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.

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.

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.

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.

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.

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.

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.

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.

A cost-effective polyethylene glycol-based method for the isolation of functional edible nanoparticles from ginger rhizomes.

Edible nanoparticles (ENPs) are nano-sized vesicles derived from edible plants. These ENPs are loaded with plant derived microRNAs, protein, lipids and phytochemicals. Recently, ginger derived ENPs was shown to prevent inflammatory bowel diseases and colon cancer, in vivo, highlighting their therapeutic potential. Conventionally, differential centrifugation with an ultra-centrifugation step is employed to purify these ENPs which imposes limitation on the cost-effectiveness of their purification. Herein, we developed polyethylene glycol-6000 (PEG6000) based ginger ENP purification (PEG-ENPs) method, which eliminates the need for expensive ultracentrifugation. Using different PEG6000 concentrations, we could recover between 60% to 90% of ENPs compared to ultracentrifugation method. PEG-ENPs exhibit near identical size and zeta potential similar to ultra-ENPs. The biochemical composition of PEG-ENPs, such as proteins, lipids, small RNAs and bioactive content is comparable to that of ultra-ENPs. In addition, similar to ultra-ENPs, PEG-ENPs are efficiently taken up by the murine macrophages and protects cells from hydrogen peroxide induced oxidative stress. Since PEG has been approved as food additive, the PEG method described here will provide a cost-effective alternative to purify ENPs, which can be directly used as a dietary supplement in therapeutic formulations.

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.

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.

Pediatric non-cirrhotic portal fibrosis: role of endoscopic management in determining long-term outcome.

AIMS: Non-cirrhotic portal fibrosis (NCPF) is a rare cause of pediatric portal hypertension. There is abundant literature in adults but paucity of data in children. We aimed to evaluate and compare the endoscopic and long-term outcomes of children with NCPF. METHODS: Consecutive children (≤ 18 years) diagnosed with NCPF evaluated for clinical and endoscopic profile and outcome. The cohort underwent 3 weekly endoscopic sessions until esophageal variceal eradication followed by 6-12 monthly endoscopic surveillance. Non-bleeders and bleeders were compared for endoscopic outcome. RESULTS: Forty-five NCPF children with median age of 14.5 (6-18) years and symptom duration 12 (1-120) months presented with spleen-related issues (78%), esophageal varices (96%), primary gastric varices (56%), and portal hypertensive gastropathy (89%). Thirty-three patients undergoing endotherapy (secondary prophylaxis n = 22, primary prophylaxis n = 11) showed primary eradication of varices after 5 (2-12) sessions. 36% showed recurrence of esophageal varices in 11 (6-42) months and secondary gastric varices developed in 12%. Overall 87% patients required endoscopic intervention at onset or follow-up. Poor outcome was observed in ten patients (n = 9 bleeders). Three children died of variceal bleeding before endoscopic eradication of esophageal varices. Three developed decompensation after a median follow-up of 48 (3-120) months and referred for liver transplantation. Four patients required surgery for portal hypertension. CONCLUSIONS: Overall, endoscopic outcome of NCPF is favorable. One-third patients have recurrence of esophageal varices. Small proportion of bleeders have poor outcome.

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.

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.

Effect of Long-term Oral Steroids on Intraocular Pressure in Children With Autoimmune Hepatitis: a Prospective Cohort Study.

OBJECTIVE: Topical corticosteroids are known to cause raised intraocular pressure (IOP). However, there is a scarcity of literature regarding systemic steroids-induced raised IOP in children. The authors aimed to evaluate the IOP in children with autoimmune hepatitis (AIH) receiving oral prednisone. METHODS: In this prospective study, children (1 to 18 y) with newly diagnosed AIH who received oral prednisone (July 2016 to December 2017) were included. Comprehensive ophthalmic check-up, including visual acuity, IOP, slit-lamp, and fundus examinations were done before and at 1, 3, and 6 months of follow-up. IOP ≥20 mm Hg was considered raised IOP. Symptomatic raised IOP patients were managed with antiglaucoma medication with a tapering dose of prednisone. RESULTS: A total of 33 children (19 boys) with a median age of 11 (3 to 18) years were analyzed. Raised IOP was observed in 20 (61%) children (19 were moderate and 1 was high responder) at 1 month, 8 (24%) at 3 months, and 1 (3%) at 6 months of treatment. Patients who developed raised IOP had a more severe liver disease in terms of decompensation and low albumin and high pediatric end-stage liver disease scores at presentation. There was no correlation between IOP and starting and cumulative dose of steroids. On multivariate analysis, decompensation at presentation was significantly associated with raised IOP (P<0.001; odds ratio, 30; 95% confidence interval, 4.2-210.6). Raised IOP returned to normal with antiglaucoma medication along with prednisone tapering in all except one. CONCLUSIONS: Systemic corticosteroids in children with AIH carry a risk for IOP elevation, especially in decompensated liver disease. A periodic ophthalmic check-up is necessary for early recognition and intervention before irreversible vision loss.