In rat adipocytes, insulin provoked rapid increases in (a) endogenous immunoprecipitable combined protein kinase C (PKC)-/ activity in plasma membranes and microsomes and (b) immunoreactive PKC-and PKCin GLUT4 vesicles. Activity and autophosphorylation of immunoprecipitable epitope-tagged PKC-and PKCwere also increased by insulin in situ and phosphatidylinositol 3,4,5-(PO 4 ) 3 (PIP 3 ) in vitro. Because phosphoinositide-dependent kinase-1 (PDK-1) is required for phosphorylation of activation loops of PKC-and protein kinase B, we compared their activation. Both RO 31-8220 and myristoylated PKC-pseudosubstrate blocked insulin-induced activation and autophosphorylation of PKC-/ but did not inhibit PDK-1-dependent (a) protein kinase B phosphorylation/activation or (b) threonine 410 phosphorylation in the activation loop of PKC-. Also, insulin in situ and PIP 3 in vitro activated and stimulated autophosphorylation of a PKC-mutant, in which threonine 410 is replaced by glutamate (but not by an inactivating alanine) and cannot be activated by PDK-1. Surprisingly, insulin activated a truncated PKC-that lacks the regulatory (presumably PIP 3 -binding) domain; this may reflect PIP 3 effects on PDK-1 or transphosphorylation by endogenous full-length PKC-. Our findings suggest that insulin activates both PKCand PKC-in plasma membranes, microsomes, and GLUT4 vesicles by a mechanism requiring increases in PIP 3 , PDK-1-dependent phosphorylation of activation loop sites in PKC-and , and subsequent autophosphorylation and/or transphosphorylation.Insulin has been reported to activate atypical forms of protein kinase C (PKC), 1 i.e. PKC-and/or PKC-, in 3T3/L1 adipocytes (1, 2), rat adipocytes (3), L6 myotubes (4), and 32D cells (5). These increases in atypical PKC enzyme activity appear to be largely dependent upon activation of phosphatidylinositol (PI) 3-kinase (1-5) and subsequent increases in D3-PO 4 polyphosphoinositides, i.e. PI 3,4,5-(PO 4 ) 3 and PI 3,4-(PO 4 ) 2 (3). Moreover, transfection studies suggest that PKC-and/or PKC-is/are required for and may be sufficient for insulin stimulation of GLUT4 translocation and subsequent glucose transport (1-4). At present, there is only limited information on the mechanism whereby D3-PO 4 polyphosphoinositides activate atypical PKCs and little or no information on the subcellular compartments in which atypical PKCs are activated or, for that matter, whether one or both atypical PKCs are activated by insulin in specific cell types. With respect to the first point, recent findings (6, 7) suggest that PI 3,4,5-(PO 4 ) 3 and PI 3,4-(PO 4 ) 2 activate, or allow access for, 3-phosphoinositide-dependent kinase-1 (PDK-1), which phosphorylates threonine 410 in the activation loop of PKC-, thereby initiating the activation of this atypical PKC. Indeed, in other studies, we have found that PDK-1 action is required for insulin-induced activation of PKC-in rat adipocytes.2 However, it is uncertain whether this requirement reflects a permissive effect of PDK-1 or whether PDK-1 mediates acute a...

Kidney failure is common in patients with Coronavirus Disease-19 (COVID-19) resulting in increased morbidity and mortality. In an international collaboration, 284 kidney biopsies were evaluated to improve understanding of kidney disease in COVID-19. Diagnoses were compared to five years of 63,575 native biopsies prior to the pandemic and 13,955 allograft biopsies to identify diseases increased in patients with COVID-19. Genotyping for APOL1 G1 and G2 alleles was performed in 107 African American and Hispanic patients. Immunohistochemistry for SARS-CoV-2 was utilized to assess direct viral infection in 273 cases along with clinical information at the time of biopsy. The leading indication for native biopsy was acute kidney injury (45.4%), followed by proteinuria with or without concurrent acute kidney injury (42.6%). There were more African American patients (44.6%) than patients of other ethnicities. The most common diagnosis in native biopsies was collapsing glomerulopathy (25.8%) which associated with high-risk APOL1 genotypes in 91.7% of cases. Compared to the five-year biopsy database, the frequency of myoglobin cast nephropathy and proliferative glomerulonephritis with monoclonal IgG deposits was also increased in patients with COVID-19 (3.3% and 1.7%, respectively), while there was a reduced frequency of chronic conditions (including diabetes mellitus, IgA nephropathy, and arterionephrosclerosis) as the primary diagnosis. In transplants, the leading indication was acute kidney injury (86.4%), for which rejection was the predominant diagnosis (61.4%). Direct SARS-CoV-2 viral infection was not identified. Thus, our multi-center large case series identified kidney diseases that disproportionately affect patients with COVID-19, demonstrated a high frequency of APOL1 high-risk genotypes within this group, with no evidence of direct viral infection within the kidney.

