Development of microsatellite markers for an outbreaking species of oak gall wasp, Zapatella davisae (Hymenoptera: Cynipidae), in the northeastern United States.

Populations of the recently described black oak gall wasp, Zapatella davisae Buffington (Hymenoptera: Cynipidae), have been identified as the cause of extensive tree damage and mortality to black oaks, Quercus velutina Lamarck (Fagales: Fagaceae), in the northeastern United States. Relatively little is known, however, about the distribution, phylogenetic placement, and lifecycle of this important tree pest. Therefore, we conducted next-generation sequencing using the Ion Torrent™ PGM (ThermoFisher Scientific, Inc.) platform to develop genomic resources for the study of Z. davisae and for other closely related species of oak gall wasps. Individual sequence reads were aligned, assembled into unique contigs, and the contigs were then utilized for the in silico isolation and development of microsatellite markers. In total, we screened 36 candidate microsatellite loci, of which 23 amplified consistently (five polymorphic and 18 monomorphic). We then examined whether the polymorphic loci could be used to infer whether populations of Z. davisae from Cape Cod and Nantucket are sexual or asexual by calculating several metrics of genetic diversity that might indicate the mode of reproduction. These included testing for statistical deviations from Hardy-Weinberg equilibrium (HWE) and for linkage disequilibrium (LD), observations for the presence of the Meselson effect, and by calculating the probability that clonal individuals are more prevalent than would be expected in a randomly mating population. While we found significant deviations from HWE and more clonal individuals than expected, our estimates of the Meselson effect were inconclusive due to limited sampling, and we found no evidence of LD. Therefore, the sexual/asexual status of Z. davisae populations remains uncertain.

The Skaergaard trough layering: sedimentation in a convecting magma chamber.

The upper parts of the floor cumulates of the Skaergaard Intrusion, East Greenland, contain abundant features known as troughs. The troughs are gently plunging synformal structures comprising stacks of crescentic modally graded layers with a sharply defined mafic base that grades upward into plagioclase-rich material. The origin of the troughs and layering is contentious, attributed variously to deposition of mineral grains by magmatic currents descending from the nearby walls, or to in situ development by localised recrystallisation during gravitationally-driven compaction. They are characterised by outcrop-scale features such as mineral lineations parallel to the trough axis, evidence of erosion and layer truncation associated with migration of the trough axis, and disruption of layering by syn-magmatic slumping. A detailed microstructural study of the modal trough layers, using electron backscatter diffraction together with geochemical mapping, demonstrates that these rocks do not record evidence for deformation by either dislocation creep or dissolution-reprecipitation. Instead, the troughs are characterised by the alignment of euhedral plagioclase crystals with unmodified primary igneous compositional zoning. We argue that the lineations and foliations are, therefore, a consequence of grain alignment during magmatic flow. Post-accumulation amplification of the modal layering occurred as a result of differential migration of an unmixed immiscible interstitial liquid, with upwards migration of the Si-rich conjugate into the plagioclase-rich upper part of the layers, whereas the Fe-rich immiscible conjugate remained in the mafic base. Both field and microstructure evidence support the origin of the troughs as the sites of repeated deposition from crystal-rich currents descending from the nearby chamber walls.

Asymmetric hybridization between non-native winter moth, Operophtera brumata (Lepidoptera: Geometridae), and native Bruce spanworm, Operophtera bruceata, in the Northeastern United States, assessed with novel microsatellites and SNPs.

The European winter moth, Operophtera brumata, is a non-native pest in the Northeastern USA causing defoliation of forest trees and crops such as apples and blueberries. This species is known to hybridize with O. bruceata, the Bruce spanworm, a native species across North America, although it is not known if there are hybrid generations beyond F1. To study winter moth population genetics and hybridization with Bruce spanworm, we developed two sets of genetic markers, single nucleotide polymorphisms (SNPs) and microsatellites, using genomic approaches. Both types of markers were validated using samples from the two species and their hybrids. We identified 1216 SNPs and 24 variable microsatellite loci. From them we developed a subset of 95 species-diagnostic SNPs and ten microsatellite loci that could be used for hybrid identification. We further validated the ten microsatellite loci by screening field collected samples of both species and putative hybrids. In addition to confirming the presence of F1 hybrids reported in previous studies, we found evidence for multi-generation asymmetric hybridization, as suggested by the occurrence of hybrid backcrosses with the winter month, but not with the Bruce spanworm. Laboratory crosses between winter moth females and Bruce spanworm males resulted in a higher proportion of viable eggs than the reciprocal cross, supporting this pattern. We discuss the possible roles of population demographics, sex chromosome genetic incompatibility, and bacterial symbionts as causes of this asymmetrical hybridization and the utility of the developed markers for future studies.

