A novel setup for simultaneous two-photon functional imaging and precise spectral and spatial visual stimulation in Drosophila.
Motion vision has been extensively characterised in Drosophila melanogaster, but substantially less is known about how flies process colour, or how spectral information affects other visual modalities. To accurately dissect the components of the early visual system responsible for processing colour, we developed a versatile visual stimulation setup to probe combined spatial, temporal and spectral response properties. Using flies expressing neural activity indicators, we tracked visual responses in the medulla, the second visual neuropil, to a projected colour stimulus. The introduction of custom bandpass optical filters enables simultaneous two-photon imaging and visual stimulation over a large range of wavelengths without compromising the temporal stimulation rate. With monochromator-produced light, any spectral bandwidth and centre wavelength from 390 to 730 nm can be selected to produce a narrow spectral hue. A specialised screen material scatters each band of light across the visible spectrum equally at all locations of the screen, thus enabling presentation of spatially structured stimuli. We show layer-specific shifts of spectral response properties in the medulla correlating with projection regions of photoreceptor terminals.


Fundamental unpredictability in multispecies competition.
One of the central goals of ecology is to predict the distribution and abundance of organisms. Here, we show that, in ecosystems of high biodiversity, the outcome of multispecies competition can be fundamentally unpredictable. We consider a competition model widely applied in phytoplankton ecology and plant ecology in which multiple species compete for three resources. We show that this competition model may have several alternative outcomes, that the dynamics leading to these alternative outcomes may exhibit transient chaos, and that the basins of attraction of these alternative outcomes may have an intermingled fractal geometry. As a consequence of this fractal geometry, it is impossible to predict the winners of multispecies competition in advance.


Factors influencing the clinical diagnosis of sinusitis.
This study was conducted to determine the influence of various clinical factors on the physician's diagnosis of sinusitis. Although the "clinical impression" appears to be a valid method of diagnosis in the literature, the specific factors that influence it have not been explored. Seven factors selected from the literature were incorporated into 128 simulated case histories of patients with possible sinusitis. These cases were presented to 40 local family physicians who were asked to assess whether sinusitis was present and whether antibiotic therapy would be prescribed. Logistic regression was performed to determine the influence of each factor. These results were compared with a 30-item Likert-type scale that asked the physicians to rate a variety of factors in terms of their importance to the diagnosis of sinusitis. The diagnosis of sinusitis and the decision to prescribe antibiotics were strongly influenced by the number of factors present. Physicians placed the most emphasis on purulent drainage seen on examination and sinus tenderness, and the least emphasis on smoking history and the failure to respond to decongestants. Transillumination and x-ray examinations were less useful to these physicians in establishing the diagnosis. The surveyed physicians based their diagnosis of sinusitis on the number of symptoms present, especially if the presentation included purulent drainage seen on examination or sinus tenderness. These results are consistent with studies that have approached the question using other designs.


[Magnetic resonance angiography].
MR angiography (MRA) is a group of MRI techniques capable of producing projectional images of arteries and veins without the use of ionising radiation or catheters. This review describes two of the main techniques, time-of-flight (TOF) MRA and phase contrast MRA, that provide high signal-intensity vessels due to flow phenomena, and a third main technique, contrast-enhanced MRA, which relies on intravenous injection of contrast media. Contrary to the iodine-containing x-ray contrast media, the contrast media for MRA are not nephrotoxic, making contrast-enhanced MRA a good alternative for patients with reduced renal function. MRA is indicated in several vascular areas, especially head and neck arteries and veins, the large thoracic and abdominal arteries and veins, and the lower extremity arteries. For several indications, the sensitivity and specificity of MRA is approaching those of the gold standard, x-ray angiography. There are reasons to believe that most diagnostic angiographic procedures will soon be performed as MRA.


Analysis of mean platelet volume and red blood cell distribution width in recurrent epistaxis.
Recurrent epistaxis is one of the most common causes of emergency department visits. Although several localized and systemic conditions has been described, the exact cause is unknown in the majority of cases. In our study, we aimed to determine the effect of mean platelet volume (MPV) and red blood cell distribution width (RDW) levels on recurrent epistaxis. One hundred and thirty six patients with recurrent epistaxis and 170 healthy cases as control group were included in the study. Demographic data, vital signs and the results of complete blood counts were recorded. The patients who had clinical conditions that might affect the levels of MPV or RDW, were excluded. MPV and RDW levels were compared between the two groups. The median level of MPV was 7.6 fL (IQR25-75%: 7.1-8.4) in the study group and 8.2 fL (IQR25-75%: 7.8-8.9) in the control group (p < 0.001). The median levels of RDW were found in the patient and control groups in order %15.4 (IQR25-75%: 14.5-15.4) and %14.3 (IQR25-75%: 13.4-15.4) (p < 0.001). Systolic blood pressure, leukocyte count, age, sex, RDW and MPV levels that were variables with p levels<0.2, were included in the multivariate analyses. It was determined that high RDW levels increased epistaxis (p < 0.001; OR:1.89 [95% CI:1.53-2.33]) and high MPV levels decreased epistaxis (p < 0.001; OR:0.54 [95% CI:0.39-0.72]). Low MPV and high RDW levels caused an increased bleeding tendency in patients with recurrent epistaxis. Although exact mechanism is not known, referring those patients for the assessment of etiologic causes would be proper.


Induction of successive follicular waves by gonadotropin-releasing hormone and prostaglandin F(2α) to improve fertility of high-producing cows during the summer and autumn.
Reduced conception rate during the hot summer and subsequent autumn is a well-documented phenomenon. Evaporative cooling systems greatly increase milk production but only slightly improve reproductive performance; hence, additional approaches to improving fertility during the hot season are required. The purpose of the present study was to examine whether the combination of an efficient cooling system and hormonal manipulation (GnRH+PGF(2α)) might improve fertility during the summer and autumn. The experiment was conducted from July to December in 2 commercial herds in Israel and included 382 healthy Holstein cows. Cows (50 to 60 d in milk) were hormonally treated to induce 3 consecutive 9-d follicular waves, with GnRH administration followed by PGF(2α) injection 7 d later. Both control (n=187) and treated (n=195) cows were inseminated following estrus, and pregnancy was determined by palpation 45 d post-insemination. Data revealed an interaction between treatment and primiparous cows, reflected by a 16% increase in conception rate [odds ratio (OR) 2.32, 95% confidence interval (CI): 0.96-5.61] and 14% increase in pregnancy rate at 120 d in milk (OR 3.16, 95% CI: 0.93-10.47). Interaction between treatment and high body condition score was reflected by a 14% increase in pregnancy rate at 90 d in milk (OR 3.02, 95% CI: 1.14-7.96). About 60% of the treated cows expressed estrus at the expected time (normal response within 5 d following the third PGF(2α) injection); the remaining 40% that manifested estrus later (late response) had higher milk yield and lower body condition score. Additional analyses indicated that treatment interacted with normal response to raise conception rates and pregnancy rates of primiparous cows and cows with high body condition score. On the other hand, treatment by late-response interaction lowered conception rate during the summer. Implementation of such hormonal treatment in combination with an efficient cooling system may improve reproductive performance of dairy cows during the summer and subsequent autumn.


A chronic fatigue syndrome model demonstrates mechanical allodynia and muscular hyperalgesia via spinal microglial activation.
Patients with chronic fatigue syndrome (CFS) and fibromyalgia syndrome (FMS) display multiple symptoms, such as chronic widespread pain, fatigue, sleep disturbance, and cognitive dysfunction. Abnormal pain sensation may be the most serious of these symptoms; however, its pathophysiology remains unknown. To provide insights into the molecular basis underlying abnormal pain in CFS and FMS, we used a multiple continuous stress (CS) model in rats, which were housed in a cage with a low level of water (1.5 cm in depth). The von Frey and Randall-Seritto tests were used to evaluate pain levels. Results showed that mechanical allodynia at plantar skin and mechanical hyperalgesia at the anterior tibialis (i.e., muscle pain) were induced by CS loading. Moreover, no signs of inflammation and injury incidents were observed in both the plantar skin and leg muscles. However, microglial accumulation and activation were observed in L4-L6 dorsal horn of CS rats. Quantification analysis revealed a higher accumulation of microglia in the medial part of Layers I-IV of the dorsal horn. To evaluate an implication of microglia in pain, minocycline was intrathecally administrated (via an osmotic pump). Minocycline significantly attenuated CS-induced mechanical hyperalgesia and allodynia. These results indicated that activated microglia were involved in the development of abnormal pain in CS animals, suggesting that the pain observed in CFS and FMS patients may be partly caused by a mechanism in which microglial activation is involved.


Stabilization of an intermolecular i-motif by lipid modification of cytosine-oligodeoxynucleotides.
This paper describes the stabilization of an intermolecular i-motif by lipophilic modification on the 3'-terminus of oligonucleotides. The hydrophobic aliphatic chain connected at the 3'-terminus of a trinucleotide (dC)3 promoted the formation of an i-motif and significantly enhanced the quadruplex's stability. The impact of lipophilic modification on i-motif's thermal stability was studied by UV-thermal denaturation melting experiments and isothermal titration calorimetry. We found that alkyl chains containing more than 14 carbon atoms could elevate the i-motif structure's stability in a wide range of pH and concentrations.


