonomic function at rest, during maximal exercise and the recovery phase in prepubertal children. METHODS: Five hundred and twelve healthy children 7-11 years of age performed a Léger test. A heart RR-interval monitor recorded the heart data and a specific software analysed the cardiac autonomic response through HR and HR variability (HRV). It analysed HR before the test (resting HR, RHR), during the test (HRpeak) and HR recovery (HRR) in the first minute (HRR1) and the fifth minute (HRR5). The values are mean ± SD. RESULTS: Collectively, 91.2% of girls and 92.3% of boys were within the recommended ranges regarding RHR. The average HRpeak was 199 ± 10.83 b.p.m. and 96.8% of girls and 95.3% of boys were within the minimum threshold value recommended (180 b.p.m.). Boys showed lower values of RHR than girls (p < 0.001) and larger values of HRR 1 and HRR5 (p < 0.001). CONCLUSIONS: This study comprehensively provides a reference set of data for the most important HR variables that can be obtained during exercise testing in prepubertal children regarding age and sex and in a field setting. IMPACT: This is the first study to provide reference values of autonomic cardiac function at rest, during maximal exercise and during the recovery period in prepubertal children aged 7-11 years. Despite the early age of participants, cardiorespiratory fitness, RHR and HRR are different according to sex. Aerobic performance and HRpeak have a negative correlation with body mass index and cardiometabolic risk.

Female reproductive energetics in mantled howler monkeys (Alouatta palliata): A follow-up study.

OBJECTIVE: Reproduction entails several challenges to primate females, among which energetic costs are remarkable at certain stages of the reproductive cycle. Still, females may use behavioral and physiological strategies to cope with those challenges. We had previously reported covariation between female energetic condition through the reproductive cycle and time-budget adjustments in mantled howler monkeys (Alouatta palliata). Accordingly, we suggested that behavioral flexibility allowed coping with the energetic challenges of reproduction. Subsequent evidence from the same population, however, suggested otherwise, so we performed a follow-up study on the variation in female reproductive energetics based on a larger sample of females. METHODS: We studied 48 free-ranging adult females at Los Tuxtlas (Mexico). We assessed energy balance via urinary C-peptide concentrations (2717 urine samples), behavioral energy intake and expenditure (5728 sampling hours), and physiological energy expenditure via fecal triiodothyronine metabolites (fTH3; 3138 fecal samples). RESULTS: We found that energy balance varied among reproductive states: (a) cycling was a period of low C-peptide concentrations; (b) the highest C-peptide concentrations occurred during gestation; and (c) the beginning of lactation marked a notable decrease in C-peptide concentrations, which then improved at mid-lactation to again decline at lactation offset. These peaks and valleys in energy balance did not seem to be associated with variation in energy acquisition but were rather mirrored by activity levels and fTH3 during lactation. DISCUSSION: Energy balance was not preserved through the reproductive cycle, supporting previous contentions that the reproductive performance of female mantled howler monkeys may be energetically constrained. The contrast between these and results that we have previously reported, highlights the importance of conducting follow-up studies to continually improve our understanding of the reproductive energetics of primate females.

Vocal and movement responses of mantled howler monkeys (Alouatta palliata) to natural loud calls from neighbors.

Loud calls (i.e., long-range acoustic signals) regulate resource competition among neighboring groups of conspecifics in several nonhuman primate species. Ultimate explanations for primate loud calls include mate, offspring, and food defense. Additionally, loud calls may provide valuable information pertaining to the identity and health status of callers, their competitive abilities, and their spatial location. The loud calls of howler monkeys (Alouatta) have been thoroughly studied and seem to play an important function in the defense of valuable resources in a variety of socioecological contexts. Here, we examined whether the behavioral responses of mantled howler monkeys (Alouatta palliata) to natural loud calls from neighbors are linked to three factors: food availability, familiarity, and distance between groups. We studied three groups of mantled howler monkeys at La Flor de Catemaco (Los Tuxtlas, Mexico) for 15 months (1817 observation hours), during which we recorded 236 neighbor loud calls. Food availability per se did not influence the behavior of groups receiving loud calls, although males produced longer vocal responses toward unfamiliar neighbors when food availability decreased. Groups vocalized quicker and both vocalized and moved for longer after loud calls from unfamiliar neighbors. Additionally, groups vocalized and moved for longer at shorter distances from unfamiliar neighbors compared with familiar neighbors. Finally, groups usually moved away from calling neighbors that were closer. These results indicate that the behavioral responses of mantled howler monkeys to loud calls from neighbor groups are associated with the integration of information pertaining to caller identity as well as to their ecological and spatial context.

