Genome-wide association study identifies novel loci and candidate genes for rust resistance in wheat (Triticum aestivum L.).

BACKGROUND: Wheat rusts are important biotic stresses, development of rust resistant cultivars through molecular approaches is both economical and sustainable. Extensive phenotyping of large mapping populations under diverse production conditions and high-density genotyping would be the ideal strategy to identify major genomic regions for rust resistance in wheat. The genome-wide association study (GWAS) population of 280 genotypes was genotyped using a 35 K Axiom single nucleotide polymorphism (SNP) array and phenotyped at eight, 10, and, 10 environments, respectively for stem/black rust (SR), stripe/yellow rust (YR), and leaf/brown rust (LR). RESULTS: Forty-one Bonferroni corrected marker-trait associations (MTAs) were identified, including 17 for SR and 24 for YR. Ten stable MTAs and their best combinations were also identified. For YR, AX-94990952 on 1A + AX-95203560 on 4A + AX-94723806 on 3D + AX-95172478 on 1A showed the best combination with an average co-efficient of infection (ACI) score of 1.36. Similarly, for SR, AX-94883961 on 7B + AX-94843704 on 1B and AX-94883961 on 7B + AX-94580041 on 3D + AX-94843704 on 1B showed the best combination with an ACI score of around 9.0. The genotype PBW827 have the best MTA combinations for both YR and SR resistance. In silico study identifies key prospective candidate genes that are located within MTA regions. Further, the expression analysis revealed that 18 transcripts were upregulated to the tune of more than 1.5 folds including 19.36 folds (TraesCS3D02G519600) and 7.23 folds (TraesCS2D02G038900) under stress conditions compared to the control conditions. Furthermore, highly expressed genes in silico under stress conditions were analyzed to find out the potential links to the rust phenotype, and all four genes were found to be associated with the rust phenotype. CONCLUSION: The identified novel MTAs, particularly stable and highly expressed MTAs are valuable for further validation and subsequent application in wheat rust resistance breeding. The genotypes with favorable MTA combinations can be used as prospective donors to develop elite cultivars with YR and SR resistance.

Functional insight into multi-omics-based interventions for climatic resilience in sorghum (Sorghum bicolor): a nutritionally rich cereal crop.

MAIN CONCLUSION: The article highlights omics-based interventions in sorghum to combat food and nutritional scarcity in the future. Sorghum with its unique ability to thrive in adverse conditions, has become a tremendous highly nutritive, and multipurpose cereal crop. It is resistant to various types of climatic stressors which will pave its way to a future food crop. Multi-omics refers to the comprehensive study of an organism at multiple molecular levels, including genomics, transcriptomics, proteomics, and metabolomics. Genomic studies have provided insights into the genetic diversity of sorghum and led to the development of genetically improved sorghum. Transcriptomics involves analysing the gene expression patterns in sorghum under various conditions. This knowledge is vital for developing crop varieties with enhanced stress tolerance. Proteomics enables the identification and quantification of the proteins present in sorghum. This approach helps in understanding the functional roles of specific proteins in response to stress and provides insights into metabolic pathways that contribute to resilience and grain production. Metabolomics studies the small molecules, or metabolites, produced by sorghum, provides information about the metabolic pathways that are activated or modified in response to environmental stress. This knowledge can be used to engineer sorghum varieties with improved metabolic efficiency, ultimately leading to better crop yields. In this review, we have focused on various multi-omics approaches, gene expression analysis, and different pathways for the improvement of Sorghum. Applying omics approaches to sorghum research allows for a holistic understanding of its genome function. This knowledge is invaluable for addressing challenges such as climate change, resource limitations, and the need for sustainable agriculture.

Pinning-depinning transition of droplets on inclined substrates with a three-dimensional topographical defect.

