Actividad citotóxica del α-humuleno y del tras-cariofileno de Salvia officinalis en dos líneas celulares tumorales animal y humana

Background: The purpose of the present work is two-fold: the fractionation of Salvia officinalis essential oil and the cytotoxic study of this oil with its fractions “in vitro” tumor cell lines. Materials and Methods: S. officinalis essential oil was obtained by hydrodistillation and fractionated with column chromatography; the essential oil and its fractions were analyzed by gas chromatography (GC) coupled to mass spectrometry (MS). The cytotoxic activity was evaluated in cellular lines of breast cancer MCF-7, colon cancer HCT-116, and murine macrophage RAW264.7 cell lines by the MTT assay. Results: the sub-subfraction F1.1.1 of S. officinalis essential oil containing α-humulene present highest activity on RAW264.7 and HCT-116 with IC50 values of 41.9 and 77.3 μg/ml, respectively. The sub-subfraction F1.2.1 of S. officinalis essential oil with trans-caryophyllene showed less activity on RAW246.7 and HCT-116 with IC50 values of 90.5 and 145.8 μg/ml. Conclusion: This paper suggests that the α-humulene and trans-caryophyllene extracted from S.officinalis essential oil inhibit tumor cell growth.Antecedentes: Este trabajo tiene dos objetivos: el fraccionamiento del aceite esencial de la especie Salvia officinalis y la determinación de la citotoxicidad del mencionado aceite esencial con sus fracciones en líneas celulares tumorales “in vitro”. Material y Métodos: El aceite esencial de Salvia officinalis fue obtenido por hidrodestilación y fraccionado mediante cromatografía en columna; el aceite esencial y sus fracciones fueron analizadas mediante cromatografía de gases (GC) acoplada a espectrometría de masas (MS). La actividad citotóxica fue evaluada en líneas celulares de cáncer de mama MCF-7; cáncer de colon HCT-116 y en macrófago murino. RAW264.7 con el ensayo MTT. Resultados: La sub-subfracción F1.1.1 del aceite esencial de Salvia officinalis que contiene alfa-humuleno presenta la actividad mas acusada frente a las líneas celulares RAW264.7 y HCT-116, con valores de IC50 de 41,9 y de 77,3 μg/ml respectivamente. La sub-subfracción F1.2.1 del aceite esencial de Salvia officinalis con trans-cariofileno, muestra menor actividad sobre células RAW246.7 y HCT-116 con valores de IC50 de 90,5 y 145,8 μg/ml respectivamente. Conclusión: Estos resultados sugieren que el alfa-humuleno y el trans-cariofileno de los extractos del aceite esencial de Salvia officinalis inhiben el crecimiento de células tumorales

Effect of Gadolinium Chloride on Liver Regeneration Following Thioacetamide-Induced Necrosis in Rats

Gadolinium chloride (GD) attenuates drug-induced hepatotoxicity by selectively inactivating Kupffer cells. The effect of GD was studied in reference to postnecrotic liver regeneration induced in rats by thioacetamide (TA). Rats, intravenously pretreated with a single dose of GD (0.1 mmol/Kg), were intraperitoneally injected with TA (6.6 mmol/Kg). Hepatocytes were isolated from rats at 0, 12, 24, 48, 72 and 96 h following TA intoxication, and samples of blood and liver were obtained. Parameters related to liver damage were determined in blood. In order to evaluate the mechanisms involved in the post-necrotic regenerative state, the time course of DNA distribution and ploidy were assayed in isolated hepatocytes. The levels of circulating cytokine TNFα was assayed in serum samples. TNFα was also determined by RT-PCR in liver extracts. The results showed that GD significantly reduced the extent of necrosis. The effect of GD induced noticeable changes in the post-necrotic regeneration, causing an increased percentage of hepatocytes in S phase of the cell cycle. Hepatocytes increased their proliferation as a result of these changes. TNFα expression and serum level were diminished in rats pretreated with GD. Thus, GD pre-treatment reduced TA-induced liver injury and accelerated postnecrotic liver regeneration. No evidence of TNFα implication in this enhancement of hepatocyte proliferation and liver regeneration was found. These results demonstrate that Kupffer cells are involved in TA-induced liver damage, as well as and also in the postnecrotic proliferative liver states

Effectiveness of Fosfomycin for the Treatment of Multidrug-Resistant Escherichia coli Bacteremic Urinary Tract Infections

