Flexible memory controls sperm competition responses in male Drosophila melanogaster.

Males of many species use social cues to predict sperm competition (SC) and tailor their reproductive strategies, such as ejaculate or behavioural investment, accordingly. While these plastic strategies are widespread, the underlying mechanisms remain largely unknown. Plastic behaviour requires individuals to learn and memorize cues associated with environmental change before using this experience to modify behaviour. Drosophila melanogaster respond to an increase in SC threat by extending mating duration after exposure to a rival male. This behaviour shows lag times between environmental change and behavioural response suggestive of acquisition and loss of memory. Considering olfaction is important for a male's ability to assess the SC environment, we hypothesized that an olfactory learning and memory pathway may play a key role in controlling this plastic behaviour. We assessed the role of genes and brain structures known to be involved in learning and memory. We show that SC responses depend on anaesthesia-sensitive memory, specifically the genes rut and amn We also show that the γ lobes of the mushroom bodies are integral to the control of plastic mating behaviour. These results reveal the genetic and neural properties required for reacting to changes in the SC environment.

Experimental and Numerical Analysis of Initial Plasticity in P91 Steel Small Punch Creep Samples.

To date, the complex behaviour of small punch creep test (SPCT) specimens has not been completely understood, making the test hard to numerically model and the data difficult to interpret. This paper presents a novel numerical model able to generate results that match the experimental findings. For the first time, pre-strained uniaxial creep test data of a P91 steel at 600 ∘C have been implemented in a conveniently modified Liu and Murakami creep damage model in order to simulate the effects of the initial localised plasticity on the subsequent creep response of a small punch creep test specimen. Finite element (FE) results, in terms of creep displacement rate and time to failure, obtained by the modified Liu and Murakami model are in good agreement with experimental small punch creep test data. The rupture times obtained by the FE calculations which make use of the non-modified creep damage model are one order of magnitude shorter than those obtained by using the modified constitutive model. Although further investigation is needed, this novel approach has confirmed that the effects of initial localised plasticity, taking place in the early stages of small punch creep test, cannot be neglected. The new results, obtained by using the modified constitutive model, show a significant improvement with respect to those obtained by a 'state of the art' creep damage constitutive model (the Liu and Murakami constitutive model) both in terms of minimum load-line displacement rate and time to rupture. The new modelling method will potentially lead to improved capability for SPCT data interpretation.

Mini-drainage devices: the Ex-PRESS Mini-Glaucoma Device.

The Ex-PRESS Mini-Glaucoma Device is a glaucoma drainage device used to shunt aqueous from the anterior chamber into a subconjunctival reservoir that is created surgically. It has been used successfully over the last decade with approximately 60,000 implantations worldwide. In an ever-evolving microsurgical environment, the Ex-PRESS glaucoma device is on the forefront of intraocular pressure lowering technology. This chapter aims to review the surgical uses, techniques and considerations of the Ex-PRESS device as well as analyze the current literature detailing the advantages and disadvantages of the Ex-PRESS implant. Special attention will also be placed on the authors' own experience using the device.

Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood.

Species of Candida frequently cause life-threatening infections in neonates, transplant and intensive care unit (ICU) patients, and others with compromised host defenses. The successful management of systemic candidiasis depends upon early, rapid diagnosis. Blood cultures are the standard diagnostic method, but identification requires days and less than half of the patients are positive. These limitations may be eliminated by using real-time polymerase chain reaction (PCR) to detect Candida DNA in the blood specimens of patients at risk. Here, we optimized a PCR protocol to detect 5-10 yeasts in low volumes of simulated and clinical specimens. We also used a mouse model of systemic candidiasis and determined that candidemia is optimally detectable during the first few days after infection. However, PCR tests are often costly, labor-intensive, and inconvenient for routine use. To address these obstacles, we evaluated the innovative microfluidic real-time PCR platform (Advanced Liquid Logic, Inc.), which has the potential for full automation and rapid turnaround. Eleven and nine of 16 specimens from individual patients with culture-proven candidemia tested positive for C. albicans DNA by conventional and microfluidic real-time PCR, respectively, for a combined sensitivity of 94%. The microfluidic platform offers a significant technical advance in the detection of microbial DNA in clinical specimens.