Revisiting graphene-polymer nanocomposite for enhancing anticorrosion performance: a new insight into interface chemistry and diffusion model.
Graphene is impermeable to all molecules and has high chemical stability, which makes it an excellent anticorrosion coating for metals. However, current studies have indicated that galvanic coupling between graphene and a metal actually accelerates corrosion at the interface. Due to the insulating nature of polymers, graphene-polymer composite coatings with a strong interaction between the filler and the polymer matrix are an alternative means of addressing this issue. Nevertheless, such coatings require well-dispersed graphene flakes to lengthen the diffusion paths of gases or liquids, while preventing the formation of a conducting network from graphene to the metal. The difficulty in preparing such coatings was mainly due to problems with the control of the assembled phase during interfacial reactions. Herein, the interactions between the filler and the polymer were found to be a key factor governing anticorrosion performance, which has scarcely been previously reported. The advantage of graphene as a filler in anticorrosion coatings lies in its dispersibility and miscibility with both the casting solvent and the polymer. Electrochemically exfoliated graphene (EC-graphene) with appropriate surface functionalities that allow high miscibility with waterborne polyurethane (PU) and hydrophobic epoxy has been found to be an ideal filler that outperforms other graphene materials such as graphene oxide (GO) and reduced graphene oxide (rGO). Furthermore, a bilayer coating with EC-graphene additives for PU over epoxy has been found to reduce the corrosion rate (CR) to 1.81 × 10-5 mm per year. With a graphene loading of less than 1%, this represents the lowest CR ever achieved for copper and steel substrates and a diffusion coefficient that is lower by a factor of nearly 2.2 than that of the pristine polymer. Furthermore, we have shown that by controlling the amount of graphene loaded in the polymer galvanic corrosion favored by the formation of an interconnected graphene percolation network can successfully be limited. The present study, together with a facile and eco-friendly method of nanocomposite synthesis, may pave the way toward practical applications in the development of graphene-based anticorrosion coatings.


Fusion hindrance for a positive-q-value system (24)Mg+(30)Si.
Measurements of the excitation function for the fusion of (24)Mg+(30)Si (Q=17.89 MeV)have been extended toward lower energies with respect to previous experimental data. The S-factor maximum observed in this large, positive-Q-value system is the most pronounced among such systems studied thus far. The significance and the systematics of an S-factor maximum in systems with positive fusion Q values are discussed. This result would strongly impact the extrapolated cross sections and reaction rates in the carbon and oxygen burnings and, thus, the study of the history of stellar evolution.


Renal aquaporins and sodium transporters with special focus on urinary tract obstruction.
The discovery of aquaporin-1 (AQP1) by Agre and colleagues explained the long-standing biophysical question of how water specifically crosses biological membranes. These studies led to the discovery and identification of a whole new family of membrane proteins, the aquaporins. At present, at least seven aquaporins are expressed at distinct sites in the kidney and 4 members of this family (AQP1-4) have been demonstrated to play pivotal roles in the physiology and pathophysiology for renal regulation of body water balance. Osmotic equilibration via renal aquaporins is maintained by active transport of NaCl. The major sodium transporters and channels in the individual renal tubule segments have been identified and the regulation of these transporters and channels are fundamental for renal sodium reabsorption and for establishing the driving force. In this mini-review the role of renal aquaporins and sodium transporters and channels is briefly described and their key role for the impaired urinary concentrating capacity in response to urinary tract obstruction is reviewed. Thus this review updates previous detailed reviews (1-5).


[Somatotropic function of the pituitary gland in persons with growth disorders].
The authors present the results of study of the somatotropic function of the hypophysis in 18 healthy children and in 70 patients suffering from various forms of growth retardation. The blood serum level of the STH of the hypophysis was studied by the authors by standard sets for radioimmunological determination on fasting stomach and in dynamics during the standard glucose-tolerance test. The data of basal, maximal and mean STH levels were taken into consideration. Investigations revealed a reduction of the somatotropic function of the hypophysis in the patients with hypophysial, cerebral and somatotropic nanism. With the first tow forms of growth retardation the disturbances were associated with the organic affections of the hypophysis or of the hypothalamus, and in the somatogenic one- they were functional. Somatotropic function was retained in spondylo-epiphysial dysplasia and in the patients with thyrogenic nanism given displacement therapy. In the patients with the Shereshevsky-Turner syndrome the STH secretion remained unchanged, but the reaction of the hypothalamo-hypophysial system to the functional distrubances was disturbed. An increase of the mean STH level followed complex therapy.


Dietary nitrate supplementation enhances high-intensity running performance in moderate normobaric hypoxia, independent of aerobic fitness.
Nitrate-rich beetroot juice (BRJ) increases plasma nitrite concentrations, lowers the oxygen cost (V⋅O2) of steady-state exercise and improves exercise performance in sedentary and moderately-trained, but rarely in well-trained individuals exercising at sea-level. BRJ supplementation may be more effective in a hypoxic environment, where the reduction of nitrite into nitric oxide (NO) is potentiated, such that well-trained and less well-trained individuals may derive a similar ergogenic effect. We conducted a randomised, counterbalanced, double-blind placebo controlled trial to determine the effects of BRJ on treadmill running performance in moderate normobaric hypoxia (equivalent to 2500 m altitude) in participants with a range of aerobic fitness levels. Twelve healthy males (V⋅O2max ranging from 47.1 to 76.8 ml kg(-1) min(-1)) ingested 138 ml concentrated BRJ (∼15.2 mmol nitrate) or a nitrate-deplete placebo (PLA) (∼0.2 mmol nitrate). Three hours later, participants completed steady-state moderate intensity running, and a 1500 m time-trial (TT) in a normobaric hypoxic chamber (FIO2 ∼ 15%). Plasma nitrite concentration was significantly greater following BRJ versus PLA 1 h post supplementation, and remained higher in BRJ throughout the testing session (p < 0.01). Average V⋅O2 was significantly lower (BRJ: 18.4 ± 2.0, PLA: 20.4 ± 12.6 ml kg(-1) min(-1); p = 0.002), whilst arterial oxygen saturation (SpO2) was significantly greater (BRJ: 88.4 ± 2.7, PLA: 86.5 ± 3.3%; p < 0.001) following BRJ. BRJ improved TT performance in all 12 participants by an average of 3.2% (BRJ: 331.1 ± 45.3 vs. PL: 341.9 ± 46.1 s; p < 0.001). There was no apparent relationship between aerobic fitness and the improvement in performance following BRJ (r(2) = 0.05, p > 0.05). These findings suggests that a high nitrate dose in the form of a BRJ supplement may improve running performance in individuals with a range of aerobic fitness levels conducting moderate and high-intensity exercise in a normobaric hypoxic environment.


Networks of nomads: negotiating access to health resources among pastoralist women in Chad.
Health resources among pastoralist groups are strongly gendered. While certain types of health resources fall within the female domain (home-based treatment, caring and supportive roles, and knowledge surrounding particular reproductive conditions) access to most outside health practitioners, treatments and knowledge is controlled largely by men. For pastoralist women, this means that actions taken during illness episodes depend largely on the nature and quality of social support systems available, and on their ability to mobilise them effectively. These support systems include husband and other affines, male kin, and networks of female kin and friends. Factors such as position within domestic and wider social units, as well as life cycle, affect women's ability to access and mobilise these different support systems for their health needs. However, seasonal mobility interacts with gender and social support systems in complex ways that profoundly influence women's access to health resources. Most literature on nomadic peoples and health focuses on the physical barriers posed by spatial mobility to accessing health resources. However, it is suggested here that, for pastoralist women in Chad, the spatial fluidity of social networks might be a more important consideration. At certain times of the year women enjoy relatively easy access to a large range of extended kin and other social contacts, while at other times, when people are very dispersed, options become much more limited, often resulting in illness treatment being delayed. Mobility should not, though, be seen purely as a constraint. It can also be an opportunity, increasing the potential geographical and social resource base with regard to health for women.


Effect of hypokinesia on the telencephalic blood vessels of rabbits.
Experiments were carried out in 37 animals (33 test animals and 4 controls). Hypokinesia was produced by keeping the animals in small cages. After 1, 2, 3, 4, 5, 6, 8, 12, 16 weeks of restricted mobility the animals were killed. The brain blood vessels were injected with dye-stuffs and examined in cleared sections. In 9 animals (2 controls and 7 test animals) the brain blood vessels were studied histochemically. Hypokinesia was stated to change significantly the angio-architectonics of the telencephalon. During first three weeks of restricted mobility, the blood vessels change but a little. By that time, together with unchanged arteries and veins, narrow arteries and a great number of dilated veins are encountered. The density of the blood vessel network is altered. Later on, after 4 weeks, more profound changes in the blood vessels occur: they become tortuous, deformed with disturbance of their usual orientation of course. These changes are specific for all the parts of the telencephalon. They increase with the time of hypokinesia.


Clinical response to arginine vasopressin therapy after paediatric cardiac surgery.
To describe the haemodynamic response of children who receive arginine vasopressin for haemodynamic instability after cardiac surgery and to identify clinical variables associated with a favourable response. Materials and Methods We reviewed patients less than or equal to 6 years undergoing open heart surgery in our institution between January, 2009 and July, 2010 who received arginine vasopressin during the first 7 days post operation. Favourable responders were defined as those in whom blood pressure was increased or maintained and catecholamine score was decreased, or blood pressure was increased by greater than or equal to 10% of baseline and catecholamine score was unchanged at 6 hours following arginine vasopressin initiation. Of the 34 patients identified, 17 (50%) patients responded favourably to arginine vasopressin. At 6 hours, the mean blood pressure was increased by 32.2% in responders as compared with 4.6% in non-responders, with a p-value less than 0.001. The mean catecholamine score decreased by 30.1% in responders and increased by 7.6% in non-responders, with a p-value less than 0.001. Anthropometric, demographic, and intra-operative variables were similar in both groups, as was maximum dose of arginine vasopressin. The median time after arrival to the intensive care unit at which arginine vasopressin was initiated, however, was later in those who responded, 20 hours as compared with those who did not, 6 hours, with a p-value equal to 0.032. Arginine vasopressin therapy led to haemodynamic improvement in only half of the children in this study, and improvement was more likely to occur if arginine vasopressin was initiated after the post-operative night.


Neuronal and cardiovascular responses to ANF microinjected into nucleus ambiguous.
Effects of microinjection of atrial natriuretic factor (ANF) into cardioinhibitory sites in the nucleus ambiguous (NA) or on single vagal cardioinhibitory neurons (VCN) were investigated in urethan-anesthetized rats. Sites containing cardioinhibitory neurons were identified by observing a marked and reproducible bradycardia in response to microiontophoretically applied (20-40 nA) or microinjected (20 nl) 0.1 M L-glutamate. In 35 of the 40 (87.5%) cardioinhibitory sites identified by microinjection of glutamate, ANF (20 nl of 10(-7) M) decreased heart rate (HR; -47.1 +/- 2.5 beats/min). No responses were elicited in the other five sites. In animals paralyzed and artificially ventilated, the HR effects of ANF were not significantly different before and after muscle paralysis. Microinjections of 10 nl of 10(-7) M ANF caused excitation of 19 of 21 VCN (90%), which was followed by a decrease in HR (-20.8 +/- 2.3 beats/min); no neuronal or cardiovascular responses were elicited by ANF in the remaining two VCN. Bilateral vagotomy or atropine sulfate (1 mg/kg iv) abolished cardiac slowing without affecting neuronal activation, whereas propranolol (2 mg/kg iv) did not affect either response to ANF. These results suggest that ANF is a neuromediator involved in the excitation of cardioinhibitory neurons in the NA.