Abstract-Multiple lines of evidence have suggested that alternative pathways to the angiotensin-converting enzyme (ACE) exists for angiotensin II (Ang II) generation in the heart, large arteries, and the kidney. In vitro studies in intact tissues, homogenates, or membrane isolates from the heart and large arteries have repeatedly demonstrated such pathways, but the issue remains unresolved because the approaches used have not made it possible to extrapolate from the in vitro to the in vivo situation. For our in vivo model, we studied young and healthy human volunteers, for the most part white and male; when these subjects achieved balance on a low salt diet to activate the renin system, the response of renal perfusion to pharmacological interruption of the renin system was studied. With this approach, we studied the renal vasodilator response to 3 ACE inhibitors, 2 renin inhibitors, and 2 Ang II antagonists at the top of their respective dose-response relationships. When these studies were initiated, our premise was that a kinin-dependent mechanism contributed to the renal hemodynamic response to ACE inhibition; therefore, the renal vasodilator response to ACE inhibition would exceed the alternatives. To our surprise, both renin inhibitors and both Ang II antagonists that were studied induced a renal vasodilator response of 140 to 150 mL/min/1.73 m 2 , Ϸ50% larger than the maximal renal hemodynamic response to ACE inhibition, which was 90 to 100 mL/min/1.73 m 2 . In light of the data from in vitro systems, our findings indicate that in the intact human kidney, virtually all Ang II generation is renin-dependent but at least 40% of Ang I is converted to Ang II by pathways other than ACE, presumably a chymase, although other enzyme pathways exist. Preliminary data indicate that the non-ACE pathway may be substantially larger in disease states such as diabetes mellitus. One implication of the studies is that at the tissue level, Ang II antagonists have much greater potential for blocking the renin-angiotensin system than does ACE inhibition-with implications for therapeutics. (Hypertension. 1998;32:387-392.) Key Words: renin-angiotensin system Ⅲ angiotensin II Ⅲ angiotensin-converting enzyme T he renin-angiotensin-aldosterone system is one of the longest recognized hormonal systems, having been described about 100 years ago in remarkable detail.1 Few would argue about the role of pharmacological interruption of the renin-angiotensin system (RAS) in the rekindling of interest in the renin system over the past 2 decades.2 Fewer still would debate the contribution of angiotensin-converting enzyme (ACE) inhibition to therapeutics in processes ranging from hypertension to congestive heart failure, and from ventricular remodeling after myocardial infarction to diabetic nephropathy. The use of ACE inhibitors has provided unambiguous benefit, well beyond even the most enthusiastic imagining 2 decades ago. Despite this extraordinary record of success, there is an undeniable basic fact that emerges from pharmacological princi...

Promoting self-awareness of wellbeing in beginning teachers will contribute to their longevity and productivity. As the profession ages we are faced with the fact that many teachers are retiring, creating some shortages; increased numbers are leaving the profession prematurely; fewer applicants are entering teacher training; and some argue that the current workforce of teachers cannot relate to current students’ life worlds. For beginning teachers, this raises issues related to their wellbeing and ability to transition and cope, and also justifiably provides challenges and opportunities for school reform, leadership and curriculum renewal. Additionally, teachers must be able to ‘connect’ with children and young people if they are to make a difference in their learning, health and wellbeing. In this paper we argue that the retention of teachers is dependent on having a wellbeing strategy in place that clearly identifies inhibiting and enabling strategies. Beginning teachers need to be able to identify such strategies to remain well – for themselves and the future of our children and young people.