Travoprost (TRV) is an efficient prostaglandin (PG) analogue, which is hydrolyzed by corneal esterases to its
active acid form (TRV acid) after absorption. Till now no spectrofluorimetric methods have been reported for
detection of TRV acid in biological fluids. Due to high sensitivity, simplicity and rapidity; a novel and ultrasensitive
spectrofluorometric method with an efficient solid phase extraction (SPE) technique was developed
for determination of TRV acid in aqueous humor (AH). The study relies on the fluorescence enhancement of
nitrogen-doped carbon dots (N-CDs), which were prepared via a hydrothermal treatment of triphenylamine and
citric acid. The formed nanoprobe responds with very high sensitivity to trace amounts of TRV acid through
hydrogen bonding formation, which is accompanied by an enhancement in fluorescence intensity due to N-CDs
aggregation. The proposed method showed excellent sensitivity for analyzing TRV acid in the range of 1–200 ng/
mL, with a very low limit of detection (0.29 ng/mL). Additionally, the method’s selectivity for analyzing TRV
acid in the presence of common ions and anticipated co-administered drugs was assessed. The structural and
chemical properties of the prepared N-CDs were extensively characterized using X-ray diffraction (XRD), Fourier
transform infrared (FT-IR) spectroscopy, Transmission electron microscopy (TEM), High resolution transmission

Tuberculosis (TB) remains one of the deadliest infectious diseases globally. Although the approved human Bacille-Calmette-Guérin (BCG) vaccines provide limited protection, a vaccine based on Mycobacterium tuberculosis (Mtb) has yet to be approved. Our previous findings demonstrated that s.c. immunization with heat-killed Mtb significantly increased the number of monocytic myeloid-derived suppressor cells (M-MDSC) in mice. Thus, we hypothesized that the defense against a subsequent BCG infection would be compromised in Mtb-immunized mice. Surprisingly, mice vaccinated with Mtb were protected against BCG infection and exhibited elevated frequencies and activation of dendritic cells (DC) and mycobacteria-specific T cells, despite high frequencies and suppressor activity of M-MDSC. Genetic ablation of CCR2+ monocytic cells or pharmacological intervention with all-trans retinoic acid (ATRA) reduced the frequency of Mtb-induced M-MDSC, enhanced the frequencies and activation of DC and CD4+ T cells, and resulted in decreased bacterial loads in the lungs and spleen. These findings provide new insights into TB vaccination using heat-killed Mtb despite the concurrent unwanted effects of vaccine-induced M-MDSC. M-MDSC depletion via ATRA further shifts the balance toward immunity and should be considered an adjunct host-directed therapy alongside TB vaccines in humans.

Tumor-draining lymph nodes (TDLNs) are sites of anti-tumor immune priming as well as metastases. Here, we examined how the cellular networks within TDLNs are reorganized in triple-negative breast cancer (TNBC). We found that the frequencies of programmed death ligand 1 high (PD-L1hi) monocytes increased in TDLNs of metastatic TNBC mouse tumors. Fibroblastic reticular cell (FRC) subtypes heightened the expression of the chemokines CCL2 and CCL7, supporting the homing of CCR2+ monocytes. These monocytes suppressed T cells in vitro via PD-L1 and inducible nitric oxide synthase (iNOS). Spatial transcriptomics revealed immunosuppressive FRC-monocyte niches in vascularized and T cell areas. Tumor-associated Toll-like receptor (TLR) 4 ligands induced CCL2 and CCL7 expression by FRCs to promote monocyte recruitment. Localized TLR4 inhibition in combination with anti-programmed cell death protein 1 (αPD-1) therapy reduced monocyte homing and boosted T cell function, ultimately attenuating lung metastases. Monocytes accumulate in human TNBC TDLNs, with evidence of a FRCmonocyte axis, and a TLR4 ligand signature is predictive of poor survival outcomes in TNBC patients. Thus, metastatic TNBC can reprogram lymph nodes (LNs) to facilitate PD-L1-mediated immune evasion and metastasis.

Cellular metabolism must adapt rapidly to environmental alterations and adjust nutrient uptake. Low glucose availability activates the AMP-dependent kinase (AMPK) pathway. We demonstrate that activation of AMPK or the downstream Unc-51-like autophagy-activating kinase (ULK1) inhibits receptor-mediated endocytosis. Beyond limiting dextran uptake, this activation prevents endocytic uptake of human pathogenic enveloped and non-enveloped, positive- and negative-stranded RNA viruses, such as yellow fever, dengue, tick-borne encephalitis, chikungunya, polio, rubella, rabies lyssavirus, and SARS-CoV-2, not only in mammalian and insect cells but also in precision-cut lung slices and neuronal organoids. ULK1 activation inhibited enveloped viruses but not EV71. However, receptor presentation at the cytoplasmic membrane remained unaffected, indicating that receptor binding was unchanged, while later stages of endocytosis were targeted via two distinct pathways. Drug-induced activation of the AMPK pathway reduced early endocytic factor TXNIP by suppressing translation. In contrast, the amounts of Rab5 and the late endosomal marker Rab7 decreased due to translation inactivation and ULK1-dependent proteasome activation within minutes. Furthermore, activation of AMPK hindered the late replication steps of SARS-CoV-2 by reducing viral RNAs and proteins and the endo-lysosomal markers LAMP1 and GRP78, suggesting a reduction in early and late endosomes and lysosomes. Inhibition of the PI3K and mTORC2 pathways, which sense amino acid and growth factor availability, promotes AMPK activity and blocks viral entry. Our results indicate that AMPK and ULK1 emerge as restriction factors of cellular endocytosis, impeding the receptor-mediated endocytic entry of enveloped and non-enveloped RNA viruses.