Fixed nuclei as alternative template of BIOMED-2 multiplex polymerase chain reaction for immunoglobulin gene clonality testing in B-cell malignancies.
Evaluation of immunoglobulin (Ig) gene rearrangements with BIOMED-2 multiplex PCR has become a standard detection of clonality in mature B cell malignancies. Conventionally, this method is relatively labor-intensive and time-consuming, as it requires DNA isolation from bone marrow aspirates (BM) or peripheral blood (PB) in patients with BM or PB involvement. On the other hand, fluorescence in situ hybridization (FISH) is routinely used as genetic screening in B cell malignancies, but the surplus fixed nuclei initially prepared for FISH usually turn useless afterwards. We sought to use these surplus nuclei after FISH as a template to perform PCR-based Ig gene clonality testing. Templates of 12 patients with mature B cell malignancies, which consisted of both DNA isolated with commercial DNA isolation kit from fresh BM or PB (DNA group) and the fixed nuclei initially prepared for FISH (nuclei group) from the same individuals, were subjected to PCR with BIOMED-2 primer sets for immunoglobulin heavy chain and kappa light chain under recommended conditions. Our result, for the first time, showed a high consistency between the two groups in detecting B cell clonality, which indicates that nuclei for FISH can function as a reliable template comparable to fresh tissue-isolated DNA in PCR based Ig clonality testing. This offers a simple, rapid and more economical alternative to standard Ig testing based on regular DNA.


[Hypospadias--not only a misplaced urinary meatus].
Hypospadias is a congenital malformation with a complex and unclear etiology; both hormonal disturbances and genetic factors may contribute. It is discussed internationally whether the incidence of the malformation has increased during the last decades. There are many operation methods for treatment of hypospadias. Incidence, possible etiologic factors and the clinical presentation of hypospadias is discussed, based on personal experience and review of literature retrieved from PubMed. Principles of treatment are outlined. The diagnosis is usually made during a routine examination after birth. The less severe cases of hypospadias can be overlooked. Most of them ( 2 / 3 ) are of a distal type and lack foreskin coverage of the ventral side of the penis head and there is often a ventral curvature of the penis. Many currently recommend the operation to be taken from 6 to 18 months of age, but it may be technically more challenging to operate such young boys than to postpone it to when they are older. The operational method is adapted to the type of hypospadias and the aim is to achieve a satisfactory cosmetic and functional result and a low incidence of postoperative complications.


Kinetic, isotherm, and thermodynamic studies of the adsorption of dyes from aqueous solution by cellulose-based adsorbents.
In this study, a highly efficient and eco-friendly porous cellulose-based aerogel was synthesized by grafting polyethyleneimine onto quaternized cellulose (PQC) to remove the anionic dye Congo Red (CR). The prepared aerogel had a good flexibility and formability. The adsorbents were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and elemental analysis. The results showed that there were many amino groups on CE/PQC aerogel and the structure was porous, which increased the adsorption capacity. The effects of initial concentration, adsorbent dose, contact time, temperature, and pH on the dye sorption were all investigated. The adsorption mechanism was also explored, including adsorption kinetics, adsorption isotherms and thermodynamic studies of adsorption. The results showed that the adsorption kinetics and isotherms fitted the pseudo-second-order kinetic model and Langmuir isotherm, respectively. The Langmuir isotherm revealed that the maximum theoretical adsorption capacity of the aerogels for CR was 518.403 mg g-1. The thermodynamic parameters including Gibbs free energy change (ΔG0), enthalpy change (ΔH0) and entropy change (ΔS0), showed the adsorption process was exothermic and spontaneous. These results imply that this new absorbent can be universally and effectively used for the removal of dyes from industrial textile wastewater.


Pilot study of a brief intervention based on the theory of planned behaviour and self-identity to increase chlamydia testing among young people living in deprived areas.
This study sought to estimate the effects of a novel intervention, compared with usual chlamydia testing promotion, on chlamydia test uptake and intentions among young people living in deprived areas. The intervention was based on the theory of planned behaviour, augmented with self-identity, and targeted the significant predictors of chlamydia testing intentions identified in the previous research. Cluster randomization was used to allocate college tutor groups (intervention n = 10; control n = 11) to the intervention or control group. The sample comprised 253 participants (intervention n = 145, control n = 108). The primary outcome was test offer uptake at the end of the session. Other outcomes measured at immediate follow-up were intention, attitude, subjective norm, perceived behavioural control, and self-identity. Generalized estimating equations, controlling for cluster effects and sexual activity, found a small but non-significant effect of condition on test offer uptake, OR = 1.65 (95% CI 0.70, 3.88) p = .25, with 57.5% of intervention participants accepting the offer of a test compared with 40.2% of control participants. Using the same analysis procedure, small-to-medium intervention effects were found on other outcome variables, including a significant effect on attitudes towards chlamydia testing, OR = 1.37 (95% CI 1.00, 1.87), p = .05. The results provide encouraging initial evidence that this theory-based intervention, targeting the key determinants of chlamydia testing, may help to improve chlamydia testing uptake in a high-risk group. They support the conduct of a larger trial to evaluate the effectiveness of the intervention. What is already known on this subject? Young people living in areas of increased socio-economic deprivation have been identified as a high-risk group for chlamydia. Previous research within an extended model of the theory of planned behaviour (TPB) found that attitude, subjective norm, perceived behavioural control, and self-identity all significantly predicted chlamydia testing intentions in this high-risk group. What does this study add? Development and testing of a novel, TPB-based intervention targeting predictors of chlamydia testing intentions. The intervention led to significantly more positive attitudes towards chlamydia testing. Preliminary indication that a TPB-based intervention may help to improve chlamydia testing in a high-risk group.


Role of posture in mechanics of the lumbar spine in compression.
Biomechanics of the entire lumbar spine L1-S1 in erect postures was investigated under physiological axial compression loads of up to 800 N with and without right axial torques of up to 10 Nm using a nonlinear finite-element model. The sagittal curvature is varied from an initial value of 46 degrees by +15 degrees to model more lordotic posture and by -7.5 degrees as well as -15 degrees to model flattened lumbar spines. The primary/coupled displacements, disc pressures, ligament forces, facet forces, disc fiber strains, along with the required stabilizing sagittal/lateral moments, are computed at different levels, loadings, and lordotic postures. The role of facetectomy and partial nucleotomy at some levels is also studied for a few cases. The primary/coupled motions, disc pressures, facet loads, and disc fiber strains are all affected by postural changes and the addition of 10-Nm axial torque. The primary responses are stiffened when axial compression and axial torque are applied together rather than separately. Flexion and left lateral moments are required to stabilize the lumbar spine in compression. The changes in lordosis markedly affect the stabilizing sagittal moments. The lateral moments, however, are significantly influenced only in the presence of axial torque. Axial compression generates small disc fibers that decrease from the innermost layer to outer ones at all disc levels. The contact forces are largest at the L5-S1 facets. These forces cause large differences in rotation between L5 anterior and posterior bony structures, suggesting the high stresses on corresponding pedicles and pars interarticularis. Finally, the stabilizing flexion moments are generated primarily by the off-centeredness of the gravity load. Relatively small muscle forces are required to balance the remaining portion.


Restriction of transpiration rate under high vapour pressure deficit and non-limiting water conditions is important for terminal drought tolerance in cowpea.
Drought stress is a major constraint on cowpea productivity, since the crop is grown under warm conditions on sandy soils having low water-holding capacity. For enhanced performance of crops facing terminal drought stress, like cowpea, water-saving strategies are crucial. In this work, the growth and transpiration rate (TR) of 40 cowpea genotypes with contrasting response to terminal drought were measured under well-watered conditions across different vapour pressure deficits (VPD) to investigate whether tolerant and sensitive genotypes differ in their control of leaf water loss. A method is presented to indirectly assess TR through canopy temperature (CT) and the index of canopy conductance (Ig). Overall, plants developed larger leaf area under low than under high VPD, and there was a consistent trend of lower plant biomass in tolerant genotypes. Substantial differences were recorded among genotypes in TR response to VPD, with tolerant genotypes having significantly lower TR than sensitive ones, especially at times with the highest VPD. Genotypes differed in TR response to increasing VPD, with some tolerant genotypes exhibiting a clear VPD breakpoint at about 2.25 kPa, above which there was very little increase in TR. In contrast, sensitive genotypes presented a linear increase in TR as VPD increased, and the same pattern was found in some tolerant lines, but with a smaller slope. CT, estimated with thermal imagery, correlated well with TR and Ig and could therefore be used as proxy for TR. These results indicate that control of water loss discriminated between tolerant and sensitive genotypes and may, therefore, be a reliable indicator of terminal drought stress tolerance. The water-saving characteristics of some genotypes are hypothesised to leave more soil water for pod filling, which is crucial for terminal drought adaptation.


