Bacteriophages are increasingly explored as antibacterial agents for therapeutic applications and pathogen biocontrol. Rigorous quality control and removal of microbial growth byproducts is essential for safety and compliance with regulatory standards for the use of phages; for phages isolated from Gram-negative bacterial lysates, this involves removal of endotoxin (lipopolysaccharide/LPS) and contaminating host proteins. Here, we describe the development of weakly hydrophobic hydroxyl-functional monolithic chromatography (CIMmultus-OH) as a single-step purification process for five tailed anti-pseudomonal phages, including myoviral and siphoviral morphologies, as well as extra-large bacteriophages so-called "jumbo" phages, yielding therapeutically compliant samples suitable for human intravenous administration. Using a salt-driven preferential exclusion chromatography with a potassium phosphate gradient, we achieved near-complete removal of endotoxin (99.98-≥99.99% depletion) and protein impurities (99.12-99.83% depletion). We found that phage recovery post-purification was inversely associated with tail length and effective particle size (hydrodynamic diameter), with shorter-tailed and smaller phages demonstrating greater recovery. We conclude that single-step CIMmultus OH-monolithic chromatography is scalable, reproducible, and free from hazardous chemicals, supporting its integration into industrial phage purification processes.

Oxcarbazepine is a widely prescribed anticonvulsant used in the management of epilepsy through modulation of abnormal neuronal excitability. The objective of the present study was to synthesize oxcarbazepine in the laboratory and develop a reliable, stability-indicating, and environmentally conscious reverse-phase high-performance liquid chromatography (RP-HPLC) method for its quality assessment, including the quantification of impurities and degradation products. Oxcarbazepine was synthesized and characterized using infrared spectroscopy (IR), nuclear magnetic resonance (NMR), and high-performance liquid chromatography (HPLC) techniques. Chromatographic separation was achieved using a Supelco Discovery C18 column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of 75% potassium phosphate monobasic buffer (pH 5.0) and 25% acetonitrile at a flow rate of 1.0 mL/min. Detection was performed at 257 nm, and oxcarbazepine exhibited a retention time of approximately 11 min. The developed RP-HPLC method was validated according to ICH, USP, and FDA guidelines. Forced degradation studies were conducted under acidic, basic, oxidative, thermal, and photolytic conditions to evaluate the stability-indicating capability of the method. In addition, the environmental impact of the analytical procedure was assessed using Green Analytical Chemistry (GAC) metrics, including the Analytical Eco-Scale, GAPI, and AGREE tools. The developed method demonstrated excellent specificity, linearity, accuracy, precision, robustness, and sensitivity. All degradation products were well resolved from the parent oxcarbazepine peak, confirming the stability-indicating nature of the method. Excellent linearity was observed over the concentration range of 0.24-2000 ppm with a correlation coefficient (R2) of 0.9995. Accuracy values ranged from 99.4% to 101.5%, while the limit of detection (LOD) and limit of quantification (LOQ) were 0.3 ppm and 0.9 ppm, respectively, indicating high sensitivity suitable for trace-level analysis. Robustness studies confirmed the reliability of the method under deliberate variations in chromatographic conditions. Furthermore, greenness assessment revealed an Analytical Eco-Scale score of approximately 75, a GAPI pictogram with predominantly green and yellow zones, and an AGREE score of 0.57/1.0, indicating moderate environmental friendliness. Overall, the validated RP-HPLC method provides a reliable, precise, sensitive, stability-indicating, and environmentally conscious analytical approach for routine quality control, impurity profiling, and stability studies of oxcarbazepine in raw materials and pharmaceutical formulations.