Eltrombopag as steroid sparing therapy for immune thrombocytopenic purpura in systemic lupus erythematosus.

BACKGROUND: Eltrombopag activates the thrombopoietin (TPO) surface receptor on the megakaryocyte, which increases the production of platelets, and rapidly improves circulating platelet numbers in patients with immune thrombocytopenic purpura (ITP). This allows for rapid tapering and/or cessation of corticosteroid therapy. Less is known about the platelet response to this drug in ITP associated with systemic lupus erythematosus (SLE). METHODS: A retrospective review was performed of the clinical course of three consecutive patients, each with SLE-associated ITP who were initially treated with corticosteroids or other immunomodulatory therapy. These patients were treated with eltrombopag at the DMC Center for Bleeding Disorders and Thrombosis. Eltrombopag was administered according the package insert, with an initial dose of 50 mg daily, with weekly, then monthly monitoring of platelet counts and dose adjustments. Some immunomodulatory agents (e.g. hydroxychloroquine) were continued to control non hematologic SLE manifestations. RESULTS: All three patients maintained acceptable platelet counts (>50,000/mm(3) for >3 years) following tapering and cessation of corticosteroids. The drug was well-tolerated and there were no adverse events, and specifically no thrombotic events. CONCLUSION: Eltrombopag is effective as a rapidly acting corticosteroid sparing therapy for patients with ITP associated with SLE. This is important in reducing corticosteroid related side effects and morbidities in treating SLE patients with ITP. Larger studies are needed to ascertain safety and efficacy of eltrombopag in SLE patients with ITP, particularly those with coexisting antiphospholipid antibodies.

Armored scale insect endosymbiont diversity at the species level: genealogical patterns of Uzinura diasipipdicola in the Chionaspis pinifoliae-Chionaspis heterophyllae species complex (Hemiptera: Coccoidea: Diaspididae).

Armored scale insects and their primary bacterial endosymbionts show nearly identical patterns of co-diversification when viewed at the family level, though the persistence of these patterns at the species level has not been explored in this group. Therefore we investigated genealogical patterns of co-diversification near the species level between the primary endosymbiont Uzinura diaspidicola and its hosts in the Chionaspis pinifoliae-Chionaspis heterophyllae species complex. To do this we generated DNA sequence data from three endosymbiont loci (rspB, GroEL, and 16S) and analyzed each locus independently using statistical parsimony network analyses and as a concatenated dataset using Bayesian phylogenetic reconstructions. We found that for two endosymbiont loci, 16S and GroEL, sequences from U. diaspidicola were broadly associated with host species designations, while for rspB this pattern was less clear as C. heterophyllae (species S1) shared haplotypes with several other Chionaspis species. We then compared the topological congruence of the phylogenetic reconstructions generated from a concatenated dataset of endosymbiont loci (including all three loci, above) to that from a concatenated dataset of armored scale hosts, using published data from two nuclear loci (28S and EF1α) and one mitochondrial locus (COI-COII) from the armored scale hosts. We calculated whether the two topologies were congruent using the Shimodaira-Hasegawa test. We found no significant differences (P = 0.4892) between the topologies suggesting that, at least at this level of resolution, co-diversification of U. diaspidicola with its armored scale hosts also occurs near the species level. This is the first such study of co-speciation at the species level between U. diaspidicola and a group of armored scale insects.

The effects of 12 weeks of beta-hydroxy-beta-methylbutyrate free acid supplementation on muscle mass, strength, and power in resistance-trained individuals: a randomized, double-blind, placebo-controlled study.

