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Article ; Online: The Amount of Cross-Linker Influences Affinity and Selectivity of NanoMIPs Prepared by Solid-Phase Polymerization Synthesis.

Testa, Valentina / Anfossi, Laura / Cavalera, Simone / Di Nardo, Fabio / Serra, Thea / Baggiani, Claudio

2024 Volume 16, Issue 4

Abstract: The cross-linker methylene-bis-acrylamide is usually present in nanoMIPs obtained by solid-phase polymerization synthesis at 2 mol% concentration, with very few exceptions. Here, we studied the influence of variable amounts of methylene-bis-acrylamide in ...

Abstract	The cross-linker methylene-bis-acrylamide is usually present in nanoMIPs obtained by solid-phase polymerization synthesis at 2 mol% concentration, with very few exceptions. Here, we studied the influence of variable amounts of methylene-bis-acrylamide in the range between 0 (no cross-linker) and 50 mol% concentration on the binding properties of rabbit IgG nanoMIPs. The binding parameters were determined by equilibrium binding experiments and the results show that the degree of cross-linking defines three distinct types of nanoMIPs: (i) those with a low degree of cross-linking, including nanoMIPs without cross-linker (0-05 mol%), showing a low binding affinity, high density of binding sites, and low selectivity; (ii) nanoMIPs with a medium degree of cross-linking (1-18 mol%), showing higher binding affinity, low density of binding sites, and high selectivity; (iii) nanoMIPs with a high degree of cross-linking (32-50 mol%), characterized by non-specific nanopolymer-ligand interactions, with low binding affinity, high density of binding sites, and no selectivity. In conclusion, the results are particularly relevant in the synthesis of high-affinity, high-selectivity nanoMIPs as they demonstrate that a significant gain in affinity and selectivity could be achieved with pre-polymerization mixtures containing quantities of cross-linker up to 10-20 mol%, well higher than those normally used in this technique.
Language	English
Publishing date	2024-02-16
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527146-5
ISSN	2073-4360 ; 2073-4360
ISSN (online)	2073-4360
ISSN	2073-4360
DOI	10.3390/polym16040532
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Article: Post-pregnancy recurrent biliary colic with intraoperative diagnosis of limy bile syndrome.

Migliore, Marco / Giraudo, Giorgio / Gianotti, Laura / Testa, Valentina / Borghi, Felice

International journal of surgery case reports

2021 Volume 83, Page(s) 105976

Abstract: Introduction: Limy bile syndrome (LBS) is an unusual condition in which gallbladder and/or bile ducts are filled with paste-like radiopaque material with a high calcium carbonate content. It can be rarely associated with PTH disorder and hypercalcemia.!# ...

Abstract	Introduction: Limy bile syndrome (LBS) is an unusual condition in which gallbladder and/or bile ducts are filled with paste-like radiopaque material with a high calcium carbonate content. It can be rarely associated with PTH disorder and hypercalcemia. Presentation of case: A 35-year-old woman presented with epigastric and right hypochondrium pain since a few hours. Similar attacks occurred in the past months soon after a pregnancy with vaginal delivery. Laboratory findings were not significant. The abdominal ultrasound highlighted a micro-lithiasis of gallbladder without complications. Considering the recurrent biliary attacks, laparoscopic cholecystectomy was performed with intraoperative diagnosis of LBS. A subsequent endocrinological screening highlighted a normocalcemic hyperparathyroidism associated with Vitamin D deficiency, likely related to the recent pregnancy and not to LBS. Discussion: LBS is a rare condition with not clear etiology, frequently associated with cholelithiasis, of which it shares clinical presentation and potential complications. Diagnosis of LBS is based on abdominal X-ray/computed tomography scan, or it could be an intraoperative finding. The gold standard treatment is represented by laparoscopic cholecystectomy. The pregnancy with its related cholestatic phenotype could facilitate the LBS manifestation. An endocrinological screening should be performed to rule out a concomitant calcium metabolism disorder. Conclusion: Knowledge of this rare condition could help general surgeons handle it properly.
Language	English
Publishing date	2021-05-13
Publishing country	Netherlands
Document type	Journal Article
ISSN	2210-2612
ISSN	2210-2612
DOI	10.1016/j.ijscr.2021.105976
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Article ; Online: A semi-quantitative visual lateral flow immunoassay for SARS-CoV-2 antibody detection for the follow-up of immune response to vaccination or recovery.

