Antiproliferative effect of indeno[1,2-d]thiazolo[3,2-a]pyrimidine analogues on IL-6 mediated STAT3 and role of the apoptotic pathway in albino Wistar rats of ethyl carbamate-induced lung carcinoma: In-silico, In-vitro, and In-vivo study.

Lung cancer (LC) ranks second most prevalent cancer in females after breast cancer and second in males after prostate cancer. Based on the GLOBOCAN 2020 report, India represented 5.9% of LC cases and 8.1% of deaths caused by the disease. Several clinical studies have shown that LC occurs because of biological and morphological abnormalities and the involvement of altered level of antioxidants, cytokines, and apoptotic markers. In the present study, we explored the antiproliferative activity of indeno[1,2-d]thiazolo[3,2-a]pyrimidine analogues against LC using in-vitro, in-silico, and in-vivo models. In-vitro screening against A549 cells revealed compounds 9B (8-methoxy-5-(3,4,5-trimethoxyphenyl)-5,6-dihydroindeno[1,2-d]thiazolo[3,2-a]pyrimidine) and 12B (5-(4-chlorophenyl)-5,6-dihydroindeno[1,2-d]thiazolo[3,2-a]pyrimidine) as potential pyrimidine analogues against LC. Compounds 9B and 12B were docked with different molecular targets IL-6, Cyt-C, Caspase9, and Caspase3 using AutoDock Vina 4.1 to evaluate the binding affinity. Subsequently, in-vivo studies were conducted in albino Wistar rats through ethyl-carbamate (EC)- induced LC. 9B and 12B imparted significant effects on physiological (weight variation), and biochemical (anti-oxidant [TBAR's, SOD, ProC, and GSH), lipid (TC, TG, LDL, VLDL, and HDL)], and cytokine (IL-2, IL-6, IL-10, and IL-1ß) markers in EC-induced LC in albino Wistar rats. Morphological examination (SEM and H&E) and western blotting (IL-6, STAT3, Cyt-C, BAX, Bcl-2, Caspase3, and caspase9) showed that compounds 9B and 12B had antiproliferative effects. Accordingly, from the in-vitro, in-silico, and in-vivo experimental findings, we concluded that 9B and 12B have significant antiproliferative potential and are potential candidates for further evaluation to meet the requirements of investigation of new drug application.

From Causatum to Erratum, all in a name.

BACKGROUND: This study highlights how a trivial mistake in collecting timed blood samples of parathyroid hormone (PTH) during parathyroidectomy (PTX) can potentially become a serious error affecting surgical closure. METHODS: For the measurement of serum PTH, the intact PTH (iPTH) test was used to obtain baseline, preoperative, intraoperative, and postoperative samples of PTH, to guide the surgical team regarding adequacy of PTX. RESULTS: Due to the lack of proper guidelines, all types of samples for PTH are labeled as iPTH by the Laboratory Information Services (LIS) software. Due to a human error in marking the PTH vacutainers generated for different time point samples by LIS, samples were swapped. The values in the lab revealed a spurious rise in PTH post-PTX. The laboratory physician carefully observed the tubes and identified the reason for this mistake. The timely action therefore led to surgical closure, otherwise it could have led to unwarranted extended PTX. CONCLUSIONS: In cases where timed samples are mandatory, having a common code for all requisitions can invariably lead to pre-analytical error, therefore proper discriminative measures need to be introduced to avoid these mistakes.

Development and validation of most efficient RNA isolation method from buffalo bull spermatozoa.

BACKGROUND: Being highly fragmented and low in concentration, isolation of good quality RNA from sperm cells is a big challenge. Attempts have been made to evaluate various sperm RNA isolation methods from purified buffalo bull sperm cells. METHODS: Both, non-membrane and membrane-based methods have been evaluated for isolating RNA from Murrah buffalo sperms and compared for their respective efficacies. The traditional TRIzol, TRIzol-heat lysed (H-TRIzol) and cocktail of TCEP-RLT lysis buffer (Qiagen RNeasy mini kit)-TRIzol (C-TRIzol) based isopropanol isolation methods have been evaluated. RESULTS: H-TRIzol yielded best results among conventional methods. The combined T-RLT RNA isolation protocol yielded best quality and quantity compared to other membrane-based methods, due to high lytic property of cocktail of lysis reagents, necessary for complete breakdown of sperm membrane and RNA binding membrane for RNA isolation. Combined lysis performed by treatment with RLT-T and T-RLT differing in order of reagents used were also evaluated. T-RLT combination giving better results compared to RLT-T due to high gDNA contamination and membrane clogging in later protocol steps. CONCLUSION: Overall, in terms of total RNA quantity and quality per million spermatozoa, the heat-lysed TRIzol method (H-TRIzol) performs best among RNA separation techniques employed and is also quite easy to perform. This comparative evaluation of sperm RNA isolation protocols can be useful in deciding the best protocol for isolation of good quality and high concentration sperm RNA from buffalo semen, for transcriptome and other downstream studies.