The M4 insulator, the TM2 matrix attachment region, and the double copy of the heavy chain gene contribute to the enhanced accumulation of the PHB-01 antibody in tobacco plants.

The expression of recombinant proteins in plants is a valuable alternative to bioreactors using mammalian cell systems. Ease of scaling, and their inability to host human pathogens, enhance the use of plants to generate complex therapeutic products such as monoclonal antibodies. However, stably transformed plants expressing antibodies normally have a poor accumulation of these proteins that probably arise from the negative positional effects of their flanking chromatin. The induction of boundaries between the transgenes and the surrounding DNA using matrix attachment regions (MAR) and insulator elements may minimize these effects. With the PHB-01 antibody as a model, we demonstrated that the insertion of DNA elements, the TM2 (MAR) and M4 insulator, flanking the transcriptional cassettes that encode the light and heavy chains of the PHB-01 antibody, increased the protein accumulation that remained stable in the first plant progeny. The M4 insulator had a stronger effect than the TM2, with over a twofold increase compared to the standard construction. This effect was probably associated with an enhancer-promoter interference. Moreover, transgenic plants harboring two transcriptional units encoding for the PHB-01 heavy chain combined with both TM2 and M4 elements enhanced the accumulation of the antibody. In summary, the M4 combined with a double transcriptional unit of the heavy chain may be a suitable strategy for potentiating PHB-01 production in tobacco plants.

Disorders of Sex Development and Malignant Germ Cell Tumors.

A woman, aged 44 years, presented at the general oncology outpatient clinic with bloating, abdominal pain, and significant unintended weight loss. Her past medical history included a bilateral inguinal hernia surgical repair at age 6, and primary amenorrhea since age 15. The patient never underwent additional studies to identify the cause of the primary amenorrhea.

Burden of disease: Closing the gaps in the burden of end-stage kidney disease in Latin America.

End-stage kidney disease (ESKD) represents a major challenge for Latin America (LA). The Latin American Dialysis and Renal Transplantation Registry (LADRTR) has published several reports, and its continuity has implied a sustained effort of the nephrology community to improve care of ESKD in the region; this paper summarizes results of the year 2014. Methods have been reported previously; participant countries complete annual surveys collecting data on incident and prevalent patients undergoing renal replacement therapy (RRT) in all modalities. 20 countries participated in the surveys (more than 90% of the region). Prevalence of treated ESKD in RRT increased from 119 patients per million population (pmp) in 1991 to 709 pmp in 2014; hemodialysis continues to be the treatment of choice in the region (68%). A wide variation was observed in the incidence rate, from 421 pmp in Jalisco (Mexico) to 23 pmp in Paraguay; diabetes was the cause of 36% of incident cases. Additionally, great heterogeneity was observed in the number of nephrologists by country, from 2 pmp in Colombia to 53 pmp in Uruguay. Heterogeneity, or even absence of registries in some LA countries, is concordant with inequities in RRT access as well as with the limited availability of qualified personnel. The LADRTR is leading the personnel training to develop and strengthen national dialysis and transplant registries in LA within the frame of the Pan-American Health Organization (PAHO) - Sociedad Latinoamericana de Nefrología e Hipertension (SLANH) cooperation program.

Sex differences in leadership during group movement in mantled howler monkeys (Alouatta palliata).

Benefits of group life depend in large part on whether animals remain cohesive, which often requires collective decisions about where and when to move. During a group movement, the leader may be considered as the individual occupying the vanguard position of the group progression, when its movement evokes following by other group members. In nondespotic societies, individuals with greater incentives to move frequently are leaders. During 15 months of observations (1,712 contact hours), we investigated two mantled howler monkey (Alouatta palliata) groups at La Flor de Catemaco (Los Tuxtlas, Mexico) to examine whether sex and female reproductive state influenced leadership likelihood in two contexts: movements toward feeding trees; movements associated with loud calls, a group-defense behavior used by males of this genus. Females led and occupied forward positions during group movements toward feeding trees more often than adult males. Adult females led these movements more frequently when they were gestating than when they were lactating or cycling. There were no differences between sexes in the leadership of group movements associated with loud calls. Leadership by gestating females is perhaps the result of their higher nutritional/energetic needs when compared with cycling females, and of their greater mobility when compared with lactating females carrying dependent offspring. Female leadership during movements toward feeding trees may be a mechanism to optimize access to food resources in mantled howler monkeys.