Cavalera, Simone / Di Nardo, Fabio / Serra, Thea / Testa, Valentina / Baggiani, Claudio / Rosati, Sergio / Colitti, Barbara / Brienza, Ludovica / Colasanto, Irene / Nogarol, Chiara / Cosseddu, Domenico / Guiotto, Cristina / Anfossi, Laura

Journal of materials chemistry. B

2024 Volume 12, Issue 8, Page(s) 2139–2149

Abstract: The lateral flow immunoassay (LFIA) technique is largely employed for the point-of-care detection of antibodies especially for revealing the immune response in serum. Visual LFIAs usually provide the qualitative yes/no detection of antibodies, while ... ...

Abstract	The lateral flow immunoassay (LFIA) technique is largely employed for the point-of-care detection of antibodies especially for revealing the immune response in serum. Visual LFIAs usually provide the qualitative yes/no detection of antibodies, while quantification requires some equipment, making the assay more expensive and complicated. To achieve visual semi-quantification, the alignment of several lines (made of the same antigen) along a LFIA strip has been proposed. The numbering of the reacting lines has been used to correlate with the quantity of some biomarkers in serum. Here, we designed the first semiquantitative LFIA for detecting antibodies and applied it to classify the immune response to SARS-CoV-2 raised by vaccination or natural infection. We used a recombinant spike receptor-binding domain (RBD) as the specific capture reagent to draw two test lines. The detection reagent was selected among three possible ligands that are able to bind to anti-spike human antibodies: the same RBD, staphylococcal protein A, and anti-human immunoglobulin G antibodies. The most convenient detector, adsorbed on gold nanoparticles, was chosen based on the highest correlation with an antibody titre of 171 human sera, measured by a reference serological method, and was the RBD (Spearman's rho = 0.84). Incorporated into the semiquantitative LFIA, it confirmed the ability to discriminate high- and low-titre samples and to classify them into two classes (Dunn's test,
MeSH term(s)	Humans ; SARS-CoV-2 ; Follow-Up Studies ; Gold ; COVID-19/diagnosis ; Metal Nanoparticles ; Immunoassay ; Vaccination ; Antibodies ; Immunity
Chemical Substances	Gold (7440-57-5) ; Antibodies
Language	English
Publishing date	2024-02-21
Publishing country	England
Document type	Journal Article
ZDB-ID	2702241-9
ISSN	2050-7518 ; 2050-750X
ISSN (online)	2050-7518
ISSN	2050-750X
DOI	10.1039/d3tb02895j
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Article ; Online: Effect of Surfactants on the Binding Properties of a Molecularly Imprinted Polymer.

Testa, Valentina / Anfossi, Laura / Cavalera, Simone / Chiarello, Matteo / Di Nardo, Fabio / Serra, Thea / Baggiani, Claudio

Polymers

2022 Volume 14, Issue 23

Abstract: In molecularly imprinted polymers, non-specific interactions are generally based on weak forces between the polymer surface and the sample matrix. Thus, additives able to interfere with such interactions should be able to significantly reduce any non- ... ...

Abstract	In molecularly imprinted polymers, non-specific interactions are generally based on weak forces between the polymer surface and the sample matrix. Thus, additives able to interfere with such interactions should be able to significantly reduce any non-specific binding effect. Surfactants represent an interesting class of substances as they are cheap and easily available. Here, we present a study of the effect of three surfactants (the anionic sodium dodecylsulphate, SDS, the cationic cetyltrimethylammonium bromide (CTAB) and the non-ionic polyoxyethylene-(20)-sorbitan monolaurate Tween 20) on the binding affinity of a 2,4,5-trichlorophenoxyacetic acid (2,4,5-T)-imprinted polymer for the template and its analogue 2,4-dichlorophenoxyacetic acid (2,4-D). The experimental results indicate that increasing amounts of surfactant decrease the binding affinity for the ligands strongly for the ionic ones, and more weakly for the non-ionic one. This effect is general, as it occurs for both 2,4,5-T and 2,4-D and for both the imprinted and the not-imprinted polymers. It also proves that the magnitude of this effect mainly depends on the presence or absence of an ionic charge, and that the hydrophobic "tail" of surfactants plays only a minor role.
Language	English
Publishing date	2022-11-30
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527146-5
ISSN	2073-4360 ; 2073-4360
ISSN (online)	2073-4360
ISSN	2073-4360
DOI	10.3390/polym14235210
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Article: Robotic approach to large tailgut cyst with malignant transformation: A case report.