The Polybasic Cleavage Site in SARS-CoV-2 Spike Modulates Viral Sensitivity to Type I Interferon and IFITM2.
The cellular entry of severe acute respiratory syndrome-associated coronaviruses types 1 and 2 (SARS-CoV-1 and -2) requires sequential protease processing of the viral spike glycoprotein. The presence of a polybasic cleavage site in SARS-CoV-2 spike at the S1/S2 boundary has been suggested to be a factor in the increased transmissibility of SARS-CoV-2 compared to SARS-CoV-1 by facilitating maturation of the spike precursor by furin-like proteases in the producer cells rather than endosomal cathepsins in the target. We investigate the relevance of the polybasic cleavage site in the route of entry of SARS-CoV-2 and the consequences this has for sensitivity to interferons (IFNs) and, more specifically, the IFN-induced transmembrane (IFITM) protein family that inhibit entry of diverse enveloped viruses. We found that SARS-CoV-2 is restricted predominantly by IFITM2, rather than IFITM3, and the degree of this restriction is governed by route of viral entry. Importantly, removal of the cleavage site in the spike protein renders SARS-CoV-2 entry highly pH and cathepsin dependent in late endosomes, where, like SARS-CoV-1 spike, it is more sensitive to IFITM2 restriction. Furthermore, we found that potent inhibition of SARS-CoV-2 replication by type I but not type II IFNs is alleviated by targeted depletion of IFITM2 expression. We propose that the polybasic cleavage site allows SARS-CoV-2 to mediate viral entry in a pH-independent manner, in part to mitigate against IFITM-mediated restriction and promote replication and transmission. This suggests that therapeutic strategies that target furin-mediated cleavage of SARS-CoV-2 spike may reduce viral replication through the activity of type I IFNs.IMPORTANCE The furin cleavage site in the spike protein is a distinguishing feature of SARS-CoV-2 and has been proposed to be a determinant for the higher transmissibility between individuals, compared to SARS-CoV-1. One explanation for this is that it permits more efficient activation of fusion at or near the cell surface rather than requiring processing in the endosome of the target cell. Here, we show that SARS-CoV-2 is inhibited by antiviral membrane protein IFITM2 and that the sensitivity is exacerbated by deletion of the furin cleavage site, which restricts viral entry to low pH compartments. Furthermore, we find that IFITM2 is a significant effector of the antiviral activity of type I interferons against SARS-CoV-2 replication. We suggest that one role of the furin cleavage site is to reduce SARS-CoV-2 sensitivity to innate immune restriction, and thus, it may represent a potential therapeutic target for COVID-19 treatment development.


[Fluorescein angiography findings after retinotomy and retinectomy].
Drainage-retinotomies and relaxing retinectomies are helpful techniques in extreme vitreoretinal surgery. They have become established as a surgical instrument, but their possible pathophysiologic effects on the retina, pigment epithelium and choroid have not been investigated as yet. 30 Patients with retinotomies or retinectomies after vitrectomy with silicone oil filling for advanced proliferative vitreoretinopathy were studied angiographically. Fluorescein angiographic findings revealed 1. a predominant effect of endophotocoagulation in the area of the retinotomy/retinectomy with occlusion of the choriocapillaris and hyperpigmentation and window effects of the retinal pigment epithelium, but sometimes there was a localized traction effect with retinal and choroidal folds, 2. that retinal perfusion is unaffected in untreated and coagulated areas without evidence for a breakdown of the blood-retina-barrier or retinal ischemia, 3. an intact blood-retinal-barrier in areas of uncovered retinal pigment epithelium, 4. no evidence for significant reproliferations in the areas of retinotomies and retinectomies, 5. choroidal neovascular membranes which may occur postoperatively at the retinotomy site probably as a complication for intraoperative injury of Bruch's membrane, 6. a cystoid macular edema, an optic atrophy, pigment-fallout, depigmented tracks and choroidal folds as unspecific findings post retinal detachment surgery. The angiographic data support the clinical impression, that retinotomies and retinectomies performed in otherwise untreatable cases cause no significant damage in the attached retina.


Ureteroscopy: The standard of care in the management of upper tract urolithiasis in children.
Advances in technology and the continued evolution in the design of ureteroscopes now permit a primary endoscopic approach to the upper urinary tract of pediatric patients on a routine basis to treat a diverse group of conditions that include urolithiasis, hematuria and strictures. The purpose of this review article is to demonstrate that ureteroscopic lithotripsy is now to be considered the standard of care in the management of upper tract urolithiasis in the pediatric patient, replacing shockwave lithotripsy (ESWL) as the first line of therapy. Additionally, the article will discuss the available endoscopic equipment and the lessons learned over the years to optimize the success of these procedures in children. A systematic review of articles written about ureteroscopy (URS) in the contemporary urological literature (1990-2009) on PubMed was undertaken. The success rates and complications of pediatric ureteroscopic procedures were abstracted from the identified publications and the results were tabulated and compared with the success rates of shockwave lithotripsy. In over 832 URS cases, there was a 5.9% complication rate and a stone-free rate of 93.4%. The stone-free rates of URS are superior to those obtained with the published success rates with ESWL of 80.3% in 1,839 cases. The safety and outcomes of ureteroscopic lithotripsy in the management of pediatric urolithiasis now justify that this treatment modality be considered the standard of care and first line of therapy in the management of children who present with upper tract stones.


Temporal variations in gas temperature in an atomization stage of cadmium and tellurium evaluated by using the two-line method in graphite furnace atomic absorption spectrometry.
In order to discuss the atomization process of an analyte element occurring in a graphite furnace for atomic absorption spectrometry, we measured variations in the characteristic temperature with the progress of an atomization stage, by using a two-line method under the assumption of a Boltzmann distribution. For this purpose, iron was chosen as the analyte element. Also, the atomic absorption of two iron atomic lines, Fe I 372.0 nm and Fe I 373.7 nm, was simultaneously monitored as a probe for the temperature determination. This method enables variations in the gas temperature to be directly traced, yielding a temperature distribution closely related to the diffusion behavior of the probe element in the furnace. This temperature variation was very different from the furnace wall temperatures, which were monitored in conventional temperature control for atomic absorption spectrometry. Correlations between the gas temperature and the charring/atomizing temperatures in the heating program of the furnace were investigated. The atomization of cadmium and tellurium was also investigated by a comparison between the gas temperature with the wall temperature of the furnace. The atomic absorption of cadmium or tellurium appeared to be apart from the absorption of iron while the gas temperature was still low. Therefore, the analyte atoms could be atomized through direct contact with the wall of the graphite furnace, which has a much higher temperature compared to the gas atmosphere during atomization. Their atomization would be caused by conductive heating from the furnace wall rather than by radiant heating in the furnace.


Probiotic Escherichia coli strain Nissle 1917 outcompetes intestinal pathogens during biofilm formation.
Many bacterial infections are associated with biofilm formation. Bacterial biofilms can develop on essentially all kinds of surfaces, producing chronic and often intractable infections. Escherichia coli is an important pathogen causing a wide range of gastrointestinal infections. E. coli strain Nissle 1917 has been used for many decades as a probiotic against a variety of intestinal disorders and is probably the best field-tested E. coli strain in the world. Here we have investigated the biofilm-forming capacity of Nissle 1917. We found that the strain was a good biofilm former. Not only was it significantly better at biofilm formation than enteropathogenic, enterotoxigenic and enterohaemorrhagic E. coli strains, it was also able to outcompete such strains during biofilm formation. The results support the notion of bacterial prophylaxis employing Nissle 1917 and may partially explain why the strain has a beneficial effect on many intestinal disorders.


Simplified insertion of the Port-A-Cath implantable drug-delivery system.
The increasing frequency with which antineoplastic drugs are being used demands improved vascular access for their administration. Venous access through implantable catheters facilitates treatment when veins are no longer available. Totally implantable catheters of the Port-A-Cath type are preferred because they give better performance and cosmetic results, and are more convenient for the patients. The flexibility of these catheters makes their introduction difficult. The author describes a simplified technique, using a commercially available introducer that has been used satisfactorily in thousands of pacemaker implantations. The author has used this technique routinely with a 50% decrease in operative and fluoroscopy time and without complications.


Does culture influence what and how we think? Effects of priming individualism and collectivism.
Do differences in individualism and collectivism influence values, self-concept content, relational assumptions, and cognitive style? On the one hand, the cross-national literature provides an impressively consistent picture of the predicted systematic differences; on the other hand, the nature of the evidence is inconclusive. Cross-national evidence is insufficient to argue for a causal process, and comparative data cannot specify if effects are due to both individualism and collectivism, only individualism, only collectivism, or other factors (including other aspects of culture). To address these issues, the authors conducted a meta-analysis of the individualism and collectivism priming literature, with follow-up moderator analyses. Effect sizes were moderate for relationality and cognition, small for self-concept and values, robust across priming methods and dependent variables, and consistent in direction and size with cross-national effects. Results lend support to a situated model of culture in which cross-national differences are not static but dynamically consistent due to the chronic and moment-to-moment salience of individualism and collectivism. Examination of the unique effects of individualism and collectivism versus other cultural factors (e.g., honor, power) awaits the availability of research that primes these factors.


Cladribine with prednisone versus chlorambucil with prednisone as first-line therapy in chronic lymphocytic leukemia: report of a prospective, randomized, multicenter trial.
The efficacy and toxicity of cladribine (2-CdA) + prednisone (P) versus chlorambucil (Chl) + P were compared in previously untreated patients with progressive or symptomatic chronic lymphocytic leukemia (CLL) in a randomized, multicenter prospective trial. Eligible patients were assigned to either 2-CdA 0.12 mg/kg per day in 2-hour infusions and P 30 mg/m(2) per day for 5 consecutive days or Chl 12 mg/m(2) per day and P 30 mg/m(2) per day for 7 consecutive days. Three courses were administered at 28-day intervals or longer if myelosuppression developed. The therapy was finished if complete response (CR) was achieved. Of 229 available patients 126 received 2-CdA+P and 103 received Chl+P as a first-line treatment. CR and overall response rates were significantly higher in the patients treated with 2-CdA+P (47% and 87%, respectively) than in the patients treated with Chl+P (12% and 57%, respectively) (P = .001). Progression-free survival was significantly longer in the 2-CdA-treated group (P = .01), but event-free survival was not statistically different. Thirteen percent of patients were refractory to 2-CdA+P and 43% to Chl+P (P = .001). Drug-induced neutropenia was more frequently observed during 2-CdA+P (23%) than Chl+P therapy (11%) (P = .02), but thrombocytopenia occurred with similar frequency in both groups (36% and 27%, respectively). Infections were seen more frequently in the 2-CdA+P-treated group (56%) than in the Chl+P-treated group (40%; P = .02). Death rates have so far been similar in patients treated with 2-CdA (20%) and with Chl (17%). The probability of overall survival calculated from Kaplan-Meier curves at 24 months was also similar for both groups (78% and 82%, respectively). (Blood. 2000;96:2723-2729)


Role of outcome conflict in dual-task interference.
The traditional explanation for dual-task interference is that tasks compete for scarce processing resources. Another possible explanation is that the outcome of the processing required for one task conflicts with the processing required for the other task (e.g., cross talk). To explore the contribution of outcome conflict to task interference, we manipulated the relatedness of the tasks. In Experiment 1, subjects searched concurrently for names of boys in one channel and names of cities in another channel. Responses were significantly delayed when a nontarget on one channel belonged to, or was even just related to, the category designated as the target for the other channel. No comparable effects were found when the tasks were performed in isolation. Thus, the difficulty of the individual tasks is not the only determinant of how much they will interfere when combined, and there must be substantial interactions between processes carrying out the two tasks. In Experiment 2 subjects searched one channel for specific target letters and another channel for specific target digits. The nontargets in a channel were either from the same alphanumeric category as the targets for that channel or from the opposite category (i.e., the category of the targets for the other channel). It was found that although between-category search was more efficient than within-category search in single tasks, it was less efficient in dual tasks. Thus, there appear to be significant task interactions due to the confusability emerging when the nontargets of one task belong to the same category as the targets of the concurrent task. In addition, the congruence of target presence or absence on the two channels was found to have a sizeable effect. We suggest four potential sources of outcome conflict that may contribute to dual-task interference, and we conjecture that a great deal of the residual interference might result from other sorts of outcome conflict.


