Role of Macrophage PIST Protein in Regulating Leishmania major Infection.

PDZ protein interacting specifically with Tc10 or PIST is a mammalian trans-Golgi resident protein that regulates subcellular sorting of plasma membrane receptors. PIST has recently emerged as a key player in regulating viral pathogenesis. Nevertheless, the involvement of PIST in parasitic infections remains unexplored. Leishmania parasites infiltrate their host macrophage cells through phagocytosis, where they subsequently multiply within the parasitophorous vacuole (PV). Host cell autophagy has been found to be important in regulating this parasite infection. Since PIST plays a pivotal role in triggering autophagy through the Beclin 1-PI3KC3 pathway, it becomes interesting to identify the status of PIST during Leishmania infection. We found that while macrophage cells are infected with Leishmania major (L. major), the expression of PIST protein remains unaltered; however, it traffics from the Golgi compartment to PV. Further, we identified that in L. major-infected macrophage cells, PIST associates with the autophagy regulatory protein Beclin 1 within the PVs; however, PIST does not interact with LC3. Reduction in PIST protein through siRNA silencing significantly increased parasite burden, whereas overexpression of PIST in macrophages restricted L. major infectivity. Together, our study reports that the macrophage PIST protein is essential in regulating L. major infectivity.

Cyclobutane based "overbred intermediates" and their exploration in organic synthesis.

"Overbred intermediates" have been used in organic synthesis for a long time, but these intermediates are not categorized as such. This name was given recently in Hoffman's book Elements of Synthesis Planning. These intermediates are very useful to synthesize complex carbocyclic molecules. A number of powerful and efficient strategies have been developed by using overbred intermediates through innovative fragmentative transformations. This review is based on four-member overbred intermediates (cyclobutane based) that have been used in the total synthesis of natural products from 1968 to 2020. In the initial part, we have discussed synthetic methods (photochemical, metal-mediated, and other miscellaneous transformations) for the construction of cyclobutane overbred intermediates. In the later section, we have discussed how the overbred skeleton is cleaved through numerous fragmentation methods to access the desired target structure.

Feature Selection and Validation of a Machine Learning-Based Lower Limb Risk Assessment Tool: A Feasibility Study.

Early and self-identification of locomotive degradation facilitates us with awareness and motivation to prevent further deterioration. We propose the usage of nine squat and four one-leg standing exercise features as input parameters to Machine Learning (ML) classifiers in order to perform lower limb skill assessment. The significance of this approach is that it does not demand manpower and infrastructure, unlike traditional methods. We base the output layer of the classifiers on the Short Test Battery Locomotive Syndrome (STBLS) test used to detect Locomotive Syndrome (LS) approved by the Japanese Orthopedic Association (JOA). We obtained three assessment scores by using this test, namely sit-stand, 2-stride, and Geriatric Locomotive Function Scale (GLFS-25). We tested two ML methods, namely an Artificial Neural Network (ANN) comprised of two hidden layers with six nodes per layer configured with Rectified-Linear-Unit (ReLU) activation function and a Random Forest (RF) regressor with number of estimators varied from 5 to 100. We could predict the stand-up and 2-stride scores of the STBLS test with correlation of 0.59 and 0.76 between the real and predicted data, respectively, by using the ANN. The best accuracies (R-squared values) obtained through the RF regressor were 0.86, 0.79, and 0.73 for stand-up, 2-stride, and GLFS-25 scores, respectively.

Lead phytoremediation potentials of four aquatic macrophytes under hydroponic cultivation.

Lead (Pb) is a major toxicological concern of the present day that demands immediate attention. The use of aquatic macrophytes with high Pb tolerance and accumulation may be a very convenient and economically viable solution for remediating Pb. We examined the ability of Salvinia cucullata, Alternanthera sessilis, Lemna minor, and Pistia stratiotes to remove 0.12 mM, 0.24 mM, 0.36 mM, and 0.48 mM Pb for 96-h under hydroponic cultivation system. The plants accumulated variable amounts of Pb: S. cucullata > A. sessilis > P. stratiotes > L. minor, with low mobility of Pb from root to shoot. Lead uptake kinetics were monitored up to 96-h. After 96-h, the uptake efficiency for S. cucullata (98-99%), A. sessilis (79-96%), L. minor (45-79%), and P. stratiotes (40-76%) was noted. For S. cucullata and A. sessilis, an extremely high uptake rate was seen within the initial 24-h of trials, followed by slower uptake till 96-h. P. stratiotes and L. minor worked best at 0.12 mM Pb. Pb-Phytotoxicity became prominent at 0.48 mM exposure with biomass loss and morphological changes. The plants had a quick growth rate, extensive root system, high biomass yield, and the ability to tolerate and accumulate Pb that made them suitable for phytoremediation purposes. NOVELTY STATEMENT: Lead phytoremediation potential of four aquatic macrophytes found in Indian waters was evaluated. These macrophytes, often considered as weeds, could be used for phytoremediation purposes that would turn out to be a sustainable means of the utilization of natural resources in developing countries like India. In this study, not only metal accumulation by plants but also the lead uptake kinetics at several time intervals and valuable growth attributes were estimated to establish the suitability of these plants as probable lead phytoremediators. Two of the plant species, Salvinia cucullata, and Alternanthera sessilis, showed excellent Pb accumulation capacities that had not been reported earlier, to the best of our knowledge. The work is all the more significant as there have been needs for identifying Pb-phytoremediators well suited to native climate and growth conditions that could take up large amounts of metal from the substratum.

