Simultaneously Targeting Two Coupled Signalling Molecules in the Mesenchymal Stem Cell Support Efficiently Sensitises the Multiple Myeloma Cell Line H929 to Bortezomib.

Several studies have shown that diverse components of the bone marrow (BM) microenvironment play a central role in the progression, pathophysiology, and drug resistance in multiple myeloma (MM). In particular, the dynamic interaction between BM mesenchymal stem cells (BM-MSC) and MM cells has shown great relevance. Here we showed that inhibiting both PKC and NF-κB signalling pathways in BM-MSC reduced cell survival in the MM cell line H929 and increased its susceptibility to the proteasome inhibitor bortezomib. PKC-mediated cell survival inhibition and bortezomib susceptibility induction were better performed by the chimeric peptide HKPS than by the classical enzastaurin inhibitor, probably due to its greatest ability to inhibit cell adhesion and its increased capability to counteract the NF-κB-related signalling molecules increased by the co-cultivation of BM-MSC with H929 cells. Thus, inhibiting two coupled signalling molecules in BM-MSC was more effective in blocking the supportive cues emerging from the mesenchymal stroma. Considering that H929 cells were also directly susceptible to PKC and NF-κB inhibition, we showed that treatment of co-cultures with the HKPS peptide and BAY11-7082, followed by bortezomib, increased H929 cell death. Therefore, targeting simultaneously connected signalling elements of BM-MSC responsible for MM cells support with compounds that also have anti-MM activity can be an improved treatment strategy.

Regional variation in multiple paternity in the brown smooth-hound shark Mustelus henlei from the northeastern Pacific.

Multiple paternity (MP) in the brown smooth-hound shark (Mustelus henlei) was assessed in 15 litters (15 mothers and 97 embryos) collected in the northern Gulf of California of which 86.7% were sired by more than one male (i.e., from 2 to 4 sires). When taken together with results from previous studies, this record indicates that there is regional variation in MP in M. henlei in the northeastern Pacific. This pattern is associated with variations in the reproductive traits of each population (e.g., female size and litter size). In the Gulf of California, the results of a generalized linear model (GLZ) indicated that the litters of larger females had a higher probability of MP compared to those of smaller females.

Lymphocyte subsets early predict mortality in a large series of hospitalized COVID-19 patients in Spain.

The role of lymphocytes and their main subsets as prognostic factors of death in SARS-CoV-2-infected patients remains unclear, with no information obtained from patients outside China. We aimed to assess whether measuring lymphocyte subpopulations added clinical value to the total lymphocyte counting regarding mortality when they were simultaneously tested at hospital admission. Peripheral blood was analysed in 701 polymerase chain reaction (PCR)-confirmed consecutive patients by lysed-no washed flow cytometry. Demographic and clinical features were registered in electronic medical records. Statistical analysis was performed after a 3-month follow-up. The 112 patients who died were older and had significantly higher frequencies of known co-morbidities than survivor COVID-19 patients. A significant reduction in total lymphocytes, CD3+ , CD4+ , CD8+ and CD19+ counts and CD3+ percentage was found in the group of deceased patients (P < 0·001), while the percentage of CD56+ /CD16+ natural killer (NK) cells was significantly higher (P < 0·001). Multivariate logistic regression analysis showed a significantly increased risk of in-hospital death associated to age [odds ratio (OR) = 2·36, 95% confidence interval (CI) = 1·9-3·0 P < 0·001]; CD4+  T counts ≤ 500 cells/µl, (OR = 2·79, 95% CI = 1·1-6·7, P = 0·021); CD8+  T counts ≤ 100 cells/µl, (OR = 1·98, 95% CI = 1·2-3·3) P = 0·009) and CD56+ /CD16+ NK ≥ 30%, (OR = 1·97, 95% CI = 1·1-3·1, P = 0·002) at admission, independent of total lymphocyte numbers and co-morbidities, with area under the curve 0·85 (95% CI = 0·81-0·88). Reduced counts of CD4+ and CD8+ T cells with proportional expansion of NK lymphocytes at admission were prognostic factors of death in this Spanish series. In COVID-19 patients with normal levels of lymphocytes or mild lymphopenia, imbalanced lymphocyte subpopulations were early markers of in-hospital mortality.

