Synthesis and hypoglycemic activity of 7-n-butyl-3-methyl-8-thioxanthine derivatives

According to the International Diabetes Federation in 2015 were registered 59.8 million patients with diabetes in Europe. Synthetic drugs are widely applied in addition to a variety of insulins to normalize blood glucose level. It should be noted that the oral anti-diabetic drugs are the common therapeutic agents for the treatment of diabetes mellitus type II, and therefore the search for new non-toxic hypoglycemic agents is one of the most urgent problems of modern pharmaceutical science. It is known that 7,8-disubstituted xanthine derivatives exhibit hypoglycemic activity. The aim of this work lies in developing unique method to synthesize undocumented in other scientific papers 7-n-butyl-3-methyl-8-thioxanthine derivatives and also studying of their hypoglycemic activity. Acute toxicity of synthesized compounds has been studied with the application of Prozorovsky’s method. The glucose homeostasis characteristic has been performed on carbohydrate tolerance that has been determined by the glucose load test on the control group and on the intact rats. The reactions of 7-n-butyl-3-methyl-8-thioxanthine with benzylchlorides, bromoketones, esters and an amide of chloroacetic acid have been studied. Reactions of mentioned syntons proceed smoothly in aqueous propanol-2 environment and lead to the formation of the corresponding 8-benzyl-, benzoylmethylthioxanthines and xanthinyl-8-thioacetic acid derivatives. Accessible laboratory method has been elaborated to synthesize unspecified in scientific papers earlier 8-thiosubstituted 7-n-butyl-3-methylxanthine by reacting 7-n-butyl-3-methyl-8-thioxanthine with benzylchlorides, bromoketones, esters and an amide of chloroacetic acid, their structure having been proved by NMR-spectroscopy data. The acute toxicity of synthesized compounds has been studied. It has been established that LD50 is in the range 820–2477 mg/kg, i. e. obtained substances are low-toxic and practically non-toxic according to Sidorov’s classification. Also the study of hypoglycemic activity of synthesized compounds has been carried out. A significant perspective for further research to find the original antidiabetic agents has been shown by bioassay results. It has been found that 7-n-butyl-3-methylxanthinyl-8-thioacetamide is more active than reference substances.

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Study of the Hypoglycemic Activity of Derivatives of Isoflavones fromCicer arietinumL.

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2017/8746823 ◽

2017 ◽

Vol 2017 ◽

pp. 1-18 ◽

Cited By ~ 2

Author(s):

Ying Wei ◽

Pengshou Li ◽

Bo Li ◽

Jiaqi Gao ◽

Dongchao Wang ◽

...

Keyword(s):

Cell Model ◽

Positive Control ◽

Hypoglycemic Activity ◽

Hypoglycemic Agents ◽

Control Group ◽

Insulin Resistant ◽

The People ◽

H4iie Cells ◽

New Ideas ◽

Derivatives Of

The chickpea, a food and medicine used by the people of Xinjiang, has a beneficial hypoglycemic effect. To better utilize this national resource and develop hypoglycemic agents from components of the chickpea, a series of new derivatives of isoflavone compounds from the chickpea were synthesized. An insulin-resistant (IR) HepG2 cell model was used to screen the hypoglycemic activities of these compounds. And the structure-activity relationships of these compounds were explored. Additionally, several combinations of these compound displayed higher hypoglycemic activity than any single compound, and they had similar hypoglycemic activity to that of the positive control group (p>0.05). In addition, combination 3 and combination 6 exerted different effects on the insulin sensitivity of H4IIE cells stimulated with resistin. And the results indicated that combination 3 would have higher hypoglycemic activity. These findings demonstrate the characteristics of multiple components and targets of Chinese herbal medicine. This evidence may provide new ideas for the development of hypoglycemic drugs.