Understanding the Cryoglobulinemias.

PURPOSE OF THE REVIEW: Cryoglobulins are immunoglobulins with the ability to precipitate at temperatures <37 °C. They are related to hematological disorders, infections [especially hepatitis C virus (HCV)], and autoimmune diseases. In this article, the state of the art on Cryoglobulinemic Vasculitis (CV), in a helpful and schematic way, with a special focus on HCV related Mixed Cryoglobulinemia treatment are reviewed. RECENT FINDINGS: Direct - acting antivirals (DAA) against HCV have emerged as an important key in HCV treatment to related Cryoglobulinemic Vasculitis, and should be kept in mind as the initial treatment in non-severe manifestations. On the other hand, a recent consensus panel has published their recommendations for treatment in severe and life threatening manifestations of Mixed Cryoglobulinemias. HCV-Cryoglobulinemic vasculitis is the most frequent form of CV. There are new treatment options in HCV-CV with DAA, with an important number of patients achieving complete response and sustained virologic response (SVR). In cases of severe forms of CV, treatment with Rituximab and PLEX are options. The lack of data on maintenance therapy could impulse future studies in this setting.

Maternal condition and maternal investment during lactation in mantled howler monkeys.

OBJECTIVES: Lactation is a costly reproductive state for females, as it may entail decreased body condition, increased morbidity, and increased mortality risk. To offset these costs, mothers should allocate investment in infant care as a function of their physical condition. Here, we assessed the relationship between maternal condition and maternal investment in wild mantled howler monkeys (Alouatta palliata) to determine: (1) whether maternal physical condition varied as a function of food availability, activity levels and feeding effort throughout lactation; (2) whether maternal investment was affected by maternal condition and offspring age. MATERIALS AND METHODS: We studied six adult females while rearing their immature offspring. We determined weekly food availability, C-peptide levels (as a measure of energy balance) in urine samples (306 samples), mothers' time budgets, feeding rates, time spent vigilant, and time spent carrying their offspring (for two years; 600 observation hours). RESULTS: C-peptide levels were positively related to food availability. Maternal care in terms of vigilance and offspring carrying peaked at early lactation and decreased progressively with offspring age. Carrying was positively related to C-peptide levels and was also influenced by variation in maternal condition throughout lactation. DISCUSSION: These results converge with previous theoretical and empirical studies on this topic and contribute to the growing evidence that nonhuman primate mothers allocate maternal care based on their current condition to maximize lifetime reproductive success.

Energetic constraints on the reproduction of female mantled howlers.

The reproductive process of female primates is energetically constrained. However, while there is extensive evidence about factors that influence the maintenance of gestation and lactation, less is known about energetic correlates of cycling and the occurrence of conception in wild primates. We examined how female physical condition affected the occurrence of conception and interbirth intervals (IBI) in female mantled howler monkeys, a species in which females experience long non-conceptive periods. For 6 years we followed 7 females at La Flor de Catemaco (Los Tuxtlas, Mexico). In addition to noting the presence of dependent immatures, we collected fecal samples that were analyzed for thyroid hormone and glucocorticoid metabolite concentrations (N = 1,460). Conception was more likely following months when females were not lactating and had higher thyroid hormone and lower glucocorticoid levels. IBIs were shorter when females had higher thyroid hormone and lower glucocorticoid levels, and when the infant at the beginning of the interval died. Due to their obvious impact on female fecundity and fertility, gestation and lactation are the stages of the reproductive process that have traditionally received more attention in studies of female reproductive performance. Still, the cycling stage accounts for a large proportion of IBIs in many primate species. Here we demonstrate that the reproductive rates of female mantled howler monkeys are energetically constrained: both the occurrence of conception and IBI are positively affected by female energetic condition.

The Influence of Leaf Consumption on Time Allocation in Black Howler Monkeys (Alouatta pigra).

The analysis of factors that determine variation in time budgets is important to understand the interactions between environment, behaviour and fitness. We tested the hypothesis that changes in the dietary patterns of black howler monkeys (Alouatta pigra) caused by a decrease in the availability of preferred foods are a main determinant of variation in time budgets. We predicted that individuals would trade off travel time for resting time (i.e., minimize energy expenditure) as the diet included more leaves. We conducted our study in the Mexican state of Campeche between 2005 and 2008, where we studied the behaviour of 28 adult males and 32 adult females belonging to 14 different groups for a total of 3,747.2 focal sampling hours. Study groups lived in forest fragments with variation in habitat quality. Individuals showed different rest:travel trade-offs in response to leaf consumption according to the quality of the forest fragments they lived in. Individuals that lived in high-quality fragments increased resting time under more folivorous regimes, whereas those living in low-quality fragments increased travel time. Our results suggest that howler monkeys living in low-quality fragments spend more time foraging to compensate for the low quality of the available resources.

