In silico and in vitro study of FLT3 inhibitors and their application in acute myeloid leukemia.

Acute myeloid leukemia (AML) is the most common hematological cancer in the adult population worldwide. Approximately 35% of patients with AML present internal tandem duplication (ITD) mutations in the FMS­like tyrosine kinase 3 (FLT3) receptor associated with poor prognosis, and thus, this receptor is a relevant target for potential therapeutics. Tyrosine kinase inhibitors (TKIs) are used to treat AML; however, their molecular interactions and effects on leukemic cells are poorly understood. The present study aimed to gain insights into the molecular interactions and affinity forces of four TKI drugs (sorafenib, midostaurin, gilteritinib and quizartinib) with the wild­type (WT)­FLT3 and ITD­mutated (ITD­FLT3) structural models of FLT3, in its inactive aspartic acid­phenylalanine­glycine motif (DFG­out) and active aspartic acid­phenylalanine­glycine motif (DFG­in) conformations. Furthermore, the present study evaluated the effects of the second­generation TKIs gilteritinib and quizartinib on cancer cell viability, apoptosis and proliferation in the MV4­11 (ITD­FLT3) and HL60 (WT­FLT3) AML cell lines. Peripheral blood mononuclear cells (PBMCs) from a healthy volunteer were included as an FLT3­negative group. Molecular docking analysis indicated higher affinities of second­generation TKIs for WT­FLT3/DFG­out and WT­FLT3/DFG­in compared with those of the first­generation TKIs. However, the ITD mutation changed the affinity of all TKIs. The in vitro data supported the in silico predictions: MV4­11 cells presented high selective sensibility to gilteritinib and quizartinib compared with the HL60 cells, whereas the drugs had no effect on PBMCs. Thus, the current study presented novel information about molecular interactions between the FLT3 receptors (WT or ITD­mutated) and some of their inhibitors. It also paves the way for the search for novel inhibitory molecules with potential use against AML.

Pelargonic acid's interaction with the auxin transporter PIN1: A potential mechanism behind its phytotoxic effects on plant metabolism.

Pelargonic acid (PA) is a saturated fatty acid commonly found in several organisms, that is known for its phytotoxic effect and its use as bioherbicide for sustainable weed management. Although PA is already commercialised as bioherbicide, its molecular targets and mode of action is unknown according to the Herbicide Resistance Action Committee. Therefore, the aim of this work was focusing on the way this natural active substance impacts the plant metabolism of the model species Arabidopsis thaliana. PA caused increase of secondary and adventitious roots, as well as torsion, loss of gravitropism and phytotoxic effects. Moreover, PA altered the cellular arrangement and the PIN proteins activity. Computational simulations revealed that the intermolecular interactions between PA and the polar auxin transporter protein PIN1 are very similar to those established between the natural auxin IAA and PIN1. However, under intracellular conditions, the PA-PIN1 binding is more energetically stable than the IAA-PIN1. These results suggest that PA could act as an auxin-mimics bioherbicide. The exogenous application of PA would be responsible for the alterations observed both at structural and ultrastructural levels, which would be caused by the alteration on the transport of auxins into the plant, inducing root inhibition and ultimately total stop of root growth.

Successful Transvenous Extraction of Adult Dirofilaria immitis Parasites from a Naturally Infected Domestic Ferret (Mustela putorius furo).

Heartworm disease caused by Dirofilaria immitis is a serious and underdiagnosed cardiovascular condition in domestic ferrets. Hemodynamic changes caused by parasitization in ferrets cause a potentially fatal syndrome, but its clinical findings and treatment have not yet been standardized. The objective of this study was to describe the clinical case of a successful surgical extraction in a ferret infected by D. immitis. The patient was a 2-year-old, 1.5 kg asymptomatic male domestic ferret. The infection was diagnosed using a commercial test for the detection of D. immitis antigens. Subsequently, their clinical status was evaluated using serological and imaging diagnostic tests, and it was finally decided to perform surgical extraction of the adult worms. The ferret was anesthetized and placed in the left lateral decubitus position to perform a venotomy in the right jugular vein. Endoscopic extraction basket-shaped devices were used in the right atrial cavity under fluoroscopic guidance following the Seldinger endovascular surgery technique. With careful handling, two adult parasites were manually removed. A transthoracic echocardiogram performed after the procedure confirmed the absence of heartworms. The ferret recovered without complications and was discharged within 24 h. In the clinical review, 30 days after surgery, no notable alterations or symptoms were observed. This case report describes the first complete surgical removal of adult D. immitis parasites in a naturally infected ferret.