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Evaluation of the Hypoglycemic Potential of Leaves Extract of Spondias pinnata (L.f.) Kurz. from Nepal

The Scientific World JOURNAL ◽

10.1155/2021/3230351 ◽

2021 ◽

Vol 2021 ◽

pp. 1-6

Author(s):

Kusum Sai ◽

Sumit Bahadur Baruwal Chhetri ◽

Shankar Raj Devkota ◽

Deepa Khatri

Keyword(s):

Blood Glucose ◽

Blood Glucose Level ◽

Glucose Level ◽

Hypoglycemic Activity ◽

Hypoglycemic Agents ◽

Deciduous Tree ◽

Control Group ◽

Oral Glucose ◽

Phytochemical Screening ◽

Traditional Use

Spondias pinnata (L.f.) Kurz. (family: Anacardiaceae) is a wild deciduous tree indigenous to southeast Asian countries. Different parts of this plant are used traditionally for the treatment and cure of various disorders and illnesses. S. pinnata leaves are used to prevent and treat diabetes in traditional Balinese medicine. However, scientific study on the antihyperglycemic effect of its leaves has not been reported yet. Therefore, this study aims to perform phytochemical screening and investigate the hypoglycemic potential of S. pinnata leaves extract. Preliminary phytochemical screening of the hydroethanolic extract was performed following the standard tests. In vivo hypoglycemic activity of the leaves extract was evaluated using normal and glucose-loaded rats. The results displayed the presence of phytochemical constituents such as saponins, phenolic compounds, flavonoids, and terpenoids. S. pinnata (500 mg/kg) and metformin (100 mg/kg) exhibited a significant ( p < 0.05 ) decrease in blood glucose level at 1, 2, and 3 h in normal rats when compared to the control group. Metformin- (100 mg/kg)- and S. pinnata- (500 mg/kg)- treated groups showed a maximum decrease in the blood glucose level at 3 h after single-dose administration in the oral glucose tolerance test (OGTT). In conclusion, S. pinnata leaves possess a significant hypoglycemic activity in the animal model and thus support its traditional use to treat diabetes. Therefore, a detailed mechanism-based study and isolation of bioactive compounds from S. pinnata leaves would be beneficial in the future for the search of new hypoglycemic agents.

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Acute toxicity of Potentilla anserina L. extract in mice

Zeitschrift für Naturforschung C ◽

10.1515/znc-2020-0019 ◽

2020 ◽

Vol 75 (5-6) ◽

pp. 129-134 ◽

Cited By ~ 1

Author(s):

Dul Dram ◽

Cui-Zhu Zhao ◽

Qin-Ge Ma ◽

Jun-Wei He ◽

Jia-Jie Duo ◽

...

Keyword(s):

Acute Toxicity ◽

Biochemical Parameters ◽

Maximum Dose ◽

Histopathological Examination ◽

Oral Dosage ◽

Control Group ◽

Acute Toxicity Study ◽

Organ Index ◽

Potentilla Anserina ◽

Safety Range

AbstractPotentilla anserina L. is not only a medicinal plant, but also a traditional cuisine. Hence, an acute toxicity study was performed to confirm its safety profile. Forty Kunming mice were randomly divided into two groups: control group and P. anserina L. extract group. Using the maximum dosage method, the P. anserina L. extract group was given the maximum dose within 12 h, equivalent to 345.6 g/kg crude drug. The control group was given distilled water. After administration, toxicity symptoms of mice were observed, body weight and food intake were recorded. After 14 days, blood was collected to measure biochemical parameters, autopsy was carried out to observe the changes of organs, and the vital organs were separated, weighed, and preserved for histopathological examination. The results showed that P. anserina L. extract group had no toxic symptoms. The activity, weight, and diet of mice were normal, and no abnormality was found in organ index, renal function, liver function, anatomical observation, and histopathological examination. Therefore, the maximum oral dosage (345.6 g/kg) of P. anserina L. was good safety. This study indicated that P. anserina L. had a large safety range and the clinical application was safe.

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PENGARUH TERAPI FAMILY PSYCHOEDUCATION (FPE) TERHADAP KECEMASAN DAN BEBAN KELUARGA DALAM MERAWAT ANGGOTA KELUARGA DENGAN SKIZOFRENIA DI KECAMATAN BOLA KABUPATEN SIKKA, NUSA TENGGARA TIMUR

Jurnal Kesehatan Mesencephalon ◽

10.36053/mesencephalon.v3i2.48 ◽

2017 ◽

Vol 3 (2) ◽

Author(s):

Adelheid Riswanti Herminsih ◽

Wisnu Barlianto ◽

Rinik Eko Kapti

Keyword(s):