Marano, Alessandra / Giuffrida, Maria Carmela / Peluso, Chiara / Testa, Valentina / Bosio, Paolo / Borghi, Felice

International journal of surgery case reports

2020 Volume 77S, Page(s) S57–S60

Abstract: Introduction: Tailgut cysts (TGC) are rare congenital retrorectal lesions deriving from vestiges of the hindgut. The risk of malignant transformation is relatively high. Thus early diagnosis and a complete removal of the mass are of paramount importance. ...

Abstract	Introduction: Tailgut cysts (TGC) are rare congenital retrorectal lesions deriving from vestiges of the hindgut. The risk of malignant transformation is relatively high. Thus early diagnosis and a complete removal of the mass are of paramount importance. We present a case of successful robotic exeresis of a large TGC adenocarcinoma. Presentation of case: A healthy 35-year-old woman was found to have a retrorectal TGC with suspected malignant transformation. The patient underwent complete robotic resection of the mass and pathology confirmed a TGC adenocarcinoma. The patient's post-operative course was unremarkable. At 24 months, the patient has not experienced any recurrence nor complained functional disorders. Discussion: The current case highlights the importance of TGC as a differential diagnosis of presacral masses, underscoring that malignant transformation can occur and may result in mortality and morbidity if radical surgery is not performed. Diagnosing TGC adenocarcinoma can be challenging because of its relatively low incidence. Biopsy is not recommended while magnetic resonance imaging (MRI) is indispensable in the evaluation of this lesion as the presence of intracystic vegetations could indicate a malignant transformation. Surgery is the mainstay of treatment and minimally invasive techniques have the potential to minimize morbidity and enhance recovery. Amongst these, the robotic approach can make the dissection of large lesions easier, reducing their manipulation and preventing nerve injuries. Conclusion: Robot-assisted resection of a TGC adenocarcinoma is safe and feasible. This approach can be particularly useful in the pelvis providing acceptable functional and oncological outcomes.
Language	English
Publishing date	2020-09-24
Publishing country	Netherlands
Document type	Case Reports
ISSN	2210-2612
ISSN	2210-2612
DOI	10.1016/j.ijscr.2020.09.025
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Article ; Online: Experimental design for the development of a multiplex antigen lateral flow immunoassay detecting the Southern African Territory (SAT) serotypes of foot-and-mouth disease virus.

Cavalera, Simone / Alladio, Eugenio / Foglia, Efrem Alessandro / Grazioli, Santina / Colitti, Barbara / Rosati, Sergio / Nogarol, Chiara / Di Nardo, Fabio / Serra, Thea / Testa, Valentina / Baggiani, Claudio / Maccabiani, Giampietro / Brocchi, Emiliana / Anfossi, Laura

Mikrochimica acta

2023 Volume 191, Issue 1, Page(s) 9

Abstract: Antigenic lateral flow immunoassays (LFIAs) rely on the non-competitive sandwich format, including a detection (labelled) antibody and a capture antibody immobilised onto the analytical membrane. When the same antibody is used for the capture and the ... ...

