Evaluation of simplified ester-linked fatty acid analysis (ELFA) for phospholipid fatty acid (PLFA) analysis of bacterial population.

Phospholipid fatty acid (PLFA) analysis is used for characterizing microbial communities based on their lipid profiles. This method avoids biases from PCR or culture, allowing data collection in a natural state. However, PLFA is labor-intensive due to lipid fractionation. Simplified ester-linked fatty acid analysis (ELFA), which skips lipid fractionation, offers an alternative. It utilizes base-catalyzed methylation to derivatize only lipids, not free fatty acids, and found glycolipid and neutral lipid fractions are scarcely present in most bacteria, allowing lipid fractionation to be skipped. ELFA method showed a high correlation to PLFA data (r = 0.99) and higher sensitivity than the PLFA method by 1.5-2.57-fold, mainly due to the higher recovery of lipids, which was 1.5-1.9 times higher than with PLFA. The theoretical limit of detection (LOD) and limit of quantification (LOQ) for the ELFA method indicated that 1.54-fold less sample was needed for analysis than with the PLFA method. Our analysis of three bacterial cultures and a simulated consortium revealed the effectiveness of the ELFA method by its simple procedure and enhanced sensitivity for detecting strain-specific markers, which were not detected in PLFA analysis. Overall, this method could be easily used for the population analysis of synthetic consortia.

B cell senescence promotes age-related changes in oral microbiota.

In recent years, there has been increasing attention towards understanding the relationship between age-related alterations in the oral microbiota and age-associated diseases, with reports emphasizing the significance of maintaining a balanced oral microbiota for host health. However, the precise mechanisms underlying age-related changes in the oral microbiota remain elusive. We recently reported that cellular senescence of ileal germinal center (GC) B cells, triggered by the persistent presence of commensal bacteria, results in diminished IgA production with aging and subsequent alterations in the gut microbiota. Consequently, we hypothesize that a similar phenomenon may occur in the oral cavity, potentially contributing to age-related changes in the oral microbiota. Examination of p16-luc mice, wherein the expression of the senescent cell marker p16INK4a can be visualized, raised under specific pathogen-free (SPF) or germ-free (GF) conditions, indicated that, unlike ileal GC B cells, the accumulation of senescent cells in GC B cells of cervical lymph nodes increases with age regardless of the presence of commensal bacteria. Furthermore, longitudinal studies utilizing the same individual mice throughout their lifespan revealed concurrent age-related alterations in the composition of the oral microbiota and a decline in salivary IgA secretion. Further investigation involving Rag1-/- mice transplanted with B cells from wild-type or p16INK4a and p21Waf1/Cip1 -double knockout mice unveiled that B cell senescence leads to reduced IgA secretion and alteration of the oral microbiota. These findings advance our understanding of the mechanism of age-associated changes in the oral microbiota and open up possibilities of their control.

Positive effect of phasin in biohydrogen production of non polyhydroxybutyrate-producing Clostridium acetobutylicum ATCC 824.

In this study, the goal was to enhance the tolerance of Clostridium acetobutylicum ATCC 824 to biomass-based inhibitory compounds for biohydrogen production and evaluate various known genes that enhance the production of biochemicals in various hosts. The introduction of phaP, the major polyhydroxyalkanoate granule-associated protein that has been reported as a chaperone-like protein resulted in increased tolerance to inhibitors and leads to higher levels of hydrogen production, cell growth, and glucose consumption in the presence of these inhibitors. It was observed that the introduction of phaP led to an increase in the transcription of the hydrogenase gene, whereas transcription of the chaperone functional genes decreased compared to the wild type. Finally, the introduction of phaP could significantly enhance biohydrogen production by 2.6-fold from lignocellulosic hydrolysates compared to that of wild type. These findings suggested that the introduction of phaP could enhance growth and biohydrogen production, even in non-polyhydroxyalkanoate-producing strains.

A radioactive CRISPR interference system using 89Zr-labeled LbCas12a.

Recently, CRISPR proteins have been recognized as promising candidates for drug development. However, there is still a lack of substances with the appropriate sensitivity and stability for targeted drug delivery systems. 89Zr is a radioactive isotope that emits positrons, allowing real-time in vivo tracking with proven safety. In this study, we confirmed that labeling with 89Zr did not compromise the functionality of CRISPR proteins during in vivo behavioral imaging. Furthermore, we demonstrated the therapeutic efficacy of the CRISPR interference system in a mouse model of liver fibrosis, highlighting the theragnostic potential of isotope-labeled CRISPR proteins. The findings of this research could contribute to various aspects of ongoing clinical studies exploring the in vivo applications of CRISPR proteins.