INTRODUCTION: Studies utilizing beta-hydroxy-beta-methylbutyrate (HMB) supplementation in trained populations are limited. No long-term studies utilizing HMB free acid (HMB-FA) have been conducted. Therefore, we investigated the effects of 12 weeks of HMB-FA supplementation on skeletal muscle hypertrophy, body composition, strength, and power in trained individuals. We also determined the effects of HMB-FA on muscle damage and performance during an overreaching cycle. METHODS: A three-phase double-blind, placebo- and diet-controlled randomized intervention study was conducted. Phase 1 was an 8-week-periodized resistance-training program; Phase 2 was a 2-week overreaching cycle; and Phase 3 was a 2-week taper. Muscle mass, strength, and power were examined at weeks 0, 4, 8, and 12 to assess the chronic effects of HMB-FA; and assessment of these, as well as cortisol, testosterone, and creatine kinase (CK) was performed at weeks 9 and 10 of the overreaching cycle. RESULTS: HMB-FA resulted in increased total strength (bench press, squat, and deadlift combined) over the 12-week training (77.1 ± 18.4 vs. 25.3 ± 22.0 kg, p < 0.001); a greater increase in vertical jump power (991 ± 168 vs. 630 ± 167 W, p < 0.001); and increased lean body mass gain (7.4 ± 4.2 vs. 2.1 ± 6.1 kg, p < 0.001) in HMB-FA- and placebo-supplemented groups, respectively. During the overreaching cycle, HMB-FA attenuated increases in CK (-6 ± 91 vs. 277 ± 229 IU/l, p < 0.001) and cortisol (-0.2 ± 2.9 vs. 4.5 ± 1.7 µg/dl, p < 0.003) in the HMB-FA- and placebo-supplemented groups, respectively. CONCLUSIONS: These results suggest that HMB-FA enhances hypertrophy, strength, and power following chronic resistance training, and prevents decrements in performance following the overreaching.

Sticky platelet syndrome: an unusual presentation of arterial ischemia.

A 48-year-old woman presented with bilateral lower extremity critical limb ischemia. In addition to this, her work-up revealed multiple other thromboembolic insults including cerebral and visceral emboli. Initial laboratory findings were significant for an indeterminate platelet count, secondary to platelet clumping. After appropriate emergent surgical treatment including bilateral lower extremity embolectomy, the patient was empirically anticoagulated with a direct thrombin inhibitor. Further embolic work-up discovered bilateral renal and splenic infarctions as well as a large mobile mitral vegetation. Finally, an upper extremity duplex revealed left axillary, left subclavian, and right internal jugular acute deep vein thromboses. Mitral valve replacement was performed to remove the septic source. A series of hypercoagulability studies was done, and results were positive for lupus anticoagulants. Months after her recovery, the patient was tested and found to be positive for sticky platelet syndrome.

Human factor VIII: morphometric analysis of purified material in solution.

Study of purified human factor VIII in buffer by freeze-etch electron microscopy reveals rounded, rod-shaped particles measuring 22 by 42 nanometers. When thrombin was added to purified normal factor VIII, there was a rapid loss of rod-shaped particles during the first 15 minutes of incubation at 37 degrees C. Purified plasma from two patients with severe hemophilia contained spherical particles measuring 10 to 50 nanometers in diameter, with no evidence of significant numbers of rod-shaped forms. Negatively stained and unstained air-dried samples of factor VIII corroborate the relative shape and size differences between normal and hemophiliac material.

The subunit structure of normal and hemophilic factor VIII.

Human factor VIII from normals and hemophiliacs was partially purified by ethanol and polyethylene glycol precipitations. Final purification was achieved by gel filtration on 2 or 4% agarose or ion exchange chromatography on diethylaminoethyl cellulose. Comparable amounts of highly purified protein were obtained from normal and hemophilic plasma following the agarose chromatography step. Highly purified factor VIII was not dissociated by 6 M guanidine hydrochloride or 1% sodium dodecyl sulfate. However, when reduced by beta-mercaptoethanol and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, a single subunit species with an estimated 195,000 molecular weight was found for both normal and hemophilic factor VIII. By sedimentation equilibrium analysis, the normal factor VIII subunit was homogeneous and had an estimated molecular weight of 202,000. The subunit polypeptides from normal or hemophilic factor VIII contained carbohydrate. Each was homogeneous by isoelectric focusing. Immunodiffusion of purified normal and hemophilic factor VIII against rabbit antiserum to purified normal human factor VIII showed a single line of precipitation. Very low concentrations of purified human thrombin initially increased the activity of normal factor VIII about threefold and then progressively destroyed activity by 3 h. Only minimal activation occurred with hemophilic factor VIII. Both the activation and inactivation of normal and hemophilic factor VIII were unaccompanied by detectable changes in subunit molecular weight. These findings may have implications for the definition of the molecular defect in hemophilic factor VIII.