IMPORTANCE The consumption of broad-spectrum drugs has increased as a consequence of the spread of multidrug-resistant (MDR) Escherichia coli. Finding alternatives for these infections is critical, for which some neglected drugs may be an option. OBJECTIVE To determine whether fosfomycin is noninferior to ceftriaxone or meropenem in the targeted treatment of bacteremic urinary tract infections (bUTIs) due to MDR E coli. DESIGN, SETTING, AND PARTICIPANTS This multicenter, randomized, pragmatic, open clinical trial was conducted at 22 Spanish hospitals from June 2014 to December 2018. Eligible participants were adult patients with bacteremic urinary tract infections due to MDR E coli; 161 of 1578 screened patients were randomized and followed up for 60 days. Data were analyzed in May 2021. INTERVENTIONS Patients were randomized 1 to 1 to receive intravenous fosfomycin disodium at 4 g every 6 hours (70 participants) or a comparator (ceftriaxone or meropenem if resistant; 73 participants) with the option to switch to oral fosfomycin trometamol for the fosfomycin group or an active oral drug or pa renteral ertapenem for the comparator group after 4 days. MAIN OUTCOMES AND MEASURES The primary outcome was clinical and microbiological cure (CMC) 5 to 7 days after finalization of treatment; a noninferiority margin of 7% was considered. RESULTS Among 143 patients in the modified intention-to-treat population (median [IQR] age, 72 [62-81] years; 73 [51.0%] women), 48 of 70 patients (68.6%) treated with fosfomycin and 57 of 73 patients (78.1%) treated with comparators reached CMC (risk difference, -9.4 percentage points; 1-sided 95% CI, -21.5 to infinity percentage points; P = .10). While clinical or microbiological failure occurred among 10 patients (14.3%) treated with fosfomycin and 14 patients (19.7%) treated with comparators (risk difference, -5.4 percentage points; 1-sided 95% CI. -infinity to 4.9; percentage points; P = .19), an increased rate of adverse event-related discontinuations occurred with fosfomycin vs comparators (6 discontinuations [8.5%] vs 0 discontinuations; P = .006). In an exploratory analysis among a subset of 38 patients who underwent rectal colonization studies, patients treated with fosfomycin acquired a new ceftriaxone-resistant or meropenem-resistant gram-negative bacteria at a decreased rate compared with patients treated with comparators (0 of 21 patients vs 4 of 17 patients [23.5%]; 1-sided P = .01). CONCLUSIONS AND RELEVANCE This study found that fosfomycin did not demonstrate noninferiority to comparators as targeted treatment of bUTI from MDR E coli; this was due to an increased rate of adverse event-related discontinuations. This finding suggests that fosfomycin may be considered for selected patients with these infections

Role of age and comorbidities in mortality of patients with infective endocarditis

[Purpose]: The aim of this study was to analyse the characteristics of patients with IE in three groups of age and to assess the ability of age and the Charlson Comorbidity Index (CCI) to predict mortality. [Methods]: Prospective cohort study of all patients with IE included in the GAMES Spanish database between 2008 and 2015.Patients were stratified into three age groups:<65 years,65 to 80 years,and ≥ 80 years.The area under the receiver-operating characteristic (AUROC) curve was calculated to quantify the diagnostic accuracy of the CCI to predict mortality risk. [Results]: A total of 3120 patients with IE (1327 < 65 years;1291 65-80 years;502 ≥ 80 years) were enrolled.Fever and heart failure were the most common presentations of IE, with no differences among age groups.Patients ≥80 years who underwent surgery were significantly lower compared with other age groups (14.3%,65 years; 20.5%,65-79 years; 31.3%,≥80 years). In-hospital mortality was lower in the <65-year group (20.3%,<65 years;30.1%,65-79 years;34.7%,≥80 years;p < 0.001) as well as 1-year mortality (3.2%, <65 years; 5.5%, 65-80 years;7.6%,≥80 years; p = 0.003).Independent predictors of mortality were age ≥ 80 years (hazard ratio [HR]:2.78;95% confidence interval [CI]:2.32–3.34), CCI ≥ 3 (HR:1.62; 95% CI:1.39–1.88),and non-performed surgery (HR:1.64;95% CI:11.16–1.58).When the three age groups were compared,the AUROC curve for CCI was significantly larger for patients aged <65 years(p < 0.001) for both in-hospital and 1-year mortality. [Conclusion]: There were no differences in the clinical presentation of IE between the groups. Age ≥ 80 years, high comorbidity (measured by CCI),and non-performance of surgery were independent predictors of mortality in patients with IE.CCI could help to identify those patients with IE and surgical indication who present a lower risk of in-hospital and 1-year mortality after surgery, especially in the <65-year group