Pan-Nox inhibitor treatment improves renal function in aging murine diabetic kidneys.

Background: Aging is a risk factor for development of chronic kidney disease and diabetes mellitus with commonly shared features of chronic inflammation and increased oxidative stress. Here, we investigated the effect of pan-Nox-inhibitor, APX-115, on renal function in aging diabetic mice. Methods: Diabetes was induced by intraperitoneal injection of streptozotocin at 50 mg/kg/day for 5 days in 52-week-old C57BL/6J mice. APX-115 was administered by oral gavage at a dose of 60 mg/kg/day for 12 weeks in nondiabetic and diabetic aging mice. Results: APX-115 significantly improved insulin resistance in diabetic aging mice. Urinary level of 8-isoprostane was significantly increased in diabetic aging mice than nondiabetic aging mice, and APX-115 treatment reduced 8-isoprostane level. Urinary albumin and nephrin excretion were significantly higher in diabetic aging mice than nondiabetic aging mice. Although APX-115 did not significantly decrease albuminuria, APX-115 markedly improved mesangial expansion, macrophage infiltration, and expression of fibrosis molecules such as transforming growth factor beta 1 and plasminogen activator inhibitor 1. Interestingly, the expression of all Nox isoforms including Nox1, Nox2, and Nox4 was significantly increased in diabetic aging kidneys, and APX-115 treatment decreased Nox1, Nox2, and Nox4 protein expression in the kidney. Furthermore, Klotho expression was significantly decreased in diabetic aging kidneys, and APX-115 restored Klotho level. Conclusion: Our results provide evidence that pan-Nox inhibition may improve systemic insulin resistance and decrease oxidative stress, inflammation, and fibrosis in aging diabetic status and may have potential protective effects on aging diabetic kidney.

Repercussion of extracellular polymeric removal by low-temperature calcium peroxide pretreatment on bacterial fragmentation for enhancing biohydrogen production.

This study envisioned attaining the percipience of effective biohydrogen production from paper mill waste-activated sludge through low-temperature calcium peroxide-mediated bacterial pretreatment (TCP-BP). Floc dissociation with limited cell destruction was attained at a calcium peroxide dosage of 0.05 g/g suspended solids (SS) at 70 °C temperature. This TCP-BP method improves bacterial fragmentation, and very high SS solubilization was achieved at 42 h, with the solubilization and solid reduction of 18.6% and 14.1%, respectively. BP-only pretreatment shows lower solubilization efficiency of 9.4% than TCP-BP pretreatment due to the presence of flocs, which inhibit the enzymatic action during bacterial fragmentation. A biohydrogen test shows a high biohydrogen potential of 94.1 mL H2/gCOD for the TCP-BP sample, which is higher than that of the BP-only and control samples. According to the findings, low-temperature calcium peroxide-mediated bacterial fragmentation is validated to be an efficient process for sludge degradation and biohydrogen production.

Green chemical and hybrid enzymatic pretreatments for lignocellulosic biorefineries: Mechanism and challenges.

The greener chemical and enzymatic pretreatments for lignocellulosic biomasses are portraying a crucial role owing to their recalcitrant nature. Traditional pretreatments lead to partial degradation of lignin and hemicellulose moieties from the pretreated biomass. But it still restricts the enzyme accessibility for the digestibility towards the celluloses and the interaction of lignin-enzymes, nonproductively. Moreover, incursion of certain special chemical treatments and other lignin sulfonation techniques to the enzymatic pretreatment (hybrid enzymatic pretreatment) enhances the lignin structural modification, solubilization of the hemicelluloses and both saccharification and fermentation processes (SAF). This article concentrates on recent developments in various chemical and hybrid enzymatic pretreatments on biomass materials with their mode of activities. Furthermore, the issues on strategies of the existing pretreatments towards their industrial applications are highlighted, which could lead to innovative ideas to overcome the challenges and give guideline for the researchers towards the lignocellulosic biorefineries.

Enhancement of biohydrogen production in Clostridium acetobutylicum ATCC 824 by overexpression of glyceraldehyde-3-phosphate dehydrogenase gene.

In the dark fermentation of hydrogen, development of production host is crucial as bacteria act on substrates and produce hydrogen. The present study aimed to improve hydrogen production through the development of Clostridium acetobutylicum as a superior biohydrogen producer. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which produces NADH/NADPH for metabolites and energy in primary pathways, was introduced to enhance hydrogen production. The strain CAC824-G containing gapC that encodes GAPDH showed a 66.3 % higher hydrogen production than the wild-type strain, with increased NADH and NADPH pools. Glucose consumption and other byproducts, such as acetone, butanol, and ethanol, were also high in CAC824-G. Overexpression of gapC resulted in increased hydrogen production with sugars obtained from different biomass, even in the presence of inhibitors such as vanillin, 5-hydroxymethylfufural, acetic acid, and formic acid. Our results imply that overexpression of gapC in Clostridium is possible to expand the production of the reported biochemicals to produce hydrogen.