Trace elements enhance biofilm formation in UASB reactor for solo simple molecule wastewater treatment.

In this paper, trace elements (TE) adding was investigated in one bench-scale UASB reactor treating solo simple molecule wastewater with the aim of evaluating its effect on enhancing biofilm formation. After adding sufficient TE (3 mL/L) in the influent, during 3 days, COD removal efficiency increased from 74% to 90% comparing to no adding TE. Over 55 days of operation, the organic loading rate (OLR) reached 11 g/L/day with COD removal efficiencies greater than 90%. While in the steady running period no effect even improvement on treatment performance was observed without any TE adding. The results illuminated that TE accounted for quick start-up of the UASB biofilm system rather than ever known biocatalyst.

Influence of hydration state and homologue composition of magnesium stearate on the physical chemical properties of liquid paraffin lipogels.

Lipogels were prepared by dispersing mixed (60:40 C(16)-C(18)) and pure (C(18)) homologue magnesium stearate (MgSt) in liquid paraffin, using three methods of preparation, i.e. addition of water at 95 °C during cooling cycle (method 1), homogenisation upon cooling (method 2) or cooling without addition of water or homogenisation (method 3). The systems were characterised by physical inspection, polarised, hot stage and scanning electron microscopy (SEM), differential scanning calorimetry (DSC), rheology, and X-ray diffraction (XRD). Systems formed stable semisolid lipogels (no syneresis), unstable solids showing syneresis or structured fluids, depending on the type of magnesium stearate used and the preparation technique. The stable semisolid lipogels containing mixed homologue MgSt (commercial-as received, anhydrous or dihydrate) prepared by methods 1 (∼ 1-2% water) and 2 contained α-crystalline lamellar structure. These were not present in the unstable solids formed with method 3 or in systems prepared from pure homologue MgSt which were generally structured fluids rather than semisolids. In addition, semisolid lipogels of pure homologue trihydrate MgSt prepared by method 3 showed plate-like crystals, implying pressure sensitivity. There is significantly more amorphous MgSt in the unstable solids compared to the stable semisolid lipogels, which are mainly crystalline (confirmed by XRD).

Bench-scale study using PVA gel as a biocarrier in a UASB reactor treating corn steep liquor wastewater.

PVA-gel beads were used as a biocarrier for treatment of corn steep liquor wastewater containing high levels of volatile fatty acids (VFA), where retention of biomass could be either solely in the porous microstructure of the gel or by granule formation using a gel bead as a nucleus. With stable COD removal efficiencies of 90% or greater, continuous treatment was demonstrated over a four month period, with organic loading rates being increased stepwise from 2.5 to 22.5 kg COD/m3 d. In addition, VFA in the effluent were, with few exceptions maintained close to zero. Gas production increased over the course of the study and reached a level of 0.38 m3/kg COD consisting of 65% methane with the remainder being mostly carbon dioxide. Biomass granules containing methane producing bacteria progressively formed around the PVA-gel beads during the study. In contrast, very few small natural granules developed apart from PVA-gel nuclei indicating that PVA gel may serve well as a seeding material to enhance granulation when natural occurrence is lacking.

A pilot-plant study of a moving-bed biofilm reactor system using PVA gel as a biocarrier for removals of organic carbon and nitrogen.

A pilot-plant study was conducted to evaluate the performance of a moving-bed biofilm reactor process using PVA-gel beads as a biocarrier. Real primary-settled wastewater was fed to the pre-denitrification system and removals of nitrogenous and organic contaminants were evaluated over a 1-year period. The results demonstrated that at a total nitrogen (TN) loading of 18 mg/L.h, a TN removal efficiency in keeping with and even exceeding the theoretical maximum efficiency based on the level of internal recycle, was possible and a nitrification rate of 15 mg/L.h was sustained with a HRT of only 2.5 h at 15 degrees C. Furthermore, soluble COD and BOD5 in the effluent of the pilot plant were reduced to levels well below most regulatory discharge limits. In addition, the possibility of using this biocarrier in a system, including the elimination of waste organic sludge, was discussed.

Physical ageing and thermal analysis of PLGA microspheres encapsulating protein or DNA.