Antimicrobial resistance and biofilm-associated infections continue to undermine standard antibiotic therapies, prompting the need for physicochemical adjuvant strategies. In this study, we evaluated the antibiofilm and antimicrobial activity of 7 biological buffer systems including citrate, acetate, sodium phosphate, potassium phosphate, borate, carbonate, and Tris-buffered saline against methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MSSA and MRSA). Using a range of biofilm, metabolic, and cytotoxicity assays coupled with bioluminescence imaging, we demonstrate that low-concentration (20 mM) buffers significantly disrupted metabolic activity and biofilm structure within minutes of exposure as indicated by rapid functional perturbations observed via bioluminescence and without cytotoxicity in human dermal fibroblasts. Among the tested solutions, citrate emerged as the most potent antibiofilm agent (>50% reduction), while acetate, borate, and phosphate systems also demonstrated significant antibiofilm activity (25-50% reduction). MRSA exhibited reduced sensitivity to buffer treatment, consistent with known stress-response adaptations. Biofilm reduction >50% was recapitulated in three functionalized brushite cement formulations prepared with selected buffer systems, supporting the feasibility of incorporating chemically active ionic additives into calcium phosphate biomaterials. These findings highlight the potential of ionic systems as non-antibiotic antimicrobial agents and therapeutic adjuvants. Strategic integration into biomaterial design or wound irrigation strategies carries significant promise as a non-antibiotic strategy to combat infection. STATEMENT OF SIGNIFICANCE: Biofilm-associated infections remain difficult to treat because bacteria in biofilms are far less responsive to antibiotics. This study shows that several simple biological buffer systems, especially citrate, can rapidly disrupt Staphylococcus aureus biofilms at low concentrations without harming human dermal fibroblasts. Beyond identifying these ionic solutions as non-antibiotic antibiofilm agents, the work also shows that their activity can be translated to calcium phosphate cements, creating biomaterials with infection-modulating properties. This is significant because it introduces a simple and scalable strategy to design antimicrobial biomaterials without relying on conventional antibiotics, metallic agents, or toxic additives, which is of broad interest to researchers developing safer materials for infection control and regenerative medicine.

Rasburicase (brand name Elitek) is a uric oxidase used to treat the high levels of uric acid that are associated with tumor lysis syndrome (TLS). Tumor lysis syndrome is a potentially life-threatening condition caused by rapid break down of tumor cells during chemotherapy. Tumor lysis syndrome is associated with the treatment of aggressive lymphoma and leukemia, but it may also occur with other tumors including solid tumors. Massive cell breakdown results in the release of potassium, phosphate, and uric acid into the circulation. Urate crystals can precipitate in the kidneys, causing acute kidney damage. Prophylaxis and treatment of TLS involve aggressive intravenous (IV) hydration and the use of drugs to lower uric acid levels. Rasburicase breaks down uric acid to a more soluble metabolite (allantoin), which is then eliminated by the kidneys. A by-product of this reaction is hydrogen peroxide, an oxidizing agent. Red blood cells that lack the enzyme glucose-6-phosphate dehydrogenase (G6PD) are sensitive to oxidative damage caused by agents like hydrogen peroxide due to a deficiency in nicotinamide adenine dinucleotide phosphate (NADPH). Once exposed, the red blood cells become rigid, trapped, and are rapidly broken down (hemolysis). This can lead to a deficiency of mature red blood cells (hemolytic anemia) and the production of red blood cells with abnormally high levels of methemoglobin (methemoglobinemia). Approximately 400 million people worldwide have G6PD deficiency. Most of these individuals are asymptomatic. However, they are at risk of life-threating hemolytic reactions and methemoglobinemia if given oxidizing drugs such as rasburicase. Rasburicase is contraindicated in individuals with G6PD deficiency. The FDA-approved drug label states that individuals at higher risk for G6PD deficiency should be screened before starting rasburicase therapy, for example, individuals of African or Mediterranean ancestry (Table 1) (1). Approximately 12% of African-Americans have G6PD deficiency. A rare cause of methemoglobinemia is a deficiency of antioxidant enzymes such as cytochrome b5 reductase 3 (CYB5R3). The drug label states it is not known whether individuals who have a deficiency of this enzyme, or another enzyme with antioxidant activity, are at increased risk of methemoglobinemia or hemolytic anemia during rasburicase therapy. The Clinical Pharmacogenetics Implementation Consortium (CPIC) has published dosing recommendations for rasburicase based on G6PD phenotype (Table 2). The CPIC states for individuals with normal G6PD phenotype, there is no reason to withhold rasburicase based on G6PD status. For individuals who are G6PD deficient, with or without hemolytic anemia, rasburicase is contraindicated. For individuals who have a variable G6PD phenotype, G6PD enzyme activity must be measured to ascertain that G6PD status is normal. For cases where rasburicase is contraindicated, alternative drugs include allopurinol (2).