Droplets on inclined substrates can depin and slide freely above a critical substrate inclination angle. Pinning can be caused by topographical defects on the substrate, and understanding the influence of defect geometry on the pinning-depinning transition is important for diverse applications such as fog harvesting, droplet-based microfluidic devices, self-cleaning surfaces, and inkjet printing. Here, we develop a lubrication-theory-based model to investigate the motion of droplets on inclined substrates with a single three-dimensional Gaussian-shaped defect that can be in the form of a bump or a dent. A precursor-film/disjoining-pressure approach is used to capture contact-line motion, and a nonlinear evolution equation is derived which describes droplet thickness as a function of the position along the substrate and time. The evolution equation is solved numerically using an alternating direction implicit finite-difference scheme to study how the defect geometry influences the critical inclination angle and the shape of a pinned droplet. It is found that the critical substrate inclination angle increases as the defect becomes taller/deeper or wider along the direction lateral to the droplet-sliding direction. However, the critical inclination angle decreases as the defect becomes wider along the sliding direction. Below the critical inclination angle, the advancing contact line of the droplet at the droplet centerline is pinned to the defect at the point having maximum negative slope. Simple scaling relations that reflect the influence of defect geometry on the droplet retention force arising from surface tension are able to account for many of the trends observed in the numerical simulations.

Rupture of thin liquid trilayer films with soluble surfactants: fundamentals and applications to droplet coalescence.

Understanding the stability of thin liquid trilayer films is of direct relevance to applications such as multilayer coatings and polymer processing. The stability of trilayer films can also be used to provide insights into emulsion dynamics, such as the rupture of the thin film formed between two droplets during coalescence. Often, emulsions are laden with surfactants and other additives, which can be present in one or both phases as well as at the interfaces between the liquids. In experimental studies, complicating factors such as variations in droplet sizes, curvatures, and collision processes make it difficult to specifically isolate the influence of surfactant transport on droplet coalescence and film rupture. The present work addresses this issue by systematic consideration of a model problem involving a thin liquid trilayer film. Surfactant is soluble in either the outer layers or the inner layer, corresponding to surfactant soluble in the droplets or the continuous phase. Rupture of the inner layer is driven by van der Waals forces. Lubrication theory is applied to derive coupled nonlinear evolution equations describing the perturbations to the interface positions and the surfactant concentrations. Our findings reveal that surfactant better stabilizes the film when soluble in the inner layer, and the stabilizing effect is more pronounced when the outer layers are thicker. These findings are consistent with experimental observations involving emulsions, where emulsions tend to be more stable when surfactant is in the continuous phase rather than in the droplets, with the distinction being more pronounced when droplets are larger.

Heterogeneity-induced retraction in viscoelastic fluids following cessation of flow.

Complex fluids including colloidal suspensions, microgels, and entangled wormlike micelles (WLMs) can develop heterogeneous flow regions under imposed steady shear. In some of these systems, the evolution to this flow state from rest is accompanied by flow reversal - when a portion of the fluid moves opposite to the imposed flow direction. Flow reversal was proposed to occur in shear startup when (1) the fluid has significant elasticity, and (2) the flow becomes heterogeneous immediately following the stress overshoot [McCauley et al., J. Rheol., 2023, 67, 661-681]. To verify this hypothesis, a new method is developed for measuring flow heterogeneity. Upon cessation of the imposed flow, elasticity and flow heterogeneity cause retraction of the fluid, which is quantified with particle tracking velocimetry. Flow is stopped at key times during shear startup in two systems: a gel-like WLM that exhibits flow reversal before heterogeneous flow and a viscoelastic, fluid-like WLM that does not. The degree of flow heterogeneity is inferred from the shape and magnitude of velocity profiles measured during retraction. Flow heterogeneity develops earlier in gel-like WLMs - supporting the proposed flow reversal criteria. For comparison, heterogeneous Couette flows described with the upper-convected Maxwell or Germann-Cook-Beris models are analyzed. These theoretical flow problems confirm that stark differences in rheological properties across the flow geometry can cause significant fluid retraction and reproduce key features of the experimentally observed retraction. This new method can be used to extract quantitative information about spatially heterogeneous flows in viscoelastic complex fluids, whether or not flow reversal occurs.

A portable EIS-based biosensor for the detection of microcystin-LR residues in environmental water bodies and simulated body fluids.

