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Abstract Overexpression of anti-apoptotic BCL-2 family members is a hallmark of many lymphoid malignancies, including chronic lymphocytic leukemia (CLL) and non-Hodgkin lymphoma (NHL) that can be targeted with small molecule inhibitors. ABT-199 is a rationally designed BCL-2 homology (BH)-3 mimetic that specifically binds to BCL-2, but not to MCL-1 and BCL-xL. Although the thrombocytopenia that occurs with navitoclax treatment has not been a problem with ABT-199, clinical trials in CLL could benefit by lowering the ABT-199 concentration through targeting other survival pathways. In this study, we investigated the mechanisms of resistance that develops to ABT-199 therapy by generating ABT-199-resistant (ABT199-R) cell lines via chronic exposure of NHL cell lines to ABT-199. Acquired resistance resulted in substantial AKT activation and upregulation of MCL-1 and BCL-xL levels that sequestered BIM. ABT199-R cells exhibited increased MCL-1 stability and failed to activate BAX in response to ABT-199. The ABT-199 acquired and inherent resistant cells were sensitized to treatment with ABT-199 by inhibitors of the PI3K, AKT, and mTOR pathways, NVP-BEZ235 and GS-1101. NVP-BEZ235, a dual inhibitor of p-AKT and mTOR, reduced MCL-1 levels causing BIM release from MCL-1 and BCL-xL, thus leading to cell death by BAX activation. The PI3Kδ inhibitor GS-1101 (idelalisib) downregulated MCL-1 and sensitized ABT199-R cells through AKT-mediated BAX activation. A genetic approach, through siRNA-mediated down-regulation of AKT, MCL-1, and BCL-xL, significantly decreased cell survival, demonstrating the importance of these cell survival factors for ABT-199 resistance. Our findings suggest a novel mechanism that modulates the expression and activity of pro-survival proteins to confer treatment resistance that could be exploited by a rational combination therapeutic regimen that could be effective for treating lymphoid malignancies.

Day-by-day increasing irrigation water scarcity requires the application of water-saving irrigation techniques to sustain agriculture production. A two-year field investigation was conducted during 2018 to 2020 to determine the effects of various mulches and irrigation volumes on the growth, leaf chemicals and soil properties of one-year-old sweet oranges (Citrus sinensis) cv. Mosambi. The study included three irrigation schedules, viz.100% ET c (I 1 ), 80% ET c (I 2 ), and 60% ET c (I 3 ), and five different mulches were used, viz. without mulch, white polythene, coriander straw, dry grass and black polythene mulches, replicated thrice. Results demonstrated that drip irrigation with 100% ET c and mulching with black polythene mulch significantly increase the plant growth attributes like height of the plant (28.64%) (30.31%), rootstock girth (36.61%) (37.90%), plant canopy spread (E-W and N-S) (EW- 63.82%, NS- 63.87%) (EW- 67.56%, NS- 67.90%) and leaf area (2.4%) (2.34%). Furthermore, plant leaf chlorophyll content (2.41 mg g -1 ) (2.41 mg g -1 ) and leaf mineral content such as N (2.39%) (2.40%), P (0.16%) (0.165%), K (1.57%) (1.59%), Ca (47.34 g kg -1 ) (47.80 g kg -1 ), Mg (4.54 g kg -1 ) (4.57 g kg -1 ), Fe (120.51 g kg -1 ) (123.15 g kg -1 ) and Zn (39.00 g kg -1 ) (37.84 g kg -1 ) were noted to be significantly (p ≤ 0.05) higher in plants that received 100% (were ET c (I 1 ) and mulching with black polythene mulch (M 1 ) treatment. Taken together, the results suggested that treatments I 1 and M 1 have the potential to maximize plant growth, leaf chemicals and soil nutrients of sweet orange (Citrus sinensis) cv. Mosambi plants., (© 2024. The Author(s).)