Immunoblastic lymphadenopathy. Evolution into a malignant lymphoma with plasmacytoid features.

In the patient described progressive lymphadenopathy, splenomegaly and interstitial pulmonary disease developed two months after the development of immunoblastic lymphadenopathy. Light microscopic examination of a lymph node biopsy specimen suggested a diagnosis of immunoblastic sarcoma. Evolution of this malignant lymphoma into a leukemic phase allowed detailed studies of the malignant cells by electron microscopy, cytochemical staining and immunologic technics. Evidence is presented that this is a case of malignant lymphoma with plasmacytoid and not lymphoid features. Review of the literature on immunoblastic sarcoma suggests that light microscopy and clinical setting are not sufficient to define a homogeneous clinicopathologic disease.

Non-Hodgkin's lymphomas in leukemic phase: clinicopathologic correlations.

A leukemic phase occurred in 30 (14 percent) of 214 patients with non-Hodgkin's lymphoma. To determine the significance of peripheral blood involvement in each type of NHL, patients were subdivided according to a modified Rappaport classification. Each histologic subtype presented a homogeneous clinical picture which differed from that seen in other histologic subtypes. Of particular note was the recognition of two distinctive cytologic and clinical subtypes within the category of nodular lymphoma, poorly differentiated lymphocytic lymphoma (NPDL). In one subtype, the predominant cells had cytologic features akin to those of lymphoblasts. In these cases, although the interval to peripheral blood involvement was variable, the median leukemic survival was only two months. In contrast in conventional NPDL the median leukemic survival was 43+ months, and peripheral blood involvement did not appear to exert an independent effect on prognosis. In diffuse large cell lymphomas the median leukemic survival was 0.5 months, with peripheral blood involvement appearing as a terminal event associated with unresponsive disease in multiple sites. The recognition of adult lymphoblastic lymphoma as a clinicopathologic entity with a high risk of leukemic conversion, 100 percent in this study, is also confirmed.

Molecular structural studies of human factor VIII.

Neither normal nor hemophilic factor VIII protein enters a 5% sosium dodecyl sulfate gel; on reduction, however, a single 195 000-molecular-weight peptide is observed. Hemophilic and normal factor VIII contain carbohydrate and appear identical in subunit molecular weight, electrical charge, and major antigenic determinants. Thrombin activation and inactivation of factor VIII does not detectably change the subunit molecular weight. Trypsin causes similar activity changes and obviously cleaves the factor VIII subunit. Human plasmin destroys factor VIII procoagulant activity and degrades the factor VIII subunit to 103 000-, 88 000-, and 17 000-molecular-weight peptides. Both normal and hemophilic factor VIII as well as thrombin-inactivated factor VIII support ristocetin-induced platelet aggregation. Purified factor VIII chromatographed on 4% agarose in 1.0 M sodium chloride shows no dissociation of the procoagulant activity from the void volume protein. Gel chromatography on 4% agarose in 0.25 M calcium chloride results in a procoagulant activity peak removed from the void volume protein; both peaks contain protein which does not enter a 5% SDS gel, but on reduction a 195 000-molecular-weight subunit band is observed for each. Both the void volume protein peak and the procoagulant activity peak from the 0.25 M calcium chloride-agarose gel column support ristocetin-induced platelet aggregation. After removal of calcium, a small amount of procoagulant activity is present only in the void volume peak. These data suggest that both the procoagulant and von Willebrand activities are on the same molecule. Thus our previous conclusion remains the same: human factor VIII is a large glycoprotein composed of identical 195 000-molecular-weight subunits jointed by disulfide bonds and is responsible for both antihemophilic and von Willebrand activities in human plasma.

The effect of maltose tetrapalmitate (MTP) on prostate cancer growth in vivo and in vitro.

