In Vitro Analysis of SARS-CoV-2 Spike Protein and Ivermectin Interaction.

The spike (S) protein of SARS-CoV-2 is a molecular target of great interest for developing drug therapies against COVID-19 because S is responsible for the interaction of the virus with the host cell receptor. Currently, there is no outpatient safety treatment for COVID-19 disease. Furthermore, we consider it of worthy importance to evaluate experimentally the possible interaction of drugs (approved by the Food and Drug Administration) and the S, considering some previously in silico and clinical use. Then, the objective of this study was to demonstrate the in vitro interaction of ivermectin with S. The equilibrium dialysis technique with UV-Vis was performed to obtain the affinity and dissociation constants. In addition, the Drug Affinity Responsive Target Stability (DARTS) technique was used to demonstrate the in vitro interaction of S with ivermectin. The results indicate the interaction between ivermectin and the S with an association and dissociation constant of Ka = 1.22 µM-1 and Kd = 0.81 µM, respectively. The interaction was demonstrated in ratios of 1:50 pmol and 1:100 pmol (S: ivermectin) by the DARTS technique. The results obtained with these two different techniques demonstrate an interaction between S and ivermectin previously explored in silico, suggesting its clinical uses to stop the viral spread among susceptible human hosts.

Hyperbaric oxygenation applied before or after mild or hard stress: effects on the redox state in the muscle tissue.

The mechanism is unclear for the reported protective effect of hyperbaric oxygen preconditioning against oxidative stress in tissues, and the distinct effects of hyperbaric oxygen applied after stress. The trained mice were divided into three groups: the control, hyperbaric oxygenation preconditioning, and hyperbaric oxygenation applied after mild (fasting) or hard (prolonged exercise) stress. After preconditioning, we observed a decrease in basal levels of nitric oxide, tetrahydrobiopterin, and catalase despite the drastic increase in inducible and endothelial nitric oxide synthases. Moreover, the basal levels of glutathione, related enzymes, and nitrosative stress only increased in the preconditioning group. The control and preconditioning groups showed a similar mild stress response of the endothelial and neuronal nitric oxide synthases. At the same time, the activity of all nitric oxide synthase, glutathione (GSH) in muscle, declined in the experimental groups but increased in control during hard stress. The results suggested that hyperbaric oxygen preconditioning provoked uncoupling of nitric oxide synthases and the elevated levels of GSH in muscle during this study, while hyperbaric oxygen applied after stress showed a lower level of GSH but higher recovery post-exercise levels in the majority of antioxidant enzymes. We discuss the possible mechanisms of the redox response and the role of the nitric oxide in this process.

Medium- and long-range transport events of Alnus pollen in western Mediterranean.

Alnus pollen has been frequently detected in the atmosphere of different airborne sampling sites of Southern Spain. However, Alnus sp. populations are very scarce and fragmented in the area, being restricted to a few river valleys in the southwest, and other further away regions of the Iberian Peninsula. This leads to think that the airborne pollen detected could be mainly the result of a medium- or long-distant transport. So, the aim of this study was to characterize the annual patterns of airborne Alnus pollen detected at three different locations of Malaga province, as well as to determine its possible origin, the pollen dispersion potential of these Alnus isolated populations, and their possible reproductive connectivity. Pollen sampling was conducted by means of three Hirst-type volumetric pollen traps. Samples were mounted and counted following the recommendations of the Spanish Aerobiology Network and the European Aeroallergen Society. The possible pollen sources were detected by means of a combination of meteorological information and backward air trajectories analysis. A high inter-annual variability in the annual pollen integrals was found in all the stations, favouring certain meteorological conditions a long-range transport and, therefore, causing the high concentrations detected in some specific days. Alnus pollen seems to have a heterogeneous origin with prevalence of the long-distant transport, which would suggest a possible reproductive connection among distant populations.

A Novel Dynamic Three-Level Tracking Controller for Mobile Robots Considering Actuators and Power Stage Subsystems: Experimental Assessment.