In recent years, there has been an increasing interest in the green synthesis of metallic nanoparticles, mostly because of the evident limitations associated with chemical and physical methods. Green synthesis, commonly referred to as "biogenic synthesis," is seen as an alternative approach to produce AgNPs (silver nanoparticles). The current work focuses on the use of Asterarcys sp. (microalga) for biological reduction of AgNO 3 to produce AgNPs. The optimal parameters for the reduction of AgNPs were determined as molarity of 3 mM for AgNO 3 and an incubation duration of 24 h at pH 9, using a 20:80 ratio of algal extract to AgNO 3 . The biosynthesized Ast-AgNPs were characterised using ultraviolet-visible spectroscopy (UV-Vis), zeta potential, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX), and high-resolution transmission electron microscopy (HR-TEM) with selected area electron diffraction (SAED) patterns. The nanoparticles exhibited their highest absorption in the UV-visible spectra at 425 nm. The X-ray diffraction (XRD) investigation indicated the presence of characteristic peaks at certain angles: 38.30° (1 1 1), 44.40° (2 0 0), 64.64° (2 2 0), and 77.59° (3 1 1) according to the JCPDS file No. 04-0783. Based on SEM and TEM, the Ast-AgNPs had an average size of 35 nm and 52 nm, respectively. The zeta potential was determined to be - 20.8 mV, indicating their stability. The highest antibacterial effectiveness is shown against Staphylococcus aureus, with a zone of inhibition of 25.66 ± 1.52 mm at 250 μL/mL conc. of Ast-AgNPs. Likewise, Ast-AgNPs significantly suppressed the growth of Fusarium sp. and Curvularia sp. by 78.22% and 85.05%, respectively, at 150 μL/mL conc. of Ast-AgNPs. In addition, the Ast-AgNPs exhibited significant photocatalytic activity in degrading methylene blue (MB), achieving an 88.59% degradation in 120 min, revealing multiple downstream applications of Ast-AgNPs., (© 2024. The Author(s).)

Waist-to-hip circumference ratio (WHR) is now recognized as among the strongest shape biometrics linked with health outcomes, although use of this phenotypic marker remains limited due to the inaccuracies in and inconvenient nature of flexible tape measurements when made in clinical and home settings. Here we report that accurate and reliable WHR estimation in adults is possible with a smartphone application based on novel computer vision algorithms. The developed application runs a convolutional neural network model referred to as MeasureNet that predicts a person's body circumferences and WHR using front, side, and back color images. MeasureNet bridges the gap between measurements conducted by trained professionals in clinical environments, which can be inconvenient, and self-measurements performed by users at home, which can be unreliable. MeasureNet's accuracy and reliability is evaluated using 1200 participants, measured by a trained staff member. The developed smartphone application, which is a part of Amazon Halo, is a major advance in digital anthropometry, filling a long-existing gap in convenient, accurate WHR measurement capabilities., (© 2023. Springer Nature Limited.)

Production of phosphorus efficient genotypes in groundnut can improve and also reduces environmental pollution. Identification of P-efficient groundnut genotypes is a need of the hour to sustain in P-deficient soils. The pot experiment showed significant differences between genotypes (G) and treatments (T) for all the traits and G × T interaction for majority of traits. The G × T × Y interaction effects were also significant for all the traits except leaf P% (LP%), leaf acid phosphatase (LAP) and root dry weight (RDW). In lysimeter experiment, the effect of G, T and G × T were significant for leaf dry weight (LDW), stem dry weight (SDW), total transpiration (TT) and transpiration efficiency (TE). For traits, LDW, SDW, TT, TE, ICGV 00351 and ICGS 76; for SDW, TT, ICGV 02266 are best performers under both P-sufficient and deficient conditions. Based on P-efficiency indices and surrogate traits of P-uptake, ICGV's 02266, 05155, 00308, 06040 and 06146 were considered as efficient P-responding genotypes. From GGE biplot, ICGV 06146 under P-deficient and TAG 24 under both P-sufficient and deficient conditions are portrayed as best performer. ICGV 06146 was identified as stable pod yielder and a promising genotype for P-deficient soils. The genotypes identified in this study can be used as a parent in developing mapping population to decipher the genetics and to devleop groundnut breeding lines suitable to P-deficient soils., (© 2022. The Author(s).)