Disease Process ◽

T Test ◽

Load Test ◽

Family Burden ◽

Control Group ◽

Family Psychoeducation ◽

Significant Difference ◽

Post Test ◽

And Control ◽

Quasi Experiment

Abstract : Schizophrenia is a disease process that affects perceptions, emotions, social behavior and the ability to accept reality correctly. Families with schizophrenics often feel anxiety and burdens associated with client care. The problem can be solved by giving FPE therapy. This study aims to explain the effect of Family Psychoeducation (FPE) therapy on anxiety and family burden in caring for family members with schizophrenia. This research uses quasi experiment research pre-post test with control group. The number of respondents in this study were 18 respondents for the control group and 18 respondents for the treatment group. The study was conducted in District Bola from 24 May to 28 June 2017. Giving therapy done by the researchers themselves who have obtained a license from nurse specializing in mental health nursing. Data analysis used in this research is dependent t test and independent t test. The result of dependent t test of anxiety and load test was obtained significance value <0,05, this result showed significant decrease of anxiety and load after FPE therapy. While the results of independent t test showed that the significance of anxiety and family burden <0.05 which means that there is a significant difference in reducing anxiety and family burden between the treatment and control group after being given FPE therapy, that is, with an average decrease in anxiety and burden For the treatment and control groups of 10.11 and 3.5, respectively. This means that FPE is more effective in reducing family anxiety. Thus it is expected that FPE can be applied as an alternative therapy in reducing the anxiety of families who care for people with schizophrenia.Keywords : family psychoeducation, anxiety, family burden Abstrak : Skizofrenia merupakan proses penyakit yang mempengaruhi persepsi, emosi, perilaku sosial dan kemampuan menerima realita dengan benar. Keluarga dengan penderita skizofrenia seringkali merasakan kecemasan dan beban yang berkaitan dengan perawatan klien. Masalah tersebut dapat diatasi dengan pemberian terapi FPE. Penlitian ini bertujuan menjelaskan pengaruh terapi Family Psychoeducation (FPE) terhadap kecemasan dan beban keluarga dalam merawat anggota keluarga dengan skizofrenia. Penelitian ini menggunakan desain penelitian quasi experiment pre-post test with control group. Jumlah responden dalam penelitian ini 18 responden untuk kelompok kontrol dan 18 responden untuk kelompok perlakuan. Penelitian dilakukan di Kecamatan Bola mulai tanggal 24 Mei-28 Juni 2017. Pemberian terapi dilakukan oleh peneliti sendiri yang telah mendapatkan lisensi dari perawat spesialis jiwa. Analisis data yang digunakan dalam penelitian ini adalah dependent t test dan independen t test. Hasil analisis dependent t test kecemasan dan beban didapatkan nilai signifikansi< 0,05, hasil ini menunjukkan penurunan kecemasan dan beban secara bermakna setelah diberikan terapi FPE. Sedangkan hasil analisis independent t test menunjukkan bahwa nilai signifikansi kecemasan dan beban keluarga < 0,05 yang berarti bahwa ada perbedaan yang bermakna dalam menurunkan kecemasan dan beban kelurga antara kelompok perlakuan dan kontrol setelah diberikan terapi FPE, yaitu dengan rata-rata penurunan kecemasan dan beban untuk kelompok perlakuan dan kontrol masing-masing yakni 10,11 dan 3,5. Hal ini berarti bahwa FPE lebih efektif dalam menurunkan kecemasan keluarga. Dengan demikian diharapkan bahwa FPE bisa diaplikasikan sebagai alternative terapi dalam menurunkan kecemasan keluarga yang merawat penderita skizofrenia.Kata Kunci : family psychoeducation, kecemasan, beban keluarga

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Histopathological Assessment of the Effect of Pregabalin Toxicity on Cerebrum and Cerebellum of Adult Albino Rats

10.21203/rs.3.rs-149289/v1 ◽

2021 ◽

Author(s):

Wael Abdou Hassan ◽

Shaimaa Shehata ◽

Ahmad ElBana

Keyword(s):