Vitiligo and Psychiatric Morbidity: A Profile from a Vitiligo Clinic of a Rural-based Tertiary Care Center of Eastern India.

BACKGROUND: Vitiligo is an idiopathic acquired progressive de/hypopigmentary disorder of skin and mucosae. In Indian skin depigmentaion is very much obvious and can cause psychological distress, low self esteem and social stigmatization. AIMS: The primary objective of this study was to evaluate the psychiatric morbidity in vitiligo patients and secondary objective was to assess the morbidity in all eight dimensions of psychosocial and physical aspects, i.e. cognitive, social, discomfort, limitations, depression, fear, embarrassment and anger. MATERIALS AND METHODS: An institution based case-control study with sixty-one patients of vitiligo and equal number of healthy age and sex matched controls was undertaken. The self-reporting questionnaire-24 (SRQ-24) and skindex (A 61-item survey questionnaire) were used to assess the psychiatric morbidity in both the groups. RESULTS: The SRQ-assessed psychiatric morbidity in the study group was 63.93%, compared with 24.59% in the control group (P<0.0001). Acral vitiligo had maximum association with psychiatric morbidity (86.67%) followed by vitiligo vulgaris (68%), mucosal vitiligo (62.5%) and others. According to the skindex, the most common psychiatric morbidity in vitiligo patients was depression (62.29%) followed by embarrassment (55.73%), social problem (54.09%), cognitive impairment (50.81%), physical limitation (47.54%), discomfort (40.98%), anger (36.06%) and fear (24.59%). The difference in Skindex scoring that marked the psychiatric morbidity among the case and control groups was statistically significant for depression, discomfort, social problem, cognitive impairment, embarrassment (P<0.0001) and physical limitation (P=0.0044). CONCLUSION: Vitiligo has a high degree of psychiatric morbidity.

Non diaphanous formin delphilin acts as a barbed end capping protein.

Formins are multi domain proteins present ubiquitously in all eukaryotes from lower fungi to higher vertebrates. Formins are characterized by the presence of formin homology domain-2 (FH2) and formin homology domain-1 (FH1). There are fifteen different formins present in mouse and human. Among these metazoan formins, Delphilin is a unique formin having two PDZ domains at the N-terminus and FH1, FH2 domain at the C-terminus respectively. In this study we observed that Delphilin binds to actin filaments, and Delphilin inhibits actin filament elongation like barbed end capping protein CapZ. In vitro, Delphilin stabilized actin filaments by inhibiting actin filament depolymerisation. Therefore, our study demonstrates Delphilin as an actin-filament capping protein.

Orientation of tyrosine side chain in neurotoxic Aß differs in two different secondary structures of the peptide.

Amyloid ß (Aß) peptide is present as a major component in amyloid plaque that is one of the hallmarks of Alzheimer's disease. The peptide contains a single tyrosine residue and Aß has a major implication in the pathology of the disease progression. Current investigation revealed that the tyrosine side chain attained two different critical stereo orientations in two dissimilar conformational states of the peptide. The extended α-helical structure of the peptide observed in an apolar solvent or methanol/water mixture became disordered in aqueous medium and the radius of gyration decreased. In aqueous medium, the torsional angle around Cα-Cß of tyrosine group became -60°. However, in its α-helical conformation in an apolar system, the measured angle was 180° and this rotameric state may be reasoned behind stronger tyrosine fluorescence compared with the disordered state of the peptide. Molecular dynamics simulation analyses and spectroscopic studies have helped us to understand the major structural changes in the secondary structure of the peptide in the two conformational states. A conformational clustering indicated that the compact state is more stable with tyrosine residue attaining the torsion angle value of -60°, whereas the native state (in HFIP/water mixture) is prevalent at a torsion angle value of -180°. High solvent accessibility has possibly stabilized the particular rotameric state (-60°) of the tyrosine residue and could be the reason behind decrease in fluorescence of the sole tyrosine residue in an aqueous buffer solution (pH 7.4) compared with its fluorescence in the α-helical structure in the micellar environment.

Comparison of dual-antiplatelet therapy durations after endovascular revascularization of infrainguinal arteries.