In order to solve the trajectory tracking task in a wheeled mobile robot (WMR), a dynamic three-level controller is presented in this paper. The controller considers the mechanical structure, actuators, and power stage subsystems. Such a controller is designed as follows: At the high level is a dynamic control for the WMR (differential drive type). At the medium level is a PI current control for the actuators (DC motors). Lastly, at the low level is a differential flatness-based control for the power stage (DC/DC Buck power converters). The feasibility, robustness, and performance in closed-loop of the proposed controller are validated on a DDWMR prototype through Matlab-Simulink, the real-time interface ControlDesk, and a DS1104 board. The obtained results are experimentally assessed with a hierarchical tracking controller, recently reported in literature, that was also designed on the basis of the mechanical structure, actuators, and power stage subsystems. Although both controllers are robust when parametric disturbances are taken into account, the dynamic three-level tracking controller presented in this paper is better than the hierarchical tracking controller reported in literature.

Apoptosis Induced by (-)-Epicatechin in Human Breast Cancer Cells is Mediated by Reactive Oxygen Species.

(-)-Epicatechin is a phenolic compound with antioxidant activity that is present in natural food and drinks, such as cocoa and red wine. Evidence suggests that (-)-epicatechin exhibits anticancer activity; however, its mechanism of action is poorly understood. Here, we investigated the anticancer effects of (-)-epicatechin and its mechanism of action in breast cancer cells. We assessed the anticancer activity by cell proliferation assays, apoptosis by DNA fragmentation and flow cytometry. The expression of proteins associated with apoptosis was analyzed by the human apoptosis array. MitoSOXTM Red and biomarkers of oxidative damage were used to measure the effect of (-)-epicatechin on mitochondrial reactive oxygen species (ROS) and cellular damage, respectively. (-)-Epicatechin treatment caused a decreasing in the viability of MDA-MB-231 and MCF-7 cells. This cell death was associated with DNA fragmentation and an apoptotic proteomic profile. Further, (-)-epicatechin in MDA-MB-231 cells upregulated death receptor (DR4/DR5), increased the ROS production, and modulated pro-apoptotic proteins. In MCF-7 cells, (-)-epicatechin did not involve death receptor; however, an increase in ROS and the upregulation of pro-apoptotic proteins (Bad and Bax) were observed. These changes were associated with the apoptosis activation through the intrinsic pathway. In conclusion, this study shows that (-)-epicatechin has anticancer activity in breast cancer cells and provides novel insight into the molecular mechanism of (-)-epicatechin to induce apoptosis.

Masseter-to-Facial Nerve Transfer for Reanimation of a Patient With Long-Term Facial Paralysis.

Facial paralysis is a condition caused by a wide variety of etiologies, including neurologic, congenital, infectious, neoplastic, systemic, and iatrogenic causes. A patient suffering from long-term facial paralysis, with minimal innervation detected through electroneurography, who was successfully reanimated by performing a masseter-to-facial nerve transfer, was presented in this study. Facial paralysis had been caused after resection of an acquired middle ear cholesteatoma more than 5 years before.

Management of Hand Burn With Pedicled Converted Anterolateral Thigh Free Flap.

The upper limb is involved in burns in a high percentage of cases and its reconstruction is extremely important, given the functional impact of this anatomical region. Among the reconstruction choices for severe and large structural defects, the pedicled anterolateral thigh flap is an available option. This case study discusses the utilization of the pedicled anterolateral thigh flap for reconstruction of a complex full-thickness hand burn, when adequate arterial perforators were not available. Complex hand burns can often present challenges for reconstructive coverage, because of the complex anatomy of the upper extremity and the need to preserve as much function as possible. The use the anterolateral thigh free flap is one option that can be utilized for coverage of these large hand defects, in the face of poor local tissue advancement options. The finding of inadequate or lack of perforator vessels necessitates intraoperative changes in the surgical approach. In these cases, different alternatives exist depending on the dimensions and characteristics of the required coverage, the dissection of a pedicled flap being one of them. The pedicled anterolateral thigh flap represents an alternative for the coverage of large hand defects in the absence of valid perforators during free-flap dissection.

Endonasal Infiltrative Anesthesia for Nasal Fracture Reduction.

