Altered metabolism and DAM-signatures in female brains and microglia with aging.

Despite Alzheimer's disease (AD) disproportionately affecting women, the mechanisms remain elusive. In AD, microglia undergo 'metabolic reprogramming', which contributes to microglial dysfunction and AD pathology. However, how sex and age contribute to metabolic reprogramming in microglia is understudied. Here, we use metabolic imaging, transcriptomics, and metabolic assays to probe age- and sex-associated changes in brain and microglial metabolism. Glycolytic and oxidative metabolism in the whole brain was determined using Fluorescence Lifetime Imaging Microscopy (FLIM). Young female brains appeared less glycolytic than male brains, but with aging, the female brain became 'male-like.' Transcriptomic analysis revealed increased expression of disease-associated microglia (DAM) genes (e.g., ApoE, Trem2, LPL), and genes involved in glycolysis and oxidative metabolism in microglia from aged females compared to males. To determine whether estrogen can alter the expression of these genes, BV-2 microglia-like cell lines, which abundantly express DAM genes, were supplemented with 17ß-estradiol (E2). E2 supplementation resulted in reduced expression of DAM genes, reduced lipid and cholesterol transport, and substrate-dependent changes in glycolysis and oxidative metabolism. Consistent with the notion that E2 may suppress DAM-associated factors, LPL activity was elevated in the brains of aged female mice. Similarly, DAM gene and protein expression was higher in monocyte-derived microglia-like (MDMi) cells derived from middle-aged females compared to age-matched males and was responsive to E2 supplementation. FLIM analysis of MDMi from young and middle-aged females revealed reduced oxidative metabolism and FAD+ with age. Overall, our findings show that altered metabolism defines age-associated changes in female microglia and suggest that estrogen may inhibit the expression and activity of DAM-associated factors, which may contribute to increased AD risk, especially in post-menopausal women.

Altered Metabolism and DAM-signatures in Female Brains and Microglia with Aging.

Despite Alzheimer's disease (AD) disproportionately affecting women, the mechanisms remain elusive. In AD, microglia undergo 'metabolic reprogramming', which contributes to microglial dysfunction and AD pathology. However, how sex and age contribute to metabolic reprogramming in microglia is understudied. Here, we use metabolic imaging, transcriptomics, and metabolic assays to probe age-and sex-associated changes in brain and microglial metabolism. Glycolytic and oxidative metabolism in the whole brain was determined using Fluorescence Lifetime Imaging Microscopy (FLIM). Young female brains appeared less glycolytic than male brains, but with aging, the female brain became 'male-like.' Transcriptomic analysis revealed increased expression of disease-associated microglia (DAM) genes (e.g., ApoE, Trem2, LPL), and genes involved in glycolysis and oxidative metabolism in microglia from aged females compared to males. To determine whether estrogen can alter the expression of these genes, BV-2 microglia-like cell lines, which abundantly express DAM genes, were supplemented with 17ß-estradiol (E2). E2 supplementation resulted in reduced expression of DAM genes, reduced lipid and cholesterol transport, and substrate-dependent changes in glycolysis and oxidative metabolism. Consistent with the notion that E2 may suppress DAM-associated factors, LPL activity was elevated in the brains of aged female mice. Similarly, DAM gene and protein expression was higher in monocyte-derived microglia-like (MDMi) cells derived from middle-aged females compared to age-matched males and was responsive to E2 supplementation. FLIM analysis of MDMi from young and middle-aged females revealed reduced oxidative metabolism and FAD+ with age. Overall, our findings show that altered metabolism defines age-associated changes in female microglia and suggest that estrogen may inhibit the expression and activity of DAM-associated factors, which may contribute to increased AD risk, especially in post-menopausal women.

Genomic insights into rapid speciation within the world's largest tree genus Syzygium.

Species radiations, despite immense phenotypic variation, can be difficult to resolve phylogenetically when genetic change poorly matches the rapidity of diversification. Genomic potential furnished by palaeopolyploidy, and relative roles for adaptation, random drift and hybridisation in the apportionment of genetic variation, remain poorly understood factors. Here, we study these aspects in a model radiation, Syzygium, the most species-rich tree genus worldwide. Genomes of 182 distinct species and 58 unidentified taxa are compared against a chromosome-level reference genome of the sea apple, Syzygium grande. We show that while Syzygium shares an ancient genome doubling event with other Myrtales, little evidence exists for recent polyploidy events. Phylogenomics confirms that Syzygium originated in Australia-New Guinea and diversified in multiple migrations, eastward to the Pacific and westward to India and Africa, in bursts of speciation visible as poorly resolved branches on phylogenies. Furthermore, some sublineages demonstrate genomic clines that recapitulate cladogenetic events, suggesting that stepwise geographic speciation, a neutral process, has been important in Syzygium diversification.

