Measuring Endoplasmic Reticulum Stress and Unfolded Protein Response in HIV-1 Infected T-Cells and Analyzing its Role in HIV-1 Replication.

Viral infections can cause Endoplasmic Reticulum (ER) stress due to abnormal protein accumulation, leading to Unfolded Protein Response (UPR). Viruses have developed strategies to manipulate the host UPR, but there is a lack of detailed understanding of UPR modulation and its functional significance during HIV-1 infection in the literature. In this context, the current article describes the protocols used in our laboratory to measure ER stress levels and UPR during HIV-1 infection in T-cells and the effect of UPR on viral replication and infectivity. Thioflavin T (ThT) staining is a relatively new method used to detect ER stress in the cells by detecting protein aggregates. Here, we have illustrated the protocol for ThT staining in HIV-1 infected cells to detect and quantify ER stress. Moreover, ER stress was also detected indirectly by measuring the levels of UPR markers such as BiP, phosphorylated IRE1, PERK, and eIF2α, splicing of XBP1, cleavage of ATF6, ATF4, CHOP, and GADD34 in HIV-1 infected cells, using conventional immunoblotting and quantitative reverse transcription polymerase chain reaction (RT-PCR). We have found that the ThT-fluorescence correlates with the indicators of UPR activation. This article also demonstrates the protocols to analyze the impact of ER stress and UPR modulation on HIV-1 replication by knockdown experiments as well as the use of pharmacological molecules. The effect of UPR on HIV-1 gene expression/replication and virus production was analyzed by Luciferase reporter assays and p24 antigen capture ELISA, respectively, whereas the effect on virion infectivity was analyzed by staining of infected reporter cells. Collectively, this set of methods provides a comprehensive understanding of the Unfolded Protein Response pathways during HIV-1 infection, revealing its intricate dynamics.

Intricacies of plants' innate immune responses and their dynamic relationship with fungi: A review.

The role of the plant innate immune system in the defense and symbiosis processes becomes integral in a complex network of interactions between plants and fungi. An understanding of the molecular characterization of the plant innate immune system is crucial because it constitutes plants' self-defense shield against harmful fungi, while creating mutualistic relationships with beneficial fungi. Due to the plant-induced awareness and their complexity of interaction with fungi, sufficient assessment of the participation of the plant innate immune system in ecological balance, agriculture, and maintenance of an infinite ecosystem is mandatory. Given the current global challenge, such as the surge of plant-infectious diseases, and pursuit of sustainable forms of agriculture; it is imperative to understand the molecular language of communication between plants and fungi. That knowledge can be practically used in diverse areas, e.g., in agriculture, new tactics may be sought after to try new methods that boost crop receptiveness against fungal pathogens and reduce the dependence on chemical management. Also, it could boost sustainable agricultural practices via enhancing mycorrhizal interactions that promote nutrient absorption and optimum cropping with limited exposure of environmental contamination. Moreover, this review offers insights that go beyond agriculture and can be manipulated to boost plant conservation, environmental restoration, and quality understanding of host-pathogen interactions. Consequently, this specific review paper has offered a comprehensive view of the complex plant innate immune-based responses with fungi and the mechanisms in which they interact.

Genomic Insights into Dementia: Precision Medicine and the Impact of Gene-Environment Interaction.

The diagnosis, treatment, and management of dementia provide significant challenges due to its chronic cognitive impairment. The complexity of this condition is further highlighted by the impact of gene-environment interactions. A recent strategy combines advanced genomics and precision medicine methods to explore the complex genetic foundations of dementia. Utilizing the most recent research in the field of neurogenetics, the importance of precise genetic data in explaining the variation seen in dementia patients can be investigated. Gene-environment interactions are important because they influence genetic susceptibilities and aid in the development and progression of dementia. Modified to each patient's genetic profile, precision medicine has the potential to detect groups at risk and make previously unheard-of predictions about the course of diseases. Precision medicine techniques have the potential to completely transform treatment and diagnosis methods. Targeted medications that target genetic abnormalities will probably appear, providing the possibility for more efficient and customized medical interventions. Investigating the relationship between genes and the environment may lead to preventive measures that would enable people to change their surroundings and minimize the risk of dementia, leading to the improved lifestyle of affected people. This paper provides a comprehensive overview of the genomic insights into dementia, emphasizing the pivotal role of precision medicine, and gene-environment interactions.