Clove essential oil and its major component, eugenol: a comparative study of their in vitro antioxidant and anticancer properties.

Syzygium aromaticum L. (clove) is a species native to subtropical countries. Its dried flower buds are rich in essential oils, which have shown insecticidal, anti-inflammatory and anaesthetic effects. This work was aimed to study the differences in antioxidant and anticancer activities between clove essential oil (CEO) and its major component, eugenol. The chemical composition of the CEO was determined by GC-MS. The physicochemical properties and antioxidant activity were determined in CEO and eugenol. Finally, anticancer activities were assayed against seven cell lines. Chemical analysis revealed that 80% of the CEO was eugenol. The density and IR were similar, and the colour was ΔE*>3. CEO had a lower phenolic content, but similar antioxidant activity to eugenol. The anticancer activity of the CEO was greater than that of eugenol in all the cell lines except for HeLa cells. These results suggest that secondary compounds in CEO enhance its antioxidant and -anticancer activities.

Evaluating the effects of azelaic acid in the metabolism of Arabidopsis thaliana seedlings through untargeted metabolomics and ionomics approaches.

The present study demonstrates that low concentrations of azelaic acid (AZA) significantly impact the metabolism of Arabidopsis thaliana seedlings, leading to imbalances in numerous minerals and metabolites due to AZA-induced stress. Untargeted metabolomic analyses were conducted on untreated and AZA-treated seedlings at two time points: 7 and 14 days after treatment initiation. The results revealed a general accumulation of sugars (e.g., glucose, mannose, xylose), amino acids (e.g., lysine, GABA, threonine, glutamine), and organic acids (e.g., glutaric acid, shikimic acid, succinic acid) in AZA treated-seedlings, suggesting that AZA triggers stress responses in Arabidopsis. Ionomic analysis revealed that AZA induces phosphorus deficiency, which plants compensate by increasing malate content in the roots. Additionally, AZA treatment induced putrescine accumulation within the root, a metabolic biomarker of potassium deficiency and plant stress. The metabolomic profile showed elevated levels of different specialized metabolites, such as nitrogen- and sulphur-containing compounds, and altered levels of various phytohormones, including jasmonates and brassinosteroids, implicated in plant protection under biotic and/or abiotic stresses. These findings support the hypothesis that AZA's mode of action is associated with an auxin imbalance, suggesting its function as an auxinic herbicide. The observed increases in starch and jasmonates, coupled with the disruptions in potassium homeostasis, are linked to the previously reported alterations in the auxin transport, root architecture and gravitropic root response. Statistical analyses were applied, including Kruskal-Wallis tests for ionomic data, as well as multifactor analysis, Principal Component Analysis, Orthogonal Partial Least Squares-Discriminant Analysis, and enrichment pathway analysis for metabolomic data, ensuring the robustness and validity of these findings.

Synergistic Strategies in Prostate Cancer Therapy: Electrochemotherapy and Electromagnetic Hyperthermia.