Abstract	Antigenic lateral flow immunoassays (LFIAs) rely on the non-competitive sandwich format, including a detection (labelled) antibody and a capture antibody immobilised onto the analytical membrane. When the same antibody is used for the capture and the detection (single epitope immunoassay), the saturation of analyte epitopes by the probe compromises the capture and lowers the sensitivity. Hence, several factors, including the amount of the probe, the antibody-to-label ratio, and the contact time between the probe and the analyte before reaching the capture antibody, must be adjusted. We explored different designs of experiments (full-factorial, optimal, sub-optimal models) to optimise a multiplex sandwich-type LFIA for the diagnosis and serotyping of two Southern African Territory (SAT) serotypes of the foot-and-mouth disease virus, and to evaluate the reduction of the number of experiments in the development. Both assays employed single epitope sandwich, so most influencing variables on the sensitivity were studied and individuated. We upgraded a previous device increasing the sensitivity by a factor of two and reached the visual limit of detection of 10
MeSH term(s)	Animals ; Foot-and-Mouth Disease Virus ; Serogroup ; Research Design ; Immunoassay ; Antigens ; Antibodies ; Epitopes
Chemical Substances	Antigens ; Antibodies ; Epitopes
Language	English
Publishing date	2023-12-06
Publishing country	Austria
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	89-9
ISSN	1436-5073 ; 0026-3672
ISSN (online)	1436-5073
ISSN	0026-3672
DOI	10.1007/s00604-023-06090-6
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Article ; Online: Development and In-House Validation of an Enzyme-Linked Immunosorbent Assay and a Lateral Flow Immunoassay for the Dosage of Tenofovir in Human Saliva.

Cavalera, Simone / Serra, Thea / Abad-Fuentes, Antonio / Mercader, Josep V / Abad-Somovilla, Antonio / Nardo, Fabio Di / D'Avolio, Antonio / De Nicolò, Amedeo / Testa, Valentina / Chiarello, Matteo / Baggiani, Claudio / Anfossi, Laura

Biosensors

2023 Volume 13, Issue 6

Abstract: Highly active antiretroviral therapy (HAART) includes very potent drugs that are often characterized by high toxicity. Tenofovir (TFV) is a widely used drug prescribed mainly for pre-exposure prophylaxis (PreP) and the treatment of human immunodeficiency ...

Abstract	Highly active antiretroviral therapy (HAART) includes very potent drugs that are often characterized by high toxicity. Tenofovir (TFV) is a widely used drug prescribed mainly for pre-exposure prophylaxis (PreP) and the treatment of human immunodeficiency virus (HIV). The therapeutic range of TFV is narrow, and adverse effects occur with both underdose and overdose. The main factor contributing to therapeutic failure is the improper management of TFV, which may be caused by low compliance or patient variability. An important tool to prevent inappropriate administration is therapeutic drug monitoring (TDM) of compliance-relevant concentrations (ARCs) of TFV. TDM is performed routinely using time-consuming and expensive chromatographic methods coupled with mass spectrometry. Immunoassays, such as enzyme-linked immunosorbent assays (ELISAs) and lateral flow immunoassays (LFIAs), are based on antibody-antigen specific recognition and represent key tools for real-time quantitative and qualitative screening for point-of-care testing (POCT). Since saliva is a non-invasive and non-infectious biological sample, it is well-suited for TDM. However, saliva is expected to have a very low ARC for TFV, so tests with high sensitivity are required. Here, we have developed and validated a highly sensitive ELISA (IC50 1.2 ng/mL, dynamic range 0.4-10 ng/mL) that allows the quantification of TFV in saliva at ARCs and an extremely sensitive LFIA (visual LOD 0.5 ng/mL) that is able to distinguish between optimal and suboptimal ARCs of TFV in untreated saliva.
MeSH term(s)	Humans ; Tenofovir/therapeutic use ; Anti-HIV Agents/therapeutic use ; HIV Infections/drug therapy ; Saliva ; Enzyme-Linked Immunosorbent Assay ; Immunoassay
Chemical Substances	Tenofovir (99YXE507IL) ; Anti-HIV Agents
Language	English
Publishing date	2023-06-20
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662125-3
ISSN	2079-6374 ; 2079-6374
ISSN (online)	2079-6374
ISSN	2079-6374
DOI	10.3390/bios13060667
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Article ; Online: Development of molecular and antigenic-based rapid tests for the identification of African swine fever virus in different tissues.

Cavalera, Simone / Colitti, Barbara / De Mia, Gian Mario / Feliziani, Francesco / Giudici, Silvia Dei / Angioi, Pier Paolo / D'Errico, Federica / Scalas, Daniela / Scollo, Annalisa / Serra, Thea / Chiarello, Matteo / Testa, Valentina / Di Nardo, Fabio / Baggiani, Claudio / Oggiano, Annalisa / Rosati, Sergio / Anfossi, Laura

Talanta

2023 Volume 258, Page(s) 124443

Abstract: African swine fever (ASF) is a severe haemorrhagic infectious disease affecting suids, thus representing a great economic concern. Considering the importance of the early diagnosis, rapid point of care testing (POCT) for ASF is highly demanded. In this ... ...

