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Background: Chronic anal fissure is a common benign anorectal condition that causes significant morbidity. Symptoms consist primarily of pain and bleeding during defecation. Fissures are predominantly located in the posterior midline, but 25% of women and 8% of men have anterior fissures. Aims and Objectives: The aim of the study is to determine the outcome of various nonsurgical therapy for chronic anal fissure and comparison with the outcome of surgical treatment. Materials and Methods: This study was a non-randomized and observational study. All patients referred to the department of general surgery between December 2012 and November 2014 for a chronic anal fissure were included in the study, above the age of 12 years, and diagnosed to have anal fissure (Both Acute and Chronic) were included in the study. Results: Seventy-nine patients (91.9%) were having symptoms of pain; this pain was persistent for hours after defecation in 67 (78%) patients. Seventy-one (82.6%) patients complained of at least one episode of bleeding per Anum, other symptoms were perianal lump (10.5%), perianal itching (23.3%), and perianal discharge (30.2%). The examination findings of these patients revealed that 81 patients (94%) had developed sentinelpiles subsequently and anal fissure with visible fibers of underlying muscle were seen in 9 (10.5%) patients. Conclusion: Chronic anal fissures can be simply and effectively treated medically without the risk of incontinence associated with sphincterotomy. Topical nifedipine and botulinum toxin injections are an excellent combination, associated with a low recurrence rate and minimal side effects.

Pharmaceuticals in the environment have the potential to be hazardous to human beings. With each passing day it is becoming a major source of concern. Due to recent findings showing the availability of pharmaceutical components in the environment, particularly in ground water bodies, pharmaceuticals have gained a growing amount of attention from worldwide health regulatory bodies and have become one of the most significant water pollutants. The aim of this article is to review the environmental hazards of pharmaceuticals that have been reported in various literature sources to promote awareness on safe usage of medicines, to increase pharmaceutical manufacturers’ knowledge on environmental safety aspects and to arrest the attention of pharmacovigilance practitioners to some of the emerging problems caused by medicines. Pharmaceutical waste has been a source of huge concern amongst environmental scientists. Pharmaceutical manufacturers and pharmacovigilance scientists should invest more attention to these increasing environmental concerns caused by medications. Ecopharmacovigilance is defined by the World Health Organization (WHO) as the science and activities concerned with the detection, assessment, understanding, and prevention of adverse events or other related problems caused by pharmaceuticals in the environment that affect people and other animal species. This review is an attempt to compile information on Ecopharmacovilance, with an emphasis on the Indian perspective.

Carbonatites are igneous rocks composed of over 50% modal carbonate minerals and their exact origin and possible mode(s) of formation are still debated. We report stable Ca isotopic compositions (δ44/40CaSRM915a) of globally distributed carbonatites (n = 46) from 22 different locations ranging in age from 2.61 Ga to recent along with their major, trace element concentrations and 87Sr/86Sr ratios. The δ44/40CaSRM915a values for carbonatites older than 300 Ma, except for four samples from the Neoproterozoic south Indian carbonatites and the Phalaborwa complex of South Africa, display limited variation (~0.18‰, n = 12) with δ44/40Ca values overlapping (given their associated analytical uncertainties) with the estimated δ44/40CaSRM915a value of the bulk silicate Earth (BSE, 0.94 ± 0.10‰). The 87Sr/86Sr(t) of these samples (0.70149–0.70394) are consistent with derivation from a depleted mantle source with a time-integrated Rb/Sr ratio of ~0.017. In contrast, carbonatites with emplacement ages younger than 300 Ma display much wider variations in δ44/40Ca values (0.63‰, n = 29) with sixteen samples displaying values lower than that for the BSE and radiogenic 87Sr/86Sr compositions (0.70313–0.71044). The low δ44/40Ca values for the younger carbonatites are consistent with the presence of recycled crustal carbonates, with compositions similar to those of 100–200 Ma and 400–500 Ma old carbonates, in their mantle source, which is also consistent with their radiogenic 87Sr/86Sr ratios. Model calculations, based on δ44/40Ca and 87Sr/86Sr(t) ratios for mantle peridotite and eclogite sources and marine carbonates of different ages, suggest that the δ44/40Ca and 87Sr/86Sr for most young carbonatites ( The contrasting Ca and Sr isotope compositions between younger ( 300 Ma) carbonatites may reflect changing geodynamic conditions. The prevalent Ca and Sr isotope signatures attributed to crustal recycling that is observed in carbonatites younger than 300 Ma most likely reflect two global-scale processes: (i) an increased amount of subduction flux due to high convergence rates subsequent to the break-up of the Pangea supercontinent, and (ii) greater amount of plume activity during the last 300 million years of Earth history. Additionally, the low δ44/40Ca values of younger carbonatites could have been accentuated by subduction of marine carbonates (aragonitic) with relatively low δ44/40Ca. In contrast to the younger carbonatites, Ca and Sr isotope compositions for the carbonatites older than 300 Ma investigated here suggest that crustal recycling was much more sporadic in the Precambrian.

