Progranulin Gene Therapy Improves Lysosomal Dysfunction and Microglial Pathology Associated with Frontotemporal Dementia and Neuronal Ceroid Lipofuscinosis.

Loss-of-function mutations in progranulin, a lysosomal glycoprotein, cause neurodegenerative disease. Progranulin haploinsufficiency causes frontotemporal dementia (FTD) and complete progranulin deficiency causes CLN11 neuronal ceroid lipofuscinosis (NCL). Progranulin replacement is a rational therapeutic strategy for these disorders, but there are critical unresolved mechanistic questions about a progranulin gene therapy approach, including its potential to reverse existing pathology. Here, we address these issues using an AAV vector (AAV-Grn) to deliver progranulin in Grn-/- mice (both male and female), which model aspects of NCL and FTD pathology, developing lysosomal dysfunction, lipofuscinosis, and microgliosis. We first tested whether AAV-Grn could improve preexisting pathology. Even with treatment after onset of pathology, AAV-Grn reduced lipofuscinosis in several brain regions of Grn-/- mice. AAV-Grn also reduced microgliosis in brain regions distant from the injection site. AAV-expressed progranulin was only detected in neurons, not in microglia, indicating that the microglial activation in progranulin deficiency can be improved by targeting neurons and thus may be driven at least in part by neuronal dysfunction. Even areas with sparse transduction and almost undetectable progranulin showed improvement, indicating that low-level replacement may be sufficiently effective. The beneficial effects of AAV-Grn did not require progranulin binding to sortilin. Finally, we tested whether AAV-Grn improved lysosomal function. AAV-derived progranulin was delivered to the lysosome, ameliorated the accumulation of LAMP-1 in Grn-/- mice, and corrected abnormal cathepsin D activity. These data shed light on progranulin biology and support progranulin-boosting therapies for NCL and FTD due to GRN mutations.SIGNIFICANCE STATEMENT Heterozygous loss-of-function progranulin (GRN) mutations cause frontotemporal dementia (FTD) and homozygous mutations cause neuronal ceroid lipofuscinosis (NCL). Here, we address several mechanistic questions about the potential of progranulin gene therapy for these disorders. GRN mutation carriers with NCL or FTD exhibit lipofuscinosis and Grn-/- mouse models develop a similar pathology. AAV-mediated progranulin delivery reduced lipofuscinosis in Grn-/- mice even after the onset of pathology. AAV delivered progranulin only to neurons, not microglia, but improved microgliosis in several brain regions, indicating cross talk between neuronal and microglial pathology. Its beneficial effects were sortilin independent. AAV-derived progranulin was delivered to lysosomes and corrected lysosomal abnormalities. These data provide in vivo support for the efficacy of progranulin-boosting therapies for FTD and NCL.

Restoring neuronal progranulin reverses deficits in a mouse model of frontotemporal dementia.

Loss-of-function mutations in progranulin (GRN), a secreted glycoprotein expressed by neurons and microglia, are a common autosomal dominant cause of frontotemporal dementia, a neurodegenerative disease commonly characterized by disrupted social and emotional behaviour. GRN mutations are thought to cause frontotemporal dementia through progranulin haploinsufficiency, therefore, boosting progranulin expression from the intact allele is a rational treatment strategy. However, this approach has not been tested in an animal model of frontotemporal dementia and it is unclear if boosting progranulin could correct pre-existing deficits. Here, we show that adeno-associated virus-driven expression of progranulin in the medial prefrontal cortex reverses social dominance deficits in Grn+/- mice, an animal model of frontotemporal dementia due to GRN mutations. Adeno-associated virus-progranulin also corrected lysosomal abnormalities in Grn+/- mice. The adeno-associated virus-progranulin vector only transduced neurons, suggesting that restoring neuronal progranulin is sufficient to correct deficits in Grn+/- mice. To further test the role of neuronal progranulin in the development of frontotemporal dementia-related deficits, we generated two neuronal progranulin-deficient mouse lines using CaMKII-Cre and Nestin-Cre. Measuring progranulin levels in these lines indicated that most brain progranulin is derived from neurons. Both neuronal progranulin-deficient lines developed social dominance deficits similar to those in global Grn+/- mice, showing that neuronal progranulin deficiency is sufficient to disrupt social behaviour. These data support the concept of progranulin-boosting therapies for frontotemporal dementia and highlight an important role for neuron-derived progranulin in maintaining normal social function.

Chronic inhibition of p38MAPK improves cardiac and endothelial function in experimental diabetes mellitus.