Cerebral Cortex ◽

Acute Toxicity ◽

Chronic Toxicity ◽

Control Group ◽

Albino Rats ◽

Histopathological Study ◽

Addictive Behaviors ◽

Group 3 ◽

Group 2 ◽

Cerebral Cortex Tissue

Abstract Background: Pregabalin (PGB) used as analgesic in treatment of neurogenic pains of chronic diseases, is considered as one of the most abused anti-epileptic drugs worldwide and it has been proved that it induces addictive behaviors. The present histopathological study aimed to identify the effect of PGB administration on cerebral cortex and cerebellar cortex, in both acute and chronic toxicity. Seventy-two male and non-pregnant female adult albino rats’ 6- to 8-week-old divided into 3 main groups of 24 rats each were studied. Group 1 represented the control group and group 2 represented the acute toxicity group, in which rats were given a single dose of PGB (5000 mg/kg) orally by gavage and after 24 hours, rats were sacrificed and examined. Group 3 represented the chronic toxicity group; were given PGB 500 mg/kg orally by gavage for 12 weeks, after which rats were sacrificed and examined. Result: Cerebral cortex tissue of acute toxicity group displayed astrocytosis and dystrophic changes, while in chronic group showed degeneration, necrosis and cellular infiltrates. The cerebellum of chronic groups showed degeneration and shrunken of Purkinje cells. Conclusion: Acute and chronic intoxication with pregabalin adversely altered the structure of cerebral cortex and cerebellum.

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Proximate and Acute Toxicity Profile of Vitex doniana (Black Plum) Fruit

Journal of Chemical Society of Nigeria ◽

10.46602/jcsn.v46i2.597 ◽

2021 ◽

Vol 46 (2) ◽

Author(s):

C. Imoisi ◽

J.U. Iyasele ◽

S.E. Okhale

Keyword(s):

Acute Toxicity ◽

Potable Water ◽

Oral Treatment ◽

Control Group ◽

Toxicity Profile ◽

North East ◽

Capability Analysis ◽

Edo State ◽

Ld50 Value ◽

Vitex Doniana

The fresh fruits of Vitex doniana sweet were collected from several randomly selected trees in a farm site in Uromi metropolis, Esan North-East Local Government Area of Edo state and then processed into an extract in form of syrup. The proximate composition and acute toxicity profile of Vitex doniana sweet fruit were investigated to ascertain its safety and nutritional capability. Analysis of the fruit showed it to be a highly nutritious food containing moisture of about, 9.90%, ash content: 21.5%, fat: 0.75%, fiber: Not detected (ND), protein: 0.006% and carbohydrate: 67.84%. Twenty mice were randomized based on body weight into five groups of four mice each. Three mice in each group were given syrup volumes of 100, 200, 300, 400 and 500 mg/ml respectively, corresponding to 1000, 2000, 3000, 4000 and 5000 mg/kg doses. In contrast, mice in the control group received potable water (10 mL/kg). The mice in all the groups were observed closely for mortality, toxicity signs and abnormalities in gross behaviour at 15 min, 30 min, 1, 2, 4 and 8 h, and then daily for 14 days. Based on the model used, the fruit was acutely safe in mice, since no death was observed within 24 hours after oral treatment and on extrapolation gave a high predictive value in humans. The LD50 value of black plum fruit was also found to be greater than 5000 mg/kg. Therefore, Vitex doniana fruit should be considered a source of edible syrup bearing in mind the safety, quantity and variety of nutrients it contains.

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Nephrolithiasis against type 2 diabetes mellitus: on the effect of hypoglycemic therapy on lithogenesis

Terapevticheskii arkhiv ◽

10.26442/terarkh201890104-64 ◽

2018 ◽

Vol 90 (10) ◽

pp. 60-64

Author(s):

S K Yarovoy ◽

E N Kareva ◽

O V Djalilov

Keyword(s):

