Carbonylative cyclization of biaryl enones with aldehydes and oxamic acids.

An oxidative radical-promoted carbonylative cyclization strategy for the synthesis of phenanthren-9-(10H)-one frameworks from biaryl enones using aldehydes as the carbonyl radical sources is disclosed. The reaction proceeds through a sequential addition of a carbonyl radical to the olefin followed by cyclization with an aryl ring. The method is further extended to carbamoyl radicals generated from oxamic acids to access the corresponding phenanthrenones with amide functionalities.

Protein disulfide isomerase uses thrombin-antithrombin complex as a template to bind its target protein and alter the blood coagulation rates.

During inflammation and situations of cellular stress protein disulfide isomerase (PDI) is released in the blood plasma from the platelet and endothelial cells to influence thrombosis. The addition of exogenous PDI makes the environment pro-thrombotic by inducing disulfide bond formation in specific plasma protein targets like vitronectin, factor V, and factor XI. However, the mechanistic details of PDI interaction with its target remain largely unknown. A decrease in the coagulation time was detected in activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) on addition of the purified recombinant PDI (175 nM). The coagulation time can be controlled using an activator (quercetin penta sulfate, QPS) or an inhibitor (quercetin 3-rutinoside, Q3R) of PDI activity. Likewise, the PDI variants that increase the PDI activity (H399R) decrease, and the variant with low activity (C53A) increases the blood coagulation time. An SDS-PAGE and Western blot analysis showed that the PDI does not form a stable complex with either thrombin or antithrombin (ATIII) but it uses the ATIII-thrombin complex as a template to bind and maintain its activity. A complete inhibition of thrombin activity on the formation of ATIII-thrombin-PDI complex, and the complex-bound PDI-catalyzed disulfide bond formation of the target proteins may control the pro- and anti-thrombotic role of PDI.

Insight into the interaction of isochroman with bovine serum albumin: extensive experimental and computational investigations.

The way therapeutic compounds interact with serum protein provides valuable information on their pharmacokinetics, toxicity, effectiveness, and even their structural-related information. Isochroman (IC) is a phytochemical compound obtained from the leaves of Olea europea plant. The derivatives of IC have various pharmacological properties including antidepressants, antihistamines, antiinflammation, anticonvulsants, appetite depressants, etc. The binding of small molecules to bovine serum albumin (BSA) is useful to ensure their efficacy. Thus, in this study, we have found out the binding mode of IC with BSA using several spectroscopic and in silico studies. UV and fluorescence spectroscopy suggested the complex formation between IC and BSA with a binding constant of 103 M-1. IC resulted in fluorescence quenching in BSA through static mechanism. The microenvironmental and conformational changes in BSA were confirmed using synchronous and three-dimensional studies. Site marker experiment revealed the IC binding in site-III of BSA. The influence of vitamins, metals and ß-cyclodextrin (ß-CD) on binding constant of IC-BSA complex was also examined. Circular dichroism spectra showed that α-helical of BSA decreased upon interaction with IC. Computational and experimental results were complimentary with one another and assisted in determining the binding sites, nature of bonds and amino acids included in the IC-BSA complex formation.Communicated by Ramaswamy H. Sarma.

Essential Oils of Some Potential Medicinal Plants and Their Wound Healing Activities.

The wound has been recognised as a deep cut or tearing of the epidermis, which is also referred to as trauma and harm to the body tissues. Healing of wounds requires a coordinated series of cellular processes, including cell attraction, proliferation, differentiation, and angiogenesis. These processes involve interactions between various cells, such as macrophages, endothelial cells, keratinocytes, fibroblasts, growth hormones, and proteases. The outcome of wounds can be fatal if not treated properly, resulting in chronic wounds, chronic pain, and even death. Wound healing is replacing missing tissue with tissue repairs and regeneration. Some local variables are the presence of tissue maceration, foreign objects, biofilm, hypoxia, ischemia, and wound infection. Sustained growth factor delivery, siRNA delivery, micro-RNA targeting, and stem cell therapy are all emerging possible therapeutic approaches for wound healing. Traditional approaches, such as Ayurveda, Siddha, and Unani medicines, are also being used for treatment. The therapeutic application of nanoformulations in wound infections has shown various beneficial effects. Several herbal medicines, especially essential oils have shown potential wound healing activities, such as lavender, tea tree, sesame, olive, etc. Various nanoparticles and their nanoformulations have been explored in wound healing therapy. The present review article highlights several aspects of essential oils for wound healing activity through a novel drug delivery system. Further, some patents on wound healing through herbal medicine have been listed.