Is decision-making based on the internet during pregnancy a predictive factor for vaccine hesitancy in pregnant women during the pandemic?
Pregnant women frequently use the Internet to get information, and the information they receive is effective in their decision-making. It is known that pregnant women get information about COVID-19 vaccines from the Internet. This study aims to determine whether decision-making based on Internet sources during pregnancy is a predictive factor for vaccine hesitancy in pregnant women during the pandemic. A descriptive, cross-sectional research design was used. The data were collected by using an online survey instrument. There was a positive, moderate, and significant relationship between the self-efficacy perception sub-dimension of the internet decision-making during pregnancy scale and the risk sub-dimension of the vaccine hesitancy in pandemics scale (r: .584, p < .05) and between the self-control sub-dimension of the internet decision-making during pregnancy scale and the risk sub-dimension of the vaccine hesitancy in pandemics scale (r: .546, p < .05). The perception of self-control (β: .291) affected the lack of confidence, and the perception of self-efficacy (β: .481) affected the risk perception more than other variables. There was a relationship between internet decision-making and COVID-19 vaccine hesitancy in pregnant women. Health professionals, information specialists, and librarians should orient people to reliable sources about vaccines.


tRNA cleavage is a conserved response to oxidative stress in eukaryotes.
Recent results have identified a diversity of small RNAs in a wide range of organisms. In this work, we demonstrate that Saccharomyces cerevisiae contains a small RNA population consisting primarily of tRNA halves and rRNA fragments. Both 5' and 3' fragments of tRNAs are detectable by Northern blot analysis, suggesting a process of endonucleolytic cleavage. tRNA and rRNA fragment production in yeast is most pronounced during oxidative stress conditions, especially during entry into stationary phase. Similar tRNA fragments are also observed in human cell lines and in plants during oxidative stress. These results demonstrate that tRNA cleavage is a conserved aspect of the response to oxidative stress.


Variation of wastewater contaminants in China and source identification using hierarchical clustering based on weighted factor scores.
Human activities have huge impact on the aquatic environment. Knowledge on sources of the contaminants provides guidelines to determine the ideal location and maintenance of monitoring stations, thus advancing environmental monitoring and pollution control. Factor analysis (FA) may be the most popular method for source identification, but the results should be affirmed. Following this logic, in this research, firstly the potential sources were determined, and secondly the contaminant concentrations in the source regions and the non-source regions were compared. To identify the potential sources, 75 meteorological, economic and social indicators were used to group the study regions. FA was used to reduce dimensionality and factor scores were calculated. The grouping was based on the weighted factor scores while the weight was variance explained by each factor respectively. Each group was supposed to correspond to a factor; that is, a potential source. The results indicated that the concentrations of chemical oxygen demand (COD), ammonia nitrogen, phosphorus and arsenic in wastewater were significantly different between groups. Animal husbandry, mining and/or energy industry were the main sources of COD, ammonia nitrogen and phosphorus; animal husbandry, mining, energy industry, and/or heavy and chemical industry were the main sources of phosphorus; humid climate and/or secondary industry were the main sources of arsenic.


Altered glycosaminoglycan production by HL-60 cells treated with 4-methylumbelliferyl-beta-D-xyloside.
Glycosaminoglycans, mainly chondroitin 4-sulfate, are located in the primary granules of human myeloid cells. These polyanionic carbohydrates are believed to play an important role in leukocyte maturation and function. To study the effect of altered chondroitin sulfate metabolism on human promyelocytic leukemia cells, we have treated HL-60 cells with 4-methylumbelliferyl-beta-D-xyloside. beta-D-Xylosides initiate the synthesis of free chondroitin sulfate chains. Cytochemical studies of treated cells demonstrated a marked increase in cytoplasmic granules stained with cationic dyes. This was confirmed by radiolabeled precursor incorporation studies that demonstrated a 344% increase in 35S-sulfate uptake into glycosaminoglycans associated with the cells and a 39% increase in incorporation into glycosaminoglycans released into the media. Chromatographic analyses of these glycosaminoglycans from treated cells demonstrated that the newly formed chondroitin sulfate chains were not attached to protein core and were of shorter length, but of greater charge density than chondroitin sulfate produced by control cells. Thus, beta-D-xyloside appears to alter the protein linkage, chain length, and sulfation of chondroitin sulfate produced by HL-60 cells, and these changes are morphologically evident. These biochemically altered cells may provide important information concerning the role of these macromolecules in myeloid development.


A retinoic acid binding cytochrome P450: CYP120A1 from Synechocystis sp. PCC 6803.
At least 35 cytochrome P450 (P450, CYP) or cytochrome P450-like genes have been identified in 10 cyanobacterial genomes yet none have been functionally characterized. CYP110 and CYP120 represent the two largest cyanobacterial P450 families with 16 and four members, respectively, identified to date. The Synechocystis sp. PCC 6803 CYP120A1 protein sequence shares high degrees of conservation with CYP120A2 from Trichodesmium erythraeum IMS101 and CYP120B1 and CYP120C1 from Nostoc punctiforme PCC 73102. In this communication, we report the cloning, expression, purification, and characterization of CYP120A1 from Synechocystis. Homology modeling predictions of the three-dimensional structure of CYP120A1 coupled with in silico screening for potential substrates and experimental spectroscopic analyses have identified retinoic acid as a compound binding with high affinity to this P450's catalytic site. These characterizations of Synechocystis CYP120A1 lay the initial foundations for understanding the basic role of cytochrome P450s in cyanobacteria and related organisms.


Ceritinib: a new tyrosine kinase inhibitor for non-small-cell lung cancer.
To review ceritinib for the treatment of anaplastic lymphoma kinase (ALK)-positive metastatic non-small-cell lung cancer (NSCLC). Literature searches were conducted in PubMed, EMBASE (1974 to July week 5, 2014), and Google Scholar using the terms ceritinib, LDK378, and non-small-cell lung cancer. One phase 1 trial and 2 abstracts were identified. Ceritinib is approved for the treatment of ALK-positive metastatic NSCLC in patients who are intolerant to or have progressed despite therapy with crizotinib. In the phase 1 clinical trial, the maximum tolerated dose was determined to be 750 mg once daily. The overall response rate (ORR) was 58% (95% CI = 48-67) in patients who received ≥400 mg daily (n = 114). In this group, the ORR was 56% (95% CI = 41-67) and 62% (95% CI = 44-78) among crizotinib-exposed and -naïve patients, respectively. The ORR was 59% (95% CI = 47-70) in patients who received 750 mg daily (n = 78). The ORR was 56% (95% CI = 41-70) in crizotinib-treated patients and 64% (95% CI = 44-81) in crizotinib-naïve patients, respectively, in this subset. The median duration of response was 8.2 months. Median progression-free survival was 7.0 months. The most common adverse reactions included diarrhea, nausea, vomiting, abdominal pain, anorexia, constipation, fatigue, and elevated transaminases. Ceritinib has activity in crizotinib-resistant and crizotinib-naïve patients and appears to be a viable alternative for ALK-positive NSCLC. Long-term data are needed to further define the role of ceritinib in the treatment of NSCLC.


Various forms of tissue damage and danger signals following hematopoietic stem-cell transplantation.
Hematopoietic stem-cell transplantation (HSCT) is the most potent curative therapy for many malignant and non-malignant disorders. Unfortunately, a major complication of HSCT is graft-versus-host disease (GVHD), which is mediated by tissue damage resulting from the conditioning regimens before the transplantation and the alloreaction of dual immune components (activated donor T-cells and recipient's antigen-presenting cells). This tissue damage leads to the release of alarmins and the triggering of pathogen-recognition receptors that activate the innate immune system and subsequently the adaptive immune system. Alarmins, which are of endogenous origin, together with the exogenous pathogen-associated molecular patterns (PAMPs) elicit similar responses of danger signals and represent the group of damage-associated molecular patterns (DAMPs). Effector cells of innate and adaptive immunity that are activated by PAMPs or alarmins can secrete other alarmins and amplify the immune responses. These complex interactions and loops between alarmins and PAMPs are particularly potent at inducing and then aggravating the GVHD reaction. In this review, we highlight the role of these tissue damaging molecules and their signaling pathways. Interestingly, some DAMPs and PAMPs are organ specific and GVHD-induced and have been shown to be interesting biomarkers. Some of these molecules may represent potential targets for novel therapeutic approaches.