Diabetes Mellitus ◽

Type 2 Diabetes ◽

Type 2 Diabetes Mellitus ◽

Negative Impact ◽

Stone Formation ◽

Hypoglycemic Agents ◽

Control Group ◽

City Hospital ◽

Increased Risk

Aim. To study the effects of oral hypoglycemic agents that can affect the probability of recurrence of nephrolithiasis. Materials and methods. The article is based on the results of examination and treatment of 315 patients suffering from recurrent nephrolithiasis and medically compensated type 2 diabetes mellitus treated at the N.A. Lopatkin Institute of Urology and Interventional Radiology - the branch of the SMRC of Radiology, Ministry of Health of Russia and D.D. Pletnev City Hospital Moscow Healthcare Department in 2012-2017. The patients were divided into three groups according to the applied tool antidiabetic: metformin, glibenclamide, canagliflozin. The control group consisted of patients receiving insulin therapy. Results and discussion. The propensity of Metformin to reduce the pH of urine, which has a negative impact in the conditions of urate nephrolithiasis, which is most common in the population of patients with type 2 diabetes mellitus. Glibenclamide, on the contrary, somewhat latches urine. But changes in the reaction of urine under the influence of the drug do not go beyond normal values and are not clinically significant. Canagliflozin increases diuresis due to medication induced glycosuria and stimulates renal excretion of uric acid and its salts. However canagliflozin does not cause significant shifts in the pH of urine that may somewhat negates the increased risk of recurrence of urate stone formation in the background of the uricosuric effect of the drug. Conclusion. Drug therapy of type 2 diabetes mellitus significantly affects the properties of urine from patients with nephrolithiasis.

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Study the Behavior of Castellated Steel Column Encasing by Different Reactive Powder Concrete Thickness with Laced Reinforcement

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.895.97 ◽

2021 ◽

Vol 895 ◽

pp. 97-109

Author(s):

Mustafa Mazin Ghazi ◽

Ahmad Jabbar Hussain Alshimmeri

Keyword(s):

Axial Load ◽

Local Buckling ◽

Axial Loading ◽

Longitudinal Axis ◽

Load Test ◽

Reactive Powder Concrete ◽

Control Group ◽

Load Carrying ◽

Steel Column ◽

Reactive Powder

Castellated columns are structural members that are created by breaking a rolled column along the center-line by flame after that rejoining the equivalent halves by welding such that for better structural strength against axial loading, the total column depth is increased by around 50 percent. The implementation of these institutional members will also contribute to significant economies of material value. The main objectives of this study are to study the enhancement of the load-carrying capacity of castellated columns with encasement of the columns by Reactive Powder Concrete (RPC) and lacing reinforcement, and serviceability of the confined castellated columns. The Castellated columns with RPC and Lacing Reinforcement improve compactness and local buckling (web and flange local buckling), as a result of steel section encasement. This study presents axial load test results for four specimens Castellated columns section encasement by Reactive powder concrete (RPC) with laced reinforcement. The encasement consists of, flanges unstiffened element height was filled with RPC for each side and laced reinforced which are used inclined continuous reinforcement of two layers on each side o0f the web of the castellated column. The inclination angle of lacing reinforcement concerning the longitudinal axis is 45o. Four specimens with four different configurations will be prepared and tested under axial load at columns. The first group was the control group (CSC1) Unconfined castellated steel column, the second group was consists of Castellated columns (web and flange) confined with 17mm of (RPC), welded web, and 6mm laced reinforcement (CSC3). While group three (CSC4) consists of a Castellated steel column same as the sample (CSC3), but without using welding between two parts of the castellated steel column. Groups four and five consist of a Castellated steel column same as sample (CSC4) encased partially with reactive powder concrete (25.5 mm) (CSC5) and full encased flange with reactive powder concrete (34mm) mm (CSC6), respectively. The tested specimens' results show that an increase in the strength of the column competitive with increasing the encased reactive powder concrete thickness. And the best sample was sample CSC6 with (34mm) mm in experimental and ABAQUS results.

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Phytochemical Identification, Acute, and Sub-Acute Oral Toxicity Studies of the Foliar Extract of Withania frutescens

Molecules ◽

10.3390/molecules25194528 ◽

2020 ◽

Vol 25 (19) ◽

pp. 4528 ◽

Cited By ~ 2

Author(s):

Abdelfattah EL Moussaoui ◽

Mohammed Bourhia ◽

Fatima Zahra Jawhari ◽

Hamza Mechchate ◽

Meryem Slighoua ◽

...