Unraveling Biohydrogen Production and Sugar Utilization Systems in the Electricigen Shewanella marisflavi BBL25.

Identification of novel, electricity-producing bacteria has garnered remarkable interest because of the various applications of electricigens in microbial fuel cell and bioelectrochemical systems. Shewanella marisflavi BBL25, an electricity-generating microorganism, uses various carbon sources and shows broader sugar utilization than the better-known S. oneidensis MR-1. To determine the sugar-utilizing genes and electricity production and transfer system in S. marisflavi BBL25, we performed an in-depth analysis using whole-genome sequencing. We identified various genes associated with carbon source utilization and the electron transfer system, similar to those of S. oneidensis MR-1. In addition, we identified genes related to hydrogen production systems in S. marisflavi BBL25, which were different from those in S. oneidensis MR-1. When we cultured S. marisflavi BBL25 under anaerobic conditions, the strain produced 427.58 ± 5.85 µl of biohydrogen from pyruvate and 877.43 ± 28.53 µl from xylose. As S. oneidensis MR-1 could not utilize glucose well, we introduced the glk gene from S. marisflavi BBL25 into S. oneidensis MR-1, resulting in a 117.35% increase in growth and a 17.64% increase in glucose consumption. The results of S. marisflavi BBL25 genome sequencing aided in the understanding of sugar utilization, electron transfer systems, and hydrogen production systems in other Shewanella species.

Inhibiting bacterial biofilm formation by stimulating c-di-GMP regulation using citrus peel extract from Jeju Island.

Biofilms consist of single or multiple species of bacteria embedded in extracellular polymeric substances (EPSs), which affect the increase in antibiotic resistance by restricting the transport of antibiotics to the bacterial cells. An alternative approach to treatment with antimicrobial agents is using biofilm inhibitors that regulate biofilm development without inhibiting bacterial growth. In this study, we found that citrus peel extract from Jeju Island (CPEJ) can inhibit bacterial biofilm formation. According to the results, CPEJ concentration-dependently reduces biofilm formation without affecting bacterial growth. Additionally, CPEJ decreased the production of extracellular polymeric substances but increased bacterial swarming motility. These results led to the hypothesis that CPEJ can reduce intracellular bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) concentration. The results showed that CPEJ significantly reduced the c-di-GMP level through increased phosphodiesterase activity. Altogether, these findings suggest that CPEJ as a biofilm inhibitor has new potential for pharmacological (e.g. drug and medication) and industrial applications (e.g. ship hulls, water pipes, and membrane processes biofouling control).

In vivo tracking of toxic diesel particulate matter in mice using radiolabeling and nuclear imaging.

Exposure to diesel particulate matter (DPM) is associated with several adverse health effects, including severe respiratory diseases. Quantitative analysis of DPM in vivo can provide important information on the behavior of harmful chemicals, as well as their toxicological impacts in living subjects. This study presents whole-body images and tissue distributions of DPM in animal models, using molecular imaging and radiolabeling techniques. The self-assembly of the 89Zr-labeled pyrene analog with a suspension of DPM efficiently produced 89Zr-incorporated DPM (89Zr-DPM). Positron emission tomography images were obtained for mice exposed to 89Zr-DPM via three administration routes: intratracheal, oral, and intravenous injection. DPM was largely distributed in the lungs and only slowly cleared after 7 days in mice exposed via the intratracheal route. In addition, a portion of 89Zr-DPM was translocated to other organs, such as the heart, spleen, and liver. Uptake values in these organs were also noticeable following exposure via the intravenous route. In contrast, most of the orally administered DPM was excreted quickly within a day. These results suggest that continuous inhalation exposure to DPM causes serious lung damage and may cause toxic effects in the extrapulmonary organs.

The lysosomal Ragulator complex activates NLRP3 inflammasome in vivo via HDAC6.