HIV-1 replication requires optimal activation of the unfolded protein response.

Several human diseases including viral infections activate the unfolded protein response (UPR) due to abnormal accumulation of unfolded/misfolded proteins. However, UPR modulation and its functional relevance in HIV-1 infection lack comprehensive elucidation. This study reveals that HIV-1 activates IRE1, PERK, and ATF6 signaling pathways of UPR. The knockdown of PERK and ATF6 reduces HIV-1 long terminal repeat (LTR)-driven gene expression, whereas the endoplasmic reticulum (ER) chaperone HSPA5 prevents proteasomal degradation of HIV-1 p24 through its chaperone activity. Interestingly, overstimulation of UPR by a chemical inducer leads to anti-HIV activity through an enhanced type-1 interferon response. Also, treatment with a chemical ER stress inhibitor reduces HIV-1 replication. These findings suggest that an optimal UPR activation is crucial for effective viral replication, as either overstimulating UPR or inhibiting ER stress leads to viral suppression.

Mitotic genome-bookmarking by nuclear hormone receptors: A novel dimension in epigenetic reprogramming and disease assessment.

Arrival of multi-colored fluorescent proteins and advances in live cell imaging has immensely contributed to our understanding of intracellular trafficking of nuclear receptors and their roles in gene regulatory functions. These regulatory events need to be faithfully propagated from progenitor to progeny cells. This is corroborated by multiple converging mechanisms that include histone modifications and lately, the phenomenon of 'mitotic genome-bookmarking' by specific transcription factors. This phenomenon refers to the retention and feed-forward transmission of progenitor's architectural blueprint of active transcription status which is silenced and preserved during mitosis. Upon mitotic exit, this phenomenon ensures accurate reactivation of transcriptome, proteome, cellular traits and phenotypes in the progeny cells. In addition to diverse modes of genome-bookmarking by nuclear receptors, a correlation between disease-associated receptor polymorphism and disruption of this phenomenon is apparent. However, breakthrough technologies shall reveal finer details of this phenomenon to help achieve normalcy in receptor-specific diseases.

Photochemical and gas adsorption studies of Keggin polyoxometalate functionalized porous melamine terephthaldehyde material.

A compound containing a microporous melamine-terephthaldehyde framework is protonated by grinding with acetic acid, resulting in a mesoporous protonated melamine-terephthaldehyde network. The Keggin polyanion [PMo12O40]3- is then immobilized into this protonated melamine-terephthaldehyde network through a solid-state reaction. The polyanion interacts with the protonated microporous organic network through electrostatic interaction. Three different Keggin-melamine-terephthaldehyde materials were synthesized by varying the Keggin anion loading of 10 wt%, 15 wt% and 20 wt%. The Keggin-melamine-terephthaldehyde materials exhibit photochromism on irradiation with sunlight. The photochromism of the POM-organic hybrid material is due to reduction of the Keggin anion. The resulting blue reduced Keggin-melamine-terephthaldehyde materials are oxidized back by treatment with hydrogen peroxide. The N2, CO2 and H2 adsorption properties of all the synthesized materials, including protonated melamine-terephthaldehyde materials, were studied. The materials were characterized by IR, PXRD, DRS, TGA, EPR spectroscopy, and FESEM electron microscopy. The elemental composition was analysed with a CHN analyser and ICP-OES analysis.

Extraction and Encapsulation of Phytocompounds of Poniol Fruit via Co-Crystallization: Physicochemical Properties and Characterization.