Maltose tetrapalmitate (MTP), a non-toxic synthetic glycolipid analog of lipid A, has been shown to have antitumor activity in tumor-transplanted animals. Its mode of action has been postulated to be as an immunoadjuvant or as an anti-angiogenesis agent. MTP has been shown to have antitumor properties in lung, bladder, mammary, colon, liver and soft tissue tumors, but its action on prostate cancer has not yet been investigated. The effect of MTP alone and in combination with hydrocortisone hemisuccinate on prostate cancer and the ability of MTP to inhibit angiogenesis were examined in this study. In vitro, MTP was minimally cytotoxic to rat prostate cancer cells and to bovine and human endothelial cells at high concentrations. In the angiogenesis inhibition assays, the MTP alone exhibited no anti-angiogenesis effect and significant anti-angiogenesis activity only when combined with hydrocortisone hemisuccinate at high doses. In vivo, however, MTP demonstrated significant inhibition of prostate cancer growth. These results suggest that MTP decreases prostate cancer growth in vivo but it is not an angiogenesis inhibitor in rat prostate cancer.

Postural responses to lateral perturbation in healthy subjects and ankle sprain patients.

The purpose of this study was to investigate postural responses of healthy subjects and patients with recent ankle sprains following a perturbation that created sway in the frontal plane. EMG data were taken from the posterior tibialis (PT) (not monitored in patients), peroneal longus (PL), and tibialis anterior muscles (TA). Subjects stood on a platform that provided a rotational perturbation (approximately 70 degrees.s-1) in the frontal plane. This perturbation had the effect of everting and loading one limb while inverting and unloading the contralateral limb. An initial response in the PT of the loaded limb and the PL of the unloaded limb was noted at approximately 50 ms following the perturbation. This was followed by a bilateral response in the TA at 60 ms. The amplitude of the TA muscle was significantly greater in the loaded limb. For ankle sprain patients a bilateral TA response and a PL response in the unloaded limb was noted at approximately 65 ms. TA response amplitude ratios between the loaded and unloaded limbs were similar to that of the healthy subjects. These data suggest that ankle sprain patients use a modified postural response following lateral perturbation as a compensation for the injury.

Ligation of the renal vein during resection of abdominal aortic aneurysm.

In resection of abdominal aortic aneurysm, ligation and division of the left renal vein may be necessary in order to expose the perirenal aorta. This manoeuvre is possible, with conservation of the left kidney function, because of the extensive venous collateral circulation of the left kidney. It is of crucial importance however, that ligation of the vein is performed close to the inferior vena cava. A case is presented where ligation of the left renal vein was performed in relation to an operation for a ruptured abdominal aortic aneurysm. After the operation there was initially dysfunction of the left kidney, and later on sepsis-induced uraemia. The renal function stabilized at a moderately reduced level. No permanent kidney damage related to the venous ligation could be demonstrated. In the literature serious renal damage has been reported in 10 cases out of 89 reported ligations of the left renal vein. Ligation of the left renal vein is thus a reasonably safe and acceptable procedure for surgical exposure in difficult aortic procedures.

Clinical fracture of the carpal scaphoid--supportive bandage or plaster cast immobilization?

In a prospective study of 108 patients with a clinical diagnosis of fracture of the carpal scaphoid, but without radiological evidence of fracture, the patients were randomised to treatment with either a supportive bandage or a dorsal plaster cast. Four patients proved to have incomplete fractures and three to have avulsions from the scaphoid tuberosity. Two of the fractures had been suspected radiologically at the primary investigation. No complete fractures of the scaphoid were seen. The average time in plaster was 15 days and in a bandage 12.2 days. The average sick leave for manual workers was 14 days in plaster and 4 days in a bandage, a difference that represents a significant loss of productivity. Since these fractures almost always heal irrespective of treatment, they may as well be treated as a soft tissue injury with a supportive bandage.

Intramuscular temperature responses in the human leg to two forms of cryotherapy: ice massage and ice bag.

