The present studies were conducted to determine whether prostaglandin F2a (PGF2a) stimulates the production of "second messengers" derived from inositol phospholipid hydrolysis and increases intracellular free Ca2+ ([Ca2+],) in isolated bovine luteal cells. PGF2. provoked rapid (10 sec) and sustained (up to 60 min) increases in the levels of inositol mono-, bis-, and trisphosphates (InsP, InsP2, and InsP3, respectively PGF2a-induced increases in [Ca2+], were also independent of extracellular calcium. These findings demonstrate that the action of PGF2a is coupled to the phospholipase C-InsP3 and diacylglycerol second messenger system in the corpus luteum.The ability of prostaglandin F2a (PGF2a) to cause a loss of function of the corpus luteum was demonstrated over 15 years ago (1,2), but the precise mechanism of action of PGF2, remains unknown. PGF2a-induced luteolysis is correlated with changes in luteal membrane lipid composition and reductions of membrane fluidity (3-6), which have been suggested to prevent gonadotropin receptor aggregation (7) and the association and coupling to adenylate cyclase (8-10). PGF2a also acts at postreceptor sites to inhibit steroidogenesis (10, 11). Behrman and coworkers (12,13)

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MR imaging of high-altitude cerebral edema shows reversible WM edema, especially in the corpus callosum and subcortical WM. Recent studies have revealed hemosiderin deposition in WM long after high-altitude cerebral edema has resolved, providing a high-altitude cerebral edema "footprint." We wished to determine whether these microbleeds are present acutely and also describe the evolution of all MR imaging findings. In 8 patients with severe high-altitude cerebral edema, we obtained 26 studies: 18 with 3T and 8 with 1.5T scanners, during the acute stage, recovery, and follow-up in 7 patients and acutely in 1 patient. Imaging confirmed reversible cytotoxic and vasogenic WM edema that unexpectedly worsened the first week during clinical improvement before resolving. The 3T SWI, but not 1.5T imaging, showed extensive microbleeds extending beyond areas of edema seen acutely, which persisted and with time coalesced. These findings support cytotoxic and vasogenic edema leading to capillary failure/leakage in the pathophysiology of high-altitude cerebral edema and provide imaging correlation to the clinical course.ABBREVIATIONS: HACE ϭ high-altitude cerebral edema; HAPE ϭ high-altitude pulmonary edema; MB ϭ microbleed

Cholesteryl ester (CE)-loaded macrophages (foam cells) are a prominent feature of atherosclerotic plaques. Previous studies have shown that human monocytes or resident mouse peritoneal macrophages accumulate CE in response to low density lipoprotein (LDL) only when the LDL has been appropriately chemically modified. By contrast, we report here that J774 macrophages accumulate large amounts of CE when incubated with unmodified LDL. The CE is stored in oil red 0-positive droplets, which have the typical appearance of foam cell inclusions by electron microscopy. (3), and LDL has been implicated as the major atherogenic lipoprotein (1); thus, the possibility exists that, under certain circumstances not yet elucidated, LDL could lead to CE accumulation. Investigations into the mechanism of CE accumulation in macrophages, which are thought to be the major source of atheromatous foam cells (3), have primarily utilized two models-mouse peritoneal macrophages and human monocytes (4). These studies have revealed that CE accumulation can be induced by LDL only when the LDL is modified by negative charges [e.g., acetyl-LDL and malondialdehyde-LDL) via the "scavenger receptor" (4)]. Although the studies with modified LDL suggest that LDL may have to be altered in order to realize its atherogenic potential, it is not known whether peritoneal macrophages or blood-borne monocytes are appropriate models for foam cell formation or whether sufficient quantities of the modified lipoproteins exist in vivo to result in significant macrophage CE accumulation. We report here that a macrophage cell line, the mouse J774 cell, does accumulate large amounts of CE when incubated with native LDL, providing an example of unmodified LDL inducing foam cell formation. MATERIALS AND METHODS -Cells. Monolayer cultures of the J774 macrophage-like cell line (5), obtained from Jay Unkeless (The Rockefeller University), were maintained in Dulbecco's modified Eagle's medium (DME medium) containing 10%o heat-inactivated (560C for 30 min) fetal bovine serum (medium A). The cells were demonstrated to have the following macrophage-like characteristics; adherence to plastic, phagocytosis of latex particles and antibody-coated sheep erythrocytes, positive staining for peroxidase and nonspecific esterase, and positive enzyme assays for B3-glucuronidase and acid phosphatase. Several tests of the culture for mycoplasma contamination were negative. Normal newborn human fibroblasts (frozen in the sixth passage), kindly provided by Joseph Cornicelli, (Columbia University) were grown in DME medium containing 10% fetal bovine serum. Mouse peritoneal macrophages and P388D1 cells (5), obtained from John Wunderlich (National Institutes of Health), were grown in medium A. HL 60 cells (5), obtained from T. R. Breitman (National Institutes of Health), were maintained in RPM1 1640 medium supplemented with 10% heat-inactivated fetal bovine serum; the cells were induced to differentiate into mature macrophage-like cells by treatment with 16 nM phorbol 12-myristate 1...

We have chosen to test the safety of human intracerebroventricular (ICV) brain injections of autologous non-geneticallymodified adipose-derived stromal vascular fraction (ADSVF). In this IRB-approved trial, 24 patients received ICV ADSVF via an implanted reservoir between 5/22/14 and 5/22/17. Seven others were injected via their ventriculo-peritoneal shunts. Ten patients had Alzheimer's disease (AD), 6 had amyotrophic lateral sclerosis (ALS), 6 had progressive multiple sclerosis (MS-P), 6 had Parkinson's "Plus" (PD+), 1 had spinal cord injury, 1 had traumatic brain injury, and 1 had stroke. Median age was 74 (range 41-83). Injections were planned every 2-3 months. Thirty-one patients had 113 injections. Patients received SVF injection volumes of 3.5-20 cc (median:4 cc) containing 4.05 × 10 5 to 6.2 × 10 7 cells/cc, which contained an average of 8% hematopoietic and 7.5% adipose stem cells. Follow-up ranged from 0 to 36 months (median: 9.2 months). MRIs post injection(s) were unchanged, except for one AD patient whose hippocampal volume increased from < 5th percentile to 48th percentile (NeuroQuant ® volumetric MRI). Of the 10 AD patients, 8 were stable or improved in tests of cognition. Two showed improvement in P-tau and ß-amyloid levels. Of the 6 MS-P patients all are stable or improved. Four of 6 ALS patients died of disease progression. Twelve of 111 injections (11%) led to 1-4 days of transient meningismus, and mild temperature elevation, which resolved with acetaminophen and/or dexamethasone. Two (1.8% of injections) required hospitalization for these symptoms. One patient (0.9% of injections) had his reservoir removed and later replaced for presumed infection. In this Phase 1 safety trial, ADSVF was safely injected into the human brain ventricular system in patients with no other treatment options. Secondary endpoints of clinical improvement or stability were particularly promising in the AD and MS-P groups. These results will be submitted for a Phase 2 FDA-approved trial.