The cellular activation of the NLRP3 inflammasome is spatiotemporally orchestrated by various organelles, but whether lysosomes contribute to this process remains unclear. Here, we show the vital role of the lysosomal membrane-tethered Ragulator complex in NLRP3 inflammasome activation. Deficiency of Lamtor1, an essential component of the Ragulator complex, abrogated NLRP3 inflammasome activation in murine macrophages and human monocytic cells. Myeloid-specific Lamtor1-deficient mice showed marked attenuation of NLRP3-associated inflammatory disease severity, including LPS-induced sepsis, alum-induced peritonitis, and monosodium urate (MSU)-induced arthritis. Mechanistically, Lamtor1 interacted with both NLRP3 and histone deacetylase 6 (HDAC6). HDAC6 enhances the interaction between Lamtor1 and NLRP3, resulting in NLRP3 inflammasome activation. DL-all-rac-α-tocopherol, a synthetic form of vitamin E, inhibited the Lamtor1-HDAC6 interaction, resulting in diminished NLRP3 inflammasome activation. Further, DL-all-rac-α-tocopherol alleviated acute gouty arthritis and MSU-induced peritonitis. These results provide novel insights into the role of lysosomes in the activation of NLRP3 inflammasomes by the Ragulator complex.

Elucidation of microbial interactions, dynamics, and keystone microbes in high pressure anaerobic digestion.

High-pressure anaerobic digestion (HPAD) is a promising technology for producing biogas enriched with high methane content in a single-step process. To enhance HPAD performance, a comprehensive understanding of microbial community dynamics and their interactions is essential. For this, mesophilic batch high-pressurized anaerobic reactors were operated under 3 bars (H3) and 6 bars (H6). The experimental results showed that the effect of high-pressure (up to 6 bar) on acidification was negligible while methanogenesis was significantly delayed. Microbial analysis showed the predominance of Defluviitoga affiliated with the phylum Thermotogae and the reduction of Thiopseudomonas under high-pressure conditions. In addition, the microbial cluster pattern in H3 and H6 was significantly different compared to the CR, indicating a clear shift in microbial community structure. Moreover, Methanobacterium, Methanomicrobiaceae, Alkaliphilus, and Petrimonas were strongly correlated in network analysis, and they could be identified as keystone microbes in the HPAD reactor.

Theranostics through Utilizing Cherenkov Radiation of Radioisotope Zr-89 with a Nanocomposite Combination of TiO2 and MnO2.

Cherenkov radiation (CR) derived from the decay of diagnostic and therapeutic radionuclides is currently being studied by the scientific community to determine if these emissions can be harnessed for cancer detection and therapy. While Cherenkov luminescence imaging (CLI) has been studied in the preclinical and clinical settings, Cherenkov radiation-induced cancer therapy (CRICT) is a relatively new area of research that harnesses the emitted photons to kill cancer cells through free radical generation and DNA damage. Nanoparticles seem well suited for developing a theranostic platform that would allow researchers to visualize therapy delivery and also generate the reactive oxygen species necessary to kill cancer cells. Herein, we report the preparation of an 89Zr-TiO2-MnO2 nanocomposite that incorporates transferrin onto the nanoparticle surface to enhance cancer cell growth inhibition. The incorporation of the positron emission tomography (PET) radioisotope 89Zr (half-life: 3.3 days) allowed for the detection of the nanoparticle using PET and for the creation of Cherenkov emissions that interacted with the nanoparticle surface to generate free radicals for therapy delivery. After preparation, these systems were observed to be stable in various media and provided excellent tumor growth control after being intratumorally injected into mice bearing CT-26 tumors. These results demonstrate that a therapeutically efficient CRICT platform can be generated using commercially available and affordable materials.

Technological advances in the production of carotenoids and their applications- A critical review.

Carotenoids are naturally occurring pigments that are widely distributed in algae, fungi, bacteria, and plants. Carotenoids play a significant role in the food, feed, cosmetic, nutraceutical, and pharmaceutical industries. These pigments are effectively considered as a health-promoting compounds, which are widely used in our daily diet to reduce the risk of chronic diseases such as cardiovascular diseases, cancer, acute lung injury, cataracts, neural disorders, etc. In this context, this review paper demonstrates the synthesis of carotenoids and their potential application in the food and pharmaceutical industries. However, the demand for carotenoid production is increasing overtime, and the extraction and production are expensive and technically challenging. The recent developments in carotenoid biosynthesis, and key challenges, bottlenecks, and future perspectives were also discussed to enhance the circular bioeconomy.

Enhanced tolerance of Cupriavidus necator NCIMB 11599 to lignocellulosic derived inhibitors by inserting NAD salvage pathway genes.