Abstract	African swine fever (ASF) is a severe haemorrhagic infectious disease affecting suids, thus representing a great economic concern. Considering the importance of the early diagnosis, rapid point of care testing (POCT) for ASF is highly demanded. In this work, we developed two strategies for the rapid onsite diagnosis of ASF, based on Lateral Flow Immunoassay (LFIA) and Recombinase Polymerase Amplification (RPA) techniques. The LFIA was a sandwich-type immunoassay exploiting a monoclonal antibody directed towards the p30 protein of the virus (Mab). The Mab was anchored onto the LFIA membrane to capture the ASFV and was also labelled with gold nanoparticles for staining the antibody-p30 complex. However, the use of the same antibody for capturing and as detector ligand showed a significant competitive effect for antigen binding, so required an experimental design to minimize reciprocal interference and maximize the response. The RPA assay, employing primers to the capsid protein p72 gene and an exonuclease III probe, was performed at 39 °C. The limit of detection of the method was assessed using a plasmid encoding the target gene and resulted in 5 copy/μL. The new LFIA and RPA were applied for ASFV detection in the animal tissues usually analysed by conventional assays (i.e., real-time PCR), such as kidney, spleen, and lymph nodes. A simple and universal virus extraction protocol was applied for sample preparation, followed by DNA extraction and purification for the RPA. The LFIA only required the addition of 3% H2O2 to limit matrix interference and prevent false positive results. The two rapid methods (25 min and 15 min were needed to complete the analysis for RPA and LFIA, respectively) showed high diagnostic specificity (100%) and sensitivity (93% and 87% for LFIA and RPA, respectively) for samples with high viral load (Ct < 27). False negative results were observed for samples with low viral load (Ct > 28) and/or also containing specific antibodies to ASFV, which decreased antigen availability and were indicative of a chronic, poorly transmissible infection. The simple and rapid sample preparation and the diagnostic performance of the LFIA suggested its large practical applicability for POC diagnosis of ASF.
MeSH term(s)	Swine ; Animals ; African Swine Fever Virus/genetics ; African Swine Fever/diagnosis ; Gold ; Hydrogen Peroxide ; Metal Nanoparticles ; Recombinases ; Antibodies, Monoclonal
Chemical Substances	Gold (7440-57-5) ; Hydrogen Peroxide (BBX060AN9V) ; Recombinases ; Antibodies, Monoclonal
Language	English
Publishing date	2023-03-14
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1500969-5
ISSN	1873-3573 ; 0039-9140
ISSN (online)	1873-3573
ISSN	0039-9140
DOI	10.1016/j.talanta.2023.124443
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Article ; Online: Investigation of the "Antigen Hook Effect" in Lateral Flow Sandwich Immunoassay: The Case of Lumpy Skin Disease Virus Detection.

Cavalera, Simone / Pezzoni, Giulia / Grazioli, Santina / Brocchi, Emiliana / Baselli, Stefano / Lelli, Davide / Colitti, Barbara / Serra, Thea / Nardo, Fabio Di / Chiarello, Matteo / Testa, Valentina / Rosati, Sergio / Baggiani, Claudio / Anfossi, Laura

Biosensors

2022 Volume 12, Issue 9

Abstract: Lumpy skin disease (LSD) is an infectious disease affecting bovine with severe symptomatology. The implementation of effective control strategies to prevent infection outbreak requires rapid diagnostic tools. Two monoclonal antibodies (mAbs), targeting ... ...

