OBJECTIVES/GOALS: This study investigates whether localization of high mobility group box 1 (HMGB1) controls inflammatory signaling and DNA damage response in human keratinocytes, the cell of origin for squamous cell carcinoma (SCC). SCC is especially metastatic in chronic wounds, burns, and in patients with recessive dystrophic epidermolysis bullosa (RDEB). METHODS/STUDY POPULATION: We used CRISPR/Cas9 gene-editing to knock out HMGB1 in a keratinocyte line, p16INK4a-negative keratinocytes immortalized by ectopic hTERT expression (N/TERT-2G [46,XY]). Following gene editing, clonal keratinocyte populations were screened for knockout by PCR followed by TIDE analysis (Tracking of Indels by Decomposition) to identify indels that would result in a frameshift mutation. Total cell lysates for each clonal population were analyzed by immunoblot and immunofluorescence for HMGB1 protein. These cells will be used to assay for DNA damage sensitivity in the presence of genotoxic agents (etoposide, ultraviolet radiation, γ-irradiation). A lentiviral vector will then be used to express mutant forms of HMGB1 that localize to the nucleus (C23/45S) or cytoplasm (C106S) and DNA damage assays repeated. RESULTS/ANTICIPATED RESULTS: We have confirmed by sequencing, immunoblot, and immunofluorescence that HMGB1 is knocked out in a clonal population of N/TERT-2G human keratinocyte cells. We anticipate that cells with a complete absence of HMGB1 will have high sensitivity to DNA damaging agents, but little change in inflammatory signaling. We also expect that cells expressing mutant HMGB1 that localizes exclusively to the cytoplasm will demonstrate an increased sensitivity to DNA damage relative to wild-type controls, while mutant HMGB1 that localizes exclusively to the nucleus will be protected from DNA damage caused by exposure to genotoxic agents. DISCUSSION/SIGNIFICANCE: HMGB1 is a nuclear protein and damage associated molecular pattern that is elevated systemically in patients with RDEB, many of whom will go on to develop metastatic squamous cell carcinoma. The experiments described here investigate whether HMGB1 plays a mechanistic role in skin carcinogenesis via regulation of the DNA damage response.

Objectives: Antimicrobial resistance (AMR) has emerged as a major concern in Vietnam, mainly due to the inappropriate use of antibiotics. Appropriate antibiotic management enables us to minimize the likelihood of antibiotic resistance and the spread of resistant bacteria. We evaluated vancomycin and colistin resistance and related factors in the intensive care unit (ICU) of Hue Central Hospital, a national hospital in central Vietnam. Methods: Using a cross-sectional descriptive study, we enrolled 362 patients who were prescribed antibiotics and were admitted to the ICU in 2019. Pathogens isolated from 473 routine clinical samples were subjected to antimicrobial susceptibility testing following the recommendations in the Clinical & Laboratory Standards Institute M100, 28th Edition. Colistin testing was performed using the broth microdilution method. Statistical significance was determined using the Fisher exact test. Results: The most commonly identified microorganisms were Acinetobacter baumannii (31.5%), Klebsiella pneumoniae (31.2%), Pseudomonas aeruginosa (12%), and Staphylococcus aureus (8.9%). All isolates of A. baumannii, K. pneumoniae, and P. aeruginosa tested with colistin were nonresistant. Moreover, >65% of A. baumannii isolates were resistant to all antibiotics except colistin. S. aureus had the highest resistance rate to erythromycin (80.6%), but no vancomycin-resistant isolates were identified. Factors associated with resistance to at least 1 antibiotic tested included length of stay (OR, 5.32; 95% CI, 1.47–19.17; P = .017), duration of antibiotics therapy (OR, 5.25; 95% CI, 1.46–18.95; P = .017), and the use of tracheal intubation and ventilator (OR, 3.08; 95% CI, 1.09–8.72; P = .038). Conclusions: These data indicated that although the vancomycin and colistin resistance rate is low, patients with longer length of stay, longer time on antibiotics, and invasive ventilation were at higher risk of AMR infection. Decreasing device use and strong antibiotic stewardship program at the hospital would help to reduce AMR infections.

As the third decade of the twenty-first century begins, Vietnam embarks on a more advanced phase of national development and international integration, with a greater emphasis on foreign policy as part of the country's overall national defence and development strategy. This informs greater expectations about shifts in Vietnam's foreign policy perception and discourse in pursuit of national interests and the regime's legitimacy amidst major domestic and international developments. This article analyses the making, in terms of processes and actors, and the evolution, in terms of themes and directions, of Vietnam's foreign policy under the Thirteenth National Congress of the Communist Party of Vietnam, which was held in early 2021. The article argues that while embedding continuity with what the country has been pursuing since its renovation process (known as Doi Moi) started in the mid-1980s, Vietnam's foreign policy under the Thirteenth Party Congress is crafted on a broader base of domestic consensus and features new dimensions, implying stronger domestic support for Hanoi's conduct of foreign affairs and a Vietnamese nation brand with greater visibility and contribution in the regional and global arenas in the coming years.

We prove that any nef $b$-divisor class on a projective variety defined over an algebraically closed field of characteristic zero is a decreasing limit of nef Cartier classes. Building on this technical result, we construct an intersection theory of nef $b$-divisors, and prove several variants of the Hodge index theorem inspired by the work of Dinh and Sibony. We show that any big and basepoint-free curve class is a power of a nef $b$-divisor, and relate this statement to the Zariski decomposition of curves classes introduced by Lehmann and Xiao. Our construction allows us to relate various Banach spaces contained in the space of $b$-divisors which were defined in our previous work.