Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Intolerance of a non-diffractive extended-depth-of-focus IOL with mini-monovision

Purpose: The use of a non-diffractive extended-depth-of-focus (EDOF) intraocular lens (IOL) with slight myopia of −0.5 D on the non-dominant eye increases the spectacle independence and has good subjective tolerance with optical phenomena comparable to those of a monofocal IOL. This case report describes the course of a myopic patient who underwent refractive lens exchange, didn't tolerate mini-monovision and received IOL exchange therefore. Observations: A healthy, 62-year-old male with myopia of approximately −5 D underwent refractive lens exchange with a non-diffractive EDOF-IOL on both eyes with slight myopia on the non-dominant left eye (mini-monovision). The operation was performed without any complications, postoperative treatment was due to the clinic's standard procedure. Two weeks postoperative the patient presented with uncorrected distance visual acuity of 0.0 logMAR, a subjective refraction of −0.25/−0.25/142° and corrected distance visual acuity of 0.1 logMAR on the right eye. On the left eye, distance visual acuity was 0.4 logMAR with a subjective refraction of −0.5/−0.75/9° (intended mini-monovision) and corrected distance visual acuity of 0.0 logMAR. Binocular distance visual acuity was 0.0 logMAR. The patient complained about the occurrence of optical phenomena at dim light while driving a car and subjective reduced visual acuity. After an IOL exchange on the left eye with the implantation of the same type of non-diffractive EDOF-IOL aimed for emmetropia, the patient was symptom-free and reported no more subjective complaints. Conclusions: Despite the satisfying subjective and objective visual outcome which is proven in multiple studies, the subjective perception of a mini-monovision with a non-diffractive EDOF-IOL can vary individually. A preoperative assessment of the patient's needs and tolerance of a mini-monovision is crucial for a satisfying postoperative outcome

Pharmacological HIF-inhibition attenuates postoperative adhesion formation

Abstract Peritoneal adhesions represent a common complication of abdominal surgery, and tissue hypoxia is a main determinant in adhesion formation. Reliable therapeutic options to reduce peritoneal adhesions are scarce. We investigated whether the formation of postsurgical adhesions can be affected by pharmacological interference with hypoxia-inducible factors (HIFs). Mice were treated with a small molecule HIF-inhibitor, YC-1 (3-[5′-Hydroxymethyl-2′-furyl]-1-benzyl-indazole), or vehicle three days before and seven days after induction of peritoneal adhesions or, alternatively, once during induction of peritoneal adhesions. Pretreatment or single intraperitoneal lavage with YC-1 significantly reduced postoperative adhesion formation without prompting systemic adverse effects. Expression analyses of cytokines in peritoneal tissue and fluid and in vitro assays applying macrophages and peritoneal fibroblasts indicated that this effect was cooperatively mediated by various putatively HIF-1α-dependent mechanisms, comprising attenuated pro-inflammatory activation of macrophages, impaired recruitment and activation of peritoneal fibroblasts, mitigated epithelial-mesenchymal-transition (EMT), as well as enhanced fibrinolysis and impaired angiogenesis. Thus, this study identifies prevention of postsurgical peritoneal adhesions as a novel and promising field for the application of HIF inhibitors in clinical practice

Inhibition of HIF-prolyl hydroxylases improves healing of intestinal anastomoses

Anastomotic leakage (AL) accounts for a major part of in-house mortality in patients undergoing colorectal surgery. Local ischemia and abdominal sepsis are common risk factors contributing to AL and are characterized by upregulation of the hypoxia-inducible factor (HIF) pathway. The HIF pathway is critically regulated by HIF-prolyl hydroxylases (PHDs). Here, we investigated the significance of PHDs and the effects of pharmacologic PHD inhibition (PHI) during anastomotic healing. Ischemic or septic colonic anastomoses were created in mice by ligation of mesenteric vessels or lipopolysaccharide-induced abdominal sepsis, respectively. Genetic PHD deficiency (Phd1–/–, Phd2+/–, and Phd3–/–) or PHI were applied to manipulate PHD activity. Pharmacologic PHI and genetic PHD2 haplodeficiency (Phd2+/–) significantly improved healing of ischemic or septic colonic anastomoses, as indicated by increased bursting pressure and reduced AL rates. Only Phd2+/– (but not PHI or Phd1–/–) protected from sepsis-related mortality. Mechanistically, PHI and Phd2+/– induced immunomodulatory (M2) polarization of macrophages, resulting in increased collagen content and attenuated inflammation-driven immune cell recruitment. We conclude that PHI improves healing of colonic anastomoses in ischemic or septic conditions by Phd2+/–-mediated M2 polarization of macrophages, conferring a favorable microenvironment for anastomotic healing. Patients with critically perfused colorectal anastomosis or abdominal sepsis could benefit from pharmacologic PHI