Ultrafast plasma dynamics play a pivotal role in the relativistic high harmonic generation, a phenomenon that can give rise to intense light fields of attosecond duration. Controlling such plasma dynamics holds key to optimize the relevant sub-cycle processes in the high-intensity regime. Here, we demonstrate that the optimal coherent combination of two intense ultrashort pulses centered at two-colors (fundamental frequency, [Formula: see text] and second harmonic, [Formula: see text]) can lead to an optimal shape in relativistic intensity driver field that yields such an extraordinarily sensitive control. Conducting a series of two-dimensional (2D) relativistic particle-in-cell (PIC) simulations carried out for currently achievable laser parameters and realistic experimental conditions, we demonstrate that an appropriate combination of [Formula: see text] along with a precise delay control can lead to more than three times enhancement in the resulting high harmonic flux. Finally, the two-color multi-cycle field synthesized with appropriate delay and polarization can all-optically suppress several attosecond bursts while favourably allowing one burst to occur, leading to the generation of intense isolated attosecond pulses without the need of any sophisticated gating techniques., (© 2022. The Author(s).)

Dill seeds (Anethum graveolens L.) is the most valuable medicinal seed spice crop of Apiaceae. It bears small yellow flowers in the form of umbels. Being a cross-pollinated crop, floral visitors play vital role in pollination and seed sets. Hence, the present study was conducted at the ICAR-National Research Centre on Seed Spices, Ajmer (Rajasthan), India to discover the pollinator's community, foraging behaviour and abundance of most frequent pollinators and different modes of pollination on seed yield and quality of this seed spice crop. The insect visitors community of dill seeds was composed of 28 insect species belonging to 14 families of 6 orders. Most of floral visitors started their foraging activity at 8.00 h, reached peak activity between 12.00 and 14.00 h and their activity ceased at 18.00 h. Apis florea, A. dorsata, A. mellifera, solitary bee, Halictus sp. and two unidentified species of Hymenoptera; Episyrphus balteatus (DeGeer), Episyrphus sp., Eristalis sp and two other Musca species of Diptera were identified as potential and regular floral visitors of dill seeds. The highest seed yield of 1505.63 kg/ha was recorded in the treated plots provided with only 10% jaggery solution and was at par with the open pollination. A lower seed yield of 1432.5 kg/ha was recorded in plots pollinated only with A. mellifera inside insect cages. Open pollination with 10% jaggery solution spray increased the seed yield of dill seed crop by 57%, one-thousand seed test weight by 96% and the essential oil content by 27% over control plots. These results show that managed pollination is a much better way to enhance yields and quality of dill seed crop than other treatments including only honeybee-based pollination., (© 2022. The Author(s).)

Proteoglycans are proteins that are modified with glycosaminoglycan chains. Chondroitin sulfate proteoglycans (CSPGs) are currently being exploited as targets for drug-delivery in various cancer indications, however basic knowledge on how CSPGs are internalized in tumor cells is lacking. In this study we took advantage of a recombinant CSPG-binding lectin VAR2CSA (rVAR2) to track internalization and cell fate of CSPGs in tumor cells. We found that rVAR2 is internalized into cancer cells via multiple internalization mechanisms after initial docking on cell surface CSPGs. Regardless of the internalization pathway used, CSPG-bound rVAR2 was trafficked to the early endosomes in an energy-dependent manner but not further transported to the lysosomal compartment. Instead, internalized CSPG-bound rVAR2 proteins were secreted with exosomes to the extracellular environment in a strictly chondroitin sulfate-dependent manner. In summary, our work describes the cell fate of rVAR2 proteins in tumor cells after initial binding to CSPGs, which can be further used to inform development of rVAR2-drug conjugates and other therapeutics targeting CSPGs., (© 2022. The Author(s).)