Poniol (Flacourtia jangomas) has beneficial health effects due to its high polyphenolic and good antioxidant activity content. This study aimed to encapsulate the Poniol fruit ethanolic extract to the sucrose matrix using the co-crystallization process and analyze the physicochemical properties of the co-crystalized product. The physicochemical property characterization of the sucrose co-crystallized with the Poniol extract (CC-PE) and the recrystallized sucrose (RC) samples was carried out through analyzing the total phenolic content (TPC), antioxidant activity, loading capacity, entrapment yield, bulk and traped densities, hygroscopicity, solubilization time, flowability, DSC, XRD, FTIR, and SEM. The result revealed that the CC-PE product had a good entrapment yield (76.38%) and could retain the TPC (29.25 mg GAE/100 g) and antioxidant properties (65.10%) even after the co-crystallization process. Compared to the RC sample, the results also showed that the CC-PE had relatively higher flowability and bulk density, lower hygroscopicity, and solubilization time, which are desirable properties for a powder product. The SEM analysis showed that the CC-PE sample has cavities or pores in the sucrose cubic crystals, which proposed that the entrapment was better. The XRD, DSC, and FTIR analyses also showed no changes in the sucrose crystal structure, thermal properties, and functional group bonding structure, respectively. From the results, we can conclude that co-crystallization increased sucrose's functional properties, and the co-crystallized product can be used as a carrier for phytochemical compounds. The CC-PE product with improved properties can also be utilized to develop nutraceuticals, functional foods, and pharmaceuticals.

Exploiting the bioactive properties of essential oils and their potential applications in food industry.

Fruits are an abundant source of minerals and nutrients. High nutritional value and easy-to-consume property have increased its demand. In a way to fulfil this need, farmers have increased production, thus making it available for consumers in various regions. This distribution of fruits to various regions deals with many associated problems like deterioration and spoilage. In a way, the common practices that are being used are stored at low temperatures, preservation with chemicals, and many more. Recently, edible coating has emerged as a promising preservation technique to combat the above-mentioned problems. Edible coating stands for coating fruits with bioactive compounds which maintains the nutritional characteristics of fruit and also enhances the shelf life. The property of edible coating to control moisture loss, solute movement, gas exchange, and oxidation makes it most suitable to use. Preservation is uplifted by maintaining the nutritional and physicochemical properties of fruits with the effectiveness of essential oils. The essential oil contains antioxidant, antimicrobial, flavor, and probiotic properties. The utilization of essential oil in the edible coating has increased the property of coating. This review includes the process of extraction, potential benefits and applications of essential oils in food industry.

Light-induced dissolution and concomitant crystallization of a Keggin-type polyoxometalate mimicking a naturally occurring phenomenon.

A suspension of a yellow polycrystalline compound [PPh4]3[PMoVI12O40] in N-methylformanilide (NMF) (in which it is not soluble), on irradiation with sunlight, initiates dissolution via its reduction followed by its crystallization leading to the isolation of single crystals of compound [PPh4]4[PMoVMoVI11O40]·3CH3(C6H5)NCHO (1). Compounds [PPh4]3[PMoVI12O40]·1.75 CH3(C6H5)NCHO (2) and [PPh4]3[PMoVI12O40]·2CH3(C6H5)NCHO (3), each containing an oxidized Keggin anion, are obtained at two different temperatures when the corresponding mother liquor is kept in the dark.

Incidence and Correlates of Early Cognitive Impairment following Intracerebral Haemorrhage.

OBJECTIVES: To determine the incidence of early cognitive impairment following intracerebral haemorrhage. METHODS: A total of 30 adult patients (>18 years) with intracerebral hemorrhage were enrolled in the study. Demographic profile, clinical and radiological profile of the patients was noted. Cognitive status at discharge was assessed using Montreal Cognitive Assessment (MoCA). Data was analyzed using Chi-square and Independent samples 't'-test. RESULTS: Mean age of patients was 63.53±12.11 years. Majority of patients were males (56.7%). At discharge, all the patients had cognitive impairment - majority (76.7%) had moderate cognitive impairment followed by severe impairment (16.7%) and mild impairment (6.7%) respectively. Among different clinicodemographic and radiological factors, only history of tobacco use showed a significant association with severe cognitive impairment. CONCLUSIONS: At discharge mild to moderate cognitive impairment is quite frequent among intracerebral hemorrhage patients irrespective of the demographic, clinical and radiological profile. Further studies on a larger sample size are recommended.