Proteins sample an ensemble of conformers under physiological conditions, having access to a spectrum of modes of motions, also called intrinsic dynamics. These motions ensure the adaptation to various interactions in the cell, and largely assist in, if not determine, viable mechanisms of biological function. In recent years, machine learning frameworks have proven uniquely useful in structural biology, and recent studies further provide evidence to the utility and/or necessity of considering intrinsic dynamics for increasing their predictive ability. Efficient quantification of dynamics-based attributes by recently developed physics-based theories and models such as elastic network models provides a unique opportunity to generate data on dynamics for training ML models towards inferring mechanisms of protein function, assessing pathogenicity, or estimating binding affinities.

The World Health Organization (WHO) defines PV as the science and activities related to the detection, assessment, understanding, and prevention of adverse effects or any other possible drug-related problems (WHO 2004). In 1968, during the 16th World Assembly the 16.36 resolution called for “a systematic collection of information on serious adverse drug reactions during the development and particularly after medicines have been made available for public use”. This led to the formation of the WHO Programme for International Drug Monitoring (PIDM) in 1968. PV systems should include all entities and resources that protect the public from medicines-related harm (adverse reactions, poor product quality, medication errors, and therapeutic ineffectiveness), whether in personal healthcare or public health services. The PV system safeguards the public through efficient and timely identification, collection, and assessment of medicine-related adverse events and by communicating risks and benefits. The WHO has provided technical and normative leadership on PV since the development of the first voluntary notification scheme in 1961. As of January 2016, 123 countries have joined the WHO PIDM, and in addition 28 associate members are awaiting full membership. WHO has defined norms and guidelines for PV and has allowed information sharing among the participant countries. Another WHO PV-related activity is the work of the Council for International Organizations of Medical Sciences (CIOMS) which was established jointly by WHO and UNESCO in 1949. Starting with the publication of the Suspect Adverse Reaction Report Form (CIOMS Form I) by the CIOMS working group II, other CIOMS publications have greatly shaped the direction of PV. CIOMS publications have also greatly influenced the development of International Conference on Harmonization of Technical requirements for Registration of Pharmaceuticals for Human Use (ICH) E2A-E2F guidelines in drug safety. The standard

Hemovigilance is defined as the surveillance procedures covering the blood transfusion chain (from the collection of blood and its components to the follow-up of its recipients), intended to collect and assess information on unexpected or undesirable effects. The word hemovigilance is derived from the word pharmacovigilance, which encompasses activities and systems to collect information useful in supervising medicinal products, with particular reference to adverse drug reactions in human beings, and to evaluate such information scientifically. An adverse drug reaction is a response to a drug which is noxious and unintended and which occurs at doses normally used in human for prophylaxis, diagnosis, or therapy of disease or for the modification of physiologic function. The aim of hemovigilance is to detect and to analyze all untoward effects of blood transfusion in order to correct their cause and to prevent recurrence, and to improve the safety of blood transfusion. Although several reports have been published on adverse events, including transfusion-associated deaths, the relative risk based on the number of actual cases divided by the number of units of blood products issued or transfused, is relatively low. Scope of different hemovigilance systems varies due to differences in spectrum of reporting. Ideally, the hemovigilance system should cover processes throughout the entire transfusion chain, from blood donation, processing, and transfusion to patients for the monitoring, reporting, and investigation of adverse events and reactions and near misses related to blood transfusion. Keywords: Serious Hazards of Transfusion (SHOT), European Hemovigilance Network (EHN), Pharmacovigilance Program of India (PVPI), Hemovigilance Program of India (HvPI), International Hemovigilance Network (IHN).