Inverse method based on 3D nonlinear physically constrained minimisation in the framework of traction force microscopy.

Traction force microscopy is a methodology that enables to estimate cellular forces from the measurement of the displacement field of an extracellular matrix (ECM)-mimicking hydrogel that a cell is mechanically interacting with. In this paper, a new inverse and physically-consistent methodology is developed and implemented in the context of 3D nonlinear elasticity. The proposed method searches for a displacement field that approximates the measured one, through the imposition of fulfillment of equilibrium with real and known forces acting in the hydrogel. The overall mathematical formulation leads to a constrained optimisation problem that is treated through a Lagrange operator and that is solved numerically by means of a nonlinear finite element framework. In order to illustrate the potential and enhanced accuracy of the proposed inverse method, it is applied to a total of 5 different real cases of cells cultured in a 3D hydrogel that is considered to behave as a nonlinear elastic material. Different error indicators are defined in order to compare ground truth simulated displacements and tractions to the ones recovered by the new inverse as well as by the forward method. Results indicate that the evaluation of displacement gradients leads to errors, in terms of recovered tractions, that are more than three times lower (on average) for the inverse method compared to the forward method. They highlight the enhanced accuracy of the developed methodology and the importance of appropriate inverse methods that impose physical constraints to traction and stress recovery in the context of traction force microscopy.

Isolation and characterization of high affinity and highly stable anti-Chikungunya virus antibodies using ALTHEA Gold Libraries™.

BACKGROUND: More than 3 million infections were attributed to Chikungunya virus (CHIKV) in the 2014-2016 outbreak in Mexico, Central and South America, with over 500 deaths directly or indirectly related to this viral disease. CHIKV outbreaks are recurrent and no vaccine nor approved therapeutics exist to prevent or treat CHIKV infection. Reliable and robust diagnostic methods are thus critical to control future CHIKV outbreaks. Direct CHIKV detection in serum samples via highly specific and high affinity anti-CHIKV antibodies has shown to be an early and effective clinical diagnosis. METHODS: To isolate highly specific and high affinity anti-CHIKV, Chikungunya virions were isolated from serum of a patient in Veracruz, México. After purification and characterization via electron microscopy, SDS-PAGE and binding to well-characterized anti-CHIKV antibodies, UV-inactivated particles were utilized as selector in a solid-phase panning in combination with ALTHEA Gold Libraries™, as source of antibodies. The screening was based on ELISA and Next-Generation Sequencing. RESULTS: The CHIKV isolate showed the typical morphology of the virus. Protein bands in the SDS-PAGE were consistent with the size of CHIKV capsid proteins. UV-inactivated CHIKV particles bound tightly the control antibodies. The lead antibodies here obtained, on the other hand, showed high expression yield, > 95% monomeric content after a single-step Protein A purification, and importantly, had a thermal stability above 75 °C. Most of the antibodies recognized linear epitopes on E2, including the highest affinity antibody called C7. A sandwich ELISA implemented with C7 and a potent neutralizing antibody isolated elsewhere, also specific for E2 but recognizing a discontinuous epitope, showed a dynamic range of 0.2-40.0 mg/mL of UV-inactivated CHIKV purified preparation. The number of CHIKV particles estimated based on the concentration of E2 in the extract suggested that the assay could detect clinically meaningful amounts of CHIKV in serum. CONCLUSIONS: The newly discovered antibodies offer valuable tools for characterization of CHIKV isolates. Therefore, the strategy here followed using whole viral particles and ALTHEA Gold Libraries™ could expedite the discovery and development of antibodies for detection and control of emergent and quickly spreading viral outbreaks.

[Extubation failure after Norwood procedure in hypoplastic left heart syndrome patients]. / Extubación fallida postoperación de Norwood en pacientes con síndrome de hipoplasia de ventrículo izquierdo.