A safe and effective vaccine against COVID-19 is urgently needed in quantities that are sufficient to immunize large populations. Here we report the preclinical development of two vaccine candidates (BNT162b1 and BNT162b2) that contain nucleoside-modified messenger RNA that encodes immunogens derived from the spike glycoprotein (S) of SARS-CoV-2, formulated in lipid nanoparticles. BNT162b1 encodes a soluble, secreted trimerized receptor-binding domain (known as the RBD-foldon). BNT162b2 encodes the full-length transmembrane S glycoprotein, locked in its prefusion conformation by the substitution of two residues with proline (S(K986P/V987P); hereafter, S(P2) (also known as P2 S)). The flexibly tethered RBDs of the RBD-foldon bind to human ACE2 with high avidity. Approximately 20% of the S(P2) trimers are in the two-RBD 'down', one-RBD 'up' state. In mice, one intramuscular dose of either candidate vaccine elicits a dose-dependent antibody response with high virus-entry inhibition titres and strong T-helper-1 CD4 + and IFNγ + CD8 + T cell responses. Prime-boost vaccination of rhesus macaques (Macaca mulatta) with the BNT162b candidates elicits SARS-CoV-2-neutralizing geometric mean titres that are 8.2-18.2× that of a panel of SARS-CoV-2-convalescent human sera. The vaccine candidates protect macaques against challenge with SARS-CoV-2; in particular, BNT162b2 protects the lower respiratory tract against the presence of viral RNA and shows no evidence of disease enhancement. Both candidates are being evaluated in phase I trials in Germany and the USA 1-3 , and BNT162b2 is being evaluated in an ongoing global phase II/III trial (NCT04380701 and NCT04368728).

Fluorescent pH biosensors have gained importance owing to their low cost utilization in real time monitoring of biological and food samples in comparison to conventional pH meters. The research reports a novel method of ultrasonic atomization for developing a fluorescent pH sensor for real-time analysis made of Fluorescein isothiocyanate (FITC)-dextran/FITC-dextran-Tris (2, 2'-bipyridyl) dichlororuthenium (II) hexahydrate as indicator and reference fluorophores, respectively. The process of ultrasonic atomization ensures formation of monodisperse dye immobilized alginate microspheres ensuring efficient pH sensing. The developed biosensor was tested on milk samples, which has a short life span and shows a significant fall in pH with time due to microbial spoilage. The proposed biosensor showed a linear range of pH 4-8 (R 2 between 0.96-0.99 for different single/dual fluorophore biosensors) which suitably cover the pH of milk during the entire storage period and spoilage. The % recovery for predicted pH falls between 90-110% compared against standard pH meter, indicating a good accuracy of estimation and low turnaround time (10 min). Thus, real-time monitoring using fluorescent pH biosensor for milk samples may profoundly improve the economics of losses occurring in processing and storage with capability of in-package continuous quality assessment.

High-dose radiation is the main component of glioblastoma therapy. Unfortunately, radio-resistance is a common problem and a major contributor to tumor relapse. Understanding the molecular mechanisms driving response to radiation is critical for identifying regulatory routes that could be targeted to improve treatment response. We conducted an integrated analysis in the U251 and U343 glioblastoma cell lines to map early alterations in the expression of genes at three levels: transcription, splicing, and translation in response to ionizing radiation. Changes at the transcriptional level were the most prevalent response. Downregulated genes are strongly associated with cell cycle and DNA replication and linked to a coordinated module of expression. Alterations in this group are likely driven by decreased expression of the transcription factor FOXM1 and members of the E2F family. Genes involved in RNA regulatory mechanisms were affected at the mRNA, splicing, and translation levels, highlighting their importance in radiation-response. We identified a number of oncogenic factors, with an increased expression upon radiation exposure, including BCL6, RRM2B, IDO1, FTH1, APIP, and LRIG2 and lncRNAs NEAT1 and FTX. Several of these targets have been previously implicated in radio-resistance. Therefore, antagonizing their effects post-radiation could increase therapeutic efficacy. Our integrated analysis provides a comprehensive view of early response to radiation in glioblastoma. We identify new biological processes involved in altered expression of various oncogenic factors and suggest new target options to increase radiation sensitivity and prevent relapse.