Tricalcium silicate (TCS) is a clinically established calcium silicate-based material. However, its highly alkaline hydration environment limits cellular compatibility. This study investigated whether direct incorporation of phosphate solution during TCS hydration promotes calcium-deficient hydroxyapatite (CDHAp) formation, modulates solution chemistry, and enhances osteogenic biological responses. TCS was mixed with KH2PO4 solutions at 0, 5, 10, 15, and 20 wt% (TP0-TP20) at a fixed liquid-to-powder ratio. Crystalline phase evolution was characterised by X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) after simulated body fluid (SBF) immersion for up to 28 days. Surface morphology was examined by scanning electron microscopy (SEM). Solution pH, solubility, and ion release (Ca2+, Si4+, PO43-) were quantified by ICP-OES. Osteogenic gene expression (RUNX2, OSX, COL1A1, OPN) was evaluated in MC3T3-E1 pre-osteoblasts by RT-qPCR. Cell viability and migration under physiological perfusion were assessed using a custom microfluidic Lab-on-a-Chip system. XRD and XPS confirmed concentration-dependent CDHAp formation in TP15 and TP20 from day 1, with peak intensity increasing progressively through day 28. TP0 showed CDHAp formation was insufficient to be clearly detected by XRD. Higher phosphate content progressively reduced solution pH and suppressed free Ca2+ accumulation, maintaining levels within 2-6 mM at early time points while elevating Si4+ release. TP15 and TP20 significantly upregulated RUNX2, OSX, and COL1A1 compared with TP0 (p < 0.05). Under microfluidic system, TP0 supported no viable cells, whereas TP20 demonstrated directional cell migration toward the specimen surface. Incorporating phosphate solution during TCS hydration accelerates CDHAp crystallisation, moderates excessive alkalinity, and enhances early osteogenic differentiation and cell migration under flow. These findings establish phosphate-modified TCS as a strategy to enhance HAp formation with enhanced chemical and biological properties.

The purpose of this research is to develop a fast, environmentally friendly, precise, sensitive, and specific high-performance liquid chromatography (HPLC) method for the simultaneous determination of dissolution rate for valsartan and sacubitril in valsartan/sacubitril tablets. The optimized mobile phase flow rate was 1 mL min[-1], with the mobile phase consisting of 400 mL of 0.025 M potassium phosphate adjusted to pH 3.5 and 600 mL of ethyl alcohol. The range of linearity used for valsartan and sacubitril was 1.25-50 µg mL[-1] for each medicinal substance. The wavelength used to analyze the samples was 254 nm; the sample size was 20 µL; and the analysis time was <6 minutes. The ultimate scores are as follows: 0.76 for AGREE, 77.78 for AGSA, 80 for BAGI, 70 for CACI, 92 for CaFRI, 88 for AES, 78 for MoGAPI, and 75.6% for the A letter score regarding whiteness. These scores indicate the high sustainability, greenness, whiteness, and economic wisdom of the current method. All validation characteristics have followed the international ICH and US FDA validation guidelines. Compared to complicated systems such as UPLC-MS, it offers benefits including shorter retention times, less reagent usage, simultaneous analysis, and cheaper prices.

Collection of hematopoietic progenitor cells and mononuclear cells by leukocytapheresis is a critical step of procuring cellular starting materials for future therapeutic use. Leukocytapheresis requires use of anticoagulant to prevent blood clotting in the extracorporeal circuit during collection. A commonly used anticoagulant is Acid Citrate Dextrose Solution A (ACD-A). ACD-A primarily prevents blood clotting by reversibly chelating free ionized calcium, which is essential for the clotting cascade. Electrolytes frequently affected by ACD-A include calcium, magnesium, potassium, phosphate, and bicarbonate. Risks for development of electrolyte derangements include procedure-related parameters as well as patient comorbidities. Symptoms range from mild paresthesias to life-threatening cardiac arrhythmias. Recognition of related signs and symptoms is important for prompt intervention with procedure modification and electrolyte replacement to optimize patient safety. This paper will provide an enhanced understanding of the underlying pathophysiology and assist in timely identification and management of electrolyte abnormalities associated with cellular therapy collections.

