Breaking Cycles: Saponification-Enhanced NMR Fingerprint Matching for the Identification and Stereochemical Evaluation of Cyclic Lipodepsipeptides from Natural Sources.

We previously described NMR based fingerprint matching with peptide backbone resonances as a fast and reliable structural dereplication approach for Pseudomonas cyclic lipodepsipeptides (CLiPs). In combination with total synthesis of a small library of configurational CLiP congeners this also allows unambiguous determination of stereochemistry, facilitating structure-activity relationship studies and enabling three-dimensional structure determination. However, the on-resin macrocycle formation in the synthetic workflow brings considerable burden and limits universal applicability. This drawback is here removed altogether by also transforming the native CLiP into a linearized analogue by controlled saponification of the ester bond. This eliminates the need for macrocycle formation, limiting the synthesis effort to linear peptide analogues. NMR fingerprints of such linear peptide analogues display a sufficiently distinctive chemical shift fingerprint to act as effective discriminators. The approach is developed using viscosin group CLiPs and subsequently demonstrated on putisolvin, leading to a structural revision, and tanniamide from Pseudomonas ekonensis COR58, a newly isolated lipododecapeptide that defines a new group characterized by a ten-residue large macrocycle, the largest to date in the Pseudomonas CLiP portfolio. These examples demonstrate the effectiveness of the saponification- enhanced approach that broadens applicability of NMR fingerprint matching for the determination of the stereochemistry of CLiPs.

Functional characterization of a high-affinity iron transporter (PiFTR) from the endophytic fungus Piriformospora indica and its role in plant growth and development.

Iron (Fe) is a micronutrient required for plant growth and development; however, most Fe forms in soil are not readily available to plants, resulting in low Fe contents in plants and, thereby, causing Fe deficiency in humans. Biofortification through plant-fungal co-cultivation might be a sustainable approach to increase crop Fe contents. Therefore, we aimed to examine the role of a Piriformospora indica Fe transporter on rice Fe uptake under low Fe conditions. A high-affinity Fe transporter (PiFTR) from P. indica was identified and functionally characterized. PiFTR fulfilled all criteria expected of a functional Fe transporter under Fe-limited conditions. Additionally, PiFTR expression was induced when P. indica was grown under low Fe conditions, and PiFTR complemented a yeast mutant lacking Fe transport. A knockdown (KD) P. indica strain was created via RNA interference to understand the physiological role of PiFTR. We observed that the KD-PiFTR-P. indica strain transported a significantly lower amount of Fe to colonized rice (Oryza sativa) than the wild type (WT) P. indica. WT P. indica-colonized rice plants were healthier and performed significantly better than KD-PiFTR-P. indica-colonized rice plants. Our study offers potential avenues for an agronomically sound amelioration of plant growth in low Fe environments.

Can urodynamic diagnosis of dysfunctional voiding/external sphincter nonrelaxation be made on two-channel pressure-flow study without video-urodynamics, electromyography, or urethral pressure profilometry? Plateau detrusor pattern in perspective.

AIMS: urodynamic diagnosis of dysfunctional voiding/external-sphincter nonrelaxation (DV/EUSD) needs assistance of specialized testing namely urethral pressure profilometry (UPP), electromyography (EMG), and/or videofluoroscopy (VUDS). We aimed to find a predictive model based on standard pressure-flow study without need for specialized testing. MATERIAL AND METHODS: In this retrospective study (2017-2021), clinical and urodynamic data of adult men and women presenting with voiding dysfunction was collected. Mandatory inclusion criteria were availability of all-(1) findings of clinical examination and neurological status, (2) a valid filling cystometry and pressure-flow study (with active detrusor contraction), (3) a final clinic-urodynamic diagnosis. Voiding cystourethrography (VCUG) was performed to confirm the location of obstruction. RESULTS: Data of 218 participants (178♂, 40♀) was eligible. Plateau detrusor contraction pattern was observed in 89.0% of men and 86% of women with DV/EUSD; whereas only 7.5% men and no women with other obstructions demonstrated this pattern. Forward likelihood Logistic regression analysis revealed presence of plateau pattern, lower bladder outlet obstruction index (BOOI), and smaller difference between Pdetmax and PdetQmax highly predictive of presence of DV/EUSD in men as per the following equation-Y = -9.900 + (0.085 × BOOI) + (0.123 × pdetmax - pdetQmax) + (4.061 × detrusor pattern). A kattan-type nomogram was constructed based on the above equation. In women, presence of plateau pattern alone was highly predictive of DV/EUSD. CONCLUSION: Diagnosis of DV/EUSD can be accurately predicted using parameters of three-channel urodynamics (plateau pattern, BOOI, Pdetmax-pdetQmax) minimizing need for specialized testing.

