Insulin-degrading enzyme binds to the nonglycosylated precursor of varicella-zoster virus gE protein found in the endoplasmic reticulum.

Insulin degradation enzyme (IDE) is a 110-kDa zinc metalloprotease found in the cytosol of all cells. IDE degrades insulin and a variety of small proteins including amyloid-beta. Recently, IDE has been proposed as the receptor for varicella-zoster virus (VZV) attachment. During our reassessment, some of the original studies were repeated and expanded in scope. We first confirmed that IDE antibody reduced VZV spread. For additional controls, we repeated the same experiments with herpes simplex virus (HSV)-infected cells as well as uninfected cells. There was a visible reduction in HSV spread but less than seen in the VZV system. Of greater importance, IDE antibody also inhibited the growth of uninfected cells. Second, we repeated the coprecipitation assays. We confirmed that antibodies to VZV gE (open reading frame 68) coprecipitated IDE and that anti-IDE antibody coprecipitated gE. However, the detected gE protein was not the mature 98-kDa form; rather, it was a precursor 73-kDa gE form found in the endoplasmic reticulum. Additional control experiments included VZV-infected cell cultures treated with tunicamycin to block gE glycosylation in the endoplasmic reticulum; again, the anti-IDE antibody coprecipitated a 73-kDa gE product. Finally, Orbitrap mass spectrometry analysis of a chromatographically purified gE sample revealed four cellular proteins associated with the unfolded protein response: BiP (HSPA5), HSPA8, HSPD1, and PPIA (peptidyl-propyl cis-trans isomerase). We conclude that IDE protease binds to the 73-kDa gE precursor and that this event occurs in the cytosol but not as a receptor/ligand interaction.

Quality of mass-reared codling moth (Lepidoptera: Tortricidae) after long-distance transportation: 1. Logistics of shipping procedures and quality parameters as measured in the laboratory.

The sterile insect technique (SIT) is a proven effective control tactic against lepidopteran pests when applied in an areawide integrated pest management program. The construction of insect mass-rearing facilities requires considerable investment and moth control strategies that include the use of sterile insects could be made more cost-effective through the importation of sterile moths produced in other production centers. For codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), this is an attractive option because mating studies have confirmed the absence of mating barriers between codling moth populations from geographically different areas. To assess the feasibility of long-distance transportation of codling moths, pupae and adult moths were transported in 2004 from Canada to South Africa in four shipments by using normal commercial transport routes. The total transport time remained below 67 h in three of the consignments, but it was 89 h in the fourth consignment. Temperature in the shipping boxes was fairly constant and remained between -0.61 and 0.16 degrees C for 76.8-85.7% of the time. The data presented indicate that transporting codling moths as adults and pupae from Canada to South Africa had little effect on moth emergence, longevity, and ability to mate, as assessed in the laboratory. These results provide support to the suggestion that the STT for codling moth in pome fruit production areas might be evaluated and implemented by the importation of irradiated moths from rearing facilities in a different country or hemisphere.

Irradiation biology of male brown marmorated stink bugs: is there scope for the sterile insect technique?

PURPOSE: Brown marmorated stink bugs, Halyomorpha halys Stål (Hemiptera: Pentatomidae), are regularly intercepted, but there are few eradication tools. Currently, no sterile insect technique program exists for Hemiptera. MATERIALS AND METHODS: Adult males were irradiated at 4-60 Gy, mated and their progeny reared for two generations, with mortality assessed at F1 egg, F1 adult and F2 egg stages. RESULTS: The F1 eggs showed a dose response to irradiation between 4 and 36 Gy, with 97% sterility at 16 Gy, and higher doses producing complete egg mortality. Only rare F1 survivors had progeny, but the F2 generation showed identical responses between maternal and paternal lines; most egg batches showed either very low or very high mortality. Irradiation with 16 Gy resulted in 98.5% sterility, cumulative over F1 and F2. CONCLUSIONS: Lack of a dose response at the F2 generation precludes the use of irradiation-induced inherited sterility. The conventional sterile insect technique appears possible by irradiation of males from ∼12 to 16 Gy. The effect of radiation dose on females is not known, thus we cannot conclude whether bi-sex release is feasible so for now the release of males only is recommended. More work is needed on the competitive fitness of irradiated males, and logistics such as mass rearing or field collection, in order to determine the feasibility of the approach.