Prostate cancer is a significant global health problem, being the second most common cancer and the fifth leading cause of death in men worldwide. Standard chemotherapy, though effective, often lacks selectivity for tumor cells, resulting in dose-limiting side effects. To address this, innovative biomedical approaches such as electrochemotherapy and electromagnetic hyperthermia have emerged. Electrochemotherapy improves drug delivery by facilitating electroporation, thereby increasing intracellular concentrations of chemotherapeutic agents. This approach reduces dosages and associated adverse effects. Meanwhile, electromagnetic hyperthermia raises the temperature of tumor cells, enhancing their sensitivity to chemotherapy. While previous research has demonstrated the inhibitory effects of magnetic hyperthermia on prostate cancer cell growth both in vitro and in vivo, and its synergy with chemotherapy has shown enhanced tumor remission, limited studies have focused on electrochemotherapy alone or in combination with hyperthermia in prostate cancer models. This study aims to assess the synergistic effects of electromagnetic hyperthermia, with superparamagnetic iron oxide nanoparticles (SPIONs) and electrochemotherapy, with electroporation and the chemotherapeutic drugs bleomycin and cisplatin, on the prostate cancer-derived cell line DU-145/GFP and prostate-derived cell line RWPE-1. Results indicate enhanced cytotoxicity with both treatments (bleomycin and cisplatin) by adding electroporation, demonstrating a particularly pronounced effect with bleomycin. Combining electroporation with hyperthermia significantly augments cytotoxicity. Moreover, electroporation effectively reduced the time of exposure to electromagnetic hyperthermia while magnifying its cytotoxic effects. Future research in in vivo trials may reveal additional insights into the combined effects of these therapies.

Sequential physical and cognitive training disrupts cocaine-context associations via multi-level stimulation of adult hippocampal neurogenesis.

Cocaine-related contextual cues are a recurrent source of craving and relapse. Extinction of cue-driven cocaine seeking remains a clinical challenge, and the search for adjuvants is ongoing. In this regard, combining physical and cognitive training is emerging as a promising strategy that has shown synergistic benefits on brain structure and function, including enhancement of adult hippocampal neurogenesis (AHN), which has been recently linked to reduced maintenance of maladaptive drug seeking. Here, we examined whether this behavioral approach disrupts cocaine-context associations via improved AHN. To this aim, C57BL/6J mice (N = 37) developed a cocaine-induced conditioned place preference (CPP) and underwent interventions consisting of exercise and/or spatial working memory training. Bromodeoxyuridine (BrdU) was administered during early running sessions to tag a subset of new dentate granule cells (DGCs) reaching a critical window of survival during spatial learning. Once these DGCs became functionally mature (∼ 6 weeks-old), mice received extinction training before testing CPP extinction and reinstatement. We found that single and combined treatments accelerated CPP extinction and prevented reinstatement induced by a low cocaine priming (2 mg/kg). Remarkably, the dual-intervention mice showed a significant decrease of CPP after extinction relative to untreated animals. Moreover, combining the two strategies led to increased number and functional integration of BrdU+ DGCs, which in turn maximized the effect of spatial training (but not exercise) to reduce CPP persistence. Together, our findings suggests that sequencing physical and cognitive training may redound to decreased maintenance of cocaine-context associations, with multi-level stimulation of AHN as a potential underlying mechanism.

Experiences of women sexual assault survivors with police in the early aftermath of assault: Results from a large-scale prospective study.

Over 100,000 women present for emergency care after sexual assault (SA) annually in the United States. To our knowledge, no large prospective studies have assessed SA survivor experiences with police. Women SA survivors enrolled at 13 sites (n = 706), and 630 survivors reported on their police interactions. Most women were interested in speaking with police, spoke with police, and reported positive experiences. Latinas and women with lower education and income were less likely to speak with police. Trauma and posttraumatic stress symptoms were associated with more negative experiences. Qualitative comments provide key points for police to consider when speaking with survivors.

Hyperalgesic Effect Evoked by il-16 and its Participation in Inflammatory Hypernociception in Mice.