Comparison of organ-specific endothelial cells in terms of microvascular formation and endothelial barrier functions.
Every organ demonstrates specific vascular characteristics and functions maintained by interactions of endothelial cells (ECs) and parenchymal cells. Particularly, brain ECs play a central role in the formation of a functional blood brain barrier (BBB). Organ-specific ECs have their own morphological features, and organ specificity must be considered when investigating interactions between ECs and other cell types constituting a target organ. Here we constructed angiogenesis-based microvascular networks with perivascular cells in a microfluidic device setting by coculturing ECs and mesenchymal stem cells (MSCs). Furthermore, we analyzed endothelial barrier functions as well as fundamental morphology, an essential step to build an in vitro BBB model. In particular, we used both brain microvascular ECs (BMECs) and human umbilical vein ECs (HUVECs) to test if organ specificity of ECs affects the formation processes and endothelial barrier functions of an engineered microvascular network. We found that microvascular formation processes differed by the source of ECs. HUVECs formed more extensive microvascular networks compared to BMECs while no differences were observed between BMECs and HUVECs in terms of both the microvascular diameter and the number of pericytes peripherally associated with the microvasculatures. To compare the endothelial barrier functions of each type of EC, we performed fluorescence dextran perfusion on constructed microvasculatures. The permeability coefficient of BMEC microvasculatures was significantly lower than that of HUVEC microvasculatures. In addition, there were significant differences in terms of tight junction protein expression. These results suggest that the organ source of ECs influences the properties of engineered microvasculature and thus is a factor to be considered in the design of organ-specific cell culture models.


EWOD (electrowetting on dielectric) digital microfluidics powered by finger actuation.
We report finger-actuated digital microfluidics (F-DMF) based on the manipulation of discrete droplets via the electrowetting on dielectric (EWOD) phenomenon. Instead of requiring an external power supply, our F-DMF uses piezoelectric elements to convert mechanical energy produced by human fingers to electric voltage pulses for droplet actuation. Voltage outputs of over 40 V are provided by single piezoelectric elements, which is necessary for oil-free EWOD devices with thin (typically <1 μm) dielectric layers. Higher actuation voltages can be provided using multiple piezoelectric elements connected in series when needed. Using this energy conversion scheme, we confirmed basic modes of EWOD droplet operation, such as droplet transport, splitting and merging. Using two piezoelectric elements in series, we also successfully demonstrated applications of F-DMF for glucose detection and immunoassay. Not requiring power sources, F-DMF offers intriguing paths for various portable and other microfluidic applications.


Chronic exposure to simulated space conditions predominantly affects cytoskeleton remodeling and oxidative stress response in mouse fetal fibroblasts.
Microgravity and cosmic rays as found in space are difficult to recreate on earth. However, ground-based models exist to simulate space flight experiments. In the present study, an experimental model was utilized to monitor gene expression changes in fetal skin fibroblasts of murine origin. Cells were continuously subjected for 65 h to a low dose (55 mSv) of ionizing radiation (IR), comprising a mixture of high‑linear energy transfer (LET) neutrons and low-LET gamma-rays, and/or simulated microgravity using the random positioning machine (RPM), after which microarrays were performed. The data were analyzed both by gene set enrichment analysis (GSEA) and single gene analysis (SGA). Simulated microgravity affected fetal murine fibroblasts by inducing oxidative stress responsive genes. Three of these genes are targets of the nuclear factor‑erythroid 2 p45-related factor 2 (Nrf2), which may play a role in the cell response to simulated microgravity. In addition, simulated gravity decreased the expression of genes involved in cytoskeleton remodeling, which may have been caused by the downregulation of the serum response factor (SRF), possibly through the Rho signaling pathway. Similarly, chronic exposure to low-dose IR caused the downregulation of genes involved in cytoskeleton remodeling, as well as in cell cycle regulation and DNA damage response pathways. Many of the genes or gene sets that were altered in the individual treatments (RPM or IR) were not altered in the combined treatment (RPM and IR), indicating a complex interaction between RPM and IR.


Double staining immunofluorescence procedure for enumeration of cells containing cytoplasmic immunoglobulin in human bone marrow: 40 selected cases.
A double staining immunofluorescence method was developed that allowed the reliable differential counting of cells containing cytoplasmic immunoglobulin in human bone marrow. These cells were stained with a polyspecific antihuman immunoglobulin serum conjugated with rhodamine. In separate preparations subpopulations of cells containing each heavy chain class and light chain type were identified by prior staining with isotype specific fluorescein conjugated antihuman immunoglobulin sera. A standardised counting procedure was adopted to indicate the degree of plasma cell infiltration and to establish the percentage of cells containing each type of immunoglobulin. A series of 122 patients requiring haematological investigation of the bone marrow for various disorders but without evidence of paraproteinaemia was tested to provide results for a nonmyelomatous reference group. These results were compared with those from 40 patients with paraproteins of various classes in their serum. The procedure described here clearly differentiated the two groups. When combined with serum paraprotein assays and conventional haematological cytology of bone marrow these counts provided useful additional information in cases of diagnostic difficulty and in the assessment of individual patients before, during, and after treatment.


Post-introduction evolution in the biological control agent Longitarsus jacobaeae (Coleoptera: Chrysomelidae).
Rapid evolution has rarely been assessed in biological control systems despite the similarity with biological invasions, which are widely used as model systems. We assessed post-introduction climatic adaptation in a population of Longitarsus jacobaeae, a biological control agent of Jacobaea vulgaris, which originated from a low-elevation site in Italy and was introduced in the USA to a high-elevation site (Mt. Hood, Oregon) in the early 1980s. Life-history characteristics of beetle populations from Mt. Hood, from two low-elevation sites in Oregon (Italian origin) and from a high-elevation site from Switzerland were compared in common gardens. The performance of low- and high-elevation populations at a low- and a high-elevation site was evaluated using reciprocal transplants. The results revealed significant changes in aestival diapause and shifts in phenology in the Mt. Hood population, compared with the low-elevation populations. We found increased performance of the Mt. Hood population in its home environment compared with the low-elevation populations that it originated from. The results indicate that the beetles at Mt. Hood have adapted to the cooler conditions by life-history changes that conform to predictions based on theory and the phenology of the cold-adapted Swiss beetles.


The obstetric care subsidy policy in Burkina Faso: what are the effects after five years of implementation? Findings of a complex evaluation.
Burkina Faso, like many low and middle income countries, has been taking a range of actions to address its poor maternal and neonatal health indicators. In 2006 the government introduced an innovative national subsidy scheme for deliveries and emergency obstetric care in public facilities. This article reports on a complex evaluation of this policy, carried out 5 years after its introduction, which examined its effects on utilisation, quality of care, equity and the health system as a whole, as well as its cost and sustainability. The evaluation was carried out in six purposively selected districts, as well as at national level, using a case study approach. Data sources included: national and district routine and survey data, household interviews with women who had recently given birth, data extraction from hospital and medical records, and key informant and health worker interviews. The underlying secular trend of a 1% annual increase in the facility-based delivery rate (1988-2010) was augmented by an additional 4% annual increase from 2007 onwards (after the policy was introduced), especially in rural areas and amongst women from poor households. The absence of baseline quality of care data made it difficult to assess the impact of the policy on quality of care, but hospitals with the best level of implementation of the subsidy offered higher quality of care (as measured by health care near-misses), so there is no evidence of a negative impact on quality (as is often feared). Similarly, there is little evidence of unintended negative effects on untargeted services. Household payments for facility-based deliveries have reduced significantly, compared with payments before the policy, and the policy as a whole is affordable, costing about 2% of total public health expenditure. Concerns include that the amounts paid by households are higher than the rates set by the policy, and also that 7% of households still say that they cannot afford to pay. Wealthier women have higher utilisation of services, as before, and the policy of fully exempting indigents is not being put into practice. These findings highlight the importance of maintaining the subsidy policy, given the evidence of positive outcomes, but they also point out areas where attention is needed to ensure the poor and most vulnerable population benefit fully from the policy.


Measurement of verapamil concentrations in plasma by gas chromatography and high pressure liquid chromatography.
This study was carried out to compare gas chromatography (GC) and high-pressure liquid chromatography (HPLC) procedures in the measurement of plasma levels of verapamil. Other analytic methods previously reported are not widely available (mass fragmentography) or are subject to interference from drug metabolites (spectrophotofluorometry). A single extraction and derivatization procedure was developed to prepare samples for either GC or HPLC analysis. The GC procedure used a nitrogen-specific detector; the HPLC, a fluorescence detector. In a 1--500 ng/ml range of verapamil concentrations, both methods resulted in good separation of verapamil from a major metabolite, norverapamil, and from compound D517, used as an internal standard. Intraassay variation was similar for both procedures, with only slightly higher interassay variability found for the GC technique. Excellent correlation was found during analysis of the same unknown samples by both methods (r = 0.97; p less than 0.001). Either assay procedure appears satisfactory for use in measurement of verapamil levels in plasma.


Sulfosulfuron incorporated in micelles adsorbed on montmorillonite for slow release formulations.
Slow release formulations of the anionic herbicide sulfosulfuron (SFS) were prepared by incorporating it in micelles of an organic cation octadecyltrimethylammonium, which adsorb on the clay-mineral montmorillonite. The fraction of SFS adsorbed on the micelle-clay complex reached 98%, whereas for monomer-clay complexes, its adsorption was insignificant. Fluorescence studies showed surface contact between the micelles and the clay surface. The rate of SFS release from the micelle-clay formulations in aqueous suspensions was slow (<1%, 72 h). Spraying SFS formulations on a thin soil layer in a funnel, followed by irrigations (50 mm), resulted in complete elution of SFS from the commercial formulation (dispersible granular) versus 4% from the micelle-clay formulation. A plant bioassay in Rehovot soil showed that these respective formulations yielded 23 and 65% of shoot growth inhibition of foxtail. Consequently, the slow release micelle-clay formulations of SFS yield significantly reduced leaching and enhanced biological activity, thus providing environmental and agricultural advantages.


Production of poly(3-hydroxybutyrate)/poly(lactic acid) from industrial wastewater by wild-type Cupriavidus necator H16.
The massive production of urban and industrial wastes has created a clear need for alternative waste management processes. One of the more promising strategies is to use waste as raw material for the production of biopolymers such as polyhydroxyalkanoates (PHAs). In this work, a lactate-enriched stream obtained by anaerobic digestion (AD) of wastewater (WW) from a candy production plant was used as a feedstock for PHA production in wild-type Cupriavidus necator H16. Unexpectedly, we observed the accumulation of poly(3-hydroxybutyrate)/poly(lactic acid) (P(3HB)/PLA), suggesting that the non-engineered strain already possesses the metabolic potential to produce these polymers of interest. The systematic study of factors, such as incubation time, nitrogen and lactate concentration, influencing the synthesis of P(3HB)/PLA allowed the production of a panel of polymers in a resting cell system with tailored lactic acid (LA) content according to the GC-MS of the biomass. Further biomass extraction suggested the presence of methanol soluble low molecular weight molecules containing LA, while 1 % LA could be detected in the purified polymer fraction. These results suggested that the cells are producing a blend of polymers. A proteomic analysis of C. necator resting cells under P(3HB)/PLA production conditions provides new insights into the latent pathways involved in this process. This study is a proof of concept demonstrating that LA can polymerize in a non-modified organism and paves the way for new metabolic engineering approaches for lactic acid polymer production in the model bacterium C. necator H16.