Adverse Drug Reaction (ADR) is a noxious and unintended response to a medicine which occurs at doses normally used in human being. Pharmacovigilance (PV) is the process to monitor patient safety in practical condition and analyse adverse events during the entire life cycle of a drug beginning from its trial phase to post-marketing of the drug. Before medicines get authorised, they are tested in controlled trial conditions within a smaller population for a limited length of time. Post authorization, drugs are used by a larger population across various geographies without any control in place. So, it is very important to monitor adverse effect caused by these medicines throughout their life cycle in order to identify and minimize risks associated with the use of drug and enhance the knowledge of healthcare professionals. Under reporting is a major obstacle in the PV system. Research shows that the major causes of under reporting are lack of knowledge and awareness among healthcare professionals and also absence of established ADR reporting system in various part of the world. Some other reasons of under-reporting included large number of patients, lower doctor-patient ratio and common practice of self-medication. This article reviews the research done on ADR under-reporting scenario with a focus on the factors leading to this obstacle in PV. Keywords: ADRAdverse Drug Reaction, PVPharmacovigilance, WHOWorld Health Organisation, UMCUppsala Monitoring Centre, AMCADRs Monitoring Centres, AIIMSAll India Institute of Medical, Sciences, CDSCOCentral Drugs Standard, Control Organization, ICSR Individual Case Safety Report, FDA Food and Drug administration, ICH International Conference on Harmonisation, EMAEuropean Medicines Agency, PRAC Pharmacovigilance Risk, Assessment Committee, PMDA Pharmaceuticals and medical, Device Agency.

The emergence of SARS-CoV-2 variants of concern (VOCs) requires the development of next-generation biologics that are effective against a variety of strains of the virus. Herein, we characterize a human VH domain, F6, which we generated by sequentially panning large phage displayed VH libraries against receptor binding domains (RBDs) containing VOC mutations. Cryo-EM analyses reveal that F6 has a unique binding mode that spans a broad surface of the RBD and involves the antibody framework region. Attachment of an Fc region to a fusion of F6 and ab8, a previously characterized VH domain, resulted in a construct (F6-ab8-Fc) that neutralized Omicron pseudoviruses with a half-maximal neutralizing concentration (IC50) of 4.8 nM in vitro. Additionally, prophylactic treatment using F6-ab8-Fc reduced live Beta (B.1.351) variant viral titers in the lungs of a mouse model. Our results provide a new potential therapeutic against SARS-CoV-2 VOCs - including the recently emerged Omicron variant - and highlight a vulnerable epitope within the spike protein RBD that may be exploited to achieve broad protection against circulating variants.

De-novo protein design explores the untapped sequence space that is otherwise less discovered during the evolutionary process. This necessitates an efficient sequence space search engine for effective convergence in computational protein design. We propose a greedy simulated annealing-based Monte-Carlo parallel search algorithm for better sequence-structure compatibility probing in protein design. The guidance provided by the evolutionary profile, the greedy approach, and the cooling schedule adopted in the Monte Carlo simulation ensures sufficient exploration and exploitation of the search space leading to faster convergence. On evaluating the proposed algorithm, we find that a dataset of 76 target scaffolds report an average root-mean-square-deviation (RMSD) of 1.07 Å and an average TM-Score of 0.93 with the modeled designed protein sequences. High sequence recapitulation of 48.7% (59.4%) observed in the design sequences for all (hydrophobic) solvent-inaccessible residues again establish the goodness of the proposed algorithm. A high (93.4%) intra-group recapitulation of hydrophobic residues in the solvent-inaccessible region indicates that the proposed protein design algorithm preserves the core residues in the protein and provides alternative residue combinations in the solvent-accessible regions of the target protein. Furthermore, a COFACTOR-based protein functional analysis shows that the design sequences exhibit altered molecular functionality and introduce new molecular functions compared to the target scaffolds. Communicated by Ramaswamy H. Sarma

The pharmacist’s contribution will remain an important element in effective healthcare system. Pharmacists have a critical role in patient drug safety by contributing to the identification, documentation and reporting of adverse drug reactions. Pharmacists have key roles to play in establishing benefits and risks profile of a marketed medicine. Country level drug ADR monitoring and programs throughout the world differ in their way of collecting ICSR reports by healthcare professionals specially pharmacist. In contrast to North American continent countries like Canada or the US, where the majority of the reports come from licensed pharmacists, some European and Asian countries, such as UK, France, Ireland, Malaysia have the maximum contribution of ADR reports coming from practicing physicians.