Vaccines for the prevention of diarrhea due to cholera, shigella, ETEC and rotavirus.

BACKGROUND: Diarrhea is a leading cause of mortality in children under 5 years along with its long-term impact on growth and cognitive development. Despite advances in the understanding of diarrheal disorders and management strategies, globally nearly 750,000 children die annually as a consequence of diarrhea. METHODS: We conducted a systematic review of the efficacy and effectiveness studies. We used a standardized abstraction and grading format and performed meta-analyses for all outcomes. The estimated effect of cholera, shigella, Enterotoxigenic Escherichia coli (ETEC) and rotavirus vaccines was determined by applying the standard Child Health Epidemiology Reference Group (CHERG) rules. RESULTS: A total of 24 papers were selected and analyzed for all the four vaccines. Based on the evidence, we propose a 74% mortality reduction in rotavirus specific mortality, 52% reduction in cholera incidence due to their respective vaccines. We did not find sufficient evidence and a suitable outcome to project mortality reductions for cholera, ETEC and shigella in children under 5 years. CONCLUSION: Vaccines for rotavirus and cholera have the potential to reduce diarrhea morbidity and mortality burden. But there is no substantial evidence of efficacy for ETEC and shigella vaccines, although several promising vaccine concepts are moving from the development and testing pipeline towards efficacy and Phase 3 trials.

Bio-composite scaffolds containing chitosan/nano-hydroxyapatite/nano-copper-zinc for bone tissue engineering.

The current study involves fabrication and characterization of bio-composite scaffolds containing chitosan (CS), nano-hydroxyapatite (nHAp) and Cu-Zn alloy nanoparticles (nCu-Zn) by freeze drying technique. The fabricated composite scaffolds (CS/nHAp and CS/nHAp/nCu-Zn) were characterized by SEM, EDX, XRD and FT-IR studies. The addition of nCu-Zn in the CS/nHAp scaffolds significantly increased swelling, decreased degradation, increased protein adsorption, and increased antibacterial activity. The CS/nHAp/nCu-Zn scaffolds had no toxicity towards rat osteoprogenitor cells. So the developed CS/nHAp/nCu-Zn scaffolds have advantageous and potential applications over the CS-nHAp scaffolds for bone tissue engineering.

Chitosan scaffolds containing silicon dioxide and zirconia nano particles for bone tissue engineering.

A scaffold harboring the desired features such as biodegradation, biocompatibility, porous structure could serve as template for bone tissue engineering. In the present study, chitosan (CS), nano-scaled silicon dioxide (Si) and zirconia (Zr) were combined by freeze drying technique to fabricate a bio-composite scaffold. The bio-composite scaffold (CS/Si/Zr) was characterized by SEM, XRD and FT-IR studies. The scaffold possessed a porous nature with pore dimensions suitable for cell infiltration and colonization. The presence of zirconia in the CS/Si/Zr scaffold decreased swelling and increased biodegradation, protein adsorption and bio-mineralization properties. The CS/Si/Zr scaffold was also found to be non-toxic to rat osteoprogenitor cells. Thus, we suggest that CS/Si/Zr bio-composite scaffold is a potential candidate to be used for bone tissue engineering.

Preparation, characterization and antimicrobial activity of a bio-composite scaffold containing chitosan/nano-hydroxyapatite/nano-silver for bone tissue engineering.

In this study, a bio-composite scaffold containing chitosan/nano-hydroxyapatite/nano-silver particles (CS/nHAp/nAg) was developed by freeze drying technique, followed by introduction of silver ions in controlled amount through reduction phenomenon by functional groups of chitosan. The scaffolds were characterized using SEM, FT-IR, XRD, swelling, and biodegradation studies. The testing of the prepared scaffolds with Gram-positive and Gram-negative bacterial strains showed antibacterial activity. The scaffold materials were also found to be non-toxic to rat osteoprogenitor cells and human osteosarcoma cell line. Thus, these results suggested that CS/nHAp/nAg bio-composite scaffolds have the potential in controlling implant associated bacterial infection during reconstructive surgery of bone.