The systemic administration of interleukin-16 (IL-16, 3-30 ng/kg) induced thermal hyperalgesia in mice, that was prevented by the acute injection of an anti-CD4 antibody (1 µg/kg), the depletion of circulating white blood cells by cyclophosphamide or the specific reduction of circulating CD4+ cells provoked by a high dose of an anti-CD4 antibody (30 µg/mouse, 24 h before). IL-16-induced hyperalgesia was locally inhibited after intraplantar (i.pl.) administration of the non-selective cyclooxygenase (COX) inhibitor diclofenac, the COX-1 inhibitor SC-560, the COX-2 inhibitor celecoxib, the TRPV1 antagonist capsazepine or the TRPA1 antagonist HC030031, thus demonstrating that prostaglandins and TRP channels are involved in this effect. The i.pl. administration of low doses of IL-16 (0.1-1 ng) evoked local hyperalgesia suggesting the possibility that IL-16 could participate in hypernociception associated to local tissue injury. Accordingly, IL-16 concentration measured by ELISA was increased in paws acutely inflamed with carrageenan or chronically inflamed with complete Freund´s adjuvant (CFA). This augmentation was reduced after white cell depletion with cyclophosphamide or neutrophil depletion with an anti-Ly6G antibody. Immunofluorescence and flow cytometry experiments showed that the increased concentration of IL-16 levels found in acutely inflamed paws is mainly related to the infiltration of IL-16+ neutrophils, although a reduced number of IL-16+ lymphocytes was also detected in paws inflamed with CFA. Supporting the functional role of IL-16 in inflammatory hypernociception, the administration of an anti-IL-16 antibody dose-dependently reduced carrageenan- and CFA-induced thermal hyperalgesia and mechanical allodynia. The interest of IL-16 as a target to counteract inflammatory pain is suggested.

Resultados postoperatorios de las colectomías electivas abiertas y laparoscópicas por cáncer de colon en el Complejo Hospitalario Doctor Arnulfo Arias Madrid

Introducción: Metaanálisis internacionales muestran que la cirugía laparoscópica presenta mejores resultados postoperatorios que la cirugía abierta en el manejo del cáncer de colon. La tendencia en Panamá es la transición a la cirugía laparoscópica; sin embargo, no existen estudios que comparen ambas técnicas quirúrgicas. Objetivo: Comparar los resultados postoperatorios de las colectomías electivas abiertas y laparoscópicas por cáncer de colon en el Complejo Hospitalario Dr. Arnulfo Arias Madrid, de enero de 2015 a diciembre de 2019. Metodología: Se realizó un estudio observacional, transversal, correlacional causal (analítico), retrospectivo mediante la revisión de expedientes clínicos de los pacientes operados de colectomías electivas abiertas y laparoscópicas por cáncer de colon en el Complejo Hospitalario Doctor Arnulfo Arias Madrid, de enero de 2015 a diciembre de 2019. Resultados: De 87 casos estudiados, 45 correspondían a cirugía abierta y 42 a cirugía laparoscópica. Encontramos que el abordaje abierto se asocia a una mayor mortalidad intrahospitalaria (χ² 4.951, p 0.026), mortalidad a los 30 días postoperatorios (χ² 6.014, p 0.014), íleo postoperatorio (χ² 5.803, p 0.015), fuga de anastomosis (χ² 12.833, p 0.0003) y dehiscencia de herida quirúrgica (χ² 7.105, p 0.007) que el laparoscópico. El riesgo relativo de infección con abordaje abierto es 8.9 veces mayor que con abordaje laparoscópico [RR = 8.865 (95% IC = 1.058 ­ 74.286) p=0.044].  Conclusiones: La epidemiología del cáncer colorrectal en nuestra institución se asemeja a la de la literatura mundial. Nuestros resultados incentivan a implementar la cirugía mínimamente invasiva para brindar mejores resultados.   (provisto por Infomedic International)

Introduction: International meta-analyses show that laparoscopic surgery has better postoperative results than open surgery in the management of colon cancer. The trend in Panama is the transition to laparoscopic surgery; however, there are no studies comparing both surgical techniques. Objective: To compare the postoperative results of elective open and laparoscopic colectomies for colon cancer at the Complejo Hospitalario Dr. Arnulfo Arias Madrid, from January 2015 to December 2019. Methodology: An observational, cross-sectional, causal correlational (analytical), retrospective, retrospective study was conducted by reviewing clinical records of patients who underwent elective open and laparoscopic colectomies for colon cancer at the Complejo Hospitalario Doctor Arnulfo Arias Madrid, from January 2015 to December 2019. Results: Of 87 cases studied, 45 corresponded to open surgery and 42 to laparoscopic surgery. We found that the open approach was associated with higher in-hospital mortality (χ² 4.951, p 0.026), 30-day postoperative mortality (χ² 6.014, p 0. 014), postoperative ileus (χ² 5.803, p 0.015), anastomotic leak (χ² 12.833, p 0.0003) and surgical wound dehiscence (χ² 7.105, p 0.007) than laparoscopic. The relative risk of infection with open approach is 8.9 times higher than with laparoscopic approach [RR = 8.865 (95% CI = 1.058 - 74.286) p=0.044].  Conclusions: The epidemiology of colorectal cancer in our institution resembles that of the world literature. Our results encourage the implementation of minimally invasive surgery to provide better results. (provided by Infomedic International)

Change in the Distribution Pattern of Dirofilaria immitis in Gran Canaria (Hyperendemic Island) between 1994 and 2020.

