Evaluating outcomes of combined bladder neck and supramontanal sparing ejaculatory preserving transurethral resection of the prostate: Results from a prospective, randomised study.

Introduction: Although conventional transurethral resection of the prostate (TURP) is highly successful in improving urinary symptoms and flow rates, a higher incidence of loss of antegrade ejaculation has been reported. Therefore, we aimed at prospectively comparing the efficacy and outcomes of a novel dual bladder neck and supramontanal sparing TURP to conventional TURP to improve voiding and ejaculation. Material and methods: Between January 2019 and November 2020, all patients with benign prostatic hyperplasia (BPH) satisfying the eligibility criteria underwent either conventional TURP (Group 1) or combined bladder neck and supramontanal sparing TURP (Group 2) after randomisation. The groups were compared for functional outcomes including International Prostate Symptom Score (IPSS), peak flow rates, post-void residual urine, perioperative variables and postoperative complications. Ejaculation was assessed with International Index of Erectile Function-Question 9 (IIEF-9) and Ejaculation Projection score (EPS). Results: A total of 90 patients were randomised, 45 each to Group 1 and 2 respectively. The demographic profiles across both groups were comparable. Retrograde ejaculation and bladder neck contracture were significantly higher in Group 1. Both groups demonstrated significant improvement in the IPSS (26.12 ±2.88 to 4.69 ±0.87 (Group 1) vs 26.60 ±3.45 to 4.36 ±1.74 in Group 2) and Qmax (7.03 ±2.71 to 24.36 ±3.82 mL/s in Group 1 vs 6.29 ±2.64 to 25.28 ±4.33 mL/s in Group 2) at 3 months. However, a significant difference in IPSS and Qmax were recorded at 6 months. IIEF-9 score in Group 2 remained similar to preoperative profile (4.18 ±0.75) vs 2.58 ±0.86 (Group 1). EPS significantly decreased in Group 1 but remained similar to preoperative EPS in Group 2. Antegrade ejaculation was preserved in 88.89% in Group 2 as compared to 22.22% in Group 1. Conclusions: Dual bladder neck and supramontanal ejaculation preserving TURP is superior to conventional TURP in preventing retrograde ejaculation and bladder neck contractures in prostates <50 cc with comparable functional results, perioperative and postoperative morbidity.

Synthetic cytokine circuits that drive T cells into immune-excluded tumors.

Chimeric antigen receptor (CAR) T cells are ineffective against solid tumors with immunosuppressive microenvironments. To overcome suppression, we engineered circuits in which tumor-specific synNotch receptors locally induce production of the cytokine IL-2. These circuits potently enhance CAR T cell infiltration and clearance of immune-excluded tumors, without systemic toxicity. The most effective IL-2 induction circuit acts in an autocrine and T cell receptor (TCR)- or CAR-independent manner, bypassing suppression mechanisms including consumption of IL-2 or inhibition of TCR signaling. These engineered cells establish a foothold in the target tumors, with synthetic Notch-induced IL-2 production enabling initiation of CAR-mediated T cell expansion and cell killing. Thus, it is possible to reconstitute synthetic T cell circuits that activate the outputs ultimately required for an antitumor response, but in a manner that evades key points of tumor suppression.

Robust Sequence Determinants of α-Synuclein Toxicity in Yeast Implicate Membrane Binding.

Protein conformations are shaped by cellular environments, but how environmental changes alter the conformational landscapes of specific proteins in vivo remains largely uncharacterized, in part due to the challenge of probing protein structures in living cells. Here, we use deep mutational scanning to investigate how a toxic conformation of α-synuclein, a dynamic protein linked to Parkinson's disease, responds to perturbations of cellular proteostasis. In the context of a course for graduate students in the UCSF Integrative Program in Quantitative Biology, we screened a comprehensive library of α-synuclein missense mutants in yeast cells treated with a variety of small molecules that perturb cellular processes linked to α-synuclein biology and pathobiology. We found that the conformation of α-synuclein previously shown to drive yeast toxicity-an extended, membrane-bound helix-is largely unaffected by these chemical perturbations, underscoring the importance of this conformational state as a driver of cellular toxicity. On the other hand, the chemical perturbations have a significant effect on the ability of mutations to suppress α-synuclein toxicity. Moreover, we find that sequence determinants of α-synuclein toxicity are well described by a simple structural model of the membrane-bound helix. This model predicts that α-synuclein penetrates the membrane to constant depth across its length but that membrane affinity decreases toward the C terminus, which is consistent with orthogonal biophysical measurements. Finally, we discuss how parallelized chemical genetics experiments can provide a robust framework for inquiry-based graduate coursework.

The Design Principles of Biochemical Timers: Circuits that Discriminate between Transient and Sustained Stimulation.

Many cellular responses for which timing is critical display temporal filtering-the ability to suppress response until stimulated for longer than a given minimal time. To identify biochemical circuits capable of kinetic filtering, we comprehensively searched the space of three-node enzymatic networks. We define a metric of "temporal ultrasensitivity," the steepness of activation as a function of stimulus duration. We identified five classes of core network motifs capable of temporal filtering, each with distinct functional properties such as rejecting high-frequency noise, committing to response (bistability), and distinguishing between long stimuli. Combinations of the two most robust motifs, double inhibition (DI) and positive feedback with AND logic (PFAND), underlie several natural timer circuits involved in processes such as cell cycle transitions, T cell activation, and departure from the pluripotent state. The biochemical network motifs described in this study form a basis for understanding common ways cells make dynamic decisions.

TNFAIP3 maintains intestinal barrier function and supports epithelial cell tight junctions.

Tight junctions between intestinal epithelial cells mediate the permeability of the intestinal barrier, and loss of intestinal barrier function mediated by TNF signaling is associated with the inflammatory pathophysiology observed in Crohn's disease and celiac disease. Thus, factors that modulate intestinal epithelial cell response to TNF may be critical for the maintenance of barrier function. TNF alpha-induced protein 3 (TNFAIP3) is a cytosolic protein that acts in a negative feedback loop to regulate cell signaling induced by Toll-like receptor ligands and TNF, suggesting that TNFAIP3 may play a role in regulating the intestinal barrier. To investigate the specific role of TNFAIP3 in intestinal barrier function we assessed barrier permeability in TNFAIP3(-/-) mice and LPS-treated villin-TNFAIP3 transgenic mice. TNFAIP3(-/-) mice had greater intestinal permeability compared to wild-type littermates, while villin-TNFAIP3 transgenic mice were protected from increases in permeability seen within LPS-treated wild-type littermates, indicating that barrier permeability is controlled by TNFAIP3. In cultured human intestinal epithelial cell lines, TNFAIP3 expression regulated both TNF-induced and myosin light chain kinase-regulated tight junction dynamics but did not affect myosin light chain kinase activity. Immunohistochemistry of mouse intestine revealed that TNFAIP3 expression inhibits LPS-induced loss of the tight junction protein occludin from the apical border of the intestinal epithelium. We also found that TNFAIP3 deubiquitinates polyubiquitinated occludin. These in vivo and in vitro studies support the role of TNFAIP3 in promoting intestinal epithelial barrier integrity and demonstrate its novel ability to maintain intestinal homeostasis through tight junction protein regulation.