Polyhydroxybutyrate (PHB) is a bio-based, biodegradable and biocompatible plastic that has the potential to replace petroleum-based plastics. Lignocellulosic biomass is a promising feedstock for industrial fermentation to produce bioproducts such as polyhydroxybutyrate (PHB). However, the pretreatment processes of lignocellulosic biomass lead to the generation of toxic byproducts, such as furfural, 5-HMF, vanillin, and acetate, which affect microbial growth and productivity. In this study, to reduce furfural toxicity during PHB production from lignocellulosic hydrolysates, we genetically engineered Cupriavidus necator NCIMB 11599, by inserting the nicotine amide salvage pathway genes pncB and nadE to increase the NAD(P)H pool. We found that the expression of pncB was the most effective in improving tolerance to inhibitors, cell growth, PHB production and sugar consumption rate. In addition, the engineered strain harboring pncB showed higher PHB production using lignocellulosic hydrolysates than the wild-type strain. Therefore, the application of NAD salvage pathway genes improves the tolerance of Cupriavidus necator to lignocellulosic-derived inhibitors and should be used to optimize PHB production.

Phase separated pretreatment strategies for enhanced waste activated sludge disintegration in anaerobic digestion: An outlook and recent trends.

A significant ecological problem was developed on disposing the enormous amounts of waste activated sludge (WAS) produced by traditional wastewater treatment. There have been various attempts recently originated to develop innovative methods for substantial sludge treatment. The most frequently used approach for treating sludge to produces methane and reduces sludge is anaerobic treatment. The hydrolysis phase in WAS limits the breakdown of complex macrobiotic compounds. The presence of extracellular polymeric substances (EPS) in biomass prevents the substrate from being hydrolyzed. Enhancing substrate hydrolysis involves removal of EPS preceded by phase separated pretreatment. Hence, a critical assessment of various phase separated pretreatment that has a remarkable effect on the anaerobic digestion process was documented in detail. Moreover, the economic viability and energy requirement of this treatment process was also discussed. Perspectives and recommendations for methane production were also provided to attain effectual sludge management.

Theragnostic 64Cu/67Cu Radioisotopes Production With RFT-30 Cyclotron.

64Cu and 67Cu are theragnostic pair radionuclides with promising application in the nuclear medicine. 64Cu is PET nuclide for the non-invasive diagnosis and 67Cu is beta emitter for therapy of various cancers. This study discusses optimization efforts in the production of these radioactive coppers carried out with 30 MeV cyclotron. Optimized conditions include target preparation, chemical separation, and quality control. The production routes of 64Cu and 67Cu were studied based on the nuclear reactions of 64Ni(p,n)64Cu and 70Zn(p,α)67Cu. The produced 64Cu and 67Cu have >99.9% of the radionuclidic purity. The yield at the end of bombardment (EOB) of 64Cu and 67Cu is 28.5 MBq/µAh and 67Cu is 0.58 MBq/µAh, respectively.

Acceleration of lactate-utilizing pathway for enhancing biohydrogen production by magnetite supplementation in Clostridium butyricum.

A conductive metal compound can be used as a catalyst for enhancing hydrogen production by dark fermentation. This study aimed to identify mechanisms of enhanced hydrogen production by magnetite supplementation. Experiments were performed with lactate and/or magnetite supplementation to confirm that the lactate-utilizing pathway is the key cause of enhanced hydrogen production. Also, ribonucleic acid sample was collected for monitoring gene regulation under each condition. Hydrogen production was significantly enhanced by approximately 25.6% and 58.9%, respectively, via magnetite alone and with lactate. Moreover, the expression of genes involved in hydrogen production, including pyruvate ferredoxin oxidoreductase, hydrogenase, and ferredoxin, via magnetite alone and with lactate was upregulated by 0.26, 0.71, and 3.50 and 1.06, 2.14, and 1.94 times, respectively.

Surfactant induced microwave disintegration for enhanced biohydrogen production from macroalgae biomass: Thermodynamics and energetics.

This research work aimed about the enhanced bio-hydrogen production from marine macro algal biomass (Ulva reticulate) through surfactant induced microwave disintegration (SIMD). Microwave disintegration (MD) was performed by varying the power from 90 to 630 W and time from 0 to 40 min. The maximum chemical oxygen demand (COD) solubilisation of 27.9% was achieved for MD at the optimal power (40%). A surfactant, ammonium dodecyl sulphate (ADS) is introduced in optimal power of MD which enhanced the solubilisation to 34.2% at 0.0035 g ADS/g TS dosage. The combined SIMD pretreatment significantly reduce the treatment time and increases the COD solubilisation when compared to MD. Maximum hydrogen yield of 54.9 mL H2 /g COD was observed for SIMD than other samples. In energy analysis, it was identified that SIMD was energy efficient process compared to others since SIMD achieved energy ratio of 1.04 which is higher than MD (0.38).