Deciphering the nitrate sources and processes in the Ganga river using dual isotopes of nitrate and Bayesian mixing model.

The dual isotopes of dissolved NO3- (n = 43) has been used to delineate the nitrate sources and N-cycling processes in the Ganga river. The proportional contribution of nitrate from different sources has been estimated using the Bayesian mixing model. The seasonal NO3- concentration in the lower stretch of the river Ganga varied between 4.1 and 64.1 µM with higher concentration during monsoon and post-monsoon season and lower concentration during the pre-monsoon and winter season. The temporal variation in the isotopic values ranged between +0.0 and +9.6‰ for δ15NNO3- and -1.2 to +11.0‰ for δ18ONO3-. The spatial NO3- concentration during the post-monsoon season varied between 23.2 and 57.7 µM, with higher values from the middle and lower values from the lower stretch of the river Ganga. The isotopic ratio during the post-monsoon season varied between -1.0 and +11.3‰ for δ15NNO3- and -4.6 to +5.2‰ for δ18ONO3-. The temporal dataset from the lower stretch of the river Ganga showed the dominance of nitrate derived from the nitrification of soil organic matter (SOM) (average ∼53.4%). The nitrate contribution from synthetic fertilizers was observed to be higher during the post-monsoon season (34.7 ± 23.4%) compared to that in the monsoon (25.5 ± 19.5%) and pre-monsoon (22.2 ± 19.6%) season. No significant seasonal variations were observed in the nitrate input from manure/sewage (∼13.9%). Spatial samples collected during the post-monsoon season showed higher contribution of synthetic fertilizer in the lower stretch (34.6 ± 22.7%) compared to the middle stretch (21.1 ± 18.2%), which indicates greater influence of the agricultural activity in the lower stretch. The dual isotope study of dissolved NO3- established that the nitrate in the Ganga river water is mostly derived from the nitrification of incoming organic compounds and is subsequently removed via assimilatory nitrate uptake. The study also emphasises significant nitrification and assimilatory nitrate removal processes operating in the mixing zone of the Ganga river and Hooghly estuary.

Demographics and appropriateness of cellular blood component irradiation practices: Ambispective analysis from a tertiary care center.

BACKGROUND: Transfusion-associated graft versus host disease (TA-GVHD) is often underreported. There may also be lapses in TA-GVHD prevention practices due to lack of revision of some of the existing clinical guidelines as well as limited audits on practices of blood component irradiation. This study was undertaken to highlight these shortcomings, and generate data for development of institutional guidelines. METHODS/MATERIALS: Study cohort was selected from patients requiring transfusion support during June 2019 to May 2020. Transfusion history of these patients were followed, both retrospectively and prospectively till July 2021. Transfusion requisitions were categorized as IR (with request for irradiation) or NIR (with no request for irradiation) and justified or unjustified according to published international guidelines. RESULTS: Total 6963 requisitions for cellular blood components were received from 255 patients included in the study cohort. Of these, 3690 (54.9 %) were IR requisitions, while remaining 3029 (45.1 %) requisitions were NIR. Overall, 4242 (63.1 %) requisition were justified for their irradiation status as per published guidelines and 1595 (23.8 %) were found to be Unjustified while justification could not be assessed for remaining 882 (13.1 %) of the requisitions. The highest proportion of Unjustified demands in NIR requisitions was observed in patients with Severe Aplastic anemia (59.4 %). CONCLUSION: Many units were unnecessarily irradiated (7.7 %) while irradiation was missed in 16 % of the requisitions included in analysis which may be attributed to lack of institutional guidelines. We recommend that every centre should adopt a published well-researched guideline including amendments based on review of practices at their center.