Fatty acid sensing in the brain: The role of glial-neuronal metabolic crosstalk and horizontal lipid flux.

The central control of energy homeostasis is a regulatory axis that involves the sensing of nutrients, signaling molecules, adipokines, and neuropeptides by neurons in the metabolic centers of the hypothalamus. However, non-neuronal glial cells are also abundant in the hypothalamus and recent findings have underscored the importance of the metabolic crosstalk and horizontal lipid flux between glia and neurons to the downstream regulation of systemic metabolism. New transgenic models and high-resolution analyses of glial phenotype and function have revealed that glia sit at the nexus between lipid metabolism and neural function, and may markedly impact the brain's response to dietary lipids or the supply of brain-derived lipids. Glia comprise the main cellular compartment involved in lipid synthesis, lipoprotein production, and lipid processing in the brain. In brief, tanycytes provide an interface between peripheral lipids and neurons, astrocytes produce lipoproteins that transport lipids to neurons and other glia, oligodendrocytes use brain-derived and dietary lipids to myelinate axons and influence neuronal function, while microglia can remove unwanted lipids in the brain and contribute to lipid re-utilization through cholesterol efflux. Here, we review recent findings regarding glial-lipid transport and highlight the specific molecular factors necessary for lipid processing in the brain, and how dysregulation of glial-neuronal metabolic crosstalk contributes to imbalanced energy homeostasis. Furthering our understanding of glial lipid metabolism will guide the design of future studies that target horizontal lipid processing in the brain to ameliorate the risk of developing obesity and metabolic disease.

Altered substrate metabolism in neurodegenerative disease: new insights from metabolic imaging.

Neurodegenerative diseases (NDs), such as Alzheimer's disease (AD), Parkinson's disease (PD) and multiple sclerosis (MS), are relatively common and devastating neurological disorders. For example, there are 6 million individuals living with AD in the United States, a number that is projected to grow to 14 million by the year 2030. Importantly, AD, PD and MS are all characterized by the lack of a true disease-modifying therapy that is able to reverse or halt disease progression. In addition, the existing standard of care for most NDs only addresses the symptoms of the disease. Therefore, alternative strategies that target mechanisms underlying the neuropathogenesis of disease are much needed. Recent studies have indicated that metabolic alterations in neurons and glia are commonly observed in AD, PD and MS and lead to changes in cell function that can either precede or protect against disease onset and progression. Specifically, single-cell RNAseq studies have shown that AD progression is tightly linked to the metabolic phenotype of microglia, the key immune effector cells of the brain. However, these analyses involve removing cells from their native environment and performing measurements in vitro, influencing metabolic status. Therefore, technical approaches that can accurately assess cell-specific metabolism in situ have the potential to be transformative to our understanding of the mechanisms driving AD. Here, we review our current understanding of metabolism in both neurons and glia during homeostasis and disease. We also evaluate recent advances in metabolic imaging, and discuss how emerging modalities, such as fluorescence lifetime imaging microscopy (FLIM) have the potential to determine how metabolic perturbations may drive the progression of NDs. Finally, we propose that the temporal, regional, and cell-specific characterization of brain metabolism afforded by FLIM will be a critical first step in the rational design of metabolism-focused interventions that delay or even prevent NDs.

Local progression after operative treatment of metastatic kidney cancer.

The cases of 78 patients with osseous metastases from kidney cancer were reviewed to determine the rate of local progression after operative resection as compared with more traditional intralesional procedures. Group I consisted of 41 (53%) patients who were treated with intralesional procedures involving internal fixation with or without curettage or polymethylmethacrylate. Of the 41 patients, additional operations were recommended for 17 (41%) of the patients who had local osseous progression. Fourteen additional procedures including nine wide resections with reconstruction, three amputations, and two mass excisions were done. Group II consisted of 37 (47%) patients who were treated with marginal or wide resection with or without reconstruction. In this group, only one patient required additional operative intervention for local osseous progression. Median survival of patients in Group I was 20 months compared with 35 months for patients in Group IL This study shows that despite shorter average survival, patients who undergo intralesional surgery are at high risk of reoperation for local progression. Resectional surgery should be considered in patients with skeletal metastases from kidney cancer to lessen the risk of reoperation for local progression.

A novel reciprocal (10;17)(p11.2;q23) in myxoid fibrosarcoma.

Recurring reciprocal translocations have been identified as the primary chromosome aberrations in a number of neoplasms. These aberrations are often closely associated with particular morphologic or phenotypic subtypes of tumors and in some cases have prognostic implications. We have identified a novel reciprocal t(10;17)(p11.2;q23) in a case of low-grade myxoid fibrosarcoma, which may prove to be a new tumor specific chromosome aberration.