Transjugular Versus Percutaneous Liver Biopsy in Children: Indication, Success, Yield, and Complications.

BACKGROUND/OBJECTIVE: Percutaneous biopsy (PB) and transjugular liver biopsy (TJLB) are 2 main ways of obtaining liver tissue. We evaluated the indications, success rate, tissue yield, and complications of TJLB in comparison to PB in children. METHODS: Electronic records of children undergoing liver biopsy (LB) were reviewed. Clinico laboratory data including indication, type of biopsy, complications, and tissue yield (length and number of complete portal tracts [CPT]) were noted. RESULTS: Five hundred forty LB (indication: neonatal cholestasis [42.9%], chronic liver disease [43.7%], liver failure [3.7%], focal lesions [3.3%] and others [6.3%]) were done. Four hundred seventy-three were PB (317 boys, 14 [1--216] months) done by percussion (322 [68%]), real-time ultrasound guidance (125 [26.4%]), or plugged method [26 (5.5%)]. Sixty-seven (12.4%) were TJLB [38 boys, 140 (24--216) months], done in patients with contraindications for PB. Technical success (67/68 vs 473/473; P = 0.7) and complications (4 [6%]; vs 15 [3.3%]; P = 0.2) of TJLB and PB were similar. Major complications (0.5%) included supraventricular tachycardia (n = 1) in TJLB and hemoperitoneum (n = 2) in PB. Tissue yield of TJLB was poorer in terms of length (1.0 [0.2--2.0] vs 1.1 [0.4--2.1] cm; P < 0.001), CPT (4 [0--9] vs 5 [2--17]; P < 0.001) and adequacy for reporting (56/67 vs 459/473; P < 0.001). Biopsy yield of <6 CPT was predicted by cirrhosis at histology and TJLB. No factor identified risk of complications with LB. CONCLUSIONS: LB is a safe procedure and only 12% children require TJLB because of contraindications of PB. Technical success and complications are similar but tissue yield is poorer in TJLB than PB. Presence of cirrhosis and TJLB adversely affected tissue yield.

Ocular manifestations of liver disease in children: Clinical aspects and implications.

Liver and eyes are interlinked to each other in various medical conditions. There are certain ocular findings which directly indicate specific liver disorders. Thus, it becomes critical to identify disorders of liver and eyes early in the course of illness, so that prompt management may be initiated before the commencement of complications. It is highly advantageous in metabolic liver disorders as it offers prognostic value and spares the patient of unnecessary invasive and detailed work up. However, due to its silent and heterogeneous presentation, it is often unrecognized and ignored. Eye abnormalities could be due to, either direct toxic effects of abnormal metabolites, excess of normal metabolites, or by deficient energy metabolism. A number of inherited liver conditions have associated ocular lesions such as Kayser-Fleischer rings in Wilson's disease, posterior embryotoxon or optic drusen in Alagille's syndrome, and cherry-red spot in Niemann-Pick's type A. A thorough eye examination is important in distinguishing between several different forms of familial intrahepatic cholestasis which are associated with anomalies of the heart, bones, or kidneys. Early diagnosis is important, as in most cases, dietary restriction and early therapy prevents the onset of disability. The aim of this review is to sensitize and make pediatricians, hepatologists and ophthalmologists aware of specific ocular findings, suggestive of certain hepatobiliary disorders, thus helping in early referral. The pediatric and adult literature was thoroughly reviewed to organize the present review.

Clinical Profile, Response to Therapy, and Outcome of Children with Primary Intestinal Lymphangiectasia.