Near infrared (NIR) fluorophores like Pt-porphyrin along with analyte specific enzymes require co-encapsulation in biocompatible and biodegradable carriers in order to be transformed into implantable biosensors for efficient and continuous monitoring of analytes in patients. The main objective of this research is to develop natural, biodegradable, biocompatible and a novel co-encapsulated system of Pt-porphyrin encapsulated polymeric nanoparticle and nano-micro hybrid carriers. A sequential emulsification-solvent evaporation and an air driven atomization technique was used for developing above matrices and testing them for fluorescence based oxygen and glucose biosensing. The results indicate Pt-porphyrin can be efficiently encapsulated in Poly-lactic acid (PLA) nanoparticles and PLA-alginate nano-micro particles with sizes ~450 nm and 10 µm, respectively. Biosensing studies have showed a linear fluorescent response in oxygen concentrations ranging from of 0-6 mM (R 2  = 0.992). The Oxygen sensitivity was transformed into a linear response of glucose catalytic reaction in the range of 0-10 mM (R 2  = 0.968) with a response time of 4 minutes and a stability over 15 days. We believe that the investigated NIR fluorophores like Pt-Porphyrin based nano/nano-micro hybrid carrier systems are novel means of developing biocompatible biodegradable carriers for developing implantable glucose biosensors which can efficiently manage glucose levels in diabetes.

Prostate cancer (PCa) is the second leading cause of cancer deaths among American men. Unfortunately, there is no cure once the tumor is established within the bone niche. Although osteocytes are master regulators of bone homeostasis and remodeling, their role in supporting PCa metastases remains poorly defined. This is largely due to a lack of suitable ex vivo models capable of recapitulating the physiological behavior of primary osteocytes. To address this need, we integrated an engineered bone tissue model formed by 3D-networked primary human osteocytes, with conditionally reprogrammed (CR) primary human PCa cells. CR PCa cells induced a significant increase in the expression of fibroblast growth factor 23 (FGF23) by osteocytes. The expression of the Wnt inhibitors sclerostin and dickkopf-1 (Dkk-1), exhibited contrasting trends, where sclerostin decreased while Dkk-1 increased. Furthermore, alkaline phosphatase (ALP) was induced with a concomitant increase in mineralization, consistent with the predominantly osteoblastic PCa-bone metastasis niche seen in patients. Lastly, we confirmed that traditional 2D culture failed to reproduce these key responses, making the use of our ex vivo engineered human 3D bone tissue an ideal platform for modeling PCa-bone interactions.

α-Synuclein (α-Syn) is an intrinsically disordered presynaptic protein, whose aggregation is critically involved in Parkinson's disease (PD). Many of the currently available drugs for the treatment of PD are not sufficiently effective in preventing progress of the disease and have multiple side-effects. With this background, efficient drug candidates, sulfated polysaccharides from Chlamydomonas reinhardtii (Cr-SPs) were isolated and investigated for their effect on inhibition of α-Syn fibrillation and dissolution of preformed α-Syn fibrillar structures through a combination of spectroscopic and microscopic techniques. The kinetics of α-Syn fibrillation demonstrates that Cr-SPs are very effective in inhibiting α-Syn fibrillation. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis gel-image shows presence of soluble protein in the presence of Cr-SPs after completion of the fibrillation process. The morphological changes associated with fibrillation monitored by transmission electron microscopy showed that Cr-SPs efficiently bind with α-Syn and delay the conversion of α-helical intermediate into β-sheet rich structures. Cr-SPs are also effective even if onset of α-Syn fibrillation has already started and they also have the ability to dissolve pre-formed fibrils. Thus, the current work has substantial therapeutic implications towards unlocking the immense potential of algal products to function as alternative therapeutic agents against PD and other protein aggregation related disorders.

Using conditional cell reprogramming, we generated a stable cell culture of an extremely rare and aggressive neuroendocrine cervical cancer. The cultured cells contained HPV-16, formed colonies in soft agar and rapidly produced tumors in immunodeficient mice. The HPV-16 genome was integrated adjacent to the Myc gene, both of which were amplified 40-fold. Analysis of RNA transcripts detected fusion of the HPV/Myc genes, arising from apparent microhomologous recombination. Spectral karyotyping (SKY) and fluorescent-in-situ hybridization (FISH) demonstrated coordinate localization and translocation of the amplified Myc and HPV genes on chromosomes 8 and 21. Similar to the primary tumor, tumor cell cultures expressed very high levels of the Myc protein and, in contrast to all other HPV-positive cervical cancer cell lines, they harbored a gain-of-function mutation in p53 (R273C). Unexpectedly, viral oncogene knockdown had no effect on the growth of the cells, but it did inhibit the proliferation of a conventional HPV-16 positive cervical cancer cell line. Knockdown of Myc, but not the mutant p53, significantly inhibited tumor cell proliferation. On the basis of these data, we propose that the primary driver of transformation in this aggressive cervical cancer is not HPV oncogene expression but rather the overexpression of Myc.