Microwave-assisted pyrolysis of kraft lignin was used to produce highly porous carbonaceous materials for environmental applications (biochars). LignoForce hardwood (LFHL) and softwood (LFSL) lignins were mixed with 30% potassium phosphate (K3PO4) and pyrolyzed at 450 °C to produce 45-47% biochar after 25 min of microwave irradiation. A 10 L min-1 nitrogen flow was used for purging during the pyrolysis and cooling. Elemental analysis (CHNS-O) revealed that kraft lignins were highly deoxygenated, but nearly 26% of their original oxygen content remained after pyrolysis, suggesting retention of oxygenated compounds in the biochar structure. Nitrogen adsorption (BET), scanning electron microscopy (SEM), and X-ray diffraction (XRD) demonstrated the formation amorphous micro- and mesoporous carbonaceous materials with a high potential for water and soil remediation. Significant specific surface areas (142 and 202 m2 g-1 for LFHL-B and LFSL-B, respectively) were obtained for these lignin biochars, which were tested for methylene blue (MB) adsorption at pH 4.5, 6.0, 7.5, and 9.0. LignoForce lignin biochars presented an adsorption capacity of approximately 15.5 mg g-1 and a removal efficiency higher than 94%, especially at higher pH levels (>4.5). However, MB adsorption was slightly faster for LFHL-B compared to LFSL-B. These results position kraft lignin as a valuable carbon-accumulating raw material for environmental applications and establish MAP as a fast and energy-efficient route for their thermal conversion.

Slaughterhouse waste is a major source of environmental pollution. However, its rich organic content holds considerable potential for enhancing soil fertility. Herein, slaughterhouses meat waste was used to produce organic fertilizer (OF), and its effect on sugarcane growth, productivity and soil fertility were evaluated. The decomposition of the collected meat waste was carried out in a controlled fermentation pit. Initially, the sample was treated by sulfuric acid (H2SO4), followed by the addition of potassium hydroxide (KOH) and dipotassium phosphate (K2HPO4). The pH of the medium was then adjusted to neutral conditions, after that a bacterial inoculum was introduced to facilitate further biodegradation of the solid waste. The prepared organic fertilizer was characterized for its chemical compositions, physicochemical properties and microbial diversity. The chemical compositions analyses revealed that potentially toxic elements such as mercury (Hg), lead (Pb), cadmium (Cd) and arsenic (As) were detected at concentration of <0.002 mg/kg, 0.411 mg/kg, 0.004 mg/kg and 0.095 mg/kg, respectively, indicating their levels in the fertilizer were very low. The total organic carbon content was determined to be 13.90% that is significantly higher than that of most soil, where organic carbon levels typically fall below 2%. MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight) mass spectrometry was used to identify culturable bacterial isolates obtained from the fermented organic fertilizer prior to soil application. A total of 20 morphological distinct isolates were analyzed, of which Lysinibacillus sphaericus was the most frequently detected species, followed by Bacillus simplex, Bacillus cereus, Kocuria varians, and Microascus sp. Identification was accepted based on MALDI-TOF score thresholds (≥2.0 species-level and 1.7-1.99 for genus level identification). Lysinibacillus sphaericus aids nitrogen-fixation, plant growth, and decomposing organic matter, all of which enhance soil fertility and plant health. Additionally, other species such as Bacillus simplex, Bacillus cereus, Kocuria varians, and Microascuswere contribute to phosphate solubilization and pathogen suppression. The organic ferilizer was tested alone and in combination with the conventional chemical fertilizer in a pot experiment for sugarcane. The results showed that applying 5 tonnes ha-1 of organic fertilizer together with 50% of the recommended chemical fertilizer performed comparable to that of 100% recommended chemical fertilizer alone. In addition, combined organic fertilizer and recommended chemical fertilizer resulted in a significantly higher juice phosphate content compared to the sole application of 100% recommended chemical fertilizer, likely due to the contribution of the organic fertilizer. This finding suggests that halving chemical fertilizer use by supplementing with organic alternatives could offer environmental benefits while maintaining productivity.