Structural and Functional Strategies in Cenchrus Species to Combat Environmental Extremities Imposed by Multiple Abiotic Stresses.

Multiple abiotic stresses such as drought, salinity, heat, and cold stress prevailing in natural habitats affect plant growth and development. Different species modify their structural and functional traits to combat these abiotic stresses while growing in stressful environments. Cenchrus species, i.e., Cenchrus pennisetiformis, C. setiger, and C. prieurii are widely distributed grasses found growing all over the world. Samples from natural populations were collected from different ecological regions in the Punjab and Khyber Pakhtoonkhwa that were exposed to aridity, salinity, and cold, while one site was designated as normal control. In the present study, structural and functional modifications of three Cenchrus species under abiotic stresses were evaluated. It was expected that each Cenchrus species may evolve different strategies to cope with multiple abiotic stresses. All Cenchrus species responded differently whether growing in normal environment or stressful conditions. The most remarkable feature for survival in C. pennisetiformis under cold stress was increased inflorescence and increased stem and root lignification. C. prieurii showed better tolerance to saline and cold environments. C. setiger showed better development of leaf sheath anatomical traits. The structural and functional modifications in Cenchrus species such as development of mechanical tissues provided structural support, while dermal and parenchymatous tissues increased water storage capacity and minimized water loss. An increase in the concentration of organic osmolytes and ionic content aids turgor pressure maintenance and ionic content crucial for plant growth and development. It was concluded that structural and functional alterations in all Cenchrus species were very specific and critical for survival under different environmental stresses. The ecological fitness of these species relied on maintenance of growth and biomass production, and the development of mechanical, vascular, dermal and parenchyma tissues under stressful environmental conditions. Moreover, accumulation of beneficial ions (K+ and Ca2+) and organic osmolytes were critical in turgor maintenance, hence survival of Cenchrus spp.

Phytoremediation through microstructural and functional alterations in alkali weed (Cressa cretica L.) in the hyperarid saline desert.

Salt excretory halophytes are the major sources of phytoremediation of salt-affected soils. Cressa cretica is a widely distributed halophyte in hypersaline lands in the Cholistan Desert. Therefore, identification of key physio-anatomical traits related to phytoremediation in differently adapted C. cretica populations was focused on. Four naturally adapted ecotypes of non-succulent halophyte Cressa cretica L. form hyper-arid and saline desert Cholistan. The selected ecotypes were: Derawar Fort (DWF, ECe 20.8 dS m-1) from least saline site, Traway Wala Toba (TWT, ECe 33.2 dS m-1) and Bailah Wala Dahar (BWD, ECe 45.4 dS m-1) ecotypes were from moderately saline sites, and Pati Sir (PAS, ECe 52.4 dS m-1) was collected from the highly saline site. The natural population of this species was collected and carefully brought to the laboratory for different structural and functional traits. As a result of high salinity, Na+, Cl-, K+, and Ca2+ content significantly increased at root and shoot level. At root level, some distinctive modifications such as increased sclerification in vascular bundles, enlarged vascular bundles, metaxylem vessels, phloem region, and storage parenchyma (cortex) are pivotal for water storage under extreme arid and osmotic condition. At the stem level, enhanced sclerification in outer cortex and vascular bundles, stem cellular area, cortical proportion, metaxylem and phloem area, and at the leaf level, very prominent structural adaptations were thicker and smaller leaves with increased density of salt glands and trichomes at surface, few and large stomata, reduced cortical and mesophyll parenchyma, and narrow xylem vessels and phloem area represent their non-succulent nature. The ecotype collected from hypersaline environments was better adapted regarding growth traits, ion uptake and excretion, succulence, and phytoremediation traits. More importantly, structural and functional traits such as root length and biomass, accumulation of toxic ions along with K+ in root and shoot, accumulation of Ca2+ in shoot and Mg2+ in root, excretion of toxic ions were the highest in this ecotype. In conclusion, all these alterations strongly favor water conservation, which certainly contributes to ecotypes survival under salt-induced physiological drought.