To investigate the influence of p38 mitogen activated kinase (p38MAPK) on the development of diabetic cardiac and endothelial dysfunction, we assessed left ventricular and vascular function as well as inflammatory markers in diabetic rats after chronic pharmacological inhibition of p38MAPK. Diabetes mellitus was induced in rats by a single injection of streptozotocin. Rats were treated with the p38MAPK inhibitor SB 239063 (40 mg/kg/day, p.o.) or vehicle. 48 days after diabetes mellitus-induction, left ventricular function and vascular function were assessed in vivo by TIP-catheter and the autoperfused hindlimb, respectively. Cell adhesion molecules staining was quantified immunohistochemically in the heart and quadriceps muscle, respectively, as well as cardiac phosphorylation of p38MAPK by Western blot analysis. Treated and untreated diabetic groups displayed similar severe hyperglycemia. Left ventricular and endothelial function were impaired in the untreated diabetic group compared to controls (dp/dtmax: -40%, dp/dtmin: +49%, maximal vasodilatation: -57%; P < 0.05) associated with significantly increased cardiac (3-fold) and peripheral cell adhesion molecules staining, respectively. Treatment of diabetic rats with SB 239063 led to a significant reduction of diabetes-induced enhancement of p38MAPK phosphorylation associated with improved left ventricular function (dp/dtmax: +39%, dp/dtmin: +47%; P < 0.05) and peripheral endothelial function (maximal vasodilatation: +71%; P < 0.05) under diabetic conditions. This was associated with reduced cardiac and peripheral inflammation indexed by reduced adhesion molecules content. Pharmacological inhibition of p38MAPK is sufficient to mitigate the development of diabetic cardiac and endothelial dysfunction despite of hyperglycemia. Our data suggest that the anti-inflammatory properties due to p38MAPK inhibition contribute to these beneficial cardiovascular effects.

Pectoralis major muscle injuries: evaluation and management.

Pectoralis major muscle tears are relatively rare injuries that primarily occur while lifting weights, particularly when doing a bench press. Complete ruptures are most commonly avulsions at or near the humeral insertion. Ruptures at the musculo-tendinous junction and intramuscular tears usually are caused by a direct blow. The patient may hear a snap at the time of injury and report pain, weakness, swelling, or muscular deformity. Physical examination can reveal ecchymosis, a palpable defect, asymmetric webbing of the axillary fold, and weakness on resisted shoulder adduction and internal rotation. A detailed history and physical examination can be augmented by radiologic studies, including magnetic resonance imaging. Nonsurgical treatment is now recommended only for the older, sedentary patient or for proximal muscle belly tears. Surgery, whether early or delayed, consistently yields superior results compared with nonsurgical management. Prompt diagnosis and timely intervention likely will produce improved results.

The effect of intra-articular methadone on postoperative pain following anterior cruciate ligament reconstruction.

BACKGROUND: Intra-articular narcotics have proven efficacy for providing pain relief following knee arthroscopy. This effect is short-lived. Methadone, with its long serum half-life (thirty-five hours, compared with two hours for morphine) could provide improved and prolonged pain relief. The purpose of the present study was to examine the effects of an intra-articular injection of methadone on postoperative analgesia following arthroscopic anterior cruciate ligament reconstruction. METHODS: Sixty-five skeletally mature patients undergoing primary anterior cruciate ligament reconstruction were randomly assigned to one of three groups, all of which received an intra-articular injection consisting of 9.5 mL of 0.5% bupivacaine with 1:200,000 epinephrine at the completion of the procedure. In addition, the remaining 0.5 mL of the syringe was filled with one of three substances. The study group (twenty-five patients) received 5 mg of methadone, the comparison group (twenty-one patients) received 5 mg of morphine, and the control group (nineteen patients) received 0.5 mL of saline solution. All supplemental pain medications were given on an as-needed basis, recorded, and converted to morphine equivalents. Specific variables that were measured included supplemental analgesia requirements during both the inpatient period and the outpatient period (from the time of discharge to the seventh postoperative day) and pain scores. RESULTS: There was no significant difference in inpatient (p = 0.998) or outpatient (p = 0.887) supplemental analgesic requirements or pain scores between the methadone group (Group 1) and the control group (Group 3). The morphine group (Group 2) required significantly less inpatient (p = 0.014) and outpatient (p = 0.044) supplemental analgesia compared with the control group (Group 3). There were no complications. CONCLUSIONS: The present report represents the first known study of the use of intra-articular methadone and establishes that this analgesic is safe at a single dose of 5 mg. At this dose, however, methadone does not provide improved postoperative analgesia following arthroscopic anterior cruciate ligament reconstruction. In contrast, intra-articular morphine does appear to be effective for decreasing postoperative pain.