Dirofilariosis is a zoonotic disease that mainly affects dogs and cats, with a high risk to public health. The island of Gran Canaria (Spain) has been shown to be a hyperendemic area of infection and, therefore, a model for studying the evolution of the disease. The objective of this study was to track the prevalence and distribution of heartworm in dogs, cats, and residents of Gran Canaria from 1994 to 2020, using published and unpublished data. Blood samples from 5841 dogs, 1203 cats, and 1604 humans were collected in the years analyzed, considering geographical and isoclimatic factors. In 1994, a prevalence of Dirofilaria immitis of 67.02% in dogs was reported, while in 2020 it was 15.81%. In cats, the seroprevalence in 2010 was 33.03%, compared to 17.20% in 2020. The incidence of D. immitis in humans in 2008 was 18.66%, while in 2020 it was 8.27%. For all study groups, temperate cold zone (TC) and temperate mild zone (TM) climates had the highest prevalence. Throughout these 20 years, the prevalence of heartworm disease has decreased. Despite this, it continues to be a hyperendemic island. This study highlights the importance of using the "One Health" perspective and the risks of contagion of the disease.

Strategy for the analysis of lignocellulosic biomass to select a viable transformation route in the Colombian context.

Lignocellulosic biomass is widely available in the world. However, a consensus has yet to be established to evaluate the biomass valorization alternatives. The chemical composition is the primary technical limitation in selecting a transformation route to obtain value-added products. In this paper, the bagasse from non-centrifuged sugar (NCS) production and Pinus patula (PP) wood chips were analyzed in terms of complete chemical composition to establish their potential for selecting the transformation routes. A strategy to select the best route based on the chemical composition was applied and a feedstock criteria model was proposed. Schemes were obtained and compared using a bioprocess selection strategy proposed in previous works. As a result, the preliminary biorefinery schemes were finally defined. The assessment of schemes derived from the outlined strategy included technical, economic, environmental, and social analyses. The environmental evaluation was complemented with a geolocation assessment, revealing a 0.75-ton CO2-eq/yr contribution to the carbon footprint for local distribution. The sustainability index for the PP biorefinery and the bagasse from NCS production was analyzed, resulting in indices of 44.8 and 60.9, respectively. These values were primarily derived from the economic and environmental analyses of both processes.

[Translated article] Retrospective study of home antibiotic infusion therapy using elastomeric infusion pumps.

OBJECTIVES: To describe the experience of home antibiotic infusion therapy using elastomeric infusion pumps, administered to patients admitted to the Home Hospitalisation Unit of a tertiary hospital for 3 years and to analyse clinical evolution and mortality. METHOD: Retrospective observational study. The medical history of the patients included in the study was reviewed. Information was obtained on personal history, antimicrobial therapy received, and clinical evolution. Statistical analysis was performed using SPSS® 19 software. RESULTS: 81 patients were included, 61.7% men, with a mean age of 73.5±17.5 years. The most frequent comorbidities were diabetes mellitus (30.9%) and chronic kidney disease (28.4%). Patients received a mean of 11.9±8.5 days of antibiotic treatment in an elastomeric infusion pump. The main focus of infection was respiratory (27.2%), followed by bacteremia (16%) and skin and soft tissue infections (12.3%). Of the infections, 65.4% were monomicrobial, with Pseudomonas aeruginosa being the main microorganism involved (39.6%). The most commonly used antimicrobial was piperacillin/tazobactam (33.3%). The clinical course was good in 85.2% of the patients, but the mortality rate in the 30 days following the end of treatment was 24.7%. In the univariate analysis, a history of neoplasia in the last 5 years (p=.01) and having received fewer days of antibiotic therapy prior to the start of outpatient antimicrobial therapy in infusion pump (p=.04) were associated with worse clinical outcome. Age over 80 years was associated with better outcome (p=.03). The diagnosis of heart failure was associated with higher mortality (p=.026) and patients from surgical services, with lower mortality (p=.047). In the multivariate analysis, the presence of neoplasia was associated with unfavourable evolution (p=.012) and heart failure with higher mortality (p=.027). CONCLUSIONS: Outpatient antimicrobial therapy in elastomeric infusion pumps is an alternative in patients requiring prolonged intravenous treatment, and age is not a conditioning factor for inclusion in these programs. However, the presence of certain comorbidities can negatively affect the clinical course and mortality of patients.