Naturally adapted salt tolerant plants provide exceptional material for exploring adaptive mechanisms they use to confront high salt concentrations. Cressa cretica is a hypersaline hyperarid desert colonizer, which was previously underexplored. In the present study, we focused on the new insight on relationship among anatomical modifications, salt accumulation and excretion and phytoremediation potential of this rare species.

Preparation of nanoformulation of 5-fluorouracil to improve anticancer efficacy: integrated spectroscopic, docking, and MD simulation approaches.

Nanoformulations (NFs) can be used as a novel drug delivery system to treat all cancer types. One of the major drawbacks of conventional anticancer drugs is that they have poor specificity and higher toxicity towards normal cells. 5-fluorouracil (5-FU) is a well-studied anticancer drug that has a significant role in various cancers, specifically colorectal cancer therapy. This study was performed to determine the functional groups, particle size, surface charge, heterogeneity, and stability of the NF. The NFs of 5-FU were prepared through the ultrasonication technique by increasing the surfactant (Tween-80) concentrations. Among all three NFs, nanoformulated 5-FU (n5-FU) showed the most effective particle size (10.72 nm) with a zeta potential of (-4.57 mV). The cytotoxicity and apoptosis profiles confirmed that n5-FU enhanced the anticancer effect of the pure drug in HCT-116 cells, as evident from MTT assay, fluorescence microscopy, and FACS analysis. In HCT-116 cells, the IC50 values of pure and n5-FU were obtained as 41.3 µM and 18.8 µM, respectively, indicating that n5-FU was more effective against the cancer cell line. The cellular uptake study was performed to check the intake of NF in cancer cells. However, the microtubule-affinity regulating kinase-4 (MARK-4), a cancer-target protein, was purified to study the inhibition and interaction studies. The inhibition assay confirmed the inhibitory potential of 5-FU against MARK-4 protein. the multi-spectroscopic, molecular docking and MD simulation studies were performed to analyse the conformational changes, binding studies, intermolecular interactions, and stability of MARK-4 protein upon binding 5-FU. This demonstrates that NF can enhance the effectiveness of anticancer drugs.Communicated by Ramaswamy H. Sarma.

Foliar architecture differentially restrains metal sequestration capacity in wheat grains (Triticum aestivum L.) grown in hyper-chloride-contaminated soils.

Anthropogenic activities, such as industrial wastewater and use of water softeners, cause hyper-accumulation of Cl- in water sources and soils. Currently, industries have no sustainable method to remove these Cl- ions from wastewater. This study was conducted to evaluate the integrative responses of wheat cultivated in five industrial effluent-affected areas (S2-S6) by investigating soil characters and bioaccumulation of metals in wheat plants and grains. The S4 site (near the second chloride outlet) exhibited a higher concentration of CO2, SO2, NO2, Cl-, Cd, Mn, Ni, Cr, and Zn. Soil from S6 (sewage wastewater downstream getting mixed with chloride-contaminated water) had a minimum level of nutrients (Na, K, and Ca), maximum metals (Cd, Fe, Pb, Mn), and reduction in plant biomass. In site S2 (sewage wastewater upstream of the chloride factory), a higher level of minerals and metals was noted in the roots. Maximum metals in grains occurred in S6 with higher organic osmolytes. The sequestration capacity of metals in leaves was also increased by alterations in anatomical traits. Results indicated that metals and hyper-Cl- concentration employed a negative influence on the plants because of poor soil quality, extremely damaged microstructures leading to reduced yield, poor grain quality, and excessive translocation from roots to wheat grains. These findings revealed that contaminated plants used as either green forage or hay are noxious to animals and if used as grain for feed or humans can lead to serious health hazards.

Naringin binds to protein disulfide isomerase to inhibit its activity and modulate the blood coagulation rates: Implications in controlling thrombosis.