BACKGROUND: The optimal dual-antiplatelet therapy (DAPT) duration after endovascular revascularization of infrainguinal arteries is uncertain. METHODS: This study examines DAPT prescription trends and 12-month major adverse limb events (MALEs; a composite of repeat endovascular or surgical revascularization, acute vessel thrombosis, or amputation of the target limb), major adverse cardiovascular events (MACEs; all-cause mortality, nonfatal myocardial infarction [MI], stroke, or coronary revascularization), fatal bleeding events, and those requiring interruption or discontinuation of DAPT (hemorrhagic complications) for patients enrolled into the Excellence in Peripheral Artery Disease (XLPAD) registry. RESULTS: Data on 368 patients prescribed antiplatelet therapy were analyzed; 8.2% were prescribed antiplatelet monotherapy, 48.6% DAPT for ≤3 months, and 43.2% for >3 months. Patients in the >3 DAPT prescribed group were older, had preexisting coronary artery disease (CAD), and prior MI (all P < 0.001). Overall MALE in the ≤3 and >3-month DAPT prescribed groups were 22.3% and 23.9%, respectively (P = 0.541). Survival analysis showed significantly higher rates of MACE in patients prescribed >3-month DAPT (17.6% vs. 9.5%; P = 0.019). An "as-treated" analysis excluded 10 patients who were prescribed DAPT for >3 months and revealed similar rates of MALE (24.9% vs. 20.8%; P = 0.386) and MACE (12.2% vs. 14.8%; P = 0.443) in patients receiving ≤3 and >3 DAPT. Hemorrhagic complications were similar across all prescribed and "as-treated" DAPT groups. CONCLUSIONS: After infrainguinal endovascular procedures, patients with underlying CAD were prescribed longer (>3 months) duration of DAPT and experienced more cardiovascular events compared with those prescribed ≤3 months of DAPT. Adverse limb events were similar in both groups.

Sequence complexity of amyloidogenic regions in intrinsically disordered human proteins.

An amyloidogenic region (AR) in a protein sequence plays a significant role in protein aggregation and amyloid formation. We have investigated the sequence complexity of AR that is present in intrinsically disordered human proteins. More than 80% human proteins in the disordered protein databases (DisProt+IDEAL) contained one or more ARs. With decrease of protein disorder, AR content in the protein sequence was decreased. A probability density distribution analysis and discrete analysis of AR sequences showed that ∼8% residue in a protein sequence was in AR and the region was in average 8 residues long. The residues in the AR were high in sequence complexity and it seldom overlapped with low complexity regions (LCR), which was largely abundant in disorder proteins. The sequences in the AR showed mixed conformational adaptability towards α-helix, ß-sheet/strand and coil conformations.

A novel antimicrobial peptide derived from modified N-terminal domain of bovine lactoferrin: design, synthesis, activity against multidrug-resistant bacteria and Candida.

Lactoferrin (LF) is believed to contribute to the host's defense against microbial infections. This work focuses on the antibacterial and antifungal activities of a designed peptide, L10 (WFRKQLKW) by modifying the first eight N-terminal residues of bovine LF by selective homologous substitution of amino acids on the basis of hydrophobicity, L10 has shown potent antibacterial and antifungal properties against clinically isolated extended spectrum beta lactamases (ESBL), producing gram-negative bacteria as well as Candida strains with minimal inhibitory concentrations (MIC) ranging from 1 to 8 µg/mL and 6.5 µg/mL, respectively. The peptide was found to be least hemolytic at a concentration of 800 µg/mL. Interaction with lipopolysaccharide (LPS) and lipid A (LA) suggests that the peptide targets the membrane of gram-negative bacteria. The membrane interactive nature of the peptide, both antibacterial and antifungal, was further confirmed by visual observations employing electron microscopy. Further analyses, by means of propidium iodide based flow cytometry, also supported the membrane permeabilization of Candida cells. The peptide was also found to possess anti-inflammatory properties, by virtue of its ability to inhibit cyclooxygenase-2 (COX-2). L10 therefore emerges as a potential therapeutic remedial solution for infections caused by ESBL positive, gram-negative bacteria and multidrug-resistant (MDR) fungal strains, on account of its multifunctional activities. This study may open up new approach to develop and design novel antimicrobials.

Isolation and characterization of novel protein with anti-fungal and anti-inflammatory properties from Aloe vera leaf gel.

The Aloe protein of 14 kDa from the Aloe vera leaf gel was isolated by an ion exchange chromatography using DEAE-cellulose and CM-cellulose column. The purified Aloe protein exhibited a potent anti-fungal activity against Candida paraprilosis, Candida krusei and Candida albicans. In addition, the purified Aloe protein also showed an anti-inflammatory property against pure lipoxygenase and cyclooxygenase-2 with 84% and 73% inhibition, respectively, and was verified by binding with these proteins by real time method by the phenomenon of surface plasmon resonance. This Aloe protein is a novel protein possessing antifungal and anti-inflammatory properties and thus sets a platform to be used as a medicinal plant product.