This study investigated the use of endonasal infiltrative anesthesia for the management of pain associated with nasal bone fracture reduction. Fifty-two patients with nasal bone fractures were distributed in 2 groups. In the first group, topical endonasal anesthesia and external transcutaneous infiltrative anesthesia were employed. In the second group, endonasal infiltrative anesthesia was also added. Visual analog scale pain scores related to the different steps of the procedure were registered. The addition of endonasal infiltrative anesthesia was associated with a significant decrease (p < .05) in pain during reduction maneuvers (6.71 vs. 4.83) and nasal packing (5.18 vs. 3.46). Addition of endonasal infiltrative anesthesia is an effective method of pain reduction during nasal bone fracture treatment.

A Risk-Adapted Approach to Patients with Stage I Seminoma according to the Status of Rete Testis: The Fourth Spanish Germ Cell Cancer Group Study.

OBJECTIVE: The aim of this study was to assess a risk-adapted strategy for stage I seminoma guided by the presence of rete testis invasion. METHODS: Between January 2013 and December 2015, a total of 135 consecutive patients with stage I seminoma from 18 Spanish tertiary hospitals were included in a prospective multicenter study. Median patient age was 38 years (range 22-60). Preoperative beta-human chorionic gonadotropin was elevated in 9.6% of patients. Rete testis invasion was present in 47.4% of patients. After orchiectomy, subjects with rete testis invasion were treated with 2 courses of adjuvant carboplatin (area under the curve of 7, with 21-day interval). Those without this risk factor were managed by surveillance. Disease-free survival (DFS) and overall survival (OS) were estimated with the Kaplan-Meier method. RESULTS: After a median follow-up time of 33 months, only 6 relapses were recorded (5 on surveillance, 1 after carboplatin). These cases were rescued with BEP or EP chemotherapy, and all 135 patients are currently disease free without sequelae. Three-year DFS was 92.0 and 98.2% for patients on surveillance and after carboplatin, respectively. Three-year OS was 100%. CONCLUSION: A risk-adapted approach based on rete testis invasion as a single risk factor is feasible and yielded an excellent outcome with a 3-year DFS of 94.9%.

Geriatric assessment in clinical practice for patients with stage IV non-small-cell lung cancer: The Grup de Investigació I Divulgació Oncològica experience.

Therapeutic decision-making for older patients with stage IV non-small-cell lung cancer (NSCLC) with no identifiable activating mutation is complex. In this prospective study, we evaluated the usefulness of geriatric assessment (GA) in identifying frail patients. Stage IV NSCLC patients ≥70 years of age were evaluated with GA and classified according to this evaluation into three different groups: fit, vulnerable and frail. Classifications based on GA, treatment decision, toxicity and overall survival were analysed. In total, 93 patients were included. Median age was 76 (70-92) years and 90% were men. Most patients had performance status (PS) 0 or 1 (82%), unrelated to their GA (p = 0.006). GA groups were associated with overall survival (p = 0.000), treatment decision (p = 0.0001), and toxicity (p = 0.0001). Chemotherapy was delivered to 100% of fit patients, to 48% of vulnerable patients, and to only 8% of frail patients (p = 0.000). Toxicity was higher in vulnerable patients than in fit individuals (p = 0.000). Multivariable analysis showed PS (p = 0.001), active treatment (p < 0.001) and GA group (p = 0.001) to be prognostic factors related to survival. Our results suggest that GA identified patients with poor natural prognosis.

Robust Switched Tracking Control for Wheeled Mobile Robots Considering the Actuators and Drivers.

By using the hierarchical controller approach, a new solution for the control problem related to trajectory tracking in a differential drive wheeled mobile robot (DDWMR) is presented in this paper. For this aim, the dynamics of the three subsystems composing a DDWMR, i.e., the mechanical structure (differential drive type), the actuators (DC motors), and the power stage (DC/DC Buck power converters), are taken into account. The proposed hierarchical switched controller has three levels: the high level corresponds to a kinematic control for the mechanical structure; the medium level includes two controls based on differential flatness for the actuators; and the low level is linked to two cascade switched controls based on sliding modes and PI control for the power stage. The hierarchical switched controller was experimentally implemented on a DDWMR prototype via MATLAB-Simulink along with a DS1104 board. With the intention of assessing the performance of the switched controller, experimental results associated with a hierarchical average controller recently reported in literature are also presented here. The experimental results show the robustness of both controllers when parametric uncertainties are applied. However, the performance achieved with the switched controller introduced in the present paper is better than, or at least similar to, performance achieved with the average controller reported in literature.

