This study explores the extraction and characterization of proteolytic enzymes from brewer's spent grain (BSG) and their potential as sustainable coagulants in the dairy industry. BSG samples from various beer types (Blonde Ale, IPA, Kölsch, Honey, and Porter) were obtained from two artisanal breweries in Mar del Plata, Argentina. Optimization of caseinolytic activity (CA) and protein extraction was conducted using a Plackett-Burman design, followed by a Box-Behnken design. Optimal protein concentration was achieved at intermediate pH and high temperature, while CA peaked at pH 8.0. The specific caseinolytic activity (SCA) varied among the extracts, with BSG3 showing the highest activity (99.6 U mg⁻¹) and BSG1 the lowest (60.4 U mg⁻¹). Protease inhibitor assays suggested the presence of aspartic, serine, metallo, and cysteine proteases. BSG3 and BSG4 showed the highest hydrolysis rates for α-casein (70% and 78%). For κ-casein, BSG1, BSG2, and BSG3 demonstrated moderate activity (56.5%, 49%, and 55.8), while BSG4 and BSG5 exhibited the lowest activity. Additionally, the milk-clotting activity (MCA) of BSG extracts was comparable to plant-based coagulants like Cynara cardunculus and Ficus carica. These findings highlight the potential of BSG-derived proteases as alternative coagulants for cheese production, offering a sustainable link between the brewing and dairy industries.

A positive fluid balance may evolve to fluid overload and associate with organ dysfunctions, weaning difficulties, and increased mortality in ICU patients. We explored whether individualized fluid management, assessing fluid-responsiveness via passive leg-raising maneuver (PLR) before spontaneous breathing trial (SBT) associated with less extubation failure in ventilated patients with high fluid balance, admitted to the ICU after liver transplantation (LT). We recruited 15 LT patients in 2023. Postoperative fluid balance was +4476 [3697,5722] mL. PLRs were conducted at ICU admission (T1) and pre-SBT (T2). Cardiac index (CI) changes were recorded before and after SBT (T3). Seven patients were fluid responsive at T1, and 12 at T2. No significant differences occurred in hemodynamic, respiratory, and perfusion parameters between fluid-responsive and fluid-unresponsive patients at any time. Fluid-responsive patients at T1 and T2 increased their CI during SBT from 3.1 [2.8,3.7] to 3.7 [3.4,4.1] mL/min/m2 (p= 0.045). All fluid-responsive patients at T2 were extubated after SBT and consolidated extubation. Two out of three of fluid-unresponsive patients experienced weaning difficulties. We concluded that fluid-responsive patients post-LT may start weaning earlier and achieve successful extubation despite high postoperative fluid balance. This highlights the profound impact of personalized assessment of cardiovascular state on critical surgical patients.