The emergence of SARS-CoV-2 variants necessitates rational assessment of their impact on the recognition and neutralization of the virus by the host cell. We present a comparative analysis of the interactions of Alpha, Beta, Gamma, and Delta variants with cognate molecules (ACE2 and/or furin), neutralizing nanobodies (Nbs), and monoclonal antibodies (mAbs) using

Major, trace element concentrations and Nd, Sr and Ca stable isotopic compositions (δ44/40Ca and δ44/42Ca w.r.t. NIST SRM915a) of carbonatites and associated igneous silicate rocks from the ~65 Ma old Ambadongar carbonatite complex and the surrounding Phenai Mata igneous complex of western India are reported. Samples of fluorspar from Ambadongar and the Bagh Limestone and Sandstone, which are part of the country rocks at Ambadongar, have also been analysed. The Ambadongar carbonatites are primarily calcio- and ferro-carbonatites while the silicate rocks from these two complexes are alkaline and tholeiitic in composition. The δ44/40Ca values of the carbonatites (0.58–1.1‰, n = 7) and the associated igneous silicate rocks (0.50–0.92‰, n = 14) show a broad range. The low K/Ca values of the carbonatites ( 1000). Phlogopite, especially if F-rich, is stable at greater depths in the mantle compared to amphibole. Hence, the correlated δ44/40Ca and K/Rb values of the carbonatites and associated igneous silicate rocks suggest the derivation of these carbonatites from a relatively deeper mantle source compared to the silicate rocks, both within the Reunion mantle plume. The origin of the carbonatites from the F-rich phlogopite-bearing mantle is also consistent with the occurrence of large fluorspar deposits within the Ambadongar carbonatite complex.

Insertions/deletions of amino acids in the protein backbone potentially result in altered structural/functional specifications. They can either contribute positively to the evolutionary process or can result in disease conditions. Despite being the second most prevalent form of protein modification, there are no databases or computational frameworks that delineate harmful multipoint deletions (MPD) from beneficial ones. We introduce a positive unlabeled learning-based prediction framework (PROFOUND) that utilizes fold-level attributes, environment-specific properties, and deletion site-specific properties to predict the change in foldability arising from such MPDs, both in the non-loop and loop regions of protein structures. In the absence of any protein structure dataset to study MPDs, we introduce a dataset with 153 MPD instances that lead to native-like folded structures and 7650 unlabeled MPD instances whose effect on the foldability of the corresponding proteins is unknown. PROFOUND on 10-fold cross-validation on our newly introduced dataset reports a recall of 82.2% (86.6%) and a fall out rate (FR) of 14.2% (20.6%), corresponding to MPDs in the protein loop (non-loop) region. The low FR suggests that the foldability in proteins subject to MPDs is not random and necessitates unique specifications of the deleted region. In addition, we find that additional evolutionary attributes contribute to higher recall and lower FR. The first of a kind foldability prediction system owing to MPD instances and the newly introduced dataset will potentially aid in novel protein engineering endeavors.

The Viral Protein 35 (VP35), a crucial protein of the Zaire Ebolavirus (EBOV), interacts with a plethora of human proteins to cripple the human immune system. Despite its importance, the entire structure of the tetrameric assembly of EBOV VP35 and the means by which it antagonizes the autophosphorylation of the kinase domain of human protein kinase R (PKRK) is still elusive. We consult existing structural information to model a tetrameric assembly of the VP35 protein where 93% of the protein is modeled using crystal structure templates. We analyze our modeled tetrameric structure to identify interchain bonding networks and use molecular dynamics simulations and normal-mode analysis to unravel the flexibility and deformability of the different regions of the VP35 protein. We establish that the C-terminal of VP35 (VP35C) directly interacts with PKRK to prevent it from autophosphorylation. Further, we identify three plausible VP35C-PKRK complexes with better affinity than the PKRK dimer formed during autophosphorylation and use protein design to establish a new stretch in VP35C that interacts with PKRK. The proposed tetrameric assembly will aid in better understanding of the VP35 protein, and the reported VP35C-PKRK complexes along with their interacting sites will help in the shortlisting of small molecule inhibitors.