Allelic diversity at BoLA DRB3 locus and association with predisposition to clinical mastitis in indicus and crossbred cattle.

Bovine lymphocyte antigen (BoLA) DRB3 locus in healthy and mastitis affected cattle has been genotyped by a polymerase chain reaction and restriction fragment length polymorphisms (PCR-RLFP) using RsaI restriction enzyme, followed by sequencing. In 130 farm animals, 25 BoLA DRB3 alleles have been detected by PCR-RFLP. Three distinct allelic patterns significantly associated with mastitis in Karan Fries crossbred and Sahiwal indicus cattle have been identified, whereas, four other allelic patterns were significantly high in frequency among healthy animals. Sequencing of RFLP genotypes revealed 25 and 47 alleles among healthy Sahiwal and Karan Fries, respectively, while 17 and 38 patterns observed in mastitis affected Sahiwal and Karan Fries animals, respectively. From Tajima's D-test of neutrality, it was concluded that alleles associated with mastitis were expanding in the population, whereas those of healthy were under contraction. Phylogenetic analysis carried out to delineate the evolutionary relationship of the farm and field animals at DRB3 locus, differentiating allelic patterns into six different clusters. Among the phylogenetic lineages, five patterns DRB3*028:01, DRB3*011:03, DRB3*031:01, DRB3*001:01 and DRB3*043:01, were previously reported, whereas one novel allelic variant was observed in indicus and crossbred cattle. This information will help in further exploring the association between BoLA-DRB3 genetic diversity and disease resistance in distinct cattle breeds, important in designing breeding strategies for increasing the distribution of favorable alleles.

Identification of novel allelic patterns and evolutionary lineage of BoLA MHC class II DQA locus in indicine and taurine cattle.

Among different cattle types, Bos indicus are known for their ability to better resist the tropical microbial infections comparatively, wherein MHC molecules play a significant role. In this study allelic diversity at MHC locus, DQA of Bos indicus, Bos taurus and crossbred of taurine-indicus has been explored to understand the possible role of MHC region in differential immune response. Thirty nine different DQA alleles were identified, out of which 14 were novel, along with documentation of duplication of DQA alleles. Indicus cattle population presented diverse types of DQA alleles compared to crossbred and exotic. Translated amino acid sequence analysis indicated, codon 64 and 50 of peptide binding sites being highly polymorphic and most of the indicus cattle presented alanine and arginine amino acid at position 64 and 50. Within breed genetic variation found to be higher than between breeds. Because of their ability to bind and subsequently respond to a wide array of antigens, the newly identified DQA alleles with high diversity present in the form of duplicated haplotypes in different combinations in cattle populations provided significant insights into probable role of this MHC locus in better tropical disease combating ability and genetic fitness of indicus cattle.

Mild Alkaline Pretreatment of Rice Straw as a Feedstock in Microbial Fuel Cells for Generation of Bioelectricity.

The dependency on non-renewable fossil fuels as an energy source has drastically increased global temperatures. Their continuous use poses a great threat to the existing energy reserves. Therefore, the energy sector has taken a turn toward developing eco-friendly, sustainable energy generation by using sustainable lignocellulosic wastes, such as rice straw (RS). For lignocellulosic waste to be utilized as an efficient energy source, it needs to be broken down into less complex forms by pretreatment processes, such as alkaline pretreatment using NaOH. Varied NaOH concentrations (0.5%,1.0%,1.5%,2%) for alkaline pretreatment of RS were used for the holocellulose generation. Amongst the four NaOH concentrations tested, RS-1.5% exhibited higher holocellulose generation of 80.1%, whereas 0.5%, 1 5 and 2% pointed 71.9%, 73.8%, and 78.5% holocellulose generation, respectively. Further, microbial fuel cells (MFCs) were tested for voltage generation by utilizing holocellulose generated from untreated (RS-0%) and mildly alkaline pretreated RS (RS-1.5%) as a feedstock. The MFC voltage and maximum power generation using RS-0% were 194 mV and 167 mW/m2, respectively. With RS-1.5%, the voltage and maximum power generation were 556 mV and 583 mW/m2, respectively. The power density of RS-1.5% was three-fold higher than that of RS-0%. The increase in MFC power generation suggests that alkaline pretreatment plays a crucial role in enhancing the overall performance.