Absorption Capacity of Wound Dressings: A Comparative Experimental Study.

In this study, absorption capacity of the most commonly used types of wound dressings was measured with an objective, quantifiable, and easily reproducible method. The use of objective measurements allowed us to perform an objective comparison of the capacity of the different types of dressings. An experimental study was performed, in which the absorptive capacity of polyurethane foams, alginate, hidrofiber, and hydrocolloid dressings was measured by using a methodology designed by the researchers. Polyurethane foams showed a higher absorption capacity than hidrofibers, alginates, and hydrocolloids when using a similar surface of dressing. A modification of dressing size after saturation was also observed, increasing its size in the case of foams and decreasing or maintaining it in the case of hidrofibers, alginates, and hydrocolloids. The results of this study show a better wound exudate management for polyurethane foams. However, when deciding which dressing to use for a specific wound, absorption capacity is not the only quality that should be taken into account, as other properties should also be considered.

The Pitt Bacteremia Score, Charlson Comorbidity Index and Chronic Disease Score are useful tools for the prediction of mortality in patients with Candida bloodstream infection.

Candida bloodstream infection (CBI) is associated with high mortality. The aim of this study was to compare the utility of the combined use of the Pitt Bacteremia Score (PBS) and Charlson Comorbidity Index (CCI) or Chronic Disease Score (CDS) to predict mortality among patients with CBI. Thereby, all consecutive patients with CBI at our institution between 2010 and 2014 were included. The PBS was used to evaluate CBI severity and the CCI and CDS were used to assess comorbidities of patients with CBI. Logistic regression analysis was used to estimate odds ratios for 30-day mortality in models including the PBS and CCI or CDS. A total of 189 CBI episodes were identified. Logistic regression models including the PBS and either CCI or CDS showed that the combined use of a comorbidity score and a severity score significantly predicted 30-day mortality. The performance of the different models was similar. Aggregated scores of comorbidity (CCI and CDS) and disease severity (PBS) are useful for the prediction of 30-day mortality risk in patients with CBI. Their use may facilitate the analysis of risk factors for poorer outcome and the development of an index for CBI mortality.

Impact on clinical practice of the implementation of guidelines for the toxicity management of targeted therapies in kidney cancer. The protect-2 study.

BACKGROUND: The impact of such recommendations after their implementation of guidelines has not usually been evaluated. Herein, we assessed the impact and compliance with the Spanish Oncology Genitourinary Group (SOGUG) Guidelines for toxicity management of targeted therapies in metastatic renal cell carcinoma (mRCC) in daily clinical practice. METHODS: Data on 407 mRCC patients who initiated first-line targeted therapy during the year before and the year after publication and implementation of the SOGUG guideline program were available from 34 Spanish Hospitals. Adherence to SOGUG Guidelines was assessed in every cycle. RESULTS: Adverse event (AE) management was consistent with the Guidelines as a whole for 28.7% out of 966 post-implementation cycles compared with 23.1% out of 892 pre-implementation cycles (p = 0.006). Analysis of adherence by AE in non-compliant cycles showed significant changes in appropriate management of hypertension (33% pre-implementation vs. 44.5% post-implementation cycles; p < 0.0001), diarrhea (74.0% vs. 80.5%; p = 0.011) and dyslipemia (25.0% vs. 44.6%; p < 0.001). CONCLUSIONS: Slight but significant improvements in AE management were detected following the implementation of SOGUG recommendations. However, room for improvement in the management of AEs due to targeted agents still remains and could be the focus for further programs in this direction.

N-(2-hydroxyphenyl)-2-propylpentanamide, a valproic acid aryl derivative designed in silico with improved anti-proliferative activity in HeLa, rhabdomyosarcoma and breast cancer cells.