We report the Raman spectroscopy studies on the strontium and ruthenium doped pyrochlore iridate compound Eu2Ir2O7. Eu2Ir2O7 is Weyl semimetallic oxide which exhibits metal-insulator transition at around 120 K. Strontium doping at the Europium site increases the metallic behavior and the metal-insulator transition temperature also lowers systematically with increasing doping concentrations.Ruthenium doping in the iridium site turns the compound to be completely insulating. The Raman spectroscopy studies show a systematic variation of specific Raman modeswith doping which involves Ir-O-Ir bond bending, Ir-O, and Eu-O bond stretching. Clear variations of peak positions have been observed for T2g(1), A1g and T2g(4) modes. The variations of electron-phonon interactions reflected in the Raman peaks can be attributed to the different electronic ground states of the doped samples. We have also corroborated the Raman peak shifts with the structural parameters obtained from XRD data.

Calcium stable isotopic compositions (delta Ca-44/40(SRM 915a)) as well as selected major and trace element concentrations are reported for micro-drilled samples of a hand-specimen sized spheroidally weathered diabase (synonymously used as dolerite or microgabbro) from southern India. A sample of the similar to 2.37 Ga old unweathered diabase (UW) dike was also analyzed. X-ray micro-CT imaging of a representative portion of the weathered sample shows the presence of two dominant mineral phases, plagioclase and clinopyroxene, with minor proportions of an iron-bearing phase (possibly haematite or ilmenite). Two different generations of crosscutting micro-fractures are identified from the micro-CT image. The older fracture is sealed with secondary mineral deposits. The delta Ca-44/40 values of the weathered samples range from 0.42 parts per thousand to 0.84 parts per thousand with samples showing both higher and lower values than that of the un-weathered diabase dike which shows a delta Ca-44/40 value of 0.65 parts per thousand. The variation in delta Ca-44/40 in the weathered samples is significantly higher than the external reproducibility of our measurements (< 0.1 parts per thousand) based on multiple measurements of Ca isotopic standards SRM 915a, SRM 915b and seawater (NASS-6). Mass balance calculations based on Sr/Ca and delta Ca-44/40 values rule out the contributions of secondary silicate as well as carbonate minerals in causing the variability of delta Ca-44/40 in the weathered samples. The positive correlations between delta Ca-44/40 and Mg/Al, Mg/Na in the weathered samples further negate the possible contribution of secondary carbonates in causing the variability in delta Ca-44/40, as Al and Na are unlikely to be present in carbonates. The samples with lower delta Ca-44/40 show higher CIA (Chemical Index of Alteration) values and Al/Ca than the UW diabase while samples having higher delta Ca-44/40 show lower CIA and Al/Ca than the UW diabase. The weathered sample having the lowest delta Ca-44/40 (close to the delta Ca-44/40 of plagioclase) exhibits the highest value of europium anomaly (Eu/Eu*). These observations are explained by the presence of variable relative proportions of residual clinopyroxene and plagioclase in the weathered diabase, due to selective weathering of these minerals. Since plagioclase and clinopyroxene could not be separated from the diabase, we measured the Ca isotopic compositions of a clinopyroxene (delta Ca-44/40= 1.06 parts per thousand) from the San Carlos mantle peridotite and plagioclase from the Chilka anorthosite complex (delta Ca-44/40= 0.40 parts per thousand) from the Eastern Ghats and used these isotopic values as representative of the compositions of these minerals in the diabase. Based on mixing models, using delta Ca-44/40, Al/Ca and Sr/Ca of the plagioclase and clinopyroxene end-members, we estimate that the delta Ca-44/40 of the UW diabase can be explained by 31 parts per thousand clinopyroxene and 69 parts per thousand plagioclase, which is consistent with the modal proportion of these minerals in an average unweathered diabase. Based on mixing calculations, the delta Ca-44/40, Sr/Ca and Al/Ca variability of the micro-drilled samples of the weathered diabase are explained by the presence of varying proportions of residual clinopyroxene (20-65 parts per thousand) and plagioclase (80-35 parts per thousand) in the weathered rock. This study demonstrates that selective weathering of major rock-forming minerals in nature can result in significant variation in delta Ca-44/40 in weathered rocks and that selective weathering of rock-forming minerals should be considered as an additional mechanism to explain the delta Ca-44/40 variability in rivers.