We have studied here using a number of biophysical tools the effects of osmolytes, betaine, citrulline, proline and sorbitol which differ significantly in terms of their physical characteristics such as, charge distribution, polarity, H-bonding abilities etc, on the fibrillation of insulin. Among these, betaine, citrulline, and proline are very effective in decreasing the extent of fibrillation. Proline also causes a substantial delay in the onset of fibrillation in the concentration range (50-250 mM) whereas such an effect is seen for citrulline only at 250 mM, and in case of betaine this effect is not seen at all in the whole concentration range. The enthalpies of interaction at various stages of fibrillation process have suggested that the preferential exclusion of the osmolyte and its polar interaction with the protein are important in inhibition. The results indicate that the osmolytes are most effective when added prior to the elongation stage of fibrillation. These observations have significant biological implications, since insulin fibrillation is known to cause injection amyloidosis and our data may help in designing lead drug molecules and development of potential therapeutic strategies.

The heterogeneity and poor prognosis associated with gliomas, makes biomarker identification imperative. Here, we report autoantibody signatures across various grades of glioma serum samples and sub-categories of glioblastoma multiforme using Human Proteome chips containing ~17000 full-length human proteins. The deduced sets of classifier proteins helped to distinguish Grade II, III and IV samples from the healthy subjects with 88, 89 and 94% sensitivity and 87, 100 and 73% specificity, respectively. Proteins namely, SNX1, EYA1, PQBP1 and IGHG1 showed dysregulation across various grades. Sub-classes of GBM, based on its proximity to the sub-ventricular zone, have been reported to have different prognostic outcomes. To this end, we identified dysregulation of NEDD9, a protein involved in cell migration, with probable prognostic potential. Another subcategory of patients where the IDH1 gene is mutated, are known to have better prognosis as compared to patients carrying the wild type gene. On a comparison of these two cohorts, we found STUB1 and YWHAH proteins dysregulated in Grade II glioma patients. In addition to common pathways associated with tumourigenesis, we found enrichment of immunoregulatory and cytoskeletal remodelling pathways, emphasizing the need to explore biochemical alterations arising due to autoimmune responses in glioma.

RNA interference pathways use small RNAs to mediate gene silencing in eukaryotes. In addition to small interfering RNAs (siRNAs) and microRNAs, several types of endogenously produced small RNAs have important roles in gene regulation, germ cell maintenance and transposon silencing. The production of some of these RNAs requires the synthesis of aberrant RNAs (aRNAs) or pre-siRNAs, which are specifically recognized by RNA-dependent RNA polymerases to make double-stranded RNA. The mechanism for aRNA synthesis and recognition is largely unknown. Here we show that DNA damage induces the expression of the Argonaute protein QDE-2 and a new class of small RNAs in the filamentous fungus Neurospora crassa. This class of small RNAs, known as qiRNAs because of their interaction with QDE-2, are about 20-21 nucleotides long (several nucleotides shorter than Neurospora siRNAs), with a strong preference for uridine at the 5' end, and originate mostly from the ribosomal DNA locus. The production of qiRNAs requires the RNA-dependent RNA polymerase QDE-1, the Werner and Bloom RecQ DNA helicase homologue QDE-3 and dicers. qiRNA biogenesis also requires DNA-damage-induced aRNAs as precursors, a process that is dependent on both QDE-1 and QDE-3. Notably, our results suggest that QDE-1 is the DNA-dependent RNA polymerase that produces aRNAs. Furthermore, the Neurospora RNA interference mutants show increased sensitivity to DNA damage, suggesting a role for qiRNAs in the DNA-damage response by inhibiting protein translation.