OBJECTIVE: Intestinal lymphangiectasia (IL; primary or secondary) is an important cause of protein-losing enteropathy. We evaluated the clinicolaboratory profile, response to therapy, complications, and outcome of children with primary IL (PIL). METHODS: Consecutive children (≤18 years) diagnosed with PIL (clinical setting, typical small bowel histopathology, and exclusion of secondary causes) from 2007 to 2017 were evaluated. RESULTS: Twenty-eight children with PIL (16 boys, age at symptom onset-12 [1-192] months and at diagnosis 8 [1-18] years) were studied. Pedal edema (93%), chronic diarrhea (78.6%), and recurrent anasarca (64%) were the common presentations. Ascites, pleural, and pericardial effusion were seen in 64 (n-18; chylous-5, non-chylous-13), 18, and 18% cases, respectively. Hypoproteinemia, hypoalbuminemia, hypocalcemia, and lymphopenia were present in 82, 82, 75 and 39% cases, respectively. Duodenal biopsy established the diagnosis in 86% cases, while 14% required distal small bowel biopsies. Dietary therapy was given in all and 6 cases required additional therapy (octreotide-6, tranexamic acid-3, and total parenteral nutrition-1). Lymphedema (3/5 vs. 1/23), pleural effusion (4/5 vs. 1/23), and the need for additional therapy (4/5 vs. 2/23) were significantly more in patients with chylous ascites (n = 5) than those without chylous ascites (n = 23). Twenty-four cases in follow-up (39 [6-120] months) showed improvement; however, 8 required readmission (symptom recurrence-6 [25%], complication-2 [8.3%], Budd Chiari Syndrome-1, and abdominal B cell lymphoma-1). CONCLUSION: Presence of chylous ascites suggests severe disease in children with PIL. Majority of PIL children respond to dietary therapy; only 20% need additional therapy. Long-term follow-up is essential to monitor for symptoms relapse and complications.

Selective IRAK4 Inhibition Attenuates Disease in Murine Lupus Models and Demonstrates Steroid Sparing Activity.

The serine/threonine kinase IL-1R-associated kinase (IRAK)4 is a critical regulator of innate immunity. We have identified BMS-986126, a potent, highly selective inhibitor of IRAK4 kinase activity that demonstrates equipotent activity against multiple MyD88-dependent responses both in vitro and in vivo. BMS-986126 failed to inhibit assays downstream of MyD88-independent receptors, including the TNF receptor and TLR3. Very little activity was seen downstream of TLR4, which can also activate an MyD88-independent pathway. In mice, the compound inhibited cytokine production induced by injection of several different TLR agonists, including those for TLR2, TLR7, and TLR9. The compound also significantly suppressed skin inflammation induced by topical administration of the TLR7 agonist imiquimod. BMS-986126 demonstrated robust activity in the MRL/lpr and NZB/NZW models of lupus, inhibiting multiple pathogenic responses. In the MRL/lpr model, robust activity was observed with the combination of suboptimal doses of BMS-986126 and prednisolone, suggesting the potential for steroid sparing activity. BMS-986126 also demonstrated synergy with prednisolone in assays of TLR7- and TLR9-induced IFN target gene expression using human PBMCs. Lastly, BMS-986126 inhibited TLR7- and TLR9-dependent responses using cells derived from lupus patients, suggesting that inhibition of IRAK4 has the potential for therapeutic benefit in treating lupus.

Bacillus Calmette-Guérin vaccine induces a selective serotonin reuptake inhibitor (SSRI)-resistant depression like phenotype in mice.

Preclinical studies have shown that administration of Bacillus Calmette-Guérin (BCG) vaccine induces depression-like behaviors in mice; however, the effect of antidepressant drug treatment has not been reported earlier. In the present study, we induced depression-like behavior by administering BCG vaccine to BALB/c mice. BCG treatment produced robust serum sickness as shown by a decrease in body weight, reduced spontaneous locomotor activity and reduced voluntary wheel running activity. BCG treatment also elevated plasma IL6 and IFNγ levels and produced a marked activation of lung IDO activity. At a time point when serum sickness-related behaviors had fully recovered (i.e., day 14) BCG-treated mice showed a significant increase in immobility in the forced swim test (FST) and tail suspension test (TST) indicative of a pro-depressant phenotype. We observed significant increase in [(3)H]PK11195 binding in cortex and hippocampus regions of BGC-treated mice in comparison to saline-treated mice indicating prominent neuroinflammation. Pharmacological evaluation of FST behavior in BCG-treated mice demonstrated selective resistance to the selective serotonin reuptake inhibitors (SSRIs) fluoxetine and escitalopram. In contrast the tricyclic antidepressant imipramine, the dual serotonin/norepinephrine reuptake inhibitor (SNRI) duloxetine, and the dual dopamine/norepinephrine reuptake inhibitor (DNRI) nomifensine retained antidepressant efficacy in these mice. The lack of efficacy with acute treatment with SSRIs could not be explained either by differences in drug exposure or serotonin transporter (SERT) occupancy. Our results demonstrate that BCG-vaccine induced depression like behavior is selectively resistant to SSRIs and could potentially be employed to evaluate novel therapeutic agents being developed to treat SSRI-resistance in humans.