Understanding decision processes in becoming a fee-for-hire service provider: A case study on direct seeded rice in Bihar, India.

While Direct Seeded Rice (DSR) has numerous potential benefits to smallholder farmers in the Eastern Gangetic Plains of South Asia, it's out-scaling has been limited by both a lack of demand by farmers and limited supply of DSR services by machinery owners. This contrasts with the comparatively more rapid scaling of zero tillage wheat in the region. This trend is yet to be fully explored, particularly when focus has been placed almost exclusively on understanding DSR adoption though the lens of farm-level agronomic, economic and environmental performance. Given that limited DSR service provision is likely to be governed outside of these considerations, this study explores with zero tillage drill owners the decision processes they apply in deciding how to use their zero tillage drills. Respondents highlight a complex web of interrelated considerations that highlight the additional complexities of DSR as compared to existing practices. Using a novel 'Decision-making Dartboard' qualitative framework, these complexities are unpacked and a set of potential changes to the assumed theory of change for DSR scaling are identified, including considerations for selection of potential DSR service providers and responsibilities for promotion and extension of DSR to overcome the prevalent negative perceptions of DSR held broadly across the communities explored. The proposed framework and analysis process are also potentially useful for exploration of other farmer decision making processes more broadly.

Correction to: Biomethanol Production from Methane by Immobilized Co-cultures of Methanotrophs.

[This corrects the article DOI: 10.1007/s12088-020-00883-6.].

Intercomparison of crop establishment methods for improving yield and profitability in the rice-wheat system of Eastern India.

Conventionally managed rice-wheat systems of the eastern Indo-Gangetic Plains (E-IGP) that rely on soil puddling for rice and intensive tillage for wheat are low-yielding and resource-inefficient, leading to low profitability. While a host of alternative tillage and crop establishment (TCE) methods have been advocated as solutions for sustainably enhancing productivity and profitability, few systematic comparisons of these methods are reported. To address this gap, a three-year field study was conducted in Bihar, India with the goal of identifying TCE methods for rice-wheat systems that are high yielding, less resource-intensive, and more profitable. The following systems were evaluated: 1) puddled transplanted rice (PTR) followed by (fb) conventional tillage wheat (CTW) or zero-tillage wheat (ZTW); 2) machine transplanted rice in non-puddled soil (MTR) fb ZTW; 3) the system of rice intensification (SRI) fb system of wheat intensification (SWI); and 4) dry-seeded rice (DSR) fb ZTW. Rice cultivar duration (short versus medium-duration) was incorporated as a subplot treatment in all systems. Rice yields were similar with all methods, except DSR yield was 11 % lower and MTR yield was 7% higher than PTR in the third year. Cost of production was US$ 149 and 77 ha-1 lower in DSR and MTR, respectively, and US$ 84 ha-1 higher in SRI than PTR. The gross margin and benefit-cost (B:C) ratio was highest in MTR followed by DSR and lowest in SRI. In wheat, ZT resulted in a higher yield than CTW, especially when ZTW was cultivated after non-puddled rice (e.g., DSR or MTR). ZTW reduced production costs by US$ 69 ha-1, whereas SWI increased it by US$ 139 ha-1 relative to CTW. The higher yield and lower cost of production resulted in a higher gross margin (US$ 82-355 ha-1 and US$ 129-409 ha-1 higher than CTW and SWI, respectively) and a higher B:C ratio in ZTW treatments than CTW and SWI. At the system level, MTR or DSR followed by ZTW had both superior crop yields and consistently higher gross margins (US $133 to 382 ha-1) than other practices. On the other hand, the SRI fb SWI system had no yield advantage and poorer economic performance than conventional practices. In all systems, the inclusion of a medium-duration rice hybrid resulted in higher rice and system yields. These results suggest that significant gains in profitability are possible with emerging TCE practices in rice-wheat systems, but alternatives such as the SRI and SWI will likely erode farmer incomes.

Phe-140 Determines the Catalytic Efficiency of Arylacetonitrilase from Alcaligenes faecalis.