Keyword(s):

Acute Toxicity ◽

Clinical Symptoms ◽

Ethanolic Extract ◽

Control Group ◽

Phytochemical Analysis ◽

Oral Toxicity ◽

Phytochemical Composition ◽

Subacute Toxicity ◽

Repeated Doses

Withania frutescens (W. frutescens) is a medicinal plant widely used to treat several diseases. This work aims to study phytochemical composition as well as acute and subacute toxicity of W. frutescens hydroethanolic extract in mice. The phytochemical composition of W. frutescens extract was performed using gas chromatographic analysis. Acute toxicity was studied in vivo with oral administration of single doses 400 mg/kg, 1000 mg/kg, and 2000 mg/kg for 14 days. Subacute toxicity was studied with the administration of repeated doses of 400 mg/kg/day and 2000 mg/kg/day for 28 days. Phytochemical analysis of W. frutescens hydro-ethanolic extract confirmed the presence of interesting chemical compounds. Acute toxicity results showed no toxic symptoms in mice treated with an increasing dose up to a maximum of 2000 mg/kg. Alongside acute toxicity, subacute data showed no clinical symptoms nor biochemical or histological alteration in mice treated with an increasing dose up to a maximum of 2000 mg/kg compared to the control group (p < 0.05). This study shows no toxic effects in animals treated with W. frutescens extract, and, therefore, this plant can be considered safe in animals up to 2000 mg/kg under both acute and subacute toxicity conditions.

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The Biocompatibility and Security of Magnetic Nanoparticles Fe3O4-DNR Used In Hematologic Malignancies Therapy

Blood ◽

10.1182/blood.v116.21.3970.3970 ◽

2010 ◽

Vol 116 (21) ◽

pp. 3970-3970

Author(s):

Bao-An Chen ◽

Wei-wei Wu ◽

Jian Cheng ◽

Jun Wang ◽

Feng Gao ◽

...

Keyword(s):

Magnetic Nanoparticles ◽

Acute Toxicity ◽

Toxicity Testing ◽

Hematologic Malignancies ◽

Control Group ◽

Significant Difference ◽

Magnetic Nanoparticles Fe3o4 ◽

Experimental Group ◽

Fe3o4 Mnps ◽

Acute Toxicity Testing

Abstract Abstract 3970 The objective of this paper is to study the biocompatibility and security of self-prepared magnetic nanoparticles Fe3O4 (Fe3O4-MNPs) loaded with daunorubicin (DNR), which has the potential application in hematologic malignancies therapy. Hemolysis test was carried out to estimate it s blood toxicity; Fe3O4-MNPs loaded with DNR were intra peritoneally injected into mouse to calculate the LD50; micronucleus (MN) assay was reckoned to identify its genotoxicity; acute toxicity testing was evaluated its influence to mouse hepatic and renal functions. The result of hemolysis rate (HR) of Fe3O4-MNPs loaded with DNR was 2.908%, far less than 5%. Therefore, we concluded that self-prepated Fe3O4-MNPs loaded with DNR nanoparticles had no hemolytic reaction, and they consistent with the requirement of hemolytic test of biomaterials. The LD50 of Fe3O4-MNPs loaded with DNR nanoparticles to the mice was 1009.71mg/kg (relative content of DNR was 10mg/kg) and the 95% confidence interval was 769.11∼1262.40mg/kg, it had no significant difference compared with LD50 of using DNR only, which data was 8.51mg/kg and the 95% confidence interval was 6.48∼10.37mg/kg, and it had wide safety value circumscription. In micronucleus assay, compared Fe3O4-MNPs loaded with DNR nanoparticles experimental groups with negative control group, we found that the result had no significant difference (P > 0. 05) in micronucleus formation rate, while compared experimental groups with positive control group, the result had significant difference (P < 0. 05). The result indicated that Fe3O4 magnetic nanoparticles had no cacogenesis and mutagenesis. Acute toxicity testing showed that mice body weigh of control group, Fe3O4-MNPs loaded with DNR nanoparticles experimental group and isodose DNR group had no significant difference in 24h, 48h, and 72h after intra peritoneally injection; they had normal activity, eating and evacuation; toxic reactions such as instability of gait, convulsion, paralysis and respiratory depression were not been found; the alanine transarninase (ALT), blood urea nitrogen (BUN), and creatinine clearance rate (CCr) of Fe3O4-MNPs loaded with DNR nanoparticles experimental group was 66.0±28.55u/L, 9.06±1.05mmol/L, and 18.03±1.84umol/L, respectively, which had no significant difference compared with control group and isodose DNR group. From the results of our experiment, we could consider that self-prepared Fe3O4-MNPs loaded with DNR nanoparticles is a kind of high biocompatibility and security materials and perhaps is suitable for further application in hematologic malignancies therapy. Disclosures: No relevant conflicts of interest to declare.

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