Genistein decreases food intake, body weight, and fat pad weight and causes adipose tissue apoptosis in ovariectomized female mice

Genistein, an isoflavone in soybean products, has estrogenic activity and is used as a natural substitute for estrogen replacement therapy in postmenopausal women. Genistein was also shown to decrease fat pad weight in female mice. The primary objective of this study was to determine the effect of genistein on adipose tissue apoptosis in vitro and in vivo. 3T3-L1 preadipocytes and mature adipocytes were treated with 0, 1, 10, 100, and 400 micromol/L genistein and then assayed for apoptosis, whereas only mature adipocytes were assayed for viability. Mature adipocytes treated with genistein demonstrated a dose-related increase in apoptosis. Ovariectomized female mice (9 mo old) were given 0, 150, or 1,500 mg/kg genistein in the semipurified phytoestrogen-free casein-based diet for 3 wk (n=10). After mice were killed, body composition was determined by dual-energy X-ray absorptiometry analysis, and parametrial (PM), inguinal (ING), and retroperitoneal (RP) fat pads were weighed and assayed for apoptosis (% DNA fragmentation). Genistein (1500 mg/kg) reduced food intake (FI) by 14% (P<0.01) and body weight (BW) by 9% (P<0.01). Body composition was not significantly affected, but PM and ING weights were decreased 22% (P<0.05) and 19% (P<0.07), respectively, by 1,500 mg/kg genistein. Apoptosis in ING fat was increased 290% (P<0.05) by 1,500 mg/kg genistein. These findings show that oral genistein treatment can reduce BW, mobilize body fat, and induce apoptosis of adipose tissue in ovariectomized female mice. Thus, genistein may be useful in treating or preventing increased adiposity after menopause.     

Rapid TLC determination of methadyl acetate and some in vitro metabolites.

Methadyl acetate was metabolized by microsomal preparations of rat liver to yield nor-methadyl acetate and 6-(dimethylamino)-4,4-diphenyl-3-heptanol. The identification and separation of these three compounds was established by TLC, using iodoplatinate spray as a visualizing agent.     

Novel treatments for obesity and osteoporosis: targeting apoptotic pathways in adipocytes

Obesity and osteoporosis have grave consequences for human health, quality of life, and even the efficiency of the labor force and economy. However, these pathologies share a common cell progenitor, revealing a surprising target for drug research and development. Recent findings show that high adipocyte count in bone marrow is directly related to bone loss, as fat cells replace osteoblasts (or bone-forming cells). The objective of this review is to examine the importance of adipocyte apoptosis in the treatment of obesity and/or osteoporosis, with special emphasis on natural products as promising leads for drug development. We have induced in vivo adipocyte apoptosis, using leptin, ciliary neurotrophic factor (CNTF), beta adrenergic agonists and conjugated linoleic acid (CLA) in rodents. The results of leptin treatments on rats are suppressed food intake, reduced body weight, reduced body fat, adipocyte apoptosis, and elevated energy expenditure. Further, leptin treatment of leptin-deficient (ob/ob) mice increases endosteal bone formation and bone mineral density. Adipocyte apoptosis has also been induced in vitro using tumor necrosis factor-alpha (TNF-alpha), (-)-epigallocatechin gallate (EGCG) from Camellia sinensis and ajoene, from Allium sativum. Natural products have potential for inducing apoptosis of adipose tissue, inhibiting bone marrow adipogenesis and increasing the expression of osteogenic factors in bone, thereby yielding effective treatments for obesity and osteoporosis.     

Alterations of temporalis muscle contractile force and histological content from the myostatin and Mdx deficient mouse