Due to the eutrophication of water bodies around the world, there is a drastic increase in harmful cyanobacterial blooms leading to contamination of water bodies with cyanotoxins. Chronic exposure to cyanotoxins such as microcystin leads to oxidative stress, inflammation, and liver damage, and potentially to liver cancer. We developed a novel and easy-to-use electrochemical impedance spectroscopy-based immunosensor by fabricating stencil-printed conductive carbon-based interdigitated microelectrodes and immobilising them with cysteamine-capped gold nanoparticles embedded in polyaniline. It has been also coupled with a custom handheld device enabling regular on-site assessment, especially in resource-constrained situations encountered in developing countries. The sensor is able to detect microcystin-LR up to 0.1 µg L-1, having a linear response between 0.1 and 100 µg L-1 in lake and river water and in serum and urine samples. In addition to being inexpensive, easy to fabricate, and sensitive, it also has very good selectivity.

Characterization of an African swine fever virus outbreak in India and comparative analysis of immune genes in infected and surviving crossbreed vs. indigenous Doom pigs.

Since 2020, African swine fever (ASF) has affected all pig breeds in Northeast India except Doom pigs, a unique indigenous breed from Assam and the closest relatives of Indian wild pigs. ASF outbreaks result in significant economic losses for pig farmers in the region. Based on sequencing and phylogenetic analysis of the B646L (p72) gene, it has been determined that ASFV genotype II is responsible for outbreaks in this region. Recent studies have shown that MYD88, LDHB, and IFIT1, which are important genes of the immune system, are involved in the pathogenesis of ASFV. The differential expression patterns of these genes in surviving ASFV-infected and healthy Doom breed pigs were compared to healthy controls at different stages of infection. The ability of Doom pigs to withstand common pig diseases, along with their genetic resemblance to wild pigs, make them ideal candidates for studying tolerance to ASFV infection. In the present study, we investigated the natural resistance to ASF in Doom pigs from an endemic area in Northeast India. The results of this study provide important molecular insights into the regulation of ASFV tolerance genes.

In planta transformation in wheat: an improved protocol to develop wheat transformants.

BACKGROUND: Lack of efficient transformation protocol continues to be a major bottleneck for successful genome editing or transgenic development in wheat. An in planta transformation method was developed in Indian bread wheat in earlier study (Vasil et al. in Nat Biotechnol 10:667-674, 1992) which was labour-intensive and time-consuming. In the present study, in planta transformation method was improved to make it simple, efficient, less labour-intensive and time-saving. METHODS AND RESULTS: PCR-based screening for generated transformants at T0 stage was introduced in this method. Shoot apical meristem of two days old wheat seedling was inoculated with the routine active culture of Agrobacterium tumefaciens harboring plasmid pCAMBIA1300-Ubi-GFP having gene GFP under the control of Zea mays ubiquitin promoter. PCR analysis at T0 stage confirmed 27 plants to be transgene positive. These 27 plants were only taken to the next generation (T1) and the rest were discarded. At T1 generation 6 plants were analyzed to be PCR positive. Out of them, 4 plants were confirmed to have stable integration of transgene (GFP). Fluorescent microscopy at T1 stage confirmed the 4 Southern hybridization positive plants to be expressing reporter gene GFP. CONCLUSIONS: Screening at T0 stage, reduced the load of plants to be taken to T1 generation and their screening thereof at T1 with no overall loss in transformation efficiency. We successfully transformed wheat genotype HD2894 with 3.33% transformation efficiency using a simple, effective method which was less labour-intensive and less time-consuming. This method may be utilized to develop wheat transgenic as well as genome edited lines for desirable traits.

CRISPR/Cas genome editing revealed non-angiogenic role of VEGFA gene in porcine luteal cells: a preliminary report.