Hormonal correlates of energetic condition in mantled howler monkeys.

Hormones have a key role in energy allocation, so their study allows understanding individual metabolic strategies. Because different hormones convey different information on the responses of individuals to energetic demands, a simultaneous analysis of variation in multiple hormones may offer a more reliable picture of metabolic strategies than single hormone assessments. In this study we focused on determining which factors were related to variation in fecal glucocorticoid and thyroid hormone metabolites in wild mantled howler monkeys (Alouatta palliata). Over 12months, we determined fecal glucocorticoid and thyroid hormone metabolite levels of 11 adults belonging to two groups, and examined the relationship between hormone metabolites and a variety of behavioral, physiological, and ecological factors (e.g., food intake, sex/reproductive state, activity, participation in agonistic interactions). We found that glucocorticoids were elevated in gestating and lactating females compared to males and cycling females, and were also higher when individuals were more active and participated in agonistic interactions. Thyroid hormone levels were also related to sex/reproductive state and activity, but were additionally positively related to fruit intake and negatively related to young leaf intake. Our study demonstrates that the non-invasive measurement of glucocorticoid and thyroid hormones of howler monkeys allows assessing different underlying physiological processes. By combining different biomarkers, which has seldom been done with wildlife, we could also parse the influence of psychological vs. metabolic challenges for individual energetic condition, which may be instrumental for deciding which factors should be accounted for when studying different hormone-behavior interactions.

Field Trial and Molecular Characterization of RNAi-Transgenic Tomato Plants That Exhibit Resistance to Tomato Yellow Leaf Curl Geminivirus.

RNA interference (RNAi) is a widely used approach to generate virus-resistant transgenic crops. However, issues of agricultural importance like the long-term durability of RNAi-mediated resistance under field conditions and the potential side effects provoked in the plant by the stable RNAi expression remain poorly investigated. Here, we performed field trials and molecular characterization studies of two homozygous transgenic tomato lines, with different selection markers, expressing an intron-hairpin RNA cognate to the Tomato yellow leaf curl virus (TYLCV) C1 gene. The tested F6 and F4 progenies of the respective kanamycin- and basta-resistant plants exhibited unchanged field resistance to TYLCV and stably expressed the transgene-derived short interfering RNA (siRNAs) to represent 6 to 8% of the total plant small RNAs. This value outnumbered the average percentage of viral siRNAs in the nontransformed plants exposed to TYLCV-infested whiteflies. As a result of the RNAi transgene expression, a common set of up- and downregulated genes was revealed in the transcriptome profile of the plants selected from either of the two transgenic events. A previously unidentified geminivirus causing no symptoms of viral disease was detected in some of the transgenic plants. The novel virus acquired V1 and V2 genes from TYLCV and C1, C2, C3, and C4 genes from a distantly related geminivirus and, thereby, it could evade the repressive sequence-specific action of transgene-derived siRNAs. Our findings shed light on the mechanisms of siRNA-directed antiviral silencing in transgenic plants and highlight the applicability limitations of this technology as it may alter the transcriptional pattern of nontarget genes.

Behavioural and Glucocorticoid Responses of a Captive Group of Spider Monkeys to Short-Term Variation in Food Presentation.

The presentation of food may affect feeding competition and the well-being of captive social species. We hypothesized that feeding competition in a captive group of 5 black-handed spider monkeys (Ateles geoffroyi) should increase in response to certain food presentations in terms of size, distribution and quality of food, and that higher feeding competition should lead to an increase in agonism and physiological stress (measured by faecal glucocorticoid metabolites, FGCM) as well as to a decrease in affiliation, proximity among individuals and feeding activity. We used 5 experimental treatments representing different combinations of size, distribution and quality of food. We observed social interactions for 100 h, collected 6,500 proximity and feeding activity records, and gathered 226 faecal samples. When food was clumped, individuals spent less time feeding, and there was also significant individual variation in feeding activity within treatments. FGCM levels were higher when food was clumped. These results are probably linked to an increase in feeding competition when food is concentrated. At least in small groups of spider monkeys, dispersing food in two feeding stations may be sufficient to decrease differences among individuals in priority of access to food resources, hence reducing physiological stress and interindividual differences in feeding activity.