PLGA microspheres undergo physical ageing but their ageing kinetics have not been reported, nor the effect of encapsulated protein or plasmid DNA on any associated changes to the glass transition. Differential scanning calorimetry (DSC) was used to measure the rate of ageing of various PLGA microsphere formulations, with temperature-modulated DSC used to accurately measure the associated glass transition. The Cowie-Ferguson model was applied to determine the parameters describing the enthalpy relaxation kinetics. We show that encapsulated proteins had no significant effect on the glass transition of the microspheres, whereas DNA and PVA were mild antiplasticising agents, particularly with high Mw PLGA. Physical ageing occurred through a range of enthalpy relaxation times (or modes) and was independent of both encapsulated protein and surfactant used during microsphere preparation. Analysis of accelerated ageing at 35 degrees C gave calculated enthalpy relaxation times to thermal equilibrium of 280-400 h. No ageing was observed < or = 10 degrees C and at 25 degrees C estimated relaxation times were at least one order of magnitude greater than at 35 degrees C. Ageing of PLGA microspheres therefore occurs at temperatures >10 degrees C, but relaxation will be far from equilibrium unless storage times and/or temperatures are prolonged or nearing the glass transition, respectively.

Controlled drug delivery to the lung: Influence of hyaluronic acid solution conformation on its adsorption to hydrophobic drug particles.

This work reports investigations into the interaction and adsorption of the hydrophilic polymer hyaluronic acid (HA) onto the surface of the hydrophobic corticosteroid drug fluticasone propionate (FP). The eventual aim is to formulate a bioadhesive pulmonary drug delivery system with prolonged action that avoids rapid clearance from the lungs by the mucociliary escalator. Adsorption isotherms detailing the adsorption of HA from aqueous HA solution concentrations ranging from 0.14 to 0.0008% (w/v) to a fixed FP particle concentration of 0.1% (w/v) were investigated. The method of preparing FP particles with HA molecules adsorbed on their surfaces (FP/HA particles) involved suspension of the FP either in hydrated HA solution or in water followed by addition of solid HA, centrifugation of the solids to form a pellet, washing the pellet several times with water until no HA was found in the supernatant and then freeze drying the suspension obtained by dispersing the final pellet. The freeze dried powder was then analysed for adsorbed HA using a Stains-all assay. The influence of order of addition of HA to FP, time for the adsorption process, and temperature of preparation on the adsorption isotherms was investigated. The non-equilibrium adsorption isotherms produced generally followed the same trend, in that as the HA solution concentration increased, the amount of HA adsorbed increased to a maximum at a solution concentration of approximately 0.1% (w/v) and then decreased. The maxima in the adsorption isotherms were close to the change from secondary to tertiary conformation in the HA solutions. Below the maxima, adsorption occurred via interaction of FP with the hydrophobic patches along the HA chains in the secondary structures. Above the maxima, secondary HA molecules aggregate in solution to form tertiary network structures. Adsorption from tertiary structure was reduced because strong interactions between the HA molecules limited the availability of hydrophobic patches for adsorption of HA onto FP. The influence of preparation variables on adsorption was also related to the availability of hydrophobic patches for adsorption.

Defining 'full-length' recombinant factor VIII: a comparative structural analysis.

Coagulation factor VIII (FVIII) is an important glycoprotein co-factor involved in haemostasis, functioning to accelerate activation of factor X by activated factor IX. Insertion of expression vectors containing the full-length cDNA sequence of human FVIII into mammalian cell lines results in the production of recombinant factor VIII (rFVIII), typically referred to as 'full-length' rFVIII (FLrFVIII). Both FLrFVIII and plasma-derived FVIII exist primarily as heterodimeric proteins, consisting of a heterogenous light and heavy chain. The objectives of this study were to compare the structural heterogeneity of high-purity FVIII preparations and further define the term 'full length' as it refers to rFVIII protein structure. Five commercially available FVIII concentrates were characterized based on SDS-PAGE, N-terminal sequencing, and peptide and domain mapping coupled to mass spectrometry. The major heavy chain species identified in FLrFVIII included various B-domain-truncated forms of FVIII, with the predominant species terminating at Arg(1313). This study demonstrates that the use of full-sequence FVIII cDNA for the production of rFVIII does not result in a homogeneous FLrFVIII protein product. Rather, commercially available FLrFVIII represents a heterogenous mixture of various B-domain-truncated forms of the molecule, with no evidence of a contiguous, intact B-domain.