Diagnosis of oral pigmentations and malignant transformations.

BACKGROUND: Oral pigmentation is a common finding in the mouth. Pigmentation can be either normal or abnormal discoloration of oral mucous membrane. The purpose of this review mainly focuses on the main oral pigmented lesions, in order to help the clinicians establish a better approach towards the patients with pigmented oral lesions and to provide thorough knowledge regarding such lesions for patient reassurance, early definitive diagnosis and prompt treatment. METHODS: Relevant data concerning oral pigmented lesions, clinical features and the possibility of malignant transformation of such lesions were reviewed thoroughly from pubmed literature published in English. Pigmented lesions affecting the skin were not included in our review. RESULTS: Few pigmented lesions have been identified and their tendency to become malignant has been reported in the literature. The oral lesions showing malignant transformation reported were mostly case series. Unfortunately, due to lack of long-term studies, follow ups and randomized controlled studies in this respect it was difficult to draw a statistical analysis. This information is quite crucial for general dental practitioners to improve their understanding regarding oral lesions and to differentiate between normal and diseased conditions, so that they can master the skill of differential diagnosis, definitive diagnosis and prompt treatment. CONCLUSION: Oral pigmentation may present as focal, multifocal or diffused macular or tumefactive lesions. They may greatly vary in color as blue, purple, brown, gray or black depending on the quantity and site of melanin in the tissues [1]. Etiology of pigmentation can be multi factorial. Mostly pigmentation is physiologic but at times it can be a precursor of severe diseases. Lesions may be caused by localized harmless accumulations of melanin, hemosiderin or exogenous metals or they may be a sign of underlying systemic or genetic disease. A few lesions may be associated with life-threatening medical conditions that require immediate intervention. The differential diagnosis for any pigmented lesion is extensive, as it includes examples of endogenous and exogenous pigmentations. Although biopsy is a helpful and necessary aid in the diagnosis of focally pigmented lesions, with diffuse pigmentation lesions require a thorough dental and medical history and laboratory investigations.

Abernethy Malformation Masquerading as Congenital Heart Disease: A Boy With Cyanosis, Clubbing, and Hypoxia.

Abernethy malformation is an extrahepatic congenital portosystemic shunt characterized by the diversion of the portal blood away from the liver through a shunt that drains directly into the inferior vena cava. We present a case of a male child with Abernethy malformation, which was initially diagnosed as cyanotic heart disease due to pulmonary arteriovenous malformation. However, after proper clinical evaluation and investigations, the diagnosis of Abernethy malformation was established. Thereafter, the patient was successfully treated with endovascular embolization. At one year follow-up, marked relief in exertional dyspnea and improvement in physical growth was achieved with no observable complications.

Role of 5G Networks in Healthcare Management System.

The present-day healthcare system operates on a 4G network, where the data rate needed for many IoT devices is impossible. Also, the latency involved in the network does not support the use of many devices in the network. The 5G-based cellular technology promises an effective healthcare management system with high speed and low latency. The 5G communication technology will replace the 4G technology to satisfy the increasing demand for high data rates. It incorporates higher frequency bands of around 100 MHz using millimetre waves and broadband modulation schemes. It is aimed at providing low latency while supporting real-time machine-to-machine communication. It requires a more significant number of antennas, with an average base station density three times higher than 4G. However, the rise in circuit and processing power for multiple antennas and transceivers deteriorates energy efficiency. Also, the data transmission power for 5G is three times higher than for 4G technology. One of the advanced processors used in today's mobile equipment is NVIDIA Tegra, which has a multicore system on chip (SoC) architecture with two ARM Cortex CPU cores to handle audio, images, and video. The state-of-the-art software coding using JAVA or Python has achieved smooth data transmission from mobile equipment, desktop or laptop through the internet with the support of 5G communication technology. This paper discusses some key areas related to 5G-based healthcare systems such as the architecture, antenna designs, power consumption, file protocols, security, and health implications of 5G networks.