Currently used antithrombotic drugs are beset with several drawbacks which necessitates the need for new and cheaper alternatives. Protein disulfide isomerase (PDI) is secreted in the blood plasma in cellular stress conditions and initiates the thrombus formation. A screening of library of natural compounds revealed that naringin had a high binding affinity for the PDI (-8.2 kcal/mol). Recombinant PDI was purified using the affinity chromatography. Incubation of purified PDI (3 µM) with naringin (0-100 µM, pH 7.4, 25 °C) partially modulated its conformation. Consequently, the fluorescence emission spectra of the PDI binding to naringin were assessed using the Stern-Volmer equation, which indicated an association constant of 2.78 × 104 M-1 suggesting an appreciable affinity for the naringin, with a unique binding site. An insulin turbidity assay showed that PDI activity is decreased in the presence of naringin indicating inhibition. Molecular dynamic simulation studies showed the changes in the PDI structure on binding to the naringin. Incubation of naringin (80 µM) in fresh human plasma along with exogenous PDI (175 nM) showed a significant delay in the intrinsic and extrinsic coagulation pathways. We show that naringin is able to modulate the PDI conformation and activity resulting in altered blood coagulation rates.

Radical-mediated sulfonylative/thiolative cyclization of biaryl enones to phenanthrone derivatives.

An approach for the assembly of phenanthrone derivatives bearing all carbon quaternary centres has been developed through visible light-promoted tandem sulfonylation/intramolecular-arylation of biaryl enones with sulfonyl chlorides. A series of sulfonylated 10,10-dialkylphenanthrones were obtained in good yields. In addition, the approach has been extended to thiotrifluoromethyl (SCF3) and thiocyanato (SCN) radicals to obtain the corresponding phenanthrones under oxidative conditions. The synthetic utility was also illustrated by the scalability and further transformations of the product.

Genetic basis of ß-thalassemia in families of pashtun ethnicity in Dera Ismail Khan district of Khyber Pakhtun-Khwa province, Pakistan.

OBJECTIVES: The objective of current genetic research was to verify the genetic basis of ß-thalassemia and its pattern of inheritance in families of Pashtun ethnicity in District Dera Ismail Khan, Pakistan. METHODOLOGY: Blood samples from clinically diagnosed five unrelated ß-thalassemia families were collected and target Sanger Sequencing of HBB gene was done. Moreover, in silico analysis including protein modeling and Protein-Protein docking was aslo performed. RESULTS AND DISCUSSION: Clinical analysis of patients from family 1,2, 4, and 5 revealed Thalassemia Intermedia, while patient from family 3 was suffering from thalassemia major. The average Hb concentrations between the cases that were severe were found to be a little lower (6.3 mg/dl) than the patients with milder clinical manifestations (7.6 ± 1.4). Genetic analysis in family 1 identified compound heterozygous mutation of HBB (NM_000518) i.e. c.20A>T +c.92 G>A, in family 2 and 4 compound heterozygous mutations c.20A>T + c.27_28insG, in family 3 homozygous mutation c.27_28insG, while in family 5 we identified homozygous mutation c.92 + 5 G>C (IVS-1 + 5 G>C). CONCLUSION: This study offers an effective incentive to establish a mutation detection as well as prenatal diagnosis (PND) centers at a larger scale in the Pashtun ethnicity residing in District Dera Ismail Khan, Pakistan.

Comparative evaluation of treatment outcome of partial pulpotomy using different agents in permanent teeth-a randomized controlled trial.

AIM: To compare and evaluate the clinical and radiographic performance, post-operative pain, and anti-inflammatory intake after partial pulpotomy (PP) with calcium hydroxide (CH), mineral trioxide aggregate (MTA), Biodentine (BD), and Emdogain (EMD) as pulp capping agents in mature permanent molars with definitive diagnosis of reversible pulpitis. MATERIALS AND METHODS: As part of this prospective, randomized clinical trial with four parallel arms (CTRI Registration No.: CTRI/2020/11/029329 dated 24/11/2020), hundred and ten permanent molars with a clinical diagnosis of reversible pulpitis and normal apical tissues, from patients between the ages of 15 and 45 years, were recruited and randomly assigned to four groups-CH, MTA, BD, and EMD. Operative procedure was performed under local anesthesia and dental dam isolation. After carious pulpal exposure, 2 mm of superficially inflamed coronal pulp tissue was amputated and either of the four pulp capping materials was placed. The outcome assessment was carried out at 1, 3, 6, and 12 month(s) and was categorized as success (asymptomatic patients with PAI score = 1) or failure (symptomatic patients or PAI score > 1). RESULTS: There was a significant difference in post-operative pain and anti-inflammatory medication intake after partial pulpotomy with Emdogain vis-à-vis other three capping agents. No difference in both clinical and radiographic performances was observed among the four capping agents. CONCLUSION: Partial pulpotomy when performed following evidence-based guidelines results in high success rates regardless of capping agent employed. EMD can be considered a valid and suitable pulp capping agent in PP. CLINICAL RELEVANCE: Meticulous examination and removal of superficially inflamed pulp under magnification and complete asepsis lead to successful pulpal healing regardless of capping agent employed.

