Transition from hemifusion to pore opening is rate limiting for vacuole membrane fusion.

Fusion pore opening and expansion are considered the most energy-demanding steps in viral fusion. Whether this also applies to soluble N-ethyl-maleimide sensitive fusion protein attachment protein receptor (SNARE)- and Rab-dependent fusion events has been unknown. We have addressed the problem by characterizing the effects of lysophosphatidylcholine (LPC) and other late-stage inhibitors on lipid mixing and pore opening during vacuole fusion. LPC inhibits fusion by inducing positive curvature in the bilayer and changing its biophysical properties. The LPC block reversibly prevented formation of the hemifusion intermediate that allows lipid, but not content, mixing. Transition from hemifusion to pore opening was sensitive to guanosine-5'-(gamma-thio)triphosphate. It required the vacuolar adenosine triphosphatase V0 sector and coincided with its transformation. Pore opening was rate limiting for the reaction. As with viral fusion, opening the fusion pore may be the most energy-demanding step for intracellular, SNARE-dependent fusion reactions, suggesting that fundamental aspects of lipid mixing and pore opening are related for both systems.

Prospective randomized study of two cryopreservation policies avoiding embryo selection: the pronucleate stage leads to a higher cumulative delivery rate than the early cleavage stage.

OBJECTIVE: To compare the cumulative live birth rates obtained after cryopreservation of either pronucleate (PN) zygotes or early-cleavage (EC) embryos. DESIGN: Prospective randomized study. SETTING: University hospital. PATIENT(S): Three hundred eighty-two patients, involved in an IVF/ICSI program from January 1993 to December 1995, who had their supernumerary embryos cryopreserved either at the PN (group I) or EC (group II) stage. For 89 patients, cryopreservation of EC embryos was canceled because of poor embryo development (group III). Frozen-thawed embryo transfers performed up to December 1998 were considered. MAIN OUTCOME MEASURE(S): Age, oocytes, zygotes, cryopreserved and transferred embryos, damage after thawing, cumulative embryo scores, implantation, and cumulative live birth rates. RESULT(S): The clinical pregnancy and live birth rates were similar in all groups after fresh embryo transfers. Significantly higher implantation (10.5% vs. 5.9%) and pregnancy rates (19.5% vs. 10.9%; P< or = .02 per transfer after cryopreserved embryo transfers were obtained in group I versus group II, leading to higher cumulative pregnancy (55.5% vs. 38.6%; P < or = .002 and live birth rates (46.9% vs. 27.7%; P< or = .0001.Conclusion(s): The transfer of a maximum of three unselected embryos and freezing of all supernumerary PN zygotes can be safely done with significantly higher cumulative pregnancy chances than cryopreserving at a later EC stage.

Pneumocystis jirovecii genotype associated with increased death rate of HIV-infected patients with pneumonia.

Pneumocystis jirovecii dihydropteroate synthase (DHPS) mutations have been associated with failure of sulfa prophylaxis; their effect on the outcome of patients with P. jirovecii pneumonia (PCP) remains controversial. P. jirovecii DHPS polymorphisms and genotypes were identified in 112 cases of PCP in 110 HIV-infected patients by using PCR single-strand conformation polymorphism. Of the 110 patients observed, 21 died; 18 of those deaths were attributed to PCP. Thirty-three percent of the PCP cases involved a P. jirovecii strain that had 1 or both DHPS mutations. The presence or absence of DHPS mutations had no effect on the PCP mortality rate within 1 month, whereas P.jirovecii type 7 and mechanical ventilation at PCP diagnosis were associated with an increased risk of death caused by PCP. Mechanical ventilation at PCP diagnosis was also associated with an increased risk of sulfa treatment failure at 5 days.

Effects of leaf compounds, climate and natural enemies on the incidence of thrips in cassava

The objective of this study was to determine the effects of rainfall, temperature, sunlight and relative humidity, as well as predators and parasitoids, leaf chemical composition and levels of leaf nitrogen and potassium on the intensity of Scirtothrips manihoti (Thysanoptera: Thripidae) attack on cassava Manihot esculenta Crantz var. Cacau. The leaf compounds (E)-farnesene/trans-farnesol and D-friedoolean-14-en-3-one correlated significantly with the population of S. manihoti. Insect population decreased in the dry and cold season probably due to leaf senescence. Significative correlation was observed between Syrphidae with S. manihoti populations.

Diagnostic delay in Crohn's disease is associated with a complicated disease course and increased operation rate.

OBJECTIVES: The impact of diagnostic delay (a period from appearance of first symptoms to diagnosis) on the clinical course of Crohn's disease (CD) is unknown. We examined whether length of diagnostic delay affects disease outcomes. METHODS: Data from the Swiss IBD cohort study were analyzed. Patients were recruited from university centers (68%), regional hospitals (14%), and private practices (18%). The frequencies of occurrence of bowel stenoses, internal fistulas, perianal fistulas, and CD-related surgery (intestinal and perianal) were analyzed. RESULTS: A total of 905 CD patients (53.4% female, median age at diagnosis 26 (20-36) years) were stratified into four groups according to the quartiles of diagnostic delay (0-3, 4-9, 10-24, and ≥25 months, respectively). Median diagnostic delay was 9 (3-24) months. The frequency of immunomodulator and/or antitumor necrosis factor drug use did not differ