INTRODUCTION: Hypoplastic left heart syndrome (HLHS) is the main cause of mortality due to congenital heart disea se. The Norwood surgery is the first procedure of the surgical staging process towards a single ventri cle physiology or Fontan-type operation and has a mortality rate of 10% to 30%. Extubation failure during the postoperative period occurs in up to 18% of these patients and is associated with increased mortality. OBJECTIVE: To describe extubation failure rates and risk factors in pediatric patients with HLHS who underwent Norwood procedure. PATIENTS AND METHOD: Case-control study that included all the patients with HLHS managed with Norwood surgery at the Hospital Clínico de la Pontificia Universidad Catolica between January 2000 and February 2018. Cases and controls were defined as patients with extubation failure and as patients without this complication, respectively. The fo llowing variables were recorded demographic, surgical, and post-surgical ones, and univariate and multivariate analyses (logistic regression) were performed to determine risk factors associated with extubation failure. RESULTS: Out of 107 patients, 26 of them presented extubation failure (24.3%). In the univariate analysis, longer mechanical ventilation time during the postsurgical period, atelectasis, pleural effusion, chylothorax, other respiratory morbidities (i.e. apneas and tracheitis), and longer infusion times of morphine and midazolam, all were associated with a higher extubation failure rate in this population. In the multivariable analysis, chylothorax, other respiratory comorbidities, and longer infusion time of midazolam remained associated with this complication, however, it was not associated with higher mortality. CONCLUSIONS: Chylothorax, respiratory comorbidities, and longer use of Midazolam should be addressed before planning airway extubation in order to avoid failure.

Nutritional and non-nutritional agents that stimulate white adipose tissue browning.

Obesity is a public health problem present in both developed and developing countries. The white adipose tissue (WAT) is the main deposit of lipids when there is an excess of energy. Its pathological growth is directly linked to the development of obesity and to a wide number of comorbidities, such as insulin-resistance, cardiovascular disease, among others. In this scenario, it becomes imperative to develop new approaches to the treatment and prevention of obesity and its comorbidities. It has been documented that the browning of WAT could be a suitable strategy to tackle the obesity epidemic that is developing worldwide. Currently there is an intense search for bioactive compounds with anti-obesity properties, which present the particular ability to generate thermogenesis in the brown adipose tissue (BAT) or beige. The present study provide recent information of the bioactive nutritional compounds capable of inducing thermogenesis and therefore capable of generate positive effects on health.

High SNP diversity in the non-toxic indigenous Jatropha curcas germplasm widens the potential of this upcoming major biofuel crop species.

BACKGROUND AND AIMS: Jatropha curcas (jatropha) is an oil crop cultivated in (sub)tropical regions around the world, and holds great promise as a renewable energy source. However, efforts to fully commercialize jatropha are currently hampered by the lack of genetic diversity in the extant breeding germplasm, and by the toxicity of its seeds meaning that its seed cake cannot be used as a protein source in animal feed, among other constraints. In Mexico, the species' native range, there are jatropha plants whose seeds are used to prepare traditional meals. This non-toxic jatropha 'type' is considered to harbour low genetic variation due to a presumed domestication bottleneck and therefore to be of limited breeding value; yet, very little is known regarding its origin and genetic diversity. METHODS: Using genotyping-by-sequencing (GBS), we extensively genotyped both indigenous toxic and non-toxic jatropha collected along roads and home gardens throughout southern Mexico. KEY RESULTS: Single nucleotide polymorphism diversity in non-toxic jatropha is relatively high, particularly in northern Veracruz state, the probable origin of this germplasm. Genetic differences between toxic and non-toxic indigenous genotypes are overall quite small. A a genome-wide association study supported a genomic region (on LG 8, scaffold NW_012130064), probably involved in the suppression of seed toxicity. CONCLUSIONS: Conservation actions are urgently needed to preserve this non-toxic indigenous, relatively wild germplasm, having potential as a fuel feedstock, animal feed and food source among other uses. More generally, this work demonstrates the value of conservation genomic research on the indigenous gene pool of economically important plant species.

Oviductal Proteins Effect in Rooster Spermatic Cryopreservation.

BACKGROUND: Cryopreservation induces spermatic cryo capacitation, which can decrease thawed sperm fertilizing capability. OBJECTIVE: To evaluate the effect of uterus-vaginal union protein factors to inhibit sperm cryo capacitation and maintain viability and fertilizing capability of rooster spermatozoa. MATERIALS AND METHODS: Rooster spermatozoa was cryopreserved using Lake extender supplemented with different hen's uterus-vaginal junction protein concentrations, to determine spermatic viability, sperm physiological condition and fertilizing capability in vivo. RESULTS: It was possible to induce spermatic decapacitation in vitro, inhibiting cryo capacitation and allowing fertility results comparable to those obtained with fresh semen. CONCLUSION: Uterus-vaginal protein extracts induce spermatic decapacitation in vitro.