Morpho-anatomical determinants of yield potential in Olea europaea L. cultivars belonging to diversified origin grown in semi-arid environments.

Plant performance is mainly estimated based on plant architecture, leaf features and internal microstructural changes. Olive (Olea europaea L.) is a drought tolerant, oil yielding, and medium sized woody tree that shows specific structural and functional modifications under changing environment. This study was aimed to know the microstructural alteration involving in growth and yield responses of different Olive cultivars. Eleven cultivars were collected all over the world and were planted at Olive germplasm unit, Barani Agricultural Research Institute, Chakwal (Punjab) Pakistan, during September to November 2017. Plant material was collected to correlate morpho-anatomical traits with yield contributing characteristics. Overall, the studied morphological characters, yield and yield parameters, and root, stem and leaf anatomical features varied highly significantly in all olive cultivars. The most promising cultivar regarding yield was Erlik, in which plant height seed weight and root anatomical characteristics, i.e., epidermal thickness and phloem thickness, stem features like collenchymatous thickness, phloem thickness and metaxylem vessel diameter, and leaf traits like midrib thickness, palisade cell thickness a phloem thickness were the maximum. The second best Hamdi showed the maximum plant height, fruit length, weight and diameter and seed length and weight. It also showed maximum stem phloem thickness, midrib and lamina thicknesses, palisade cell thickness. Fruit yield in the studied olive cultivars can be more closely linked to high proportion of storage parenchyma, broader xylem vessels and phloem proportion, dermal tissue, and high proportion of collenchyma.

Phytoremediation potential modulated by structural and functional traits in a saline desert halophyte Fagonia indica Burm. f.

Using halophytes for phytoremediation is an environmentally friendly technique, now gaining importance all over the world. Fagonia indica Burm. f. (Indian Fagonia) is primarily distributed in salt-affected lands of the Cholistan Desert and surrounding habitats. Four populations with three replications from salt-affected habitats were collected from natural habitats to evaluate structural and functional adaptation for salinity tolerance and phytoremediation of hypersaline habitats. The populations collected from the highest saline sites Pati Sir (PS) and Ladam Sir (LS) had restricted growth habit, increased accumulation of K+ and Ca2+ along Na+ and Cl-, more excretion of Na+ and Cl-, increased cross-sectional area of root and stem, larger exodermal and endodermal cells in roots, and broad metaxylem area. Sclerification in stem was high in population. Specific modifications in leaves were reduced stomatal area and increased adaxial epidermal cell area. Important traits associated with phytoremediation potential of F. indica populations (Pati Sir and Ladam Sir) were deeper roots and taller plants, increased density of salt glands on leaf surface, and high excretion of Na+. Additionally, higher bio-concentration factor, translocation factor, and dilution factor for Na and Cl- in same Ladam Sir and Pati Sir population were identified as key phytoremediation attributes. The plants of F. indica colonizing high salinities (Pati Sir and Ladam Sir) were, therefore, more efficient in phytoremediation of saline soils as these populations accumulated and/or excrete toxic salts. Density of salt glands remarkably increased in the Pati Sir population collected from the highest salinity. This population accumulated and excreted the highest amount of Na+ and Cl-. The dilution factor of Na+ and Cl- ions was also the highest in this population. Anatomical modifications such as root and stem cross-sectional areas, proportion of storage parenchyma, and broad metaxylem vessels were the maximum in Pati Sir population. These modifications indicate not only better salt tolerance of the Pati Sir population but also better in accumulation and excretion of toxic salts. This population can potentially rehabilitate hypersaline uncultivated lands through green reclamation.

In vitro and in silico binding studies of phytochemical isochroman with calf thymus DNA using multi-spectroscopic and computational modelling techniques.