Increased phospho-AKT (Ser(473)) expression in bronchial dysplasia: implications for lung cancer prevention studies.
AKT, a downstream mediator of phosphatidylinositol 3'-kinase, is activated in non-small cell lung cancer (NSCLC), but we have not yet defined the stage of malignant transformation at which AKT is activated in the bronchial epithelium. We performed immunohistochemical analysis of activated AKT [phosphorylated (p)-AKT Ser(473)] in tissue specimens of normal bronchial epithelium, bronchial hyperplasia and squamous metaplasia ("reactive" epithelium), bronchial dysplasia, and NSCLC. Among NSCLC specimens, immunohistochemical findings were correlated with patient demographics, tumor stage, histology, and survival. We observed p-AKT expression in 12 of 44 (27.3%) normal bronchial biopsy specimens, 4 of 9 (44.4%) reactive epithelium specimens, 22 of 25 (88%) dysplastic specimens, and 25 of 76 (33%) NSCLC specimens. Among patients with resected early-stage or locally advanced NSCLC, p-AKT expression had no effect on tumor stage, histology, or survival. Of the histological groups examined, bronchial dysplasia specimens expressed p-AKT most frequently, supporting AKT activation as an early event in lung cancer progression. Given its role as a mediator of malignant transformation, p-AKT should be investigated as a potential target in future lung cancer prevention studies.


An Evaluation of a Train-the-Trainer Workshop for Social Service Workers to Develop Community-Based Family Interventions.
Evaluation studies on train-the-trainer workshops (TTTs) to develop family well-being interventions are limited in the literature. The Logic Model offers a framework to place some important concepts and tools of intervention science in the hands of frontline service providers. This paper reports on the evaluation of a TTT for a large community-based program to enhance family well-being in Hong Kong. The 2-day TTT introduced positive psychology themes (relevant to the programs that the trainees would deliver) and the Logic Model (which provides a framework to guide intervention development and evaluation) for social service workers to guide their community-based family interventions. The effectiveness of the TTT was examined by self-administered questionnaires that assessed trainees' changes in learning (perceived knowledge, self-efficacy, attitude, and intention), trainees' reactions to training content, knowledge sharing, and benefits to their service organizations before and after the training and then 6 months and 1 year later. Missing data were replaced by baseline values in an intention-to-treat analysis. Focus group interviews were conducted approximately 6 months after training. Fifty-six trainees (79% women) joined the TTT. Forty-four and 31 trainees completed the 6-month and 1-year questionnaires, respectively. The trainees indicated that the workshop was informative and well organized. The TTT-enhanced trainees' perceived knowledge, self-efficacy, and attitudes toward the application of the Logic Model and positive psychology constructs in program design. These changes were present with small to large effect size that persisted to the 1 year follow-up. The skills learned were used to develop 31 family interventions that were delivered to about 1,000 families. Qualitative feedback supported the quantitative results. This TTT offers a practical example of academic-community partnerships that promote capacity among community social service workers. Goals included sharing basic tools of intervention development and evaluation, and the TTT offered, therefore, the potential of learning skills that extended beyond the lifetime of a single program. The research protocol was registered at the National Institutes of Health (identifier number: NCT01796275).


Mutation screening of the 3q29 microdeletion syndrome candidate genes DLG1 and PAK2 in schizophrenia.
Deletion of chromosome 3q29, which is associated with mental retardation and autism, was recently identified as being present in excess or occurring de novo in schizophrenia cases, being present in approximately 1/1,000 cases and 1/40,000 unscreened controls. Of the ∼20 genes in the commonly deleted region two are prominent candidates for involvement in the behavioral features of the microdeletion syndrome: DLG1 and PAK2. We report the result of mutation screening of the entire protein coding sequence of both genes in a sample of 234 unrelated cases and 272 unrelated controls from the UK. We find no evidence for any amino acid changing genetic variants in PAK2. We observe several rare and singleton non-synonymous genetic variations at DLG1, however there is no excess of these variants in cases when compared to controls. Our sample was underpowered to detect very rare or low-penetrance disease relevant alleles in the studied genes. Therefore very rare, low-to-moderate penetrance protein coding mutations or non-coding mutations at DLG1 and/or PAK2, or a nearby gene, may reproduce the behavioral characteristics of the 3q29 microdeletion.


Autoimmunity-mediated antitumor immunity: tumor as an immunoprivileged self.
The association of autoimmunity with antitumor immunity challenges a paradigm of selective surveillance against tumors. Aided with well-characterized models of robust autoimmunity, we show that self-antigen-specific effector T (Teff) cell clones could eradicate tumor cells. However, a tumor microenvironment reinforced by Treg cells and myeloid-derived suppressor cells (MDSCs) presented a barrier to the autoimmune effectors, more so in tumors than in healthy tissues. This barrier required optimal CTLA4 expression in Teff cells. In a spontaneous model of breast cancer, subtle reductions in CTLA4 expression impeded tumor onset and progression, providing the first direct evidence that CTLA4 inhibits spontaneous tumor development. In an adoptive therapy model of lymphoma, self-antigen-specific Teff cells were potentiated by even a modest reduction of CTLA4. A subtle reduction of CTLA4 did not curtail Treg-cell suppression. Thus, Teff cells had an exquisite sensitivity to physiological levels of CTLA4 variations. However, both Treg and Teff cells were impacted by anti-CTLA4 antibody blockade. Therefore, whether CTLA4 impacts through Treg cells or Teff cells depends on its expression level. Overall, the results suggest that the tumor microenvironment represents an "immunoprivileged self" that could be overcome practically and at least partially by RNAi silencing of CTLA4 in Teff cells.


Evaluation of a high throughput toxicity biosensor and comparison with a Daphnia magna bioassay.
A high throughput toxicity biosensor has been designed and constructed using recombinant Escherichia coli cells, containing stress specific promoters (recA, fabA, or katG) or constitutive promoters (lac) fused to luciferase genes originating from Vibrio fisheri. These genetically engineered cells were immobilized in 96 well plates. By optimizing cell immobilization conditions and the strains' response specificity to toxic chemicals, bioluminescent outputs decreased or increased dose-dependently upon adding test chemicals. However, to date the toxicity data obtained using this biosensor have not been compared with the results of other toxicity tests. Phenolics were chosen to evaluate the correlation between the LD50 and the EC50 (GC2) or EC120 (DPD2540) of Daphnia magna and E. coli, respectively. Toxicity data obtained from constitutive strains by bioluminescent level decrements were compared with the results from D. magna as a standard. LD50 values were used as parameters of D. magna toxicity and EC50 of EC120 values were used for the immobilized biosensor. In the DPD2540 test, phenolics, membrane damaging toxic chemicals, for testing immobilized stress specific bacterial strains trigger dose-dependant bioluminescence increase within specific concentration. Although the stress specific responsiveness from the strains could not be compared with D. magna's LD50 values, these responses offer additional information, such as upon the mode of toxic action in the sample, in addition to the cellular toxicity results as indicated by the EC50. This novel high throughput toxicity biosensor can be implemented to investigate the toxicity of any other soluble materials, and can be used as a standardization tool for the evaluation of toxicity.


The interaction of projectiles with tissues and the management of ballistic fractures.
Wounds to the limbs are the commonest injuries seen during armed conflict and injury results from the transfer of energy from the missile to the tissues. There are a number of factors that determine the transfer of energy, and thus the extent of wounding. These include the velocity of the missile, its shape and stability, and the tissue through which the missile passes. Many of the wounds involve bone, and because of the interaction of missiles with bone, significant fractures can occur. In many previous conflicts amputation was considered the treatment of choice for many limb injuries, but with recent advances in the management of severe open fractures, many of these limbs are now salvageable. Whilst the basic principles of the initial débridement remain unchanged, techniques of fracture stabilisation and definitive soft tissue cover have changed, and it is necessary to consider these in relation to military fractures. Definitive soft tissue closure can be safely delayed until evacuation further down the medical chain, but stabilisation of the fracture must be considered at the time of initial surgery. Many of the advances in fracture management may be unsuitable for use in a military environment due to logistical constraints. In addition it is likely that wound infection will be more common with military injuries, and this will influence the treatment. This paper considers the interaction of missiles with soft tissue and bone, and discusses possible methods of fracture stabilisation in the military environment.


How a haemosporidian parasite of bats gets around: the genetic structure of a parasite, vector and host compared.
Parasite population structure is often thought to be largely shaped by that of its host. In the case of a parasite with a complex life cycle, two host species, each with their own patterns of demography and migration, spread the parasite. However, the population structure of the parasite is predicted to resemble only that of the most vagile host species. In this study, we tested this prediction in the context of a vector-transmitted parasite. We sampled the haemosporidian parasite Polychromophilus melanipherus across its European range, together with its bat fly vector Nycteribia schmidlii and its host, the bent-winged bat Miniopterus schreibersii. Based on microsatellite analyses, the wingless vector, and not the bat host, was identified as the least structured population and should therefore be considered the most vagile host. Genetic distance matrices were compared for all three species based on a mitochondrial DNA fragment. Both host and vector populations followed an isolation-by-distance pattern across the Mediterranean, but not the parasite. Mantel tests found no correlation between the parasite and either the host or vector populations. We therefore found no support for our hypothesis; the parasite population structure matched neither vector nor host. Instead, we propose a model where the parasite's gene flow is represented by the added effects of host and vector dispersal patterns.