Epigenetic alterations are associated with cancer and their targeting is a promising approach for treatment of this disease. Among current epigenetic drugs, histone deacetylase (HDAC) inhibitors induce changes in gene expression that can lead to cell death in tumors. Valproic acid (VPA) is a HDAC inhibitor that has antitumor activity at mM range. However, it is known that VPA is a hepatotoxic drug. Therefore, the aim of this study was to design a set of VPA derivatives adding the arylamine core of the suberoylanilide hydroxamic acid (SAHA) with different substituents at its carboxyl group. These derivatives were submitted to docking simulations to select the most promising compound. The compound 2 (N-(2-hydroxyphenyl)-2-propylpentanamide) was the best candidate to be synthesized and evaluated in vitro as an anti-cancer agent against HeLa, rhabdomyosarcoma and breast cancer cell lines. Compound 2 showed a better IC50 (µM range) than VPA (mM range) on these cancer cells. And also, 2 was particularly effective on triple negative breast cancer cells. In conclusion, 2 is an example of drugs designed in silico that show biological properties against human cancer difficult to treat as triple negative breast cancer.

Comparative study of bacterial translocation control with nitric oxide donors and COX2 inhibitor.

OBJECTIVE AND DESIGN: To evaluate the beneficial effects of exogenous NO and an inhibitor of the COX2, and their action levels in a model of SIRS/bacterial translocation (BT) induced by Zymosan A(®). MATERIAL AND METHODS: Ninety Wistar rats were submitted to different treatments, and after 12h and 24h they were anaesthetized in order to collect blood, mesenteric lymph nodes, and kidney for subsequent biochemical analyses and microbiological examinations. TREATMENTS: A nitric oxide donor, Molsidomine(®), was compared with a COX2 inhibitor, Celecoxib(®). METHODS: Zymosan A(®) was administered to Wistar rats. The animals were divided into 6 groups: one group for survival study, Group (1) No manipulation (BASAL); Group (2) vehicle of Zymosan A(®) given intraperitoneally (SHAM); Group I (control), with Zymosan A(®) (0.6g/kg) intraperitoneally; Group II (Molsidomine), with Molsidomine(®) (4mg/kg) through the penis dorsal vein, 30min prior to administration of the Zy(®) (0.6g/kg); Group III (Celecoxib), with Celecoxib(®) (400mg/kg) orally through a stomach tube, 6h prior to administration of the Zy (0.6g/kg). DETERMINATIONS: The parameters survival, bacterial translocation, renal function, neutrophil accumulation, oxygen free radicals (OFR), detoxifying enzymes, and cytokines were measured at different times after Zymosan administration. RESULTS: The model established induced a mortality rate of 100% and generated BT and systemic inflammatory response syndrome (SIRS) in all samples. It also significantly increased all variables, with p<.001 for MPO and all pro-inflammatory cytokines, and p<.01 for all OFR. Treatment with Molsidomine reduced mortality to 0%, decreased BT, MPO, pro-inflammatory cytokines and OFR (p<.001) significantly and increased IL-10 and IL-6 production. Moreover, the Celecoxib(®) showed a lower capacity for SIRS regulation. CONCLUSIONS: The exogenous administration of NO prevented BT and controlled SIRS. Therefore these results suggest that Molsidomine could be used as a therapeutic strategy to protect against BT.

Identification of Ligustrum lucidum pollen allergens using a proteomics approach.

BACKGROUND: Ligustrum spp. are members of the Oleaceae family, one of the most prominent allergic families worldwide. The genus Ligustrum contains approximately fifty species, including Ligustrum lucidum, which have been widely cultivated as ornamental plants, and its pollen is a source of inhalant allergens associated with respiratory allergic diseases. Little is known about the presence of allergenic proteins in L. lucidum. METHODS: The L. lucidum pollen proteins were extracted by a modified phenolic extraction method. A pool of four sera from mono sensitive patients was analyzed by 2DE immunoblotting and mass spectrometric analysis was performed on 6 immunoreactive protein spots. RESULTS: SDS-PAGE of L. lucidum pollen extract revealed proteins in ranges of 15-150 kDa. The 2DE gel profile of the L. lucidum pollen protein extract showed approximately 180 spots, and the 2DE immunoblots obtained using sera from Ligustrum monosensitive patients as the source of IgE antibodies revealed six allergen protein spots, corresponding to Profilin, Enolase, Fra e 9.01 (ß-1,3-glucanase), Pollen-specific Polygalacturonases, Alanine aminotransferase, and two ATP synthase beta subunits. CONCLUSION: We report for the first time the identification of IgE-reactive proteins from L. lucidum.