Azelaic acid can efficiently compete for the auxin binding site TIR1, altering auxin polar transport, gravitropic response, and root growth and architecture in Arabidopsisthaliana roots.

The present study investigates the phytotoxic potential of azelaic acid (AZA) on Arabidopsis thaliana roots. Effects on root morphology, anatomy, auxin content and transport, gravitropic response and molecular docking were analysed. AZA inhibited root growth, stimulated lateral and adventitious roots, and altered the root apical meristem by reducing meristem cell number, length and width. The treatment also slowed down the roots' gravitropic response, likely due to a reduction in statoliths, starch-rich organelles involved in gravity perception. In addition, auxin content, transport and distribution, together with PIN proteins' expression and localisation were altered after AZA treatment, inducing a reduction in auxin transport and its distribution into the meristematic zone. Computational simulations showed that AZA has a high affinity for the auxin receptor TIR1, competing with auxin for the binding site. The AZA binding with TIR1 could interfere with the normal functioning of the TIR1/AFB complex, disrupting the ubiquitin E3 ligase complex and leading to alterations in the response of the plant, which could perceive AZA as an exogenous auxin. Our results suggest that AZA mode of action could involve the modulation of auxin-related processes in Arabidopsis roots. Understanding such mechanisms could lead to find environmentally friendly alternatives to synthetic herbicides.

Epidemiología quirúrgica de la obstrucción intestinal por adherencias en el servicio de cirugía general del Complejo Hospitalario Dr. Arnulfo Arias Madrid. 2015-2020, Panamá

Introducción: La obstrucción intestinal supone un 20% de las consultas por dolor abdominal en urgencias y hasta un 75% se deben a adherencias por cirugías previas. A pesar de las altas tasas de resolución con el tratamiento conservador, es a menudo recurrente y amerita manejo quirúrgico. Objetivo: Determinar la epidemiología quirúrgica de la obstrucción intestinal por adherencias en el servicio de cirugía general del Complejo Hospitalario Dr. Arnulfo Arias Madrid en el período 2015-2020. Metodología: Se realizó un estudio observacional descriptivo, retrospectivo, transversal mediante revisión de expedientes de pacientes con diagnóstico de obstrucción intestinal por adherencias. Resultados: El 54% de los pacientes fueron mujeres y el 46% hombres. Con mayor frecuencia en >65 años y una edad media de 72,5 años. La mayoría de los pacientes presentaban al menos un antecedente quirúrgico abdominal previo (89%). El tiempo transcurrido desde la última cirugía fue más de 10 años (53%). La respuesta al tratamiento conservador fue del 75%. De los pacientes operados, al 77% se le realizó adherenciolisis, al 13% resección y anastomosis intestinal, al 3% ostomías y al 7% otros procedimientos. La moda del tiempo de hospitalización fue de <5 días para el manejo conservador y de 5-10 días para el manejo quirúrgico. Conclusiones: Este estudio marca el inicio de una serie de proyectos, encaminados a evaluar los riesgos y resultados del tratamiento, y decidir el momento óptimo para la intervención quirúrgica, con el objetivo primordial de disminuir la morbi-mortalidad y mejorar la calidad de vida de la población. (provisto por Infomedic International)