Can we predict early renal impairment in pediatric cirrhosis?

AIMS: Published literature on renal dysfunction (RD) in pediatric cirrhosis are limited. We aimed to detect early RD in cirrhotic children by renal resistive index (RI) and plasma aldosterone (PA). We evaluated the effects of large-volume paracentesis (LVP) and albumin infusion on the same. METHODS: Non-azotemic cirrhotic children with tense ascites (undergoing LVP with albumin infusion) were prospectively enrolled. Blood biochemistry and doppler ultrasonography for RI and PA were measured at regular intervals. RI >0.7 was considered as RD. Outcomes were noted at D90 and 1 year. Chronic liver disease children without ascites were included as controls. RESULTS: Of the 99 cirrhotic children, tense ascites (n=51) had higher baseline RI than controls (n=48) (p<0.001). Overall, baseline RD was observed in 32% and was significantly higher in tense ascites compared to controls (59% vs. 4%, p<0.001). Tense ascites with RD at admission had higher chances of acute kidney injury (AKI) (p=0.009), ascites recurrence (p=0.043), hospital readmission (p=0.048), and mortality (p=0.009) compared to patients without RD by D90. Significant reduction in RI was noted at 48 h, D7, D30, and D90 compared to baseline after LVP with albumin. Pediatric End-stage Liver Disease (PELD) score and PA had a strong positive correlation with baseline RI (R2=0.51, R2=0.47). Using multivariate analysis, PELD score and PA were predictors of AKI (odds ratio [OR]: 1.14; 95% confidence interval [CI]: 1.04-1.24; p=0.003) and mortality (OR: 1.82; 95% CI: 1.22-2.72; p=0.004), respectively. CONCLUSIONS: Abnormal baseline RI can be used as an early predictor of RD and predict long-term renal ouctomes in pediatric cirrhosis. Baseline RI correlated well with the severity of liver disease and PA. Paracentesis and albumin infusion effectively reduced RI.

Existence of ß-diketone form of curcuminoids revealed by NMR spectroscopy.

Curcumin (CUR), demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC) - the class of natural compound derived from turmeric can exist as keto-enol and ß-diketone tautomer form. The structure and dynamics of particular relevance CUR is reported in prior studies, whereas DMC and BDMC, by far, have not been scrutinized. In the present studies, we have investigated the detailed molecular structure of CUR, DMC and BDMC by employing NMR spectroscopy as a key tool. The bridging carbon as methylene in ß-diketone form and methine in keto-enol form shows significant difference in NMR spectrum. The results justified that Curcuminoids (CC) has nearly 3% of ß-diketone tautomer in DMSO solvent at 298 K. Further, results revealed that ß-diketone form was favoured in alkaline pH condition whereas acidic and neutral pH conditions favour keto-enol tautomer. However, at higher temperature equilibrium shift towards ß-diketone tautomer. Moreover, this is the first report by NMR for observing the presence of ß-diketone tautomer.

Facile NMR approach for profiling curcuminoids present in turmeric.

Curcumin, together with demethoxycurcumin and bisdemethoxycurcumin as a whole called curcuminoids, is an active phytochemical constituent present in the turmeric. When it comes to their analysis, most will rely on UV-Visible spectroscopy, HPLC and LC-MS methods. Looking to improve productivity, time and simplicity, we are proposing a 1H NMR based approach for curcuminoids analysis and its applications to different geographical regions. In the present work, sample preparation protocol is reported for the simultaneous determination of curcuminoids using 1H NMR. For the quantification of curcuminoids, 6-7 ppm vinylic proton region in the 1H NMR spectrum was used, where acetone was observed as the suitable solvent in terms of curcuminoids solubility and proper resolution of peak. The result shows that curcumin (46.8-59.50%) was major among all varieties, followed by DMC (22.15-27.70%) and BDMC (17.52-30.29%) except in Andhrapradesh variety, where BDMC (30.29%) was more than DMC (22.89%). These studies were further supported by HPLC analysis.

Proteomics of Two Thermotolerant Isolates of Trichoderma under High-Temperature Stress.