Photothermal therapeutic response of cancer cells to aptamer-gold nanoparticle-hybridized graphene oxide under NIR illumination.
The objective of this study was to synthesize a nanocomposite, aptamer-gold nanoparticle-hybridized graphene oxide (Apt-AuNP-GO), to facilitate targeted treatment of tumor cells by near-infrared (NIR) light-activatable photothermal therapy. We also investigated whether Apt-AuNP-GO with NIR illumination modulates heat shock proteins (HSPs) expression leading to therapeutic response in human breast cancer cells. These findings can provide strategies for improving the photothermal therapy efficacy of cancer. The self-assembled Apt-AuNP-GO nanocomposite could selectively target MUC1-positive human breast cancer cells (MCF-7) due to the specific interaction between the MUC1-binding-aptamer and the MUC1 (type I transmembrane mucin glycoprotein) on cell membrane. In addition, Apt-AuNP-GO has a high light-to-heat conversion capability for photoabsorption of NIR light, and it is able to exert therapeutic effects on MCF-7 cells at an ultralow concentration without inducing adverse effects in healthy cells. The Apt-AuNP-GO nanocomposites combine the advantages of GOs, AuNPs, and Apts, possess specific targeting capability, excellent biocompatibility, and tumor cell destruction ability, suggesting great potential for application in the photothermal therapy of breast cancer. Under NIR illumination, Apt-AuNP-GO induced transient increase in HSP70 expression, which decreased thereafter. This phenomenon may cause irreversible damage to Apt-AuNP-GO-treated MCF-7 cell under NIR illumination. We also demonstrated that the combination therapy of heat and HSP70 inhibitor could synergistically generate marked tumoricidal effects against breast cancer. These results suggest that the degree and duration of HSP70 protein expression are correlated with therapeutic effects against breast cancer for Apt-AuNP-GO-assisted photothermal therapy. We believe that such a nanocomposite can be readily extended to the construction of HSP70 inhibitors-loaded Apt-AuNP-GO, which could deliver both heat and HSP70 inhibitors to tumorigenic regions for the chemo-photothermal therapy.


Structure-directing and high-efficiency photocatalytic hydrogen production by Ag clusters.
H2 production by water splitting is hindered mainly by the lack of low-cost and efficient photocatalysts. Here we show that sub-nanometric silver clusters can catalyze the anisotropic growth of gold nanostructures by preferential adsorption at certain crystal planes of Au seeds, with the result that the final nanostructure can be tuned via the cluster/seed ratio. Such semiconducting Ag clusters are extremely stable and retain their electronic structure even after adsorption at the tips of Au nanorods, enabling various photocatalytic experiments, such as oxygen evolution from basic solutions. In the absence of electron scavengers, UV irradiation generates photoelectrons, which are stored within the nanorods, increasing their Au Fermi level up to the redox pinning limit at 0 V (RHE), where hydrogen evolution occurs with an estimated high efficiency of 10%. This illustrates the considerable potential of very small zerovalent, nonmetallic clusters as novel atomic-level photocatalysts.


On the kinetics of anaerobic power.
This study investigated two different mathematical models for the kinetics of anaerobic power. Model 1 assumes that the work power is linear with the work rate, while Model 2 assumes a linear relationship between the alactic anaerobic power and the rate of change of the aerobic power. In order to test these models, a cross country skier ran with poles on a treadmill at different exercise intensities. The aerobic power, based on the measured oxygen uptake, was used as input to the models, whereas the simulated blood lactate concentration was compared with experimental results. Thereafter, the metabolic rate from phosphocreatine break down was calculated theoretically. Finally, the models were used to compare phosphocreatine break down during continuous and interval exercises. Good similarity was found between experimental and simulated blood lactate concentration during steady state exercise intensities. The measured blood lactate concentrations were lower than simulated for intensities above the lactate threshold, but higher than simulated during recovery after high intensity exercise when the simulated lactate concentration was averaged over the whole lactate space. This fit was improved when the simulated lactate concentration was separated into two compartments; muscles + internal organs and blood. Model 2 gave a better behavior of alactic energy than Model 1 when compared against invasive measurements presented in the literature. During continuous exercise, Model 2 showed that the alactic energy storage decreased with time, whereas Model 1 showed a minimum value when steady state aerobic conditions were achieved. During interval exercise the two models showed similar patterns of alactic energy. The current study provides useful insight on the kinetics of anaerobic power. Overall, our data indicate that blood lactate levels can be accurately modeled during steady state, and suggests a linear relationship between the alactic anaerobic power and the rate of change of the aerobic power.


[Nitric oxide donors activated by two-photon excitation].
Nitric oxide (NO) is a key molecule in blood pressure regulation, neuromodulation, and biodefense. Since it is an unstable gas under ambient conditions, various NO donors have been developed and employed for biological studies as an alternative to direct NO application. However, many of them release NO via spontaneous decomposition, so that it is difficult to control the NO release. Therefore, NO donors from which NO release can be temporally and spatially well-controlled by means of photoexcitation offer considerable advantages. Various NO donors from which NO release is activated by photoirradiation have been reported. However, the maximum absorption of these NO donors is generally limited to the UV or short-wavelength visible light range, which does not adequately penetrate living tissues. To overcome this limitation, NO donors working via two-photon excitation (TPE) were developed by Ford and Prasad, based on NO release from Fe-nitrosyl complex. We have also developed TPE-type NO donors based on our photo-controllable 2,6-dimethylnitrobenzene derivatives, and investigated their NO-releasing properties. Here, we present an overview of photocontrollable NO donors and discuss their properties in relation to biological applications.


Structural variability and complexity of the giant Pithovirus sibericum particle revealed by high-voltage electron cryo-tomography and energy-filtered electron cryo-microscopy.
The Pithoviridae giant virus family exhibits the largest viral particle known so far, a prolate spheroid up to 2.5 μm in length and 0.9 μm in diameter. These particles show significant variations in size. Little is known about the structure of the intact virion due to technical limitations with conventional electron cryo-microscopy (cryo-EM) when imaging thick specimens. Here we present the intact structure of the giant Pithovirus sibericum particle at near native conditions using high-voltage electron cryo-tomography (cryo-ET) and energy-filtered cryo-EM. We detected a previously undescribed low-density outer layer covering the tegument and a periodical structuring of the fibres in the striated apical cork. Energy-filtered Zernike phase-contrast cryo-EM images show distinct substructures inside the particles, implicating an internal compartmentalisation. The density of the interior volume of Pithovirus particles is three quarters lower than that of the Mimivirus. However, it is remarkably high given that the 600 kbp Pithovirus genome is only half the size of the Mimivirus genome and is packaged in a volume up to 100 times larger. These observations suggest that the interior is densely packed with macromolecules in addition to the genomic nucleic acid.


The effect of adropin on lipid and glucose metabolism in rats with hyperlipidemia.
The aim of this study was to evaluate, for the first time, whether the effects of low-dose adropin administration is effective in rats with hyperlipidemia. Twenty one Wistar albino female rats were randomly divided into 3 groups and fed with high-fat diet for 4 weeks to establish the hyperlipidemia model. Meanwhile, adropin was administrated intraperitonealy (2.1 μg/kg/day), once a day for continuous 10 days. Then, body weights and serum biochemical parameters, adropin, insulin and blood glucose levels were determined. Additionally, in liver tissue, inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) mRNA gene expressions were evaluated by RT-PCR. The results showed that intraperitoneal administration of adropin to hyperlipidemic rats for 10 days were extremely effective in decreasing the levels of serum triglycerides (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine aminotransferase (ALT), and gamma glutamil transferase (GGT) and increasing the levels of high density lipoprotein cholesterol (HDL-C). It could decrease mRNA expressions of pro-inflammatory cytokines TNF-α and IL-6 via regulating the expressions of iNOS. In addition, treatment with adropin showed a significant reduction in blood glucose, serum insulin levels, HbA1c (%), and HOMA-IR, and increase in serum adropin levels. Adropin may ameliorate lipid metabolism, reduce insulin resistance, and inhibit hepatocytes inflammation. Thus, adropin had significant therapeutic benefits and could be suggested as a potential candidate agent against hyperlipidemia.


An improved in vivo deuterium labeling method for measuring the biosynthetic rate of cytokinins.
An improved method for determining the relative biosynthetic rate of isoprenoid cytokinins has been developed. A set of 11 relevant isoprenoid cytokinins, including zeatin isomers, was separated by ultra performance liquid chromatography in less than 6 min. The iP-type cytokinins were observed to give rise to a previously-unknown fragment at m/z 69; we suggest that the diagnostic (204-69) transition can be used to monitor the biosynthetic rate of isopentenyladenine. Furthermore, we found that by treating the cytokinin nucleotides with alkaline phosphatase prior to analysis, the sensitivity of the detection process could be increased. In addition, derivatization (propionylation) improved the ESI-MS response by increasing the analytes' hydrophobicity. Indeed, the ESI-MS response of propionylated isopentenyladenosine was about 34% higher than that of its underivatized counterpart. Moreover, the response of the derivatized zeatin ribosides was about 75% higher than that of underivatized zeatin ribosides. Finally, we created a web-based calculator (IZOTOP) that facilitates MS/MS data processing and offer it freely to the research community.


Influence of regional cerebral blood volume on voxel-based morphometry.
The investigation of structural brain alterations is one focus in research of brain diseases like depression. Voxel-based morphometry (VBM) based on high-resolution 3D MRI images is a widely used non-invasive tool for such investigations. However, the result of VBM might be sensitive to local physiological parameters such as regional cerebral blood volume (rCBV) changes. In order to investigate whether rCBV changes may contribute to variation in VBM, we performed analyses in a study with the congenital learned helplessness (cLH) model for long-term findings. The 3D structural and rCBV data were acquired with T2 -weighted rapid acquisition with relaxation enhancement (RARE) pulse sequences. The group effects were determined by standard statistical parametric mapping (SPM) and biological parametric mapping (BPM) and examined further using atlas-based regions. In our genetic animal model of depression, we found co-occurrence of differences in gray matter volume and rCBV, while there was no evidence of significant interaction between both. However, the multimodal analysis showed similar gray matter differences compared with the standard VBM approach. Our data corroborate the idea that two group VBM differences might not be influenced by rCBV differences in genetically different strains. Copyright © 2016 John Wiley & Sons, Ltd.