Insulin-like growth factor-1 content and pattern of expression correlates with histopathologic grade in diffusely infiltrating astrocytomas.

Studies of experimental tumorigenesis have strongly implicated signaling of the insulin-like growth factor 1 (IGF-1) as a key component in astrocytic neoplasia; however, its role in the growth of low-grade and malignant human tumors is not well understood. Correlative analyses of IGF-1, p53, and Ki-67 (MIB-1) immunohistochemistry and IGF-1 receptor (IGF-1R) mRNA expression were performed to examine the cellular pattern of IGF-1 signaling in 39 cases of astrocytoma (World Health Organization grades II-IV). Tumor cells expressing IGF-1 and IGF-1R were present in all tumor grades. The proportion of tumor cells that expressed IGF-1 correlated with both histopathologic grade and Ki-67 labeling indices, while expression of IGF-1R mRNA correlated with Ki-67 indices. In cases where stereotactic tissue sampling could be identified with a specific tumor area by neuroimaging features, the numbers of IGF-1 immunoreactive cells correlated with the tumor zones of highest cellularity and Ki-67 labeling. In glioblastomas, the localization of IGF-1 immunoreactivity was notable for several features: frequent accentuation in the perivascular tumor cells surrounding microvascular hyperplasia; increased levels in reactive astrocytes at the margins of tumor infiltration; and selective expression in microvascular cells exhibiting endothelial/pericytic hyperplasia. IGF-1R expression was particularly prominent in tumor cells adjacent to both microvascular hyperplasia and palisading necrosis. These data suggest that IGF-1 signaling occurs early in astroglial tumorigenesis in the setting of cell proliferation. The distinctive correlative patterns of IGF-1 and IGF-1R expression in glioblastomas also suggest that IGF-1 signaling has an association with the development of malignant phenotypes related to aberrant angiogenesis and invasive tumor interactions with reactive brain.

Failure of growth hormone to alter the biomechanics of fracture-healing in a rabbit model.

Standardized tibial osteotomies were created and stabilized with external fixation in twenty-seven skeletally mature rabbits. Fourteen animals were treated with a daily injection of human growth hormone (150 micrograms per kilogram of body weight), and thirteen received a daily injection of saline solution. Serial non-destructive biomechanical tests, radiography, and determinations of the levels of serum insulin-like growth-factor I were performed for all of the animals. Destructive strength-testing of the sites of osteotomy was performed at four, six, or eight weeks. Twenty-five of the twenty-seven osteotomies healed uneventfully. There were no significant differences in the serial biomechanical measurements at the sites of the healing osteotomies, in the radiographic measurements, or in the ultimate strength of the sites of the osteotomy between the two groups. The mean level of serum insulin-like growth-factor I increased 33 per cent relative to the preoperative baseline level in the group that received growth hormone and increased 10 per cent in the control group. This difference was not statistically significant. There was no significant correlation between the biomechanical properties at the sites of the osteotomies and the levels of serum insulin-like growth-factor I. Administration of growth hormone had no measurable effect on fracture-healing in this model of normal animals. It remains to be determined whether injection of growth hormone might affect healing when there is a state of deficiency of endogenous growth hormone or when there is a non-union of a fracture.

The effects of muscle fatigue on shoulder joint position sense.

Proprioception, or joint position sense, probably plays an important role in shoulder joint function. In this study, we assessed the effect of muscle fatigue on shoulder proprioception in 20 volunteers with no shoulder abnormalities. Shoulder proprioception was measured as the threshold to first detection of humeral rotation with the joint at 90 degrees of abduction and 90 degrees of external rotation. Subjects were tested while rested, exercised on a isokinetic testing machine until fatigued, and then retested in an identical fashion. Both shoulders were tested, and the order of dominant and nondominant shoulder was randomized. Shoulder proprioception was analyzed for its dependence on arm dominance, direction of rotation, and muscle fatigue. Subjects detected external rotation after significantly less movement than they did internal rotation. Overall, before exercise, motion was detected after a mean of 0.92 degrees of rotation. After exercise, this threshold to detection of movement increased to 1.59 degrees, an increase of 73%. This significant increase occurred with both internal and external rotation. The decrease in proprioceptive sense with muscle fatigue may play a role in decreasing athletic performance and in fatigue-related shoulder dysfunction. It remains to be determined if training can lessen this loss in position sense.