Apoptosis of osteocytes in glucocorticoid-induced osteonecrosis of the hip.

An increase in osteoblast and osteocyte apoptosis has been demonstrated in mice and humans receiving glucocorticoids and may be involved in the pathogenesis of the associated osteonecrosis. To examine the spatial relationship between osteocyte apoptosis and glucocorticoid-induced osteonecrosis, we determined the prevalence of osteocyte apoptosis in whole femoral heads obtained from patients who underwent prosthetic hip replacement because of osteonecrosis due to chronic glucocorticoid treatment (n = 5), alcoholism (n = 3), and trauma (n = 1) as well as in femoral neck cores from patients with sickle cell disease (n = 5). Abundant apoptotic osteocytes and cells lining cancellous bone were found juxtaposed to the subchondral fracture crescent in femurs from the patients with glucocorticoid excess. In contrast, apoptotic bone cells were absent from the specimens taken from patients with trauma or sickle cell disease and were rare with alcohol abuse. These results indicate that glucocorticoid-induced osteonecrosis is a misnomer. The bone is not necrotic; instead, it shows prominent apoptosis of cancellous lining cells and osteocytes. Glucocorticoid-induced osteocyte apoptosis, a cumulative and irreparable defect, could uniquely disrupt the mechanosensory function of the osteocyte network and thus start the inexorable sequence of events leading to collapse of the femoral head.

Orthopaedic management of chronic pain: pain management for the cancer patient.

Chronic pain is best managed through a multidisciplinary approach, which includes traditional and nontraditional therapies. Orthopaedists probably deal with chronic pain patients more than any other medical specialists. As Foley points out, it is critical that a trusting relationship between the patient and the physician is established; a relationship in which the patient believes the physician is taking the pain symptoms seriously and intends to address the pain aggressively. Although dealing with chronic pain can be a very frustrating part of orthopaedic practice, it can also be seen as a challenge.

Effect of methotrexate on distraction osteogenesis.

To assess the potential of using distraction osteogenesis to reconstruct bone deficient limbs after limb salvage for musculoskeletal sarcomas, the authors examined the effect of methotrexate on distraction osteogenesis in a rabbit tibial lengthening model. Eighteen rabbits underwent tibial corticotomy and application of a ring external fixator. Rabbits were assigned randomly to one of two groups in which either methotrexate (n = 12) or placebo (n = 6) was administered during a 21-day distraction period. Serum methotrexate levels and complete blood cell counts were monitored during distraction, and radiographs of the tibia were obtained weekly. Half of the animals from each group were sacrificed at the end of distraction, and the remaining animals were sacrificed after 6 weeks of neutral fixation when bone normally bridges the gap. Using methotrexate at serum concentrations similar to those used clinically for the treatment of human osteosarcomas, the authors were unable to show significant radiographic, histologic, or chemical differences in the effect of this antineoplastic drug on distraction osteogenesis in the rabbit model.

Evidence of an association between 6q13-21 chromosome aberrations and locally aggressive behavior in patients with cartilage tumors.

BACKGROUND: The finding of a cytogenetic-pathologic correlation between complex karyotypes and high grade cartilaginous tumors has been reported. However, few cytogenetic reports exist regarding benign or low grade lesions. A subset of low grade malignant cartilaginous tumors is characterized by locally aggressive behavior but no metastatic potential. Because the histopathologic distinction between benign, borderline, or low grade malignant cartilaginous lesions can be difficult, the finding of additional tumor markers associated with the clinical behavior of borderline cartilaginous lesions could be clinically significant. METHODS: Four cartilaginous tumors, including an osteochondroma (OC), a chondromyxoid fibroma (CF), an enchondroma (EC), and a dedifferentiated chondrosarcoma (DCS), were cultured and harvested using short term, in situ culture techniques. Chromosome analysis was performed by conventional G-banding and fluorescence in situ hybridization was used to confirm G-banding. RESULTS: The stemlines of all four tumors showed multiple chromosome anomalies that included aberrations of 6q13-21. The OC showed a t(6;16)(q21;p13.3). The CF showed a complex rearrangement between the chromosome 6 homologues, resulting in an inv(6)(p25q23)t(6;6)(q23;q13). This tumor also showed the clonal evolution of telomeric associations resulting in duplications, deletions, and the formation of a ring 15. The EC showed a der(6)t(6;15)(q13;q11)t(15;22)(q22;q13) stemline and subclones with an unstable iso 17q that subsequently fused to both ends of chromosome 16. The DCS showed a del(6)(q13), r(9), +12 stemline. CONCLUSIONS: The cytogenetic findings of this study suggest the cytogenetic-pathologic correlation of complex karyotypes found in high grade cartilage tumors may extend to lower grade tumors with complex karyotypes. These findings further suggest that chromosome aberrations in the region of 6q13-21 may be associated with locally aggressive behavior in patients with cartilage tumors.

