Productive Infection of Human Skeletal Muscle Cells by Pandemic and Seasonal Influenza A(H1N1) Viruses

Besides the classical respiratory and systemic symptoms, unusual complications of influenza A infection in humans involve the skeletal muscles. Numerous cases of acute myopathy and/or rhabdomyolysis have been reported, particularly following the outbreak of pandemic influenza A(H1N1) in 2009. The pathogenesis of these influenza-associated myopathies (IAM) remains unkown, although the direct infection of muscle cells is suspected. Here, we studied the susceptibility of cultured human primary muscle cells to a 2009 pandemic and a 2008 seasonal influenza A(H1N1) isolate. Using cells from different donors, we found that differentiated muscle cells (i. e. myotubes) were highly susceptible to infection by both influenza A(H1N1) isolates, whereas undifferentiated cells (i. e. myoblasts) were partially resistant. The receptors for influenza viruses, α2-6 and α2-3 linked sialic acids, were detected on the surface of myotubes and myoblasts. Time line of viral nucleoprotein (NP) expression and nuclear export showed that the first steps of the viral replication cycle could take place in muscle cells. Infected myotubes and myoblasts exhibited budding virions and nuclear inclusions as observed by transmission electron microscopy and correlative light and electron microscopy. Myotubes, but not myoblasts, yielded infectious virus progeny that could further infect naive muscle cells after proteolytic treatment. Infection led to a cytopathic effect with the lysis of muscle cells, as characterized by the release of lactate dehydrogenase. The secretion of proinflammatory cytokines by muscle cells was not affected following infection. Our results are compatible with the hypothesis of a direct muscle infection causing rhabdomyolysis in IAM patients.     

THE DIET OF THE CURLEW SANDPIPER AT LANGEBAAN LAGOON,SOUTH AFRICA

Puttick, G. M. 1978. The diet of the Curlew Sandpiper at Langebaan Lagoon, South Africa. Ostrich 49:158-167.

Nereid worms (mainly Ceratonereis erythraensis) and the hydrobiid gastropod Assiminea globulus were the most important prey items throughout the year for Curlew Sandpipers Calidris ferruginea feeding at Langebaan Lagoon. Other important items were the amphipod Urothoe grimaldi, the crabs Hymenosoma orbiculare and Cleistostoma edwardsii and dipteran larvae (Stratiomyidae). Immature Curlew Sandpipers took smaller prey items than adult birds. The diet of male and female birds differed in the proportion, sue and range of prey items taken.     

Safety of withholding anticoagulation in pregnant women with suspected deep vein thrombosis following negative serial compression ultrasound and iliac vein imaging

Background:

Compression ultrasonography performed serially over a 7-day period is recommended for the diagnosis of deep vein thrombosis in symptomatic pregnant women, but whether this approach is safe is unknown. We evaluated the safety of withholding anticoagulation from pregnant women with suspected deep vein thrombosis following negative serial compression ultrasonography and iliac vein imaging.

Methods:

Consecutive pregnant women who presented with suspected deep vein thrombosis underwent compression ultrasonography and Doppler imaging of the iliac vein of the symptomatic leg(s). Women whose initial test results were negative underwent serial testing on 2 occasions over the next 7 days. Women not diagnosed with deep vein thrombosis were followed for a minimum of 3 months for the development of symptomatic deep vein thrombosis or pulmonary embolism.

Results:

In total, 221 pregnant women presented with suspected deep vein thrombosis. Deep vein thrombosis was diagnosed in 16 (7.2%) women by initial compression ultrasonography and Doppler studies; none were identified as having deep vein thrombosis on serial testing. One patient with normal serial testing had a pulmonary embolism diagnosed 7 weeks later. The overall prevalence of deep vein thrombosis was 7.7% (17/221); of these, 65% (11/17) of cases were isolated to the iliofemoral veins and 12% (2/17) were isolated iliac deep vein thromboses. The incidence of venous thromboembolism during follow-up was 0.49% (95% confidence interval [CI] 0.09%–2.71%). The sensitivity of serial compression ultrasonography with Doppler imaging was 94.1% (95% CI 69.2%–99.7%), the negative predictive value was 99.5% (95% CI 96.9%–100%), and the negative likelihood ratio was 0.068 (95% CI 0.01–0.39).

Interpretation:

Serial compression ultrasonography with Doppler imaging of the iliac vein performed over a 7-day period excludes deep-vein thrombosis in symptomatic pregnant women.Over the last 2 decades, venous compression ultrasonography has become the imaging test of choice for diagnosing deep vein thrombosis in the lower extremities of men and nonpregnant women.^1–4 Although this test is highly sensitive (about 97%) for deep vein thrombosis involving the femoral and popliteal veins, compression ultrasonography is less sensitive for the detection of isolated deep vein thrombosis in the calf.⁵ Because proximal propagation of isolated calf deep vein thrombosis occurs in about 20% of cases, serial compression ultrasonography performed over a 7-day period is recommended to definitely exclude such thromboses if the results of the initial compression ultrasound are negative.⁶The use of serial compression ultrasonography in symptomatic men and nonpregnant women has been validated in prospective studies,^1,7 suggesting that withholding anticoagulation from symptomatic patients whose serial compression ultrasound results are negative is safe, with less than 2% of patients subsequently being diagnosed with deep vein thrombosis.^1,7,8 Although the use of serial compression ultrasonography has not been validated in pregnant women, this strategy is also advocated for symptomatic pregnant women.⁹The appeal of using compression ultrasonography for diagnosing deep vein thrombosis in pregnant women is obvious: it is noninvasive, widely available and does not expose the fetus to ionizing radiation. However, generalizing results from studies involving men and nonpregnant women to pregnant women is problematic because of differences in clinical presentation and anatomic distribution of deep vein thromboses.¹⁰ Compared with men and nonpregnant women, pregnant women more often present with very proximal deep vein thrombosis (including isolated iliac vein deep vein thrombosis); isolated distal calf deep vein thromboses are infrequent.¹⁰ In a recent review of the literature, we found that 62% of all deep vein thromboses in symptomatic pregnant women were in the iliofemoral veins, 17% were in the iliac vein alone, and 6% were in the calf veins.¹⁰ In contrast, in the general population, more than 80% of deep vein thromboses involved calf veins, and iliofemoral deep vein thromboses or isolated iliac veins are uncommon (< 5%).^1–4Physiologic changes associated with pregnancy might affect blood flow patterns and normal compressibility of the proximal veins, thereby affecting the diagnostic accuracy of compression ultrasonograpy. This technique cannot be used to detect isolated deep vein thromboses in the iliac vein; these veins are not compressible because of their intrapelvic location. Whether Doppler studies are sensitive for detecting deep vein thromboses in these high proximal veins (i.e., iliac veins) has not been well studied, but data suggest that this method of detection compares favourably to compression ultrasonography in men and nonpregnant women for proximal deep vein thromboses.¹¹ The use of Doppler imaging in pregnant women for the purpose of detecting iliac vein deep vein thromboses has been reported in the literature,^12,13 but it has not been adequately evaluated.Currently, the standard practice of diagnosing deep vein thrombosis in symptomatic pregnant women is by compression ultrasonography. If the results of the compression ultrasound are negative, Doppler imaging of the iliac vein (with or without vagal manoeuvres) is recommended, particularly for women with a high clinical probability of deep vein thrombosis in the iliac vein.^9,12,13 This diagnostic approach is advocated despite the absence of any prospective studies validating its use. In this study, we evaluated the diagnostic accuracy of serial compression ultrasonography and Doppler imaging of the iliac veins over a 7-day period among symptomatic pregnant women.     

Mechanisms for Stabilising Water Clathrates

Molecular dynamics computer simulations are used to examine various possible mechanisms for stabilising water clathrates. It is found that the attractive interactions between guest and host molecules are not sufficient to explain the clathrate's stability; nor is there any significant dynamic coupling between the guest and host molecules in the clathrate. Instead, it appears that the repulsive guest-host interactions provide the mechanical stability of the clathrate, and that they are even sufficient to define its structure. Some of the thermodynamic implications of these observations are discussed.     

A Molecular Dynamics Simulation of a Polyamine-Induced Conformational Change of DNA. A Possible Mechanism for the B to Z Transition

Abstract

A 75ps molecular dynamics simulation has been performed on a fully solvated complex of spermine with the B DNA decamer (dGdC)₅ · (dGdC)₅. The simulation indicates a possible mechanism by which polyamines might induce the formation of a left-handed helix, the B to Z transition. Spermine was initially located in the major groove, hydrogen bonded to the helix. During the simulation the ligand migrates deeper into the DNA, maintaining strong hydrogen bonding to the central guanine bases and destroying the Watson-Crick base pairing with their respective cytosines. Significant rotation of these and other cytosine bases was observed, in part due to interactions of the helix with the aminopropyl chains of spermine. An intermediate B_II conformation might be of importance in this process.     

A Binding Mode of A-[tris(1,10-phenanthroline)ruthenium(II)]2+ Exhibiting Preference for Purine-3′,5′-Pyrimidine Sites of DNA

Abstract

Molecular mechanics calculations and molecular dynamics simulations have been used to study the binding of the partially inserted major groove complex of A-[Ru(1,10-phenanthroline)₃]²⁺ with DNA. Energy refinements of this complex showed a clear preference for binding at purine-3′,5′-pyrimidine sites over pyrimidine-3′,5′-purine sites. The basis for this difference is shown to be a slight change in the binding orientation induced by interchanging the purine and pyrimidine bases. This in turn provides for a better secondary interaction with the helix backbone at a point beyond the immediate binding site. It is this secondary interaction that provides the additional energetic stabilisation for complexes formed at purine-3′,5′-pyrimidine sites. Molecular dynamics simulations including explicit representation of solvent support these conclusions and provide an insight into the positional stability of the ligand at a particular site. Repuckering of specific deoxyribose rings to the C3′-endo conformation seems to be an important feature of the DNA/ligand complex.     

The acute-phase protein serum amyloid A3 is expressed in the bovine mammary gland and plays a role in host defence

The serum amyloid A protein is one of the major reactants in the acute-phase response. Using representational difference analysis comparing RNA from normal and involuting quarters of a dairy cow mammary gland, we found an mRNA encoding the SAA3 protein (M-SAA3). The M-SAA3 mRNA was localized to restricted populations of bovine mammary epithelial cells (MECs). It was expressed at a moderate level in late pregnancy, at a low level through lactation, was induced early in milk stasis, and expressed at high levels in most MECs during mid to late involution and inflammation/mastitis. The mature M-SAA3 peptide was expressed in Escherichia coli, antibodies made, and shown to have antibacterial activity against E. coli, Streptococcus uberis and Pseudomonas aeruginosa. These results suggest that the mammary SAA3 may have a role in protection of the mammary gland during remodelling and infection and possibly in the neonate gastrointestinal tract.     

A variant in MCF2L is associated with osteoarthritis

Osteoarthritis (OA) is a prevalent, heritable degenerative joint disease with a substantial public health impact. We used a 1000-Genomes-Project-based imputation in a genome-wide association scan for osteoarthritis (3177 OA cases and 4894 controls) to detect a previously unidentified risk locus. We discovered a small disease-associated set of variants on chromosome 13. Through large-scale replication, we establish a robust association with SNPs in MCF2L (rs11842874, combined odds ratio [95% confidence interval] 1.17 [1.11–1.23], p = 2.1 × 10⁻⁸) across a total of 19,041 OA cases and 24,504 controls of European descent. This risk locus represents the third established signal for OA overall. MCF2L regulates a nerve growth factor (NGF), and treatment with a humanized monoclonal antibody against NGF is associated with reduction in pain and improvement in function for knee OA patients.     

Identification and characterization of an inborn error of metabolism caused by dihydrofolate reductase deficiency

Dihydrofolate reductase (DHFR) is a critical enzyme in folate metabolism and an important target of antineoplastic, antimicrobial, and antiinflammatory drugs. We describe three individuals from two families with a recessive inborn error of metabolism, characterized by megaloblastic anemia and/or pancytopenia, severe cerebral folate deficiency, and cerebral tetrahydrobiopterin deficiency due to a germline missense mutation in DHFR, resulting in profound enzyme deficiency. We show that cerebral folate levels, anemia, and pancytopenia of DHFR deficiency can be corrected by treatment with folinic acid. The characterization of this disorder provides evidence for the link between DHFR and metabolism of cerebral tetrahydrobiopterin, which is required for the formation of dopamine, serotonin, and norepinephrine and for the hydroxylation of aromatic amino acids. Moreover, this relationship provides insight into the role of folates in neurological conditions, including depression, Alzheimer disease, and Parkinson disease.     

Influence of sticky trap color and height above ground on capture of alate Elatobium abietinum (Hemiptera: Aphididae) in Sitka spruce plantations

A series of field trials were used to assess the practicality of using sticky traps to monitor populations of green spruce aphid, Elatobium abietinum (Walker), in plantations of Sitka spruce. The highest numbers of alate E. abietinum were caught on sticky traps placed in the upper third of the live canopy at 9-17 m above the ground, whereas low numbers of aphids were caught just below the live canopy or at 2 m above the ground. Trials in 2005 with sticky traps of different colors showed that significantly more alate E. abietinum were caught on yellow, red, and green sticky traps than on white, blue, and black traps. A repeat trial in 2007 resulted in significantly more alate aphids being caught on red sticky traps than on traps of any other color except for green. Attraction to red is unusual among aphids, as aphids are thought not to possess a red-sensitive photoreceptor. The attraction of E. abietinum to red-colored sticky traps suggests that conifer-feeding aphids might have a fundamentally different color response compared with aphids that live on cereals, grasses, or herbaceous plants. Alternatively, the attraction to red might be a physiological artifact related to the presence of red-screening pigments in the aphid's compound eye.