The effects of overuse combined with intrinsic or extrinsic alterations in an animal model of rotator cuff tendinosis.

An in vivo animal model was used to evaluate overuse and overuse plus intrinsic tendon injury or extrinsic tendon compression in the development of rotator cuff injury. Forty-four male Sprague-Dawley rats were divided into groups of 22. Each left shoulder received an intrinsic or extrinsic injury plus overuse (treadmill running), and each right shoulder received only overuse. Eleven rats from each group were sacrificed at 4 and 8 weeks. Supraspinatus tendons were evaluated histologically or geometrically and biomechanically. Ten rats constituted a cage-activity control group. Both supraspinatus tendons of the experimental groups had increases in cellularity and collagen disorganization and changes in cell shape compared with control tendons. Tendons with injury plus overuse exhibited a worse histologic grade than those with overuse alone. The cross-sectional area of both supraspinatus tendons of the experimental rats was significantly more than in control tendons. The area of the injury plus overuse tendons was increased on average compared with overuse-alone tendons. Biomechanically, the tissue moduli of overuse/intrinsic injury tendons at 4 weeks and those of the overuse/extrinsic injury tendons at 8 weeks were significantly lower than in control tendons. Tissue moduli of the overuse/injury tendons were significantly lower than in the overuse-alone tendons at 8 weeks. This study demonstrated that damage to the supraspinatus tendon can be caused by overuse and intrinsic injury, overuse and extrinsic compression, and overuse alone.

Biomechanics of the rotator cuff.

Thorough understanding of rotator cuff mechanics is important for effective treatment and/or prevention of cuff injuries. This understanding is achieved through knowledge of normal cuff structure and mechanics. Only then, can the effects of injuries and pathologic processes on normal cuff function be carefuly assessed. Rotator cuff structures are viewed and analyzed on a number of different levels. This article presents current knowledge of rotator cuff mechanics through review of cuff structure and anatomy, corocoacromial arch structure and biomechanics, and biomechanical models.

Fractures of the acromion process: a proposed classification system.

A review of 27 fractures of the acromion process during a 15-year period revealed five distinct types that were classified into three groups. Stress fractures are rare, do not result from acute trauma, and gain little benefit from nonoperative treatment. Type I fractures are minimally displaced. Type IA fractures are avulsion fractures and heal rapidly. Type IB fractures result from direct trauma to the extremity, and are minimally displaced. Most heal with nonoperative treatment. Type II fractures are displaced laterally, superiorly or anteriorly and do not reduce the subacromial space. Most are pain free with full motion after 6 weeks of nonoperative treatment. Type III fractures reduce the subacromial space. This may occur by an inferiorly displaced acromion fracture, or an acromion fracture associated with an ipsilateral, superiorly displaced glenoid neck fracture. Patients in this group sustained significant trauma to the involved extremity. All type III fractures treated nonoperatively develop significant limited shoulder motion with pain, suggesting that early surgical intervention may be indicated.

Biomechanical evaluation of current patella fracture fixation techniques.