Introduction: Intestinal obstruction accounts for 20% of emergency department visits for abdominal pain and up to 75% are due to adhesions from previous surgery. Despite high resolution rates with conservative treatment, it is often recurrent and requires surgical management. Objective: To determine the surgical epidemiology of intestinal obstruction due to adhesions in the general surgery service of the Complejo Hospitalario Dr. Arnulfo Arias Madrid in the period 2015-2020. Methodology: A descriptive, retrospective, cross-sectional, retrospective, observational study was carried out by reviewing the records of patients with a diagnosis of intestinal obstruction due to adhesions. Results: 54% of the patients were women and 46% men. With a higher frequency in >65 years and a mean age of 72.5 years. Most patients had at least one previous abdominal surgical history (89%). The time elapsed since the last surgery was more than 10 years (53%). The response to conservative treatment was 75%. Of the operated patients, 77% underwent adhesiolysis, 13% underwent intestinal resection and anastomosis, 3% underwent ostomies and 7% underwent other procedures. The mode of hospitalization time was <5 days for conservative management and 5-10 days for surgical management. Conclusions: This study marks the beginning of a series of projects, aimed at evaluating the risks and results of treatment, and deciding the optimal time for surgical intervention, with the primary objective of decreasing morbi-mortality and improving the quality of life of the population. (provided by Infomedic International)

Letter to the Editor Ross procedure valve function, clinical outcomes and predictors after 25 years follow up.

The following letter presents an answer of a comment of our work titled "Ross procedure: valve function, clinical outcomes and predictors after 25 years' follow-up," recently published in your journal by Rangwala et al.1 As our colleagues point out, the Ross procedure has excellent survival rates but a significant risk of valve dysfunction and therefore reintervention at follow-up. Although the survival advantage with the Ross procedure appears to be consistent compared with mechanical valve substitutes, this benefit is not as clear compared with biological valve substitutes. However, biological valve substitutes also have significant reintervention rates during follow-up. The different surgical modifications of the Ross procedure have not clearly demonstrated better results in follow-up in terms of autograft reintervention. This procedure can be performed in a medium-volume center with good results as long as adequate patient selection and adequate surgical training are carried out.

Low unspliced cell-associated HIV RNA in early treated adolescents living with HIV on long suppressive ART.

Introduction: Initiation of antiretroviral treatment (ART) in patients early after HIV-infection and long-term suppression leads to low or undetectable levels of HIV RNA and cell-associated (CA) HIV DNA and RNA. Both CA-DNA and CA-RNA, overestimate the size of the HIV reservoir but CA-RNA as well as p24/cell-free viral RNA can be indicators of residual viral replication. This study describes HIV RNA amounts and levels of cytokines/soluble markers in 40 well-suppressed adolescents who initiated ART early in life and investigated which viral markers may be informative as endpoints in cure clinical trials within this population. Methods: Forty adolescents perinatally infected with HIV on suppressive ART for >5 years were enrolled in the CARMA study. HIV DNA and total or unspliced CA-RNA in PBMCs were analyzed by qPCR/RT-qPCR and dPCR/RT-dPCR. Cell-free HIV was determined using an ultrasensitive viral load (US-VL) assay. Plasma markers and p24 were analyzed by digital ELISA and correlations between total and unspliced HIV RNA and clinical markers, including age at ART, Western Blot score, levels of cytokines/inflammation markers or HIV CA-DNA, were tested. Results: CA-RNA was detected in two thirds of the participants and was comparable in RT-qPCR and RT-dPCR. Adolescents with undetectable CA-RNA showed significantly lower HIV DNA compared to individuals with detectable CA-RNA. Undetectable unspliced CA-RNA was positively associated with age at ART initiation and Western Blot score. We found that a higher concentration of TNF-α was predictive of higher CA-DNA and CA-RNA. Other clinical characteristics like US-VL, time to suppression, or percent CD4+ T-lymphocytes were not predictive of the CA-RNA in this cross-sectional study. Conclusions: Low CA-DNA after long-term suppressive ART is associated with lower CA-RNA, in concordance with other reports. Patients with low CA-RNA levels in combination with low CA-DNA and low Western Blot scores should be further investigated to characterize candidates for treatment interruption trials. Unspliced CA-RNA warrants further investigation as a marker that can be prioritized in paediatric clinical trials where the sample volume can be a significant limitation.