Arylacetonitrilase from Alcaligenes faecalis ATCC8750 (NitAF) hydrolyzes various arylacetonitriles to the corresponding carboxylic acids. A systematic strategy of amino acid residue screening through sequence alignment, followed by homology modeling and biochemical confirmation was employed to elucidate the determinant of NitAF catalytic efficiency. Substituting Phe-140 in NitAF (wild-type) to Trp did not change the catalytic efficiency toward phenylacetonitrile, an arylacetonitrile. The mutants with nonpolar aliphatic amino acids (Ala, Gly, Leu, or Val) at location 140 had lower activity, and those with charged amino acids (Asp, Glu, or Arg) exhibited nearly no activity for phenylacetonitrile. Molecular modeling showed that the hydrophobic benzene ring at position 140 supports a mechanism in which the thiol group of Cys-163 carries out a nucleophilic attack on a cyanocarbon of the substrate. Characterization of the role of the Phe-140 residue demonstrated the molecular determinant for the efficient formation of arylcarboxylic acids.

Biomethanol Production from Methane by Immobilized Co-cultures of Methanotrophs.

Methanol production by co-culture of methanotrophs Methylocystis bryophila and Methyloferula stellata was examined from methane, a greenhouse gas. Co-culture exhibited higher methanol yield of 4.72 mM at optimum ratio of M. bryophila and M. stellata (3:2) compared to individual cultures. The immobilized co-culture within polyvinyl alcohol (PVA) showed relative efficiency of 90.1% for methanol production at polymer concentration of 10% (v/v). The immobilized co-culture cells within PVA resulted in higher bioprocess stability over free cells at different pH, and temperatures. Free and encapsulated co-cultures showed maximum methanol production of 4.81 and 5.37 mM under optimum conditions, respectively. After five cycles of reusage under batch conditions, free and encapsulated co-cultures retained methanol production efficiency of 23.8 and 61.9%, respectively. The present investigation successfully revealed the useful influence of co-culture on the methanol production over pure culture. Further, encapsulation within the polymeric matrix proved to be a better approach for the enhanced stability of the bioprocess.

Exploitation of Citrus Peel Extract as a Feedstock for Power Generation in Microbial Fuel Cell (MFC).

Microbial fuel cells (MFCs) are envisioned as an evolving cost-effective process for treating organic wastes to simultaneously generate bioelectricity. Therefore, in present study a single chambered mediator- less air cathode MFC was operated for bioelectricity generation using citrus waste (CW) as a feedstock. The MFC was operated at four organic loading conditions (OLs; 3, 6, 9 and 12 kg/m3). The voltage generation and organic content reduction demonstrated the possibility of utilizing CW as a substrate in MFC. The polarization analysis revealed a high-power generation of 71.1 mW/m2 with low OL of 3 kg/m3. The decrease in pH and high volatile fatty acids (VFAs) generation was noted at high OL. Our current findings suggest better performance of MFC, in terms of energy generation and organic reduction at high OL.

Application and Limitations of Nanocasting in Metal-Organic Frameworks.

Nanocasting can be a useful strategy to transfer the catalytic metal clusters in metal-organic frameworks (MOFs) to an all-inorganic support such as silica. The incorporation of silica in the MOF pores as a secondary support has the potential to extend the application of the highly tunable metal-based active sites in MOFs to high temperature catalysis. Here, we demonstrate the applicability of the nanocasting method to a range of MOFs that incorporate catalytically attractive hexazirconium, hexacerium, or pentanickel oxide-based clusters (UiO-66, (Ce)UiO-66, (Ce)UiO-67, (Ce)MOF-808, DUT-9, and In- and Ni-postmetalated NU-1000). We describe, in tutorial form, the challenges associated with nanocasting of MOFs that are related to their small pore size and to considerations of chemical and mechanical stability, and we provide approaches to overcome some of these challenges. Some of these nanocast materials feature the site-isolated clusters in a porous, thermally stable silica matrix, suitable for catalysis at high temperatures; in others, structural rearrangement of clusters or partial cluster aggregation occurs, but extensive aggregation can be mitigated by the silica skeleton introduced during nanocasting.

Versatile technique for assessing thickness of 2D layered materials by XPS.