OBJECTIVE: To compare the mechanical effectiveness of three different techniques for stabilization of transverse fractures of the patella. DESIGN: Cadaveric knees were used to model acute fractures of the patella. To test three treatment techniques in pairs of knees, specimen pairs were assigned randomly to a set of predetermined treatment pairs so as to provide equal numbers of paired and unpaired data sets. Results then were analyzed using a two-way analysis of variance. SETTING: The treatment techniques used are widely applicable in the clinical setting for the treatment of transverse fractures of the patella. No specialized equipment or training is required for the general or subspecialized orthopaedic surgeon. PATIENTS/PARTICIPANTS: None. INTERVENTIONS: Fractures were created in eighteen knees (nine pairs) and repaired by one of three techniques: (a) modified tension band (AO technique); (b) two parallel 4.5-millimeter interfragmentary lag screws; or (c) a new technique using four-millimeter cannulated lag screws with a tension band wired through the screws. MAIN OUTCOME MEASURES: In mechanical testing, the amount of interfragmentary separation in simulated knee extension and the maximum load to failure at 45 degrees of flexion were measured. RESULTS: Fractures stabilized with a modified tension band were found to displace significantly more than those fixed with screws alone or screws plus a tension band in simulated knee extensions (p < 0.05). The fractures fixed with the cannulated screws plus the tension band failed at higher loads (mean = 732 newtons) than those stabilized with screws alone (mean = 554 newtons, p = 0.06) or those with a modified tension band (mean = 395 newtons, p < 0.05). CONCLUSIONS: Combining interfragmentary screw fixation with the tension band principle appears to provide improved stability over the modified tension band or screws alone for transverse patella fractures. Cannulated screws allow for simple, reliable addition of a tension band to screw fixation.

Fractures of the patella.

Patellar fractures usually occur from distraction and three-point bending of the patella as well as from direct blows. Surgical treatment is necessary for fractures that are displaced more than 2 mm and may include open reduction and internal fixation, partial patellectomy, or rarely, total patellectomy. We have presented a new technique for the stabilization of a simple transverse fracture that has provided superior results in laboratory tests. Postoperative complications can be minimized by good attention to wound care, accurate fracture reduction, secure fracture fixation and an early range of motion. Despite the surgeon's best efforts, however, post-traumatic osteoarthrosis can develop and may require additional treatment.

Electrical stimulation of soft tissues.

It is well-documented that electrical potentials normally are present in biologic soft tissues. The discovery of these has generated interest in effecting a change in the biology of these tissues by externally imposing an electrical field. There is experimental evidence that ligament and tendon repair can be affected by ES, but there have been no clinical studies to demonstrate its effectiveness in human ligament or tendon injuries. The only clinical support for use of ES to enhance soft-tissue healing is for its use in chronic, open dermal wounds. ES has been shown to be effective for these chronic wounds under certain conditions. There is currently insufficient justification for the use of ES for reduction of soft-tissue edema.

Milkweed (Gentianales: Apocynaceae): a farmscape resource for increasing parasitism of stink bugs (Hemiptera: Pentatomidae) and providing nectar to insect pollinators and monarch butterflies.

In peanut-cotton farmscapes in Georgia, the stink bugs Nezara viridula (L.) and Chinavia hilaris (Say) (Hemiptera: Pentatomidae) and the leaffooted bug, Leptoglossus phyllopus (L.) (Hemiptera: Coreidae), disperse at crop-to-crop interfaces to feed on bolls in cotton. The main objective of this study was to determine whether insecticide-free tropical milkweed (Asclepias curassavica L.), a nectar-producing plant, can increase parasitism of these bugs by Trichopoda pennipes (F.) (Diptera: Tachinidae) and provide nectar to monarch butterflies and insect pollinators in these farmscapes. Peanut-cotton plots with and without flowering milkweed plants were established in 2009 and 2010. Adult T. pennipes, monarch butterflies, honey bees, and native insect pollinators readily fed on floral nectar of milkweed. Monarch larvae feeding on milkweed vegetation successfully developed into pupae. In 2009, N. viridula was the primary host of T. pennipes in cotton, and parasitism of this pest by the parasitoid was significantly higher in milkweed cotton (61.6%) than in control cotton (13.3%). In 2010, parasitism of N. viridula, C. hilaris, and L. phyllopus by T. pennipes was significantly higher in milkweed cotton (24.0%) than in control cotton (1.1%). For both years of the study, these treatment differences were not owing to a response by the parasitoid to differences in host density, because density of hosts was not significantly different between treatments. In conclusion, incorporation of milkweed in peanut-cotton plots increased stink bug parasitism in cotton and provided nectar to insect pollinators and monarch butterflies.