Defect passivation is one feasible and effective approach to minimize the charge recombination of perovskite solar cells (PSCs). However, the efficacy of conventional defect passivation using Lewis base-functionalized molecules is limited due to weak bonding and large space steric hindrance. Herein, we propose a molecular engineering approach employing ultrasmall-sized potassium diformate and potassium phosphate to synergistically passivate defects at the SnO2/perovskite buried interface. It is found that the C═O and P═O groups effectively passivate interfacial traps, reducing trap density by ∼50% relative to the control sample. The co-modification also lowers the surface energy of SnO2 layers, promoting the growth of perovskite grains with an average grain size increased from 780 to 1370 nm. Moreover, thermally diffused K[+] ions can inhibit ion migration in the perovskite lattice, thereby suppressing the hysteresis effect. According to these advantages, the optimized devices achieve a champion power conversion efficiency of 24.42% (vs 22.64% for the pristine SnO2-based counterpart). The unencapsulated co-passivated PSCs retain 92% of their initial efficiency after 2100 h under ambient conditions, substantially outperforming the control devices (74% retention).

Quantitative bone single-photon emission computed tomography (SPECT) using[99m]Tc-labeled phosphates has become increasingly feasible due to advances in SPECT/CT technology and reconstruction algorithms. The xSPECT bone algorithm further improves local anatomical fidelity by segmenting CT images into six tissue classes to generate a zone map for context-specific enhancement of spatial resolution. Therefore, when evaluating xSPECT Bone using phantom studies, bone-equivalent solutions must be prepared to ensure that CT values fall within the bone segmentation range. This study aims to clarify how two commonly used substances (dipotassium hydrogen phosphate (K2HPO4) and iodinated contrast agent) affect CT values, zone maps, attenuation coefficients (μ-map), and quantitative accuracy of xSPECT bone. The inner container of the JS 10 SPECT performance phantom was filled with a[99m]Tc solution prepared by dissolving either K2HPO4or iodinated contrast agent in 100 ml of water. Four types of inserts (with no additive or different K2HPO4or iodine concentrations) were placed in the same sectional area. Both K2HPO4and iodine exhibited strong correlations (p< 0.01) between CT values (HU) and mass densities (g ml[-1]). No significant differences in zone map orμ-map values were observed between K2HPO4and iodine. At equivalent CT values, percentage contrast was significantly higher with iodine than with K2HPO4. Furthermore, K2HPO4demonstrated stable percentage contrast near unity within 158.5-269.7 HU, indicating superior quantitative consistency. The zone maps andμ-maps were similar with K2HPO4and iodine; however, differences in attenuation during SPECT acquisition due to differences in mass density were not considered. This suggests that attenuation correction becomes overcorrected, thereby decreasing quantitative accuracy. These findings indicate that K2HPO4is more suitable than iodine for creating bone equivalent solutions in quantitative bone SPECT phantom studies.