Cryotherapy is often used to affect the muscle underlying skin and subcutaneous tissues. However, the relationship between the penetrative ability of various cryotherapy methods and treatment time is not thoroughly understood. The purpose of this study was to measure muscle temperature responses during two frequently used cryotherapy techniques. A 23-gauge hypodermic needle microprobe was inserted to one-half skin-fold thickness plus 1 cm into the medial aspect of the gastrocnemius muscle of 14 subjects. Two groups of seven subjects each were measured for temperature changes during ice bag or ice massage treatment. Each treatment consisted of a 15-minute application of the selected method. A significant difference between the two methods was observed (t = -2,157, p < or = 0.05). Ice massage achieved its lowest temperature in an average of 17.9 +/- 2.4 minutes, while ice bag reached its lowest temperature in 28.2 +/- 12.5 minutes. Ice massage appears to cool muscle more rapidly than ice bag.

Test-retest reliability of reciprocal isokinetic knee extension and flexion peak torque measurements.

Our purpose in conducting this study was to estimate the reliability of reciprocal concentric knee extension and flexion peak torque obtained in uninjured male athletes using the Biodex isokinetic dynamometer. Twenty-six male intercollegiate athletes (age=19.5+/-4.1 yr; ht=70.3+/-14.9 in; wt=212.9+/-48.5 lb) participated in this study. We used the Biodex to measure peak torque occurring during right knee extension and flexion over 3 consecutive days. Means and standard deviations were calculated for both extension and flexion on each of the 3 days. We observed a significant main effect for days for both extension and flexion. Mean peak torque for the first day was significantly higher (Turkey, p<.05) than the means for the other 2 days, which were not significantly different from each other (Turkey, p>0.05). Intraclass correlation coefficients (ICCs) were estimated for each of the six trials on each of the 3 days. Intraclass correlation coefficients (ICCs) ranged from .88 on trial 1, day 1 for both extension and flexion peak torque to .97 for extension peak torque and .98 for flexion peak torque on day 3, trial 6. The standard error of measurement for extension peak torque was 7.0 ft-lbs and for flexion peak torque was 3.0 ft-lbs. These results indicate that reliable measurements of reciprocal right knee extension and flexion peak torque can be obtained from uninjured male athletes with the Biodex isokinetic dynamometer.

[Myocardial enzymes in the serum after high and low laparotomy]. / Myokardieenzymer i serum efter øovre og nedre laparotomi.

The serum activities of ASAT, ALAT, LDH, creatine kinase (CK) and creatine kinase B (CK-B) were measured perioperatively in 20 patients in connection with vagotomy or subtotal/total gastrectomy and in 20 patients in connection with resection of the rectum. None of the patients presented clinical or serial electrocardiographic signs of acute myocardial infarction (AMI). ASAT, ALAT and LDH showed moderate increases postoperatively, the median values of which were within the reference range, and no statistical differences were found between the maximal values for these enzymes between patients with high and low laparotomies. In both patient categories, CK and CK-B showed marked postoperative increase and the maximal values for these enzymes were significantly higher in the group of patients subjected to high laparotomies, presumably on account of more pronounced muscle trauma in this group. The discriminative limits for CK-B as regards AMI diagnosis usually employed were found to be unsuitable after stomach and rectum surgery. The traditional myocardial enzymes must be interpreted with restraint after laparotomy as regards the diagnosis of AMI. A discriminative limit for CK-B% (maximal CK-B/total CK) of 6 is probably more suitable.

Implementation and evaluation of an interdisciplinary health professions core curriculum.

An interdisciplinary core curriculum has been implemented in the College of Health Professions at Armstrong Atlantic State University since spring 1996. The curriculum is designed to provide students with the knowledge, skills, and values necessary for interprofessional practice. The courses are taught by interdisciplinary teams and are offered as electives or as part of major requirements in nursing, health science, physical therapy, dental hygiene, medical technology, radiologic sciences, and respiratory therapy. In addition to ongoing evaluation methods, a survey designed to assess the student and faculty perceptions of the experience has been conducted. Both groups agreed that the experience has had a positive impact on the students' professional performances, patient interactions, understanding of the health care delivery system, and health career preparation. Faculty agreed that teaching in an interdisciplinary team was a positive experience. The collaboration among the health professions' faculty has resulted in increased respect for one another and for others' disciplines. Although the experience places an additional burden on their workload, they agreed that the experience is beneficial, their efforts are worthwhile, and they would be willing to continue to teach interdisciplinary courses.