Effect of thermal treatment on the extraction efficiency, physicochemical quality of Jatropha curcas oil, and biological quality of its proteins.

Jatropha curcas seeds are an important source of oil (5-60%), used to obtain biodiesel. The generated residual paste has a high concentration of proteins (50-55%); however, the seeds contain non-nutritional factors that limit their use. The objective of this work was to analyze the effect on the physicochemical properties of the oil obtained from J. curcas seeds subjected to different thermal treatments and to evaluate the biological quality of the proteins contained in the residual cake. The best extraction of oil (95%) was obtained after 10 h from roasted or boiled seeds. In the oil from roasted samples, the acid index increased significantly (p ≤ 0.05) with respect to the untreated sample, whereas the iodine index increased significantly (p ≤ 0.05) in the oil extracted from the boiled samples. With respect to the proximal chemical composition of the flour, roasting and boiling treatments allowed for greater oil extraction (97 and 92%), achieving, in turn, a higher content of proteins (59.56 and 58.5 g/100 g) and fiber (6.67 and 6.67 g/100 g), and lower activity of trypsin inhibitors (45 and 38%) and phytates (63 and 72%), respectively. According to the in vivo biological quality test, conducted on Wistar rats, the thermal treatments applied to the seeds improved digestibility (> 70%) and the protein efficiency index (PER). The thermal treatment allowed extracting more efficiently the oil and improved the quality of the proteins present in the residual paste.

Determination of the effect of polyamines on an oil-degrading strain of Yarrowia lipolytica using an odc minus mutant.

Yarrowia lipolytica is an ascomycetous dimorphic yeast with immense potential for industrial applications, including bioremediation of crude oil-contaminated environments. It has been shown that a dimorphic marine isolate of Y. lipolytica (var. indica) has significant capacity to degrade fatty acids and alkanes, when in its yeast morphology. It has also been demonstrated that polyamines play an important role in the yeast-to-mycelium transition of different strains of Y. lipolytica that are unable to utilize those carbon sources. To determine the role of polyamines on their capacity to utilize oils and hydrocarbons, on the dimorphic transition, and also on other characteristics of the var. indica strain of Y. lipolytica, we proceeded to obtain ornithine decarboxylase minus (odc-) mutants. These mutants behaved as yeasts independently of the concentrations of putrescine added. Further, they conserved the oil-degrading capacity of the parent strain. The odc- mutant can thus be used in fatty acid degradation, and oil spill remediation with distinct advantages.

Prevalence of functional gastrointestinal disorders in adults with systemic lupus erythematosus.

Objective (a) to assess the prevalence of functional gastrointestinal disorders (FGIDs) in female Mexican systemic lupus erythematosus (SLE) patients using the Rome III criteria and (b) to examine the effect of disease duration on FGID prevalence. Methods Female SLE outpatients aged ≥18 years with no organic gastrointestinal disorder were included. Participants were invited to upper gastrointestinal endoscopy screening and a faecal immunochemical test. FGID symptoms were evaluated using the Rome III questionnaire. Results Eighty-six SLE patients with median age of 45 (interquartile range 34-54) years were included. At least one FGID was found in 76.7% (66/88) of patients with SLE. The most prevalent domains of FGID diagnosed were functional oesophageal, gastroduodenal disorders and bowel disorders, of which functional dyspepsia (72.7%), functional heartburn (68.1%) and bloating (63.8%) were the most frequent. Fifty-nine per cent of patients had overlapping FGIDs. The most prevalent overlap was the combination of functional dyspepsia and functional heartburn. Patients with longer disease duration had a higher prevalence of FGID than those with shorter disease duration. Conclusions There was a high prevalence of FGIDs in Mexican SLE women with low disease activity. Overlapping FGIDs were frequent. Longer disease duration may be associated with FGIDs in SLE patients.

Transcriptomic analysis of basidiocarp development in Ustilago maydis (DC) Cda.