Several species of the soil borne fungus of the genus Trichoderma are known to be versatile, opportunistic plant symbionts and are the most successful biocontrol agents used in today's agriculture. To be successful in field conditions, the fungus must endure varying climatic conditions. Studies have indicated that a high atmospheric temperature coupled with low humidity is a major factor in the inconsistent performance of Trichoderma under field conditions. Understanding the molecular modulations associated with Trichoderma that persist and deliver under abiotic stress conditions will aid in exploiting the value of these organisms for such uses. In this study, a comparative proteomic analysis, using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption/time-of-flight (MALDI-TOF-TOF) mass spectrometry, was used to identify proteins associated with thermotolerance in two thermotolerant isolates of Trichoderma: T. longibrachiatum 673, TaDOR673 and T. asperellum 7316, TaDOR7316; with 32 differentially expressed proteins being identified. Sequence homology and conserved domains were used to identify these proteins and to assign a probable function to them. The thermotolerant isolate, TaDOR673, seemed to employ the stress signaling MAPK pathways and heat shock response pathways to combat the stress condition, whereas the moderately tolerant isolate, TaDOR7316, seemed to adapt to high-temperature conditions by reducing the accumulation of misfolded proteins through an unfolded protein response pathway and autophagy. In addition, there were unique, as well as common, proteins that were differentially expressed in the two isolates studied.

Clinical Profile, Liver Dysfunction and Outcome of Dengue Infection in Children: A Prospective Observational Study.

BACKGROUND: Liver involvement in dengue illness is common and can lead to acute liver failure (ALF). No single method can effectively identify patients at risk for disease progression and bad outcome. We aimed to determine the relationship between liver dysfunction, kinetics of liver function tests (LFTs) and severity of hepatitis on the outcome in pediatric dengue illness. METHODS: We conducted a prospective observational study of hospitalized children (1-12 years) with dengue infection (July 2014-July 2015). Serial monitoring of LFTs was done in confirmed dengue cases. Patients were classified into non-severe (NSD) and severe dengue (SD). Severity of hepatitis was graded: mild, moderate and severe hepatitis. Events were noted during hospitalization. RESULTS: One hundred two children (66, boys), median age 72 (48-96) months, were analyzed (NSD, n = 41; SD, n = 61). Elevated transaminases (92%) was the most common abnormality; aspartate transaminase (AST) and alanine transaminase (ALT) in 87% and 82%, respectively. Maximum abnormalities in LFTs peaked at day 5 (AST, ALT) and day 7 (Alkaline Phosphatase [ALP], gamma-glutamyl transferase) of illness. Elevated transaminases was found to be higher in SD than NSD (100% vs. 80%, P = 0.006). Severe hepatitis developed organ dysfunction such as altered sensorium (P < 0.001), ALF (P < 0.001), acute kidney injury (P < 0.001) and shock (P < 0.001), more commonly than those with mild to moderate hepatitis. Fourteen patients died, two-thirds of whom had severe hepatitis (P < 0.001). Using binary logistic regression, presence of severe hepatitis and shock at presentation was an independent predictor for ALF (odds ratio: 77; 95% confidence interval : 13-457, P < 0.001) and mortality (odds ratio: 55; 95% confidence interval: 4.6-66, P < 0.001), respectively. CONCLUSIONS: Many children with dengue have liver involvement. Severe hepatitis in dengue is associated with significant organ dysfunction and poor outcome.

R-VAPOL-phosphoric acid based 1H and 13C-NMR for sensing of chiral amines and acids.

Enantiomers have significant importance in pharmaceuticals, biology and modern chemistry and therefore distinguishing and quantifying the enantiomeric forms is of utmost importance. Herein, we propose diphenyl-3,3'-biphenanthryl-4,4'-diyl phosphate (R-VAPOL-PA) as a promising chiral solvating agent to discriminate amines and acids of poly-functional groups such as chiral amines, amino alcohols and hydroxy acids. The methodological approach involves using the nature of hydrogen bonds and ion pairs as a mode of weak interactions to form diastereomers where the probe is associated with enantiomers. The resulting diastereomer difference in the NMR spectrum enables the chiral discrimination with a complete baseline peak separation and an accurate enantiomeric excess (ee) analysis. We also carried out density functional theory (DFT) calculations to understand the complex formation to explain enantiodiscrimination by analysing the formation and stability of different chiral complexes. The binding energy differences between enantiomeric forms revealed by DFT calculations are qualitatively in agreement with the diastereomer difference in the NMR spectrum and unequivocally establishes the suggested experimental protocol of R-VAPOL-PA-based enantiomeric discrimination.