BACKGROUND: The angiogenic cytokine vascular endothelial growth factor A (VEGFA) also exerts non-angiogenic effects on endocrine functionality of porcine luteal cells critical for progesterone (P4) production. METHOD AND RESULTS: The expression dynamics of VEGFA-FLT/KDR system were investigated using RT-qPCR during luteal stages and VEGFA gene knock out (KO) porcine luteal cells were generated using CRISPR/Cas9 technology. The downstream effects of VEGFA ablation were studied using RT-qPCR, Annexin V, MTT, ELISA for P4 estimation and scratch wound assay. Bioinformatics analysis of RNA-Seq data of porcine mid-luteal stage was conducted for exploring protein-protein interaction network, KEGG pathways, transcription factors and kinase mapping for VEGFA-FLT/KDR interactomes. The VEGFA-FLT/KDR system expressed throughout the luteal stages with highest expression during mid- luteal stage. Cellular morphology, structure and oil-red-o staining for lipid droplets did not differ significantly between VEGFA KO and wild type cells, however, VEGFA KO significantly decreased (p < 0.05) viability and proliferation efficiency of edited cells on subsequent passages. Expression of apoptotic gene, CASP3 and hypoxia related gene, HIF1A were significantly (p < 0.05) upregulated in KO cells. The relative mRNA expression of VEGFA and steroidogenic genes STAR, CYP11A1 and HSD3B1 decreased significantly (p < 0.05) upon KO, which was further validated by the significant (p < 0.05) decrease in P4 output from KO cells. Bioinformatics analysis mapped VEGFA-FLT/KDR system to signalling pathways associated with steroidogenic cell functionality and survival, which complemented the findings of the study. CONCLUSION: The ablation of VEGFA gene resulted in decreased steroidogenic capability of luteal cells, which suggests that VEGFA exerts additional non-angiogenic regulatory effects in luteal cell functionality.

Trimethylation profile of histones H3 lysine 4 and 9 in late preantral and early antral caprine follicles grown in vivo versus in vitro in the presence of anethole.

This study assessed the histones methylation profile (H3K4me3 and H3K9me3) in late preantral (PA) and early antral (EA) caprine follicles grown in vivo and in vitro, and the anethole effect during in vitro culture of PA follicles. Uncultured in vivo-grown follicles (PA, n = 64; EA, n = 73) were used as controls to assess the methylation profile and genes' expression related to apoptosis cascade (BAX, proapoptotic; BCL2, antiapoptotic), steroidogenesis (CYP17, CYP19A1), and demethylation (KDM1AX1, KDM1AX2, KDM3A). The isolated PA follicles (n = 174) were cultured in vitro for 6 days in α-MEM+ in either absence (control) or presence of anethole. After culture, EA follicles were evaluated for methylation, mRNA abundance, and morphometry. Follicle diameter increased after culture, regardless of treatment. The methylation profile and the mRNA abundance were similar between in vivo-grown PA and EA follicles. Anethole treatment led to higher H3K4me3 fluorescence intensity in EA follicles. The mRNA abundances of BAX, CYP17, and CYP19A1 were higher, and BCL2 and KDM3A were lower in in vitro-grown EA follicles than in vivo-grown follicles. In conclusion, in vitro follicle culture affected H3K4me3 fluorescence intensity, mRNA abundance of apoptotic genes, and steroidogenic and demethylase enzymes compared with in vivo-grown follicles.

Capillary Filling in Open Rectangular Microchannels with a Spatially Varying Contact Angle.

Capillary flow in microchannels is important for many technologies, such as microfluidic devices, heat exchangers, and fabrication of printed electronics. Due to a readily accessible interior, open rectangular microchannels are particularly attractive for these applications. Here, we develop modifications of the Lucas-Washburn model to explore how a spatially varying contact angle influences capillary flow in open rectangular microchannels. Four cases are considered: (i) different uniform contact angles on channel sidewalls and channel bottom, (ii) contact angles varying along the channel cross section, (iii) contact angle varying monotonically along the channel length, and (iv) contact angle varying periodically along the channel length. For case (i), it is found that the maximum filling velocity is more sensitive to changes in the wall contact angle. For case (ii), the contact angles can be averaged to transform the problem into that of case (i). For case (iii), the time evolution of the meniscus position no longer follows the simple square-root law at short times. Finally, for case (iv), the problem is well described by using a uniform contact angle that is a suitable average. These results provide insights into how to design contact-angle variations to control capillary filling and into the influence of naturally occurring contact-angle variations on capillary flow.

Photochemically Induced Marangoni Patterning of Polymer Bilayers.