A wide range of therapeutic molecules uses deoxyribonucleic acid (DNA) as an intracellular target. The interaction of small molecules to DNA is a key feature in pharmacology and plays a vital role in the development of novel and more efficient drugs with increased selective activity and enhanced therapeutic effectiveness. Isochroman (IC) is a constituent of Olea europea plant, which has been shown to exhibit several beneficial pharmacological activities. At present, its interaction studies using calf thymus DNA (ct-DNA) have not been explained. A set of multi-spectroscopic techniques has been performed to determine the interaction mechanism of isochroman with ct-DNA. Absorption spectra and quenching in fluorescence studies show that isochroman and ct-DNA form a complex. The static mode of quenching was determined by the Stern-Volmer plot. The value of binding constant, Kb = 4.0 × 103 M-1 revealed moderate type of binding. Effects of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) and ionic strength were studied to examine the isochroman binding to ct-DNA. Potassium iodide (KI) quenching effects and competitive binding studies clearly showed that isochroman binds in the minor groove of ct-DNA. Circular dichroic and DNA melting experiments also confirmed these results. The experimental outputs were further corroborated via in silico computational modelling studies. Lipinski's rule of 5 and SwissADME showed drug-likeness and oral bioavailability scores. Protox ІІ online software predicts oral and organ toxicity.Communicated by Ramaswamy H. Sarma.

Structural determinants of phytoremediation capacity in saltmarsh halophyte Diplachne fusca (L.) P. Beauv. ex Roem. & Schult. subsp. fusca.

Micro and macro-morphological features contribute to plants' tolerance to a variety of environmental pollutants. The contribution of such structural modifications in the phytoremediation potential of Diplachne fusca populations collected from five saline habitats were explored when treated with 100 to 400 mM NaCl for 75 days along with control. Structural modifications in the populations from the highest salinity included development of aerenchyma in stem instead of chlorenchyma, absence of excretory hairs in stem, and exceptionally large trichomes on the leaf surface to help excretion of excess salt. Large parenchyma cells provided more space for water and solute storage, while broad metaxylem vessels were linked to better conduction water and nutrients, which ultimately excreted via glandular hairs, microhairs, and vesicular hairs. Broad metaxylem vessels and exceptionally long hairs observed in the populations collected from 52 dS m-1. In conclusion, large stem aerenchyma, exceptionally large trichomes on the leaf surface, and tightly packed outer cortical region in roots with intensive sclerification just inside the epidermis accompanied with salt excretion via glandular hairs, microhairs, and vesicular hairs were the main anatomical modifications involved in the phytoremediation potential of D. fusca in hyper-saline environments.

Morpho-anatomical characteristics of the differently adapted populations of Diplachne fusca has never been reported. In particular, structural variation in their mechanism of adaptation for salinity tolerance was investigated for the first time in current study.

Mannose 2, 3, 4, 5, 6-O-pentasulfate (MPS): a partial activator of human heparin cofactor II with anticoagulation potential.

Thromboembolic diseases are a major cause of mortality in human and the currently available anticoagulants are associated with various drawbacks, therefore the search for anticoagulants that have better safety profile is highly desirable. Compounds that are part of the dietary routine can be modified to possibly increase their anticoagulant potential. We show mannose 2,3,4,5,6-O-pentasulfate (MPS) as a synthetically modified form of mannose that has appreciable anticoagulation properties. An in silico study identified that mannose in sulfated form can bind effectively to the heparin-binding site of antithrombin (ATIII) and heparin cofactor II (HCII). Mannose was sulfated using a simple sulfation strategy-involving triethylamine-sulfur trioxide adduct. HCII and ATIII were purified from human plasma and the binding analysis using fluorometer and isothermal calorimetry showed that MPS binds at a unique site. A thrombin inhibition analysis using the chromogenic substrate showed that MPS partially enhances the activity of HCII. Further an assessment of in vitro blood coagulation assays using human plasma showed that the activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged in the presence of MPS. A molecular dynamics simulation analysis of the HCII-MPS complex showed fluctuations in a N-terminal loop and the cofactor binding site of HCII. The results indicate that MPS is a promising lead due to its effect on the in vitro coagulation rate.Communicated by Ramaswamy H. Sarma.

Strand 1A variant in neuroserpin shows increased aggregation and no loss of inhibition: implication in ameliorating polymerization to retain activity.