Modeling energy savings from urban shade trees: an assessment of the CITYgreen energy conservation module.

CITYgreen software has become a commonly used tool to quantify the benefits of urban shade trees. Despite its frequent use, little research has been conducted to validate results of the CITYgreen energy conservation module. The first objective of this study is to perform a familiar application of CITYgreen software to predict the potential energy savings contribution of existing tree canopies in residential neighborhoods during peak cooling summer months. Unlike previous studies utilizing CITYgreen, this study also seeks to assess the software's performance by comparing model results (i.e., predicted energy savings) with actual savings (i.e., savings derived directly from energy consumption data provided by the electric utility provider). Homeowners in an older neighborhood with established trees were found to use less energy for air-conditioning than homeowners in a recently developed site. Results from the assessment of model performance indicated that CITYgreen more accurately estimated the energy savings in the highly vegetated, older neighborhood.

Distribution and care of shipboard blast injuries (USS Cole DDG-67).

On October 12, 2000, the destroyer USS Cole was anchored in a foreign port and was severely damaged by explosives in a small craft adjacent to the ship at the port side waterline. Seventeen crew members were killed in the incident. The wounded were evacuated to several medical facilities for their initial care and then to the military hospital in Landstuhl, Germany, and subsequently to the Charette Health Care Center (Naval Medical Center) in Portsmouth, Virginia. There were 35 surviving patients who had sustained 81 total injuries associated with the explosion. The distribution of the injuries included orthopedic, ophthalmologic, soft tissue, otolaryngologic, burns, inhalation, and other miscellaneous injuries. Twenty-seven of the patients were discharged after 24-hour observation. This article reviews the distribution of injuries found in the fatalities and the wounded crew members and the subsequent care required to prepare the military medical community for potential future incidents of this type.

Orthopaedic injuries among survivors of USS COLE attack.

The objective of this article is to describe the range of orthopaedic injuries and outcomes of acute treatment regimens among survivors of the USS COLE terrorist attack and to reemphasize basic treatment principles for blast injuries. With the current geopolitical environment, the average community orthopaedic surgeon may be involved in treating injuries due to an explosive terrorist attack. This is a retrospective review of a consecutive series of the 39 patients who were injured during the USS COLE attack on October 12, 2000, and were received at Naval Medical Center, Portsmouth, Virginia, from the MEDEVAC (Medical Evacuation) system. The 17 casualties from the attack were not included in this study. Data were retrospectively collected from patient charts for all patients who survived the USS COLE attack. The 39 patients who survived the USS COLE attack sustained 81 injuries. Fourteen patients sustained 32 orthopaedic injuries, of which 61% were lower extremity injuries. Of the 10 patients who required hospitalization, 6 had orthopaedic injuries (60%). Three of five open fractures (60%) became infected, and two of two (100%) open fracture wounds treated with primary closure in the initial setting were infected. Lower extremity orthopaedic injuries may predominate in a shipboard blast scenario. Even minor injuries require prolonged time before patients return to active duty. Complex wounds have high infection rates and should be treated according to previously established protocols for wartime injuries. Principles of provisional fracture stabilization prior to transport, adequate wound débridement, and delayed wound closure are reviewed.

An evaluation of the histological effects of intra-articular methadone in the canine model.

Methadone hydrochloride is an opiate that has pharmacodynamic and pharmacokinetic properties that suggest it may provide longer analgesia than morphine when administered via the intra-articular route. However, no studies to date have been conducted examining the effects of intra-articular methadone hydrochloride on local tissues. Therefore, the purpose of this study was to determine the histopathologic effects of intra-articular methadone hydrochloride on local tissues in the canine knee. Nine canines, 1 to 4 years old, weighing between 20 kg and 23 kg were used. All canines had their knees randomized to receive either bupivacaine, 0.5% with epinephrine 1:200,000 (4.5 mL), and 5 mg methadone hydrochloride (0.5 mL) for the study knee, or bupivacaine, 0.5% with epinephrine 1:200,000 (4.5 mL), and 0.5 mL normal saline for the control knee. Serum methadone hydrochloride levels were obtained on all canines at 6 and 24 hours. Canines were randomly assigned to 1 of 3 groups to be euthanized at either 24 hours, 14 days, or 28 days. Following euthanization and necropsy, synovial fluid levels and tissue samples were obtained and examined for histopathologic changes. Synovial fluid samples noted a few white blood cells at 24 hours and none at 14 and 28 days. Tissue samples showed no histopathologic changes, and serum concentration levels of methadone hydrochloride were negligible.