Spatiotemporal Characteristics of the Largest HIV-1 CRF02_AG Outbreak in Spain: Evidence for Onward Transmissions

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41–6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain.Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000–2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively.Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p &lt; 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 2–79 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (tMRCA) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999–2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin.Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics

Modificación de las enzimas antioxidantes endógenas mediante ácido ursólico en el daño hepático inducido por tetracloruro en ratas y cultivos primarios de hepatocitos de rata

El objetivo de este estudio consistió en investigar los posibles efectos protectores del ácido ursólico frente a las alteraciones de las enzimas antioxidantes inducidas por el CCl4 in vivo, así como sus efectos frente a la toxicidad por CCl4, in vitro. El pretratamiento de ratas con ácido ursólico redujo significativamente los niveles séricos de glutamato-oxalato-transaminasa y glutamato-piruvato-transaminasa previamente incrementados por la administración de CCl4. El tratamiento con ácido ursólico también invirtió significativamente la disminución de las actividades superóxido dismutasa, catalasa, glutation reductasa, glutation peroxidasa y de los niveles de glutation en el hígado, ya que la concentración de glutation reducido aumentó y el contenido de glutation oxidado disminuyó en los grupos tratados con ácido ursólico. Los niveles de peroxidación lipídica fueron mayores en el grupo CCl4, pero el aumento también se redujo tras el tratamiento farmacológico (p < 0,01 para 1, 2,5 y 5 mmol/kg). Los resultados in vitro indicaron que la adición al medio de cultivo de ácido ursólico (p < 0,01 para 500 microM) dio lugar a una reducción de las actividades glutamato-oxalato-transaminasa, lactato deshidrogenasa y a una buena tasa de supervivencia de los hepatocitos sometidos al efecto del CCl4. El ácido ursólico también mejoró la peroxidación lipídica en hepatocitos primarios de rata en cultivo expuestos a CCl4, como se demostró con la reducción de la producción de malondialdehído. Además, el ácido ursólico (50-500 microM) mostró propiedades de barrido de radicales en términos de formación de hidroxilo. Los resultados obtenidos sugieren que el tratamiento con ácido ursólico puede normalizar el estado antioxidante alterado de las ratas tratadas con CCl4 manteniendo los niveles de glutation e inhibiendo la producción de malondialdehído debido a sus propiedades de eliminación de radicales.The purpose of this study was to investigate possible protective effects of ursolic acid against CCl4-induced alterations of antioxidant defence enzymes in vivo as well as its effects against CCl4-intoxication in vitro. Pre-treatment of rats with ursolic acid significantly reduced serum levels of glutamate-oxalate-transaminase and glutamate-pyruvate-transaminase previously increased by administration of CCl4. Treatment with ursolic acid also significantly reversed the decreased superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase activities and glutathione levels in the liver, as the concentration of reduced glutathione was increased and the content of oxidized glutathione decreased in ursolic acid treated groups. Levels of lipid peroxidation were higher in the CCl4 group but the increase was also reduced after drug treatment (p < 0.01 for 1, 2.5 and 5 mmol/kg). In vitro results indicated that addition to the culture medium of ursolic acid (p < 0.01 for 500 microM) resulted in a reduction of glutamate-oxalate-transaminase, lactate dehydrogenase activities and in a good survival rate for the CCl4-intoxicated hepatocytes. Ursolic acid also ameliorated lipid peroxidation in primary cultured rat hepatocytes exposed to CCl4, as demonstrated by a reduction in malondialdehyde production. Moreover, ursolic acid (50-500 microM) showed radical scavenging properties in terms of hydroxyl formation. The results obtained suggest that ursolic acid treatment can normalize the disturbed antioxidant status of rats intoxicated with CCl4 by maintaining the levels of glutathione and by inhibiting the production of malondialdehyde due to its radical scavenging properties.University Complutense MadridDepto. de Farmacología, Farmacognosia y BotánicaFac. de FarmaciaTRUEpu

Role of Kupffer Cells in Thioacetamide-Induced Cell Cycle Dysfunction

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