Neuroserpin (NS) is predominantly expressed in the brain and is the primary inhibitor of tissue plasminogen activator (tPA). NS variants are associated with the neurogenerative disease termed familial encephalopathy with neuroserpin inclusion bodies (FENIB). The disease is characterized by variable age of onset and severity. The reactive center loop (RCL) insertion-based inhibitory mechanism of NS requires a coordinated conformational change leading to a shift in the strands of the ß-sheet A and movement of helix F. Strand 1A is connected to the helix F at its C terminal end and with the strand 2A at its N terminal, both these domain move for accommodating the inserting loop; therefore, a variant that influences their movement may alter the inhibition rates. A molecular dynamic simulation analysis of a H138C NS variant from strand 1A showed a large decrease in conformational fluctuations as compared with wild-type NS. H138 was mutated, expressed, purified and a native-PAGE and transmission electron microscopy (TEM) analysis showed that this variant forms large molecular weight aggregates on a slight increase in temperature. However, a circular dichroism analysis showed its secondary structure to be largely conserved. Surprisingly, its tPA inhibition activity and complex formation remain unhindered even after the site-specific labeling of H138C with Alexa fluor C5 maleimide. Further, a helix F-strand 1A (W154C-H138C) double variant still shows appreciable inhibitory activity. Increasingly, it appears that aggregation and not loss of inhibition is the more likely cause of shutter region-based variants phenotypes, indicating that hindering polymer formation using small molecules may retain inhibitory activity in pathological variants of NS.

Nanogels as Potential Delivery Vehicles in Improving the Therapeutic Efficacy of Phytopharmaceuticals.

Nanogel is a promising drug delivery approach to improve the pharmacokinetics and pharmacodynamic prospect of phytopharmaceuticals. In the present review, phytopharmaceuticals with astonishing therapeutic utilities are being explored. However, their in vivo delivery is challenging, owing to poor biopharmaceutical attributes that impact their drug release profile, skin penetration, and the reach of optimal therapeutic concentrations to the target site. Nanogel and its advanced version in the form of nanoemulgel (oil-in-water nanoemulsion integrated gel matrix) offer better therapeutic prospects than other conventional counterparts for improving the biopharmaceutical attributes and thus therapeutic efficacy of phytopharmaceuticals. Nanoemulgel-loaded phytopharmaceuticals could substantially improve permeation behavior across skin barriers, subsequently enhancing the delivery and therapeutic effectiveness of the bioactive compound. Furthermore, the thixotropic characteristics of polymeric hydrogel utilized in the fabrication of nanogel/nanoemulgel-based drug delivery systems have also imparted improvements in the biopharmaceutical attributes of loaded phytopharmaceuticals. This formulation approach is about to be rife in the coming decades. Thus, the current review throws light on the recent studies demonstrating the role of nanogels in enhancing the delivery of bioactive compounds for treating various disease conditions and the challenges faced in their clinical translation.

Identification and validation of two alternatively spliced novel isoforms of human α-1-antichymotrypsin.

α-1-antichymotrypsin (ACT) is a serine proteinase inhibitor that controls the activity of proteases like chymotrypsin, cathepsin G and mast cell chymase. Familial variants of ACT results in liver and lung diseases, but it is also reported to be associated with several other disease conditions. ACT is mainly synthesized in the liver using four coding exons, namely E1, E2, E3 and E4 encoding a 423 amino acid protein that also includes a 23 amino acid signal peptide. It is found to be associated with amyloid plaques and is elevated during inflammatory response and modulates cytokine based signal transduction pathways, independent of its anti-protease activity. Therefore, the multispecificity of ACT and its non-inhibitory roles in diseased conditions warrants an assessment of possible existence of the other isoforms. Consequently, scanning of introns, 5' and 3' region of the ACT gene using computational tools like FGENESH and FEX did indicate the presence of coding regions. Using a combined approach of bioinformatics and molecular biology, we have found one novel exon located in the intronic region between exons E1 and E2, that splices with exon E2 and replaces N-terminal exon E1, generating an ACT isoform with a novel 151 base pair N-terminus. This isoform was found to lack the signal sequence and is smaller in size but its reactive centre loop remains intact. A truncated transcript was also confirmed with an extension of the E3 by a 12 nucleotide intronic region including a stop codon. Modelling studies show that due to removal of E4 this isoform lacks the RCL. Novel isoform ACT-N lacks E1 but has a conserved RCL. However, due to loss of strands of ß-sheet A, it may also be inactive, but with ability to bind the target proteases. The novel truncated ACT-T isoform lacks the RCL and may have a non-inhibitory role. These hypothesis will need further work for functional validation.