Surface-tension gradients created along a polymer film by patterned photochemical reactions are a powerful tool for creating surface topography. Here, we use mathematical modeling to explore a strategy for patterning photochemically inactive polymers by coupling a light-sensitive and light-insensitive polymer to form a polymer bilayer. The light-sensitive polymer forms the top layer, and the most dominant surface-tension gradients are introduced at the interface between this layer and air. Lubrication theory is used to derive nonlinear partial differential equations describing the heights of each layer, and linear analysis and nonlinear simulations are performed to characterize interface dynamics. Patterns form at both the polymer-air and polymer-polymer interfaces at early thermal annealing times as a result of Marangoni stresses but decay on prolonged thermal annealing as a result of the dissipative mechanisms of capillary leveling and photoproduct diffusion, thus setting a limit to the maximum individual layer deformation. Simulations also show that the bottom-layer features can remain "trapped", i.e., exhibit no significant decay, even while the top layer topography has dissipated. We study the effects of two key parameters, the initial thickness ratio and the viscosity ratio of the two polymers, on the maximum deformation attained in the bottom layer and the time taken to attain this deformation. We identify regions of parameter space where the maximum bottom-layer deformation is enhanced and the attainment time is reduced. Overall, our study provides guidelines for designing processes to pattern photochemically inactive polymers and create interfacial topography in polymer bilayers.

Postbiotics: an exposition on next generation functional food compounds- opportunities and challenges.

Consumer's interest for health promoting foods has reshaped the food industry to come up with novel biological compounds with diverse health benefits. Postbiotic are the cell fractions, or cell lysates which have emerged as potential functional food compounds during the last decade. The health benefits of postbiotic are well established while attempts are underway to understand their interaction, production, processing and safety. The review explore the challenges and opportunities to devise better growth mediums, cell lysis and extraction, characterization, stability and applications of postbiotics in both food and pharma industry along with the market trends, success stories and safety concerns regarding postbiotics. The scientific and commercial interest in postbiotic have resulted in extensive investigations and clinical documentation of various physiological benefits and additional bioactivity. The findings validate food and pharma application of the postbiotics and further emphasize on documentation of bioactivity and safety of these compounds.

Instability and rupture of surfactant-laden bilayer thin liquid films.

The stability of surfactant-laden bilayer thin films, where the top layer is subject to van der Waals driven breakup, is of particular relevance to applications where one thin liquid layer is spread on another, such as film-forming firefighting foams and multilayer coatings. Although there has been much prior modeling work on the stability of thin liquid bilayers, additional physical effects and assumptions were incorporated in those studies, making it difficult to isolate the influence of surfactant on the rupture of the top layer. The present work addresses this issue through application of the lubrication approximation to derive a coupled system of nonlinear evolution equations describing the perturbations to the liquid-liquid and liquid-air interfaces and the surfactant interfacial concentrations. The surfactant is assumed to be insoluble and can be present at each interface. Linear stability analysis suggests, and nonlinear simulations confirm, that by using surfactant that adsorbs to both interfaces, the rupture time can be increased by an order of magnitude relative to the surfactant-free case. However, we find it crucial to have the right amount of surfactant to generate strongly stabilizing Marangoni stresses without reducing the interfacial tension too much. Nonlinear simulations and linear stability analysis provide insight into the mechanisms of the delayed rupture and show how the direction and strength of the Marangoni stresses strongly depend on the viscosity ratio of the layers. These results can help guide the choice and design of surfactants to achieve more effective firefighting foams and more stable liquid coatings.

AMMI and GGE biplot analysis of yield under terminal heat tolerance in wheat.