Polychlorinated biphenyls and organochlorine pesticides in plasma and the embryonic development in Lake Erie water snakes (Nerodia sipedon insularum) from Pelee Island, Ontario, Canada (1999).

From three locations along a 34-km shoreline of Pelee Island, Ontario, 30 gravid female Lake Erie water snakes (Nerodia sipedon insularum) were sampled to determine the organochlorine (OC) contaminant levels in plasma and the number of live and dead embryos present in the body cavity. Plasma was analyzed for 59 polychlorinated biphenyl (PCB) congeners and 14 organochlorine pesticides. Concentrations of pesticides were low (< or =0.1 ng/g wet wt) in all snakes, but there was significant variation in mean PCB concentrations in plasma from among the sampling locations on Pelee Island. Snakes (n = 5) from the West shore and dock area of the island had significantly higher PCB concentrations (90.4 +/- 15.0 ng/g wet wt) in plasma than those from Lighthouse Point (n = 5; 34.4 +/- 13 ng/g wet wt) and the south shore of the island (n = 5; 29.4 +/- 16.3 ng/g wet wt). Body mass of the female snakes ranged from 252 to 880 g, and mean masses were not significantly different among sample sites. The number of live embryos found ranged from 13 to 46 female snakes and no dead embryos were detected. There were significant positive correlations among body mass, snout-vent length, and number of young per female. There were no significant correlations among body mass, snout-vent length, number of young per female, or per-gram body mass of female snakes and contaminant concentrations in plasma. It was concluded that an interim estimate of a no-effect level on embryonic survival in N. sipedon insularum may be a maximum average concentration of 90.4 ng/g wet wt PCBs and a maximum average concentration of 3.6 ng/g wet wt p,p'-dichloro-diphenyl-dichloroethylene in plasma.

The influence of surfactant on PLGA microsphere glass transition and water sorption: remodeling the surface morphology to attenuate the burst release.

PURPOSE: The stability of protein unloaded and loaded poly(lactic-co-glycolic acid) (PLGA) microspheres fabricated with surfactant was challenged through exposure to environmental conditions of different relative humidity. METHODS: Polyvinyl alcohol (PVA) or Triton X-100 was added to the primary emulsion of the double-emulsion solvent evaporation technique. After storage at ambient humidity and 75% relative humidity, the mechanical stability of the polymer was tested to reveal PLGA chain mobility using differential scanning calorimetry. Subsequent surface modifications were examined by atomic force microscopy (AFM), and protein release profiles were collected. RESULTS: Residual amounts of PVA and particularly Triton X-100 raised the hydrophilicity of the microspheres. When exposed to ambient humidity or 75% relative humidity, PVA and Triton X-100 had, respectively, an antiplasticizing and a plasticizing effect upon PLGA, and both led to physical aging. The high-resolution AFM imaging of microspheres containing model protein and Triton X-100 showed that the depth of the surface pores was reduced when exposed to 75% relative humidity, and the initial burst release subsequently decreased. CONCLUSION: These studies suggested that the mechanical stability of PLGA was influenced by the addition of surfactants, which, depending on the formulation, led to surface pore remodeling under high humidity, reducing the initial burst release while maintaining the spherical integrity of the microsphere.

MRI bone oedema predicts eight year tendon function at the wrist but not the requirement for orthopaedic surgery in rheumatoid arthritis.

OBJECTIVE: To investigate the role of early magnetic resonance imaging (MRI) of the wrist in predicting functional outcome in rheumatoid arthritis. METHODS: MRI scans of the dominant wrist were scored for synovitis, tendon inflammation, bone oedema, and erosion at first presentation (n = 42), at 1 year (n = 42), and at 6 years (n = 31). At 8 years, clinical reassessment (n = 28) was undertaken. Tendon function was graded 0-3 for movement, tendon sheath swelling, and pain on resistance at nine flexor and extensor tendons of the hand. Hand function was also assessed using the Sollerman grip test. The requirement for joint or tendon surgery by 8 years was determined by telephone survey in 39 of the original 42 patients. RESULTS: At 8 years, tendon function was highly correlated with hand function (Sollerman score, R = -0.51, p = 0.005) and global function (health assessment questionnaire score, R = 0.53, p = 0.004). Using a model incorporating baseline and 1 year MRI scores, the MRI bone oedema score was strongly predictive of tendon function at 8 years (chi(2)(2) = 15.3, p = 0.0005), as was the MRI bone erosion score (chi(2)(2) = 9.23, p = 0.01). Hand function was also predicted by the baseline MRI erosion score (p = 0.02). MRI variables did not predict the requirement for surgery, but patients who had surgery were more likely to show progression of MRI bone erosion scores between baseline and 1 year (p = 0.008). CONCLUSIONS: Extensive MRI bone oedema and erosions at the wrist in early rheumatoid arthritis predict tendon dysfunction and impaired hand function in the medium term but not the requirement for joint or tendon surgery.