Myostatin (GDF8) and dystrophin are critical molecules for muscle organisation. Myostatin is involved in regulating muscle growth and development, whereas dystrophin is part of the dystrophin-glycoprotein complex (DGC), which anchors the cytoskeleton to the sarcolemma. We examined temporalis muscle morphology and function in myostatin deficient and dystrophin deficient (Mdx) mice in order to determine how myostatin and dystrophin affect bite force and muscle fibre composition. Bite forces from 4-month-old myostatin-/-, dystrophin deficient (Mdx) and normal control mice were measured by load cell and field stimulation of the temporalis muscle. Tissue sections were stained with haemotoxylin and eosin (H&E) and the periodic acid-Schiff reaction (PAS) to assess morphology and fibre type differences. A positive relationship between bite force and muscle mass for both genetic models was observed. Both Mstn-/- and Mdx mice showed significant elevation in bite force and muscle mass. Histological examination revealed greater muscle fibre cross-sectional area variability in Mdx mice (ANOVA, F=5.6, P<0.01). Surprisingly, the Mstn-/- mice demonstrated a disproportionate increase in bite force at higher stimulation frequencies with comparison of regression lines for force-frequency data (ANOVA, F=3.46, P<0.07). Muscle fibre typing using a PAS staining technique revealed significantly more type IIx/b glycolytic muscle fibres in the Mstn-/- mice. Our results suggest that histopathologies associated with Mdx mice did not diminish gross temporalis structure or function, whilst the force-frequency changes associated with Mstn-/- mice were reflected in an elevation of type IIx/b fibres.     

Granulocyte colony-stimulating factor administration alters femoral biomechanical properties in C57BL/6 mice

The clinical application of G-CSF is broadening. In addition to treating neutropenia and in bone marrow transplants, it is now being considered for functional recovery after myocardial infarction and stroke. It is thus very important that the effects of extended G-CSF administration on the skeleton are investigated. To simulate this potential clinical use of G-CSF in postmyocardial infarction or cerebral vascular accident, a 2-week course of administration was selected. Ten C57BL/6 mice at 22 weeks of age were given intraperitoneal injection of saline, and another nine of the same age were given G-CSF. Four weeks later, femurs were harvested and three-point-bend tests were performed until fracture. From the load-displacement curve recorded during the test, the stiffness, Young's modulus, fracture strength of the bone, fracture energy, and the total energy to break the femur were determined. The test data show that mice treated with G-CSF have significantly lower modulus in their femurs when compared to the controlled mice treated with saline. The stiffness demonstrates the largest decrease, by as much as 25%. As its clinical use increases, G-CSF effects on the mechanical properties of the skeleton become increasingly more important because many of these diseases occur in older patients with already compromised skeleton by osteopenia or osteoporosis. How G-CSF administration achieves these alterations in skeletal biomechanical properties is unclear. Although the current findings confirm its known temporary catabolic effects on bone homeostasis, it also suggests that a transient state of higher bone compliance following the end of G-CSF administration can be achieved that may have clinical benefits.     

A role for myokines in muscle-bone interactions

This review presents the hypothesis that muscle is a source of secreted factors (myokines) that can influence bone mass in both positive and negative ways. Growth factor secretion by muscle may therefore be one pathway through which mechanical signals are transduced biologically.     

Complex Anatomic Abnormalities of the Lower Leg Muscles and Tendons Associated With Phocomelia: A Case Report

Musculoskeletal anatomy is widely known to have components that stray from the norm in the form of variant muscle and tendon presence, absence, origin, insertion, and bifurcation. Although these variant muscles and tendons might be deemed incidental and insignificant findings by most, they can be important contributors to pathologic physiology or, more importantly, an option for effective treatment. In the present case report, we describe a patient with phocomelia and Müllerian abnormalities secondary to in utero thalidomide exposure. The patient had experienced recurrent bilateral foot pain accompanied by numbness, stiffness, swelling, and longstanding pes planus. These symptoms persisted despite conservative treatment with orthotics, steroids, and nonsteroidal anti-inflammatory drugs. Radiographic imaging showed dysmorphic and degenerative changes of the ankle and foot joints. Further investigation with magnetic resonance imaging revealed complex anatomic abnormalities, including the absence of the posterior tibialis and peroneus brevis, lateralization of the peroneus longus, and the presence of a variant anterior compartment muscle. The variant structure was likely a previously described anterior compartment variant, anterior fibulocalcaneus, and might have been a source of the recurrent pain. Also, the absence of the posterior tibialis might have caused the pes planus in the present patient, considering that posterior tibialis tendon dysfunction is the most common cause of acquired pes planus. Although thalidomide infrequently affects the lower extremities, its effects on growth and development were likely the cause of this rare array of anatomic abnormalities and resulting ankle and foot pathologic features.