BACKGROUND: Wheat is an important cereal crop that helps to meet the food grain needs of people all over the world. Heat stress is one of the most significant abiotic stresses that wheat crops face during terminal growth stages in the wheat growing regions like India. It is very important to identify heat tolerant genotypes to be used as donors for breeding tolerant varieties. METHODS: Thirty-six wheat genotypes were evaluated under different sowing dates viz., Timely sown (TS), Late sown (LS) and very late sown (VLS), and the fourth was sown in the Temperature controlled phenotyping facility (TCPF) across two years. Genotypes were planted following lattice square design with two replications. Data was recorded for yield and yield contributing traits and analysed using selection indices as well AMMI and GGE biplot stability models. RESULTS: Heat stress affected all the traits under different heat environments which ranged from 1.6% (Spikelet number) to 37.2% (grain yield). Regression analysis indicated that the thousand grains weight (R2 = 0.50) contributed significantly towards grain yield under heat stress. Stress susceptibility index (SSI) found genotypes GW322, RAJ3765, Raj4037and MACS6145 as heat tolerant whereas, Stress Tolerance Index (STI) identified C306, HD2967, WH1080, WH730, DBW90, HD2932, DBW17, RAJ3765 as heat tolerant and high yielding. AMMI biplot analysis indicated stable genotypes DBW90, WH730, RAJ4083, CBW38, HD2932, NI5439, WR544, whereas GGE biplot analysis revealed stable genotypes NIAW34, NI5439, RAJ4083, DBW90, PBW590, Raj3765, HUW 510, WH730, HD2967 and UP2382. CONCLUSION: Heat stress affects significantly all yield contributing traits. Thousand grain weight was the most important trait that can be used as a selection criterion for selecting tolerant lines. Based on selection indices and both AMMI and GGE analysis, genotype RAJ3765 was identified to be highly heat tolerant with good grain yield.

Valorised polypropylene waste based reversible sensor for copper ion detection in blood and water.

Heavy metals and plastic pollutants are the two most disastrous challenges to the environment requiring immediate actions. In this work, a techno-commercially feasible approach to address both challenges is presented, where a waste polypropylene (PP) based reversible sensor is produced to selectively detect copper ions (Cu2+) in blood and water from different sources. The waste PP-based sensor was fabricated in the form of an emulsion-templated porous scaffold decorated with benzothiazolinium spiropyran (BTS), which produced a reddish colour upon exposure to Cu2+. The presence of Cu2+ was checked by naked eye, UV-Vis spectroscopy, and DC (Direct Current) probe station by measuring the current where the sensor's performance remained unaffected while analysing blood, water from different sources, and acidic or basic environment. The sensor exhibited 1.3 ppm as the limit of detection value in agreement with the WHO recommendations. The reversible nature of the sensor was determined by cyclic exposure of the sensor towards visible light turning it from coloured to colourless within 5 min and regenerated the sensor for the subsequent analysis. The reversibility of the sensor through exchange between Cu2+- Cu+ was confirmed by XPS analysis. A resettable and multi-readout INHIBIT logic gate was proposed for the sensor using Cu2+ and visible light as the inputs and colour change, reflectance band and current as the output. The cost-effective sensor enabled rapid detection of the presence of Cu2+ in both water and complex biological samples such as blood. While the approach developed in this study provides a unique opportunity to address the environmental burden of plastic waste management, it also allows for the possible valorization of plastics for use in enormous value-added applications.

Layered double hydroxides and their tailored hybrids/composites: Progressive trends for delivery of natural/synthetic-drug/cosmetic biomolecules.

Layered double hydroxides (LDHs) are well-known and important class of hydrotalcite-type anionic clays (HTs) materials that are cost-effective with additional advantages of facile synthesis, composition, tenability, and reusability. These convincing characteristics are liable for their applications in various fields related to energy, environment, catalysis, biomedical, and biotechnology. HTs/LDHs are generally synthesized from low cost abundantly available chemical precursors through the aqueous synthetic pathways under mild reaction conditions. These materials can be termed green materials based on their non-toxic nature, availability of precursors, facile and low-cost production using aqueous medium conditions with less hazardous effluents. Diverse and fascinating characteristics have been attributed to HTs/LDHs like anion exchange ability, surface basicity, biocompatibility, controlled release of the anion specific area, porosity, easy surface modification, and pH dependent biodegradability. Hence, HTs/LDHs and their modified and/or functionalized nanohybrids/nanocomposites are reported as the potential drug delivery carriers with a capability to stabilize the susceptible bioactive molecules, may enhance the solubility of poorly soluble drugs along with controlled drug/bioactive molecule release and delivery. These clay and bioactive hybrid materials have good biocompatibility, less cytotoxicity, and better site-targeting with improved cellular uptake than that of free parent biomolecules. These lamellar solids of micro/nanostructure are compatible, host-guest materials and able to fabricate with drugs/cosmeceutical/bio- or synthetic polymers without any change in their molecular structure and reactivity along with improvement in their stabilities. Other important features are facile synthesis, basicity, high stability with easy storage, and efficient administration with low bio-toxicity. This study enlightens the applications of HTs/LDHs along with their hybrids/composites in the field of drug/cosmeceutical/gene delivery systems of natural/synthetic biomolecules.