Previously, we demonstrated that when Ustilago maydis (DC) Cda., a phytopathogenic basidiomycete and the causal agent of corn smut, is grown in the vicinity of maize embryogenic calli in a medium supplemented with the herbicide Dicamba, it developed gastroid-like basidiocarps. To elucidate the molecular mechanisms involved in the basidiocarp development by the fungus, we proceeded to analyze the transcriptome of the process, identifying a total of 2002 and 1064 differentially expressed genes at two developmental stages, young and mature basidiocarps, respectively. Function of these genes was analyzed with the use of different databases. MIPS analysis revealed that in the stage of young basidiocarp, among the ca. two thousand differentially expressed genes, there were some previously described for basidiocarp development in other fungal species. Additional elements that operated at this stage included, among others, genes encoding the transcription factors FOXO3, MIG3, PRO1, TEC1, copper and MFS transporters, and cytochromes P450. During mature basidiocarp development, important up-regulated genes included those encoding hydrophobins, laccases, and ferric reductase (FRE/NOX). The demonstration that a mapkk mutant was unable to form basidiocarps, indicated the importance of the MAPK signaling pathway in this developmental process.

Changes in the membrane carbohydrates from sperm cryopreserved with dimethylsulfoxide or polyvinylpyrrolidone of red-tailed hawk (Buteo jamaicencis).

BACKGROUND: That cryopreservation can induce alterations in sperm. OBJECTIVE: The goal of this study was to evaluate sperm quality and distribution of N-acetylglucosamine, sialic acid and mannose residues in sperm cryopreserved of red-tailed hawk (Buteo jamaicensis). MATERIALS AND METHODS: We studied twenty samples of ejaculated semen for each cryoprotectant dimethylsulfoxide or polyvinylpyrrolidone. Carbohydrate identification was carried out with lectins Triticum vulgaris agglutinin to N-acetylglucosamine and sialic acid and Concanavalia ensiformis for mannose residues. Sperm viability was not altered but motility decreased significantly with both crioprotectants compared with fresh sperm. RESULTS: Neither the number of WGA positive sperm nor the distribution of N-acetylglucosamine and/or sialic acid residues were affected by the cryopreservation procedure. The sperm proportion with fluorescence associated with the presence of mannose residues was higher in thawed sperm. CONCLUSION: Values obtained with the cryopreservation technique proposed in this study by freezing drops in liquid nitrogen, were within normal parameters established for good quality fresh semen. We can say that it can be used for assisted reproduction of Buteo jamaicensis.

The difficulty of the postural control task affects multi-muscle control during quiet standing.

The aim of this study was to compare the electromyographic (EMG) coherence between the lower limb and the core muscles when carrying out two postural tasks at different difficulty levels. EMG was recorded in 20 healthy male subjects while performing two independent quiet standing tasks. The first one involved a bipedal stance with the eyes open, while the second consisted of a dominant unipedal stance also with the eyes open. The obtained EMG signals were analysed by computing estimations of EMG-EMG coherence between muscle pairs, both singly (single-pair estimations) and combined (pooled estimations). Pooled and single coherence of anterior, posterior, core, antagonist and mixed pairs of muscles were significant in the 0-5 Hz frequency band. The results indicate that core and antagonist muscle groups, such as the anterior and posterior muscles, share low-frequency neural inputs (0-5 Hz) which could be responsible of the M-modes assembly. The core muscles could therefore provide the necessary synergy to maintain spine stability during the balancing exercise. Finally, differences in EMG-EMG coherence suggest that the muscle synergies formed during unipedal stance tasks are different from those established during bipedal stance.

Self-Sensing of Damage Progression in Unidirectional Multiscale Hierarchical Composites Subjected to Cyclic Tensile Loading.

The electrical sensitivity of glass fiber/multiwall carbon nanotube/vinyl ester hierarchical composites containing a tailored electrically-percolated network to self-sense accumulation of structural damage when subjected to cyclic tensile loading-unloading is investigated. The hierarchical composites were designed to contain two architectures differentiated by the location of the multiwall carbon nanotubes (MWCNTs), viz. MWCNTs deposited on the fibers and MWCNTs dispersed within the matrix. The changes in electrical resistance of the hierarchical composites are associated to their structural damage and correlated to acoustic emissions. The results show that such tailored hierarchical composites are able to self-sense damage onset and accumulation upon tensile loading-unloading cycles by means of their electrical response, and that the electrical response depends on the MWCNT location.