Myositis ossificans: an unusual presentation in the foot.

Myositis ossificans is a non-neoplastic lesion characterized by heterotopic ossification of soft tissue. At varying stages of maturity, it shares similar histologic characteristics with sarcomatous lesions or maturing bone. Misdiagnosis can result in unnecessary radical treatment. This lesion has only rarely been reported in the foot. We present the case of a patient with plantar forefoot myositis ossificans.

Biomechanical analysis of patellar tendon allografts as a function of donor age.

We evaluated the biomechanical properties of patellar tendon allografts from donors aged 18 to 55 years. Bone-patellar tendon-bone complexes were harvested from acceptable donors and processed. Fat and soft tissue were removed, and the tendons were sectioned lengthwise leaving the central third. Area measurements were taken, and mechanical testing was performed. Specimens were pulled to failure at a rate of 10% of the initial length per second. The force at failure, tensile stress, modulus of elasticity, and percent elongation were determined for each specimen. There was no significant correlation (P > 0.05) between age and any of the mechanical properties. Load at failure ranged from 2110 to 4650 N, with a mean of 3424 N. Regression analysis showed slightly decreasing tensile stress with increasing age, but the correlation was not significant. It appears that patellar tendon allografts from donors up to age 55 have similar mechanical properties.

Pullout strength of fixation screws from polymethylmethacrylate bone cement.

Polymethylmethacrylate bone cement is often used to fill voids and increase the strength of osteoporotic and pathological bone. However, it is unclear as to which method of cement augmentation provides optimal screw fixation. This study was conducted to determine which of the current cement augmentation techniques provides the strongest construct when used in association with orthopaedic fixation screws. Pullout strength was determined for screws placed in sawbones with no cement, soft cement, doughy cement and hard cement after drilling and tapping. All cement-screw constructs were significantly stronger than the no cement group. Screws placed in doughy cement had a significantly higher pullout force than those placed in hard cement. Pullout strength of screws placed in soft cement was intermediate between the other cement techniques but not significantly different from either group.

Giant-cell tumor of bone: a rare entity in the hands of children.

Giant-cell tumor of bone is a relatively uncommon tumor in skeletally immature patients. It also occurs only infrequently in the hand. We present two cases of this tumor occurring in the hands of pediatric patients and discuss the literature regarding this disease process, including clinical presentation, pathology, and options for treatment.

Granular tricalcium phosphate grafting of cavitary lesions in human bone.

Development of an effective and safe bone grafting substitute could potentially alleviate many of the problems associated with human bone graft materials. Reported here are the results of 18 patients who were treated with a ceramic bone graft substitute, tricalcium phosphate. Twenty cavitary defects were created by intralesional curettage of a variety of benign bone neoplasms. Following complete removal of the tumor, each defect was filled with tricalcium phosphate granules. All patients were followed with periodic clinical and radiologic examinations for a minimum of 2 years (average followup of 37 months; range, 24 to 66 months). The amount of graft resorption and new bone formation were separately assessed on a percentage basis. Judged by radiographic criteria, the tricalcium phosphate implant lost its postoperative granular pattern in advance of bony trabeculae formation. Radiographic healing of defects with normal appearing bone was complete in four patients by 12 months, in an additional eight patients by 24 months, and in all except four patients by 48 months. Healing was dependent upon defect size; the larger lesions healing more slowly. Clinical parameters of healing were seen in advance of complete bone remodeling and all patients, except one, had unrestricted activity by 24 months. No adverse reactions were attributed to the graft material.

Case report 800: Skeletal fibrous dysplasia associated with intramuscular myxoma (Mazabraud's syndrome).

In summary, intramuscular myxoma associated with fibrous dysplasia of bone represents a benign disorder of uncertain etiology. Magnetic resonance imaging has proved to be a useful diagnostic tool in the evaluation of this benign disorder. In addition, MR is a valuable aid in the preoperative planning process.

Loose body of the elbow mimicking tumor.

Distinguishing between calcific soft-tissue masses and intra-articular loose bodies about the elbow can be difficult. We present the case of a 59-year-old woman with a slowly enlarging soft-tissue mass about the elbow. History, physical examination, and diagnostic studies did not establish a definite diagnosis, nor did they rule out the possibility of a neoplastic process. Excisional biopsy revealed a loose body contained in a cystic mass with elbow joint communication.