Abstract	Lumpy skin disease (LSD) is an infectious disease affecting bovine with severe symptomatology. The implementation of effective control strategies to prevent infection outbreak requires rapid diagnostic tools. Two monoclonal antibodies (mAbs), targeting different epitopes of the LSDV structural protein p32, and gold nanoparticles (AuNPs) were used to set up a colorimetric sandwich-type lateral flow immunoassay (LFIA). Combinations including one or two mAbs, used either as the capture or detection reagent, were explored to investigate the hook effect due to antigen saturation by the detector antibody. The mAb-AuNP preparations were optimized by a full-factorial design of experiment to achieve maximum sensitivity. Opposite optimal conditions were selected when one Mab was used for capture and detection instead of two mAbs; thus, two rational routes for developing a highly sensitive LFIA according to Mab availability were outlined. The optimal LFIA for LSDV showed a low limit of detection (103.4 TCID50/mL), high inter- and intra-assay repeatability (CV% < 5.3%), and specificity (no cross-reaction towards 12 other viruses was observed), thus proving to be a good candidate as a useful tool for the point-of-need diagnosis of LSD.
MeSH term(s)	Animals ; Cattle ; Antibodies, Monoclonal ; Epitopes ; Gold/chemistry ; Immunoassay ; Lumpy skin disease virus ; Metal Nanoparticles/chemistry
Chemical Substances	Antibodies, Monoclonal ; Epitopes ; Gold (7440-57-5)
Language	English
Publishing date	2022-09-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662125-3
ISSN	2079-6374 ; 2079-6374
ISSN (online)	2079-6374
ISSN	2079-6374
DOI	10.3390/bios12090739
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Article ; Online: Management of Hemorrhoidal Disease in Special Conditions: A Word of Caution.

Salusso, Paola / Testa, Valentina / Mochet, Sylvie / Arezzo, Alberto / Allaix, Marco Ettore / Salzano, Antonio / Morino, Mario / Mistrangelo, Massimiliano

Reviews on recent clinical trials

2020 Volume 16, Issue 1, Page(s) 22–31

Abstract: Background: Hemorrhoids are a common disease that is often considered an easy problem to solve. Unfortunately, some particular clinical conditions, including Inflammatory Bowel Diseases (IBD), pregnancy, immunosuppression, coagulopathy, cirrhosis with ... ...

Abstract	Background: Hemorrhoids are a common disease that is often considered an easy problem to solve. Unfortunately, some particular clinical conditions, including Inflammatory Bowel Diseases (IBD), pregnancy, immunosuppression, coagulopathy, cirrhosis with portal hypertension, and proctitis after radiotherapy, challenge hemorrhoids management and the outcomes. Methods: Research and online contents related to hemorrhoids' treatment in special conditions are reviewed in order to help colorectal surgeons in daily practice. Results: There are very limited data about the outcomes of hemorrhoids treatment in these subgroups of patients. Patients in pregnancy can be effectively treated with medical therapy, reserving surgical intervention in highly selected and urgent cases. In case of thrombosed haemorrhoids, the excision allows a fast symptoms' resolution, with a low incidence of recurrence and a long remission interval. In case of immunosuppressed patients, there is no consensus for the best treatment, even in most HIV positive patients, a surgical procedure can be safely proposed when indicated. There is no sufficient data in the literature related to transplanted patients. The surgical treatment of hemorrhoids in patients with IBD, especially Crohn's Disease, can be unsafe, although there is a paucity of literature on this topic. In case of previous pelvic radiotherapy, it must always be considered that severe complications, like abscesses and fistulas with subsequent pelvic and retroperitoneal sepsis, can occur after surgical treatment of hemorrhoids, so a conservative treatment is advocated. Moreover, caution is recommended in treating patients with coagulopathy, considering possible complications (mostly bleeding) also after outpatient treatments. In case of portal hypertension and cirrhosis, a 'conservative treatment' is recommended. Bleeding hemorrhoids can be treated with hemorrhoidectomy when they do not respond to other treatments. Conclusion: International literature is very scant about the treatment of patients affected by hemorrhoids in particular situations. A word of caution and concern even about the indication for minor outpatient procedures must be expressed in these patients, in order to avoid possible life-threatening complications. The first-line treatment is the conservative medical approach associated with the treatment of the primary disease.
MeSH term(s)	Crohn Disease ; Hemorrhoidectomy ; Hemorrhoids/complications ; Hemorrhoids/diagnosis ; Hemorrhoids/therapy ; Humans ; Inflammatory Bowel Diseases ; Recurrence ; Treatment Outcome
Language	English
Publishing date	2020-04-06
Publishing country	United Arab Emirates
Document type	Journal Article
ZDB-ID	2251879-4
ISSN	1876-1038 ; 1574-8871
ISSN (online)	1876-1038
ISSN	1574-8871
DOI	10.2174/1574887115666200406121308
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