Socioeconomic status and organ damage in Mexican systemic lupus erythematosus women.

The objective of this cross-sectional study was to determine relationships between socioeconomic status and organ damage in Mexican systemic lupus erythematosus (SLE) patients. Demographic and clinical variables were assessed. Socioeconomic status was evaluated using the Graffar method and monthly household income. Lupus activity and organ damage were measured using the SLE disease activity scale, validated for the Mexican population (Mex-SLEDAI), and the Systemic Lupus International Collaborating Clinics/American College of Rheumatology (SLICC/ACR) scale. The 143 Mexican female SLE patients included (mean age 40.1 ± 8.9 years, mean disease duration 8.9 ± 6.3 years) had a mean monthly household income of $ 407.2 ± 326.5. According to the Graffar index, 18.9%, 52.5%, and 28.7% had high/medium-high, medium, and medium-low/low socioeconomic status, respectively. Organ damage was observed in 61 patients (42.7%). Patients with organ damage had lower monthly household incomes ($241.4 ± 152.4 vs. $354.8 ± 288.3) and were more frequently unemployed (57.3% vs. 35.3%; p = 0.01) than those without. Low monthly income was not associated with lupus activity or self-reported health status. In the adjusted multivariate analysis, low monthly income ( < $300) was associated with organ damage. In conclusion, low income may be associated with organ damage in Mexican SLE patients.

Seroprevalence and risk factors associated with within-flock transmission of Leptospira interrogans in transhumant farming systems in Mexico.

A number of recent reports emphasize the risk of zoonotic diseases and the high degree of prevalence of asymptomatic animals infected with Leptospira interrogans. This report sought to assess the prevalence of antibodies to certain serovars of L. interrogans, and to describe the association between seropositivity and risk factors associated with within-flock transmission in a mountainous region of Mexico. Overall seroprevalence to L. interrogans was 54·5% (95% confidence interval 48·3-60·7); the most frequent serovar was Icterohaemorrhagiae. The accumulation of placentas and fetuses at a site close to lambing paddocks can play a significant role as a risk factor for within-flock transmission of L. interrogans in transhumant farming systems in the municipality of Xalatlaco. The high prevalence of L. interrogans antibodies supports the hypothesis that natural foci of this zoonosis are present in sheep flocks in this area. These findings emphasize the need for planning and implementation of control programmes for ovine leptospirosis in Mexico and elsewhere.

Functional near infrared spectroscopy as a potential biological assessment of addiction recovery: preliminary findings.

BACKGROUND: Addiction science has primarily utilized self-report, continued substance use, and relapse factors to explore the process of recovery. However, the entry into successful abstinence substantially reduces our assessment abilities. Advances in neuroscience may be the key to objective understanding, treating, and monitoring long-term success in addiction recovery. OBJECTIVES: To explore functional near infrared spectroscopy (fNIR) as a viable technique in the assessment of addiction-cue reactivity. Specifically, prefrontal cortex (PFC) activation to alcohol cues was explored among formally alcohol-dependent individuals, across varying levels of successful abstinence. The aim of the investigation was to identify patterns of PFC activation change consistent with duration of abstinence. METHODS: A total of 15 formally alcohol-dependent individuals, with abstinence durations ranging from 1 month to 10 years, viewed alcohol images during fNIR PFC assessment. Participants also subjectively rated the same images for affect and arousal level. RESULTS: Subjective ratings of alcohol cues did not significantly correlate with duration of abstinence. As expected, days of abstinence did not significantly correlate with neutral cue fNIR reactivity. However, for alcohol cues, fNIR results showed increased days of abstinence was associated with decreased activation within the dorsolateral and dorsomedial prefrontal cortex regions. CONCLUSIONS: The present results suggest that fNIR may be a viable tool in the assessment of addiction-cue reactivity. RESULTS also support previous findings on the importance of dorsolateral and dorsomedial PFC in alcohol-cue activation. The findings build upon these past results suggesting that fNIR-assessed activation may represent a robust biological marker of successful addiction recovery.