X-ray photoelectron spectroscopy (XPS) has been utilized as a versatile method for thickness characterization of various two-dimensional (2D) films. Accurate thickness can be measured simultaneously while acquiring XPS data for chemical characterization of 2D films having thickness up to approximately 10 nm. For validating the developed technique, thicknesses of few-layer graphene (FLG), MoS2 and amorphous boron nitride (a-BN) layer, produced by microwave plasma chemical vapor deposition (MPCVD), plasma enhanced chemical vapor deposition (PECVD), and pulsed laser deposition (PLD) respectively, were accurately measured. The intensity ratio between photoemission peaks recorded for the films (C 1s, Mo 3d, B 1s) and the substrates (Cu 2p, Al 2p, Si 2p) is the primary input parameter for thickness calculation, in addition to the atomic densities of the substrate and the film, and the corresponding electron attenuation length (EAL). The XPS data was used with a proposed model for thickness calculations, which was verified by cross-sectional transmission electron microscope (TEM) measurement of thickness for all the films. The XPS method determines thickness values averaged over an analysis area which is orders of magnitude larger than the typical area in cross-sectional TEM imaging, hence provides an advanced approach for thickness measurement over large areas of 2D materials. The study confirms that the versatile XPS method allows rapid and reliable assessment of the 2D material thickness and this method can facilitate in tailoring growth conditions for producing very thin 2D materials effectively over a large area. Furthermore, the XPS measurement for a typical 2D material is non-destructive and does not require special sample preparation. Therefore, after XPS analysis, exactly the same sample can undergo further processing or utilization.

Absorptive Hydrogen Scavenging for Enhanced Aromatics Yield During Non-oxidative Methane Dehydroaromatization on Mo/H-ZSM-5 Catalysts.

Addition of Zr metal particles to MoCx /ZSM-5 in interpellet mixtures (2:1 weight ratio) resulted in maximum single-pass methane conversion of 27 % for dehydroaromatization at 973 K (in significant excess of the equilibrium prescribed circa 10 % conversion at these conditions) and a concurrent 1.4-5.6-fold increase in aromatic product yields due to circumvention of thermodynamic equilibrium limitations by absorptive H2 removal by Zr while retaining cumulative aromatic product selectivity. The absorptive function of the polyfunctional catalyst formulation can be regenerated by thermal treatment in He flow at 973 K, yielding above-equilibrium methane conversion in successive regeneration cycles. H2 uptake experiments demonstrate formation of bulk ZrH1.75 on hydrogen absorption by Zr at 973 K. Cooperation between absorption and catalytic centers distinct in location and function enables circumvention of persistent thermodynamic challenges in non-oxidative methane dehydrogenation.

Visible light assisted hydrogen generation from complete decomposition of hydrous hydrazine using rhodium modified TiO2 photocatalysts.

Hydrogen is considered to be an ideal energy carrier, which produces only water when combined with oxygen and thus has no detrimental effect on the environment. While the catalytic decomposition of hydrous hydrazine for the production of hydrogen is well explored, little is known about its photocatalytic decomposition. The present paper describes a highly efficient photochemical methodology for the production of hydrogen through the decomposition of aqueous hydrazine using titanium dioxide nanoparticles modified with a Rh(i) coordinated catechol phosphane ligand (TiO2-Rh) as a photocatalyst under visible light irradiation. After 12 h of visible light irradiation, the hydrogen yield was 413 µmol g-1 cat with a hydrogen evolution rate of 34.4 µmol g-1 cat h-1. Unmodified TiO2 nanoparticles offered a hydrogen yield of 83 µmol g-1 cat and a hydrogen evolution rate of only 6.9 µmol g-1 cat h-1. The developed photocatalyst was robust under the experimental conditions and could be efficiently reused for five subsequent runs without any significant change in its activity. The higher stability of the photocatalyst is attributed to the covalent attachment of the Rh complex, whereas the higher activity is believed to be due to the synergistic mechanism that resulted in better electron transfer from the Rh complex to the conduction band of TiO2.

A Prussian blue/carbon dot nanocomposite as an efficient visible light active photocatalyst for C-H activation of amines.

A Prussian blue/carbon dot (PB/CD) nanocomposite was synthesised and used as a visible-light active photocatalyst for the oxidative cyanation of tertiary amines to α-aminonitriles by using NaCN/acetic acid as a cyanide source and H2O2 as an oxidant. The developed photocatalyst afforded high yields of products after 8 h of visible light irradiation at room temperature. The catalyst was recycled and reused several times without any significant loss in its activity.