Advances in Microscopy and Its Applications with Special Reference to Fluorescence Microscope: An Overview.

The microscope has revolutionized the understanding of an organism's structural details and cellular functions. With the invention of highly evolved microscopes, the diagnosis and treatment of diseases has gained momentum. Technology has immensely helped demonstrate cellular events like phagocytosis, cell movement, cell division, etc. with enhanced temporal and spatial resolution. One of these advanced inventions is the fluorescent microscope which has enabled scanning through various physiological activities of the cell. A fluorescence microscope uses the property of fluorescence to create an image. In addition to visualizing the structural details of the cells, a fluorescence microscope also aids in witnessing cellular activities. With an immunofluorescence microscope, cellular antigens can be localized. This chapter highlights the basics of microscopy, types of microscopes, principles, and types of fluorescence microscopes, and recent advances in microscopy and its application.

Visualization in Anatomy Education.

In the post-pandemic era, one of the significant challenges for anatomy teachers is to reciprocate the experience of practical exposure while teaching the subject to undergraduates. These challenges span from conducting cadaveric dissections to handling real human bones, museum specimens, and tissue sections in the histology lab. Such exposures help the instructors to develop interactive communication with their fellow students and thus help to enhance communication skills among them. Recently, anatomy teachers all over the world started using cutting-edge educational technologies to make teaching-learning experiences for students more engaging, interesting, and interactive. Utilizing such cutting-edge educational technologies was an "option" prior to the pandemic, but the pandemic has significantly altered the situation. What was previously an "option" is now a "compulsion." Despite the fact that the majority of medical schools have resumed their regular on-campus classes, body donation and the availability of cadavers remain extremely limited, resulting in a deadlock. Anatomy teachers must incorporate cutting-edge educational technologies into their teaching and learning activities to make the subject more visual. In this chapter, we have attempted to discuss various new technologies which can provide a near-realistic perception of anatomical structures as a complementary tool for dissection/cadaver, various visualization techniques currently available and explore their importance as a pedagogic alternative in learning anatomy. We also discussed the recent advancement in visualization techniques and the pros and cons of technology-based visualization. This chapter identifies the limitations of technology-based visualization as a supplement and discusses effective utilization as an adjunct to the conventional pedagogical approaches to undergraduate anatomy education.

Androgen receptor gene deficiency results in the reduction of steroidogenic potential in porcine luteal cells.

Luteal steroidogenesis is critical to implantation and pregnancy maintenance in mammals. The role of androgen receptors (AR) in the progesterone (P4) producing luteal cells of porcine corpus luteum (CL) remains unexplored. The aim of the present study was to establish AR gene knock out (KO) porcine luteal cell culture system model by CRISPR/Cas9 genome editing technology and to study the downstream effects of AR gene deficiency on steroidogenic potential and viability of luteal cells. For this purpose, genomic cleavage detection assay, microscopy, RT-qPCR, ELISA, annexin, MTT, and viability assay complemented by bioinformatics analysis were employed. There was significant downregulation (p < 0.05) in the relative mRNA expression of steroidogenic marker genes STAR, CYP11A1, HSD3B1 in AR KO luteal cells as compared to the control group, which was further validated by the significant (p < 0.05) decrease in the P4 production. Significant decrease (p < 0.05) in relative viability on third passage were also observed. The relative mRNA expression of hypoxia related gene HIF1A was significantly (p < 0.05) downregulated in AR KO luteal cells. Protein-protein interaction analysis mapped AR to signaling pathways associated with luteal cell functionality. These findings suggests that AR gene functionality is critical to luteal cell steroidogenesis in porcine.