Geochemical and isotopic investigation of Proterozoic clastic and chemical sedimentary rocks provide insights into surface conditions of the early Earth. The mid-Proterozoic Vindhyan Basin of central India, comprising ~5 km thick, mostly undeformed and unmetamorphosed clastic and chemical sediments, is one such archive which is exposed in two sub-basins, the Son Valley Vindhyan sub-basin in the east (SVV) and Chambal Valley Vindhyan sub-basin in the west (CVV). Geochemical and Nd isotopic compositions of siliciclastic sedimentary rocks from the CVV sub-basin are reported in order to understand the provenance of the Vindhyan sediments in this sub-basin and the results are compared with the provenance of the sediments exposed in the SVV sub-basin to evaluate whether the two sub-basins had shared a common provenance. Elemental ratios, such as Th/Co, La/Sc suggest that the clastic sedimentary rocks from the CVV have dominantly upper crustal silicic/felsic provenances and are broadly similar to those of equivalent stratigraphic horizons from the SVV. Initial Nd isotopic composition (eNd(t)) of the siliciclastic rocks from the Lower Vindhyans of the CVV, calculated at their respective times of deposition, suggest sediment derivation from the Banded Gneissic Complex, Berach Granitoids, 1.82 Ga continental arc granitoids of the Aravalli Range and the Hindoli Volcanics. In contrast, the eNd(t) values of the Upper Vindhyan siliciclastics suggest that the Delhi Belt granitoids were the primary source of detritus during the deposition of these sediments. Paleo-depositional environment and connectivity between the SVV and CVV sub-basins are evaluated using geochemical, radiogenic Nd, and stable Ca isotopic compositions of carbonates from both sub-basins. Non-redox sensitive REY anomalies and Y/Ho ratios of carbonates from the two sub-basins suggest deposition in an epeiric sea with limited connection to the open ocean. Neodymium isotopic compositions of the carbonate units suggest that the two sub-basins were connected during the deposition of both Lower and Upper Vindhyan sediments. Carbonates from both sub-basins record a significant shift towards more radiogenic Nd isotopic compositions around 1.6 Ga, possibly as a result of an Andean-type arc magmatism in near vicinity of the Son Valley sub-basin. The δ44/40Ca compositions of the SVV carbonates are typically lower than the CVV carbonates. This difference is likely due to greater freshwater input in the SVV sub-basin which is consistent with the existence of paleo-gradients in the Vindhyan Basin.

The proposed work aims at designing a classification system for automatic identification of A. muelleri species, grown as a potential cash crop in many Asian countries, from the DNA fingerprints of Amorphophallus genus. Four sets of 48 DNA fingerprints belonging to 37 species of the Amorphophallus genus, developed with the help of four different primers are considered for the experiment, with an objective to identify only the fingerprints of the species of interest. A second experimental setup deals with the automatic classification of species containing high amounts of glucomannan from the same set of DNA fingerprints of the Amorphophallus genus. For each set of 48 DNA fingerprints generated with a specific primer, the DNA fingerprints are preprocessed to extract a 42 dimensional feature vector which is used to generate a k-Nearest Neighbor based classifier based on the Leave One Out Cross Validation protocol. Final classification based on outputs from individual classifiers constructed with respect to the four different primers is performed according to a n-star consensus strategy. The n-star consensus predicts species A. muelleri with cent per cent accuracy while it predicts species containing glucomannan with a more modest accuracy of 81.25%.

A new system (ARTeM) for human authentication using finger vein images is described here. The developed algorithm combines 1) a fuzzy contrast enhancement algorithm with 2) a mutual information and affine transformation based registration technique and 3) a correlation coefficient based template matching algorithm, to detect the identity of a person based on the match-scores with finger vein images stored in the database. For performance assessment of the ARTeM algorithm, the benchmark SDUMLA multimodal biometric database containing 3816 images of 106 persons is used. On the complete database, up to 95.28% classification accuracy is achieved with single finger images; while up to 98.11% accuracy is observed with a consensus of two fingers. On a reduced subset of 86 persons’ database, 98.84% accuracy is achieved with single finger classification and cent percent classification is obtained using a consensus of two fingers. Comparative analyses with other works also validate the effectiveness of the developed methodology.