Metabolic dysfunction-associated steatohepatitis (MASH) is emerging as a leading cause of chronic liver disease. MTOR (mechanistic target of rapamycin kinase) complex 1 (MTORC1) is a potential therapeutic target, whereas suppression of total MTORC1 activity can lead to unwanted effects. Here, we found that byakangelicin (Bya), a natural compound, selectively inhibited MTORC1-mediated phosphorylation of TFEB (transcription factor EB), without affecting canonical MTORC1 substrates. Knockout of hepatic Tfeb blocked the alleviation effects of Bya on hepatic steatosis, inflammation, insulin resistance, and fibrosis in mice, while reintroduction of TFEB restored these effects. We identified Bya directly bound to MET370 and PHE552 of FLCN (folliculin), suppressing the function of the FLCN-FNIP1 (folliculin interacting protein 1)/FNIP2 complex, which in turn inhibited MTORC1-mediated cytoplasmic sequestration of TFEB. Mutation of FLCN (M370A and F552A) in the liver abolished Bya-induced protection against MASH. Thus, Bya is a promising therapeutic natural compound for MASH, and selective inhibition of MTORC1 is a potential approach to treat this disease.Abbreviations: aa, amino acids; AAV, adeno-associated virus; Bio, biotin; Bio-Bya, biotin-conjugated Bya; BSA, bovine serum albumin; BW, body weight; Bya, byakangelicin; CETSA, cellular thermal shift assay; CHIP-atlas, chromatin immunoprecipitation atlas; Cmax, maximum concentration; CQ, chloroquine; DARTS; drug affinity responsive target stability assay; EIF4EBP1/4E-BP1, eukaryotic translation initiation factor 4E binding protein 1; FBS, fetal bovine serum; FDA, food and drug administration; FIMO-JASPAR, find individual motif occurrences-JASPAR; FLCN, folliculin; FNIP1, folliculin interacting protein 1; GAP, GTPase-activating protein; GOT1/AST, glutamic-oxaloacetic transaminase 1; GPT/ALT, glutamic-pyruvic transaminase; GTRD, gene transcription regulatory database; GTT, glucose tolerance test; H&E, hematoxylin and eosin; Hbonds, hydrogen bonds; HFD, high-fat diet; HFHC, high-fat and high-cholesterol; HOMA-IR, homeostatic model assessment of insulin resistance; HSCs, hepatic stellate cells; IP, immunoprecipitation; ITT, insulin tolerance test; KD, dissociation constant; KEGG, kyoto encyclopedia of genes and genomes; KPBS, potassium phosphate-buffered saline; LC-MS/MS, liquid chromatography-tandem mass spectrometry; LW/BW, liver-to-body weight ratio; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; MASH, metabolic dysfunction-associated steatohepatitis; MASLD, metabolic dysfunction-associated steatotic liver disease; MCD, methionine and choline deficient; MST, microscale thermophoresis assay; MTOR, mechanistic target of rapamycin kinase; MTORC1, MTOR complex 1; ND, normal diet; NFKB/NF-κB, nuclear factor kappa B; NFKBIA/IKBA, NFKB inhibitor alpha; OP, oleate acid and palmitate acid; PBS, phosphate-buffered saline; PCA, principal component analysis; qRT-PCR, real-time quantitative PCR; RELA/p65, RELA proto-oncogene, NF-kB subunit; Res, resmetirom; Rg, radius of gyration; RMSD, root-mean-square deviation; RMSF, root-mean-square fluctuation; RPS3, ribosomal protein S3; RPS6KB1/S6K1, ribosomal protein S6 kinase B1; RRAGC, ras related GTP binding C; SASA, solvent-accessible surface area; SNRPD2, small nuclear ribonucleoprotein D2 polypeptide; SQSTM1/p62, sequestosome 1; T1/2, half-life; TFE3, transcription factor binding to IGHM enhancer 3; TFEB, transcription factor EB; TMEM192, transmembrane protein 192; VIM, vimentin; WT, wild-type.

Magnesium potassium phosphate cement (MKPC) is a promising bone repair material but suffers from excessively rapid setting time (typically within minutes) that limits clinical application. This study systematically investigates trimagnesium phosphate (TMP) as a candidate retarding additive for MKPC. TMP was used to partially replace dead-burned magnesium oxide at replacement levels of 0%, 10%, and 15% by mass. The effects of TMP content, water-to-cement ratio (0.17-0.23), and magnesium-to-phosphate molar ratio (4-10) on setting time, fluidity, hydration kinetics, compressive strength, and microstructure were comprehensively evaluated. Results show that TMP effectively extends the setting time from 9-13 min (without TMP) to 10-19 min, providing a working window that may be suitable for biomedical applications requiring extended handling time. Notably, 10% TMP incorporation enhances early compressive strength, with 1-day strength reaching 35.2 MPa compared to 28.5 MPa for control samples. Hydration heat analysis reveals TMP moderates the acid-base reaction kinetics through its slower dissolution rate compared to MgO. Microstructural characterization shows TMP promotes the formation of denser K-struvite crystals with refined microstructure. The optimal TMP dosage of 10% achieves a balanced performance, extending setting time while improving early strength and microstructural densification. These findings establish TMP as an effective retarder for developing MKPC-based materials with potential for biomedical applications, pending further biological validation.