[Polio, the long walk to the endgame]. / La polio, el largo camino hacia el final de la partida.

Although the WHO original target date for the global eradication of poliomyelitis was the year 2000 -thanks to vaccination and institutional, public and private, resources for that purpose-, in 2013 the disease remained endemic in three countries, Afghanistan, Pakistan and Nigeria, and some cases were described in five others. The circulation of wild type 1 poliovirus in Israel, Gaza and the West Bank and the cases in Syria were a wakeup call, as at that time there were polioviruses derived from the oral vaccine that are still circulating among the human population and can cause the development of the disease. Travelling "from" and "to" endemic areas are factors to consider in poliovirus exportation and in its spread when it reaches areas with poor immunogenicity. Wars, terrorism, intolerance, lack of culture and proliferation of anti-vaccine groups and the rise of the anti-vaccination movement are important factors in the maintenance and expansion of the virus and in the "non-vaccination" against it. Based on the international situation to date, the Emergency Committee of WHO met in May 2014 to address the problem. It is still necessary to enhance the knowledge of the disease and its agent. In the first case to perform a differential diagnosis of flaccid paralysis and to continue vaccination programs, and in the second case to keep studying and looking for the poliovirus in environmental samples, which is a model for the study of many other viruses.

[Anaerobic bacteria 150 years after their discovery by Pasteur]. / Las bacterias anaerobias 150años después de su descubrimiento por Pasteur.

In 2011 we celebrated the 150th anniversary of the discovery of anaerobic bacteria by Louis Pasteur. The interest of the biomedical community on such bacteria is still maintained, and is particularly focused on Clostridium difficile. In the past few years important advances in taxonomy have been made due to the genetic, technological and computing developments. Thus, a significant number of new species related to human infections have been characterised, and some already known have been reclassified. At pathogenic level some specimens of anaerobic microflora, that had not been isolated from human infections, have been now isolated in some clinical conditions. There was emergence (or re-emergence) of some species and clinical conditions. Certain anaerobic bacteria have been associated with established infectious syndromes. The virulence of certain strains has increased, and some hypotheses on their participation in certain diseases have been given. In terms of diagnosis, the routine use of MALDI-TOF has led to a shortening of time and a cost reduction in the identification, with an improvement directly related to the improvement of data bases. The application of real-time PCR has been another major progress, and the sequencing of 16srRNA gene and others is currently a reality for several laboratories. Anaerobes have increased their resistance to antimicrobial agents, and the emergence of resistance to carbapenems and metronidazole, and multi-resistance is a current reality. In this situation, linezolid could be an effective alternative for Bacteroides. Fidaxomicin is the only anti-anaerobic agent introduced in the recent years, specifically for the diarrhoea caused by C.difficile. Moreover, some mathematical models have also been proposed in relation with this species.

The association of subtypes of breast cancer with tumour characteristics and reproductive factors in 1326 Mexican women.

AIM OF THE STUDY: In breast cancer, oestrogen receptor (ER), progesterone receptor (PgR), and HER2 (HER2/Neu) expression status are used to classify neoplasms into subtypes: Luminal A, Luminal B, HER2/Neu type, and Basallike. The aim of the present study was to establish the molecular subtypes of breast cancers and their association with tumour characteristics and reproductive factors in Mexican women. MATERIAL AND METHODS: A total of 1326 biopsies of breast tumour tissues were analysed for ER, PR, and HER2/Neu by immunohistochemistry (IHC). Information regarding age, tumour characteristics, and node involvement profiles were collected. RESULTS: IHC established that the most common subtype of breast cancer was Luminal A (64.93%), followed by Basal-Like (13.88%), Luminal B (12.52%), and HER2/Neu (8.67%). T2-size tumours (> 2 cm but < 5 cm) were present in 47.59% of all patients. Univariate analysis showed that lymph node positivity (p = 0.009), stage (p = 0.013), and placement of the tumour (p = 0.001) were factors associated with breast cancer subtype. CONCLUSIONS: Our data show that IHC is useful for distinguishing different subtypes of breast cancer and that Luminal A is the most common breast cancer subtype in the Mexican population. All subtypes were associated with unfavourable clinicopathological features, suggesting that late diagnosis is an important contributor to high mortality rates in the Mexican population.