Salmeterol decreases eosinophilic cationic protein and rescue medication in patients inhaling beclomethasone dipropionate: preliminary study in mild and moderate asthma in Trinidad, West Indies.

Activated eosinophils play a critical role in asthma pathogenesis, and eosinophil cationic protein (ECP) is a useful indicator of inflammation. Inhaled corticosteroids and long-acting beta2-agonists (LABA) effectively control asthma symptoms and improve airway function. Salmeterol's anti-inflammatory efficacy as add-on therapy to inhaled corticosteroids has not been evaluated in Caribbean populations. We investigated nine non-smoking subjects (three men and six women; mean age: +/- SE, 50.7 +/- 3.82 years) with stable mild and moderate persistent asthma who were inhaling > or = 500 microg beclomethasone dipropionate (BDP) daily. This was a with-in-patient controlled laboratory blind study performed over 8 weeks. Patients received BDP for 2 weeks, add-on salmeterol 100 microg in weeks 3-6 and BDP alone in weeks 7-8. Patients recorded daily morning and night symptoms. Morning peak expiratory flow rate was measured on entry to the study and with sputum ECP at the end of weeks 2, 4, 6 and 8. Salmeterol together with BDP decreased sputum ECP from a pretreatment median value of 897.84 microg/l to 628.38 microg/l after 4 weeks, and ECP continued to decrease even after salmeterol withdrawal. Both drugs decreased the frequency of rescue medication use by approximately 50% and increased the median number of days per week without rescue salbutamol from 0 to 3 days. Salmeterol's bronchoprotective effect was maximal after 4 weeks and was sustained after its withdrawal. In conclusion, this study, performed in Trinidadian asthmatics, used ECP as a surrogate marker of bronchial inflammation and supports the recent Salmeterol Multi-center Asthma Research Trial (SMART) data recommending add-on salmeterol therapy to adequate anti-inflammatory medication such as inhaled corticosteroids for optimal asthma management. Further studies are required to evaluate the anti-inflammatory efficacy and possible tolerance to salmeterol in Caribbean patients.

Predisposition to efficient mammary tumor metastatic progression is linked to the breast cancer metastasis suppressor gene Brms1.

Tumor metastasis is one of the most important clinical aspects of neoplastic disease because patient mortality is frequently attributable to disseminated rather than primary tumors. However, it still is not possible to definitively distinguish those individuals at high risk for disseminated disease, who would benefit from aggressive adjuvant therapy, from the low-risk patients who might be spared the side effects of additional anticancer therapy. To identify factors that predispose toward metastatic disease, we have used a genetic approach. Using a highly metastatic model of mammary cancer, we identified previously inbred mouse strains (DBA/2J, NZB/B1NJ, and I/LnJ) that harbor genetic factors that significantly suppress metastatic efficiency. In this study, we report the results of four experiments to localize the genetic map locations of the metastasis efficiency modifier genes. One statistically significant locus was identified on proximal Chr 19 designated Mtes1. Secondary candidate intervals were detected on Chrs 6, 9, 13, and 17. Interestingly, Mtes1 colocalizes with the murine orthologue of the human breast cancer metastasis suppressor gene Brms1, suggesting that allelic variants of Brms1 might be responsible for the metastasis suppression observed.

Analysis of sulfated peptides using positive electrospray ionization tandem mass spectrometry.

Presented is a method for analyzing sulfated peptides, and differentiating the post-translational modification (PTM) from its isobaric counterpart phosphorylation, using quadrupole time-of-flight (Qq/TOF) mass spectrometry (MS) and positive ion nanoelectrospray MS/MS. A set of commercially available sulfo- and phosphopeptide standards was analyzed via in-source dissociation and MS/MS to generate fragmentation signatures that were used to characterize and differentiate the two modifications. All of the phosphorylated peptides retained their +80 Da modifications under collision-induced decomposition (CID) conditions and peptide backbone fragmentation allowed for the site-specific identification of the modification. In sharp contrast, sulfated peptides lost SO3 from the precursor as the collision energy (CE) was increased until only the non-sulfated form of the peptide was observed. The number of 80 Da losses indicated the number of sulfated sites. By continuing to ramp the CE further, it was possible to fragment the non-sulfated peptides and obtain detailed sequence information. It was not possible to obtain site-specific information on the location of the sulfate moieties using positive ion MS/MS as none of the original precursor ions were present at the time of peptide backbone fragmentation. This method was applied to the analysis of recombinant human B-domain deleted factor VIII (BDDrFVIII), which has six well-documented sulfation sites and several potential phosphorylation sites located in two of the sulfated regions of the protein. Seven peptides with single and multiple +80 Da modifications were isolated and analyzed for their respective PTMs. The fragmentation patterns obtained from the BDDrFVIII peptides were compared with those obtained for the standard peptides; and in all cases the peptides were sulfated. None of the potential phosphorylation sites were found to be occupied, and these results are consistent with the literature.

Diagnostic accuracy of focused appendiceal CT in clinically equivocal cases of acute appendicitis.

PURPOSE: To determine the diagnostic accuracy of modified focused appendiceal computed tomography (CT) to exclude or confirm appendicitis in patients who presented with equivocal symptoms and signs of appendicitis. MATERIALS AND METHODS: One hundred patients (age range, 14-81 years; mean age, 30.6 years) with equivocal symptoms and signs of acute appendicitis were included in this prospective study. Patients were given 30 mL of diatrizoate meglumine and diatrizoate sodium and 60 mL of sorbitol mixed in 1 L of water orally over 1 hour. CT was performed 1.5 hours after the commencement of oral contrast material administration. The criteria used for the diagnosis of appendicitis were (a) appendix greater than 6 mm in maximum diameter, (b) no contrast material in the appendiceal lumen, and (c) inflammatory changes in the periappendiceal fat. CT results were compared with histopathologic findings at appendectomy. Patients with negative CT findings were followed up by telephone or clinically. RESULTS: Of 100 cases, 30 were positive at CT and 70 were negative. There were 28 true-positive cases; two false-positive cases, one cecal diverticulitis and one pelvic peritonitis with periappendicitis; and two false-negative cases, one perforated appendix and one mucosal and submucosal inflammation of the appendix but no transmural inflammation. Sensitivity was 93%, specificity was 97%, and accuracy was 96%. CONCLUSION: Focused appendiceal CT in which oral contrast material is used alone yields high levels of accuracy in clinically equivocal cases of acute appendicitis.

Technician, friend, detective, and healer: family physicians' responses to emotional distress.

OBJECTIVE: We sought to develop a typology of physicians' responses to patients' expressed mental health needs to better understand the gap between idealized practice and actual care for emotional distress and mental health problems. STUDY DESIGN: We used a multimethod comparative case study design of 18 family practices that included detailed descriptive field notes from direct observation of 1637 outpatient visits. An immersion/crystallization approach was used to explore physicians' responses to emotional distress and apparent mental health issues. POPULATION: A total of 379 outpatient encounters were reviewed from a purposeful sample of 13 family physicians from the 57 clinicians observed. OUTCOMES MEASURED: Descriptive field notes of outpatient visits were examined for emotional content and physicians' responses to emotional distress. RESULTS: Analyses revealed a 3-phase process by which physicians responded to emotional distress: recognition, triage, and management. The analyses also uncovered a 4-quadrant typology of management based on the physician's philosophy (biomedical vs holistic) and skill level (basic vs more advanced). CONCLUSIONS: Physicians appear to manage mental health issues by using 1 of 4 approaches based on their philosophy and core set of skills. Physician education and practice improvement should be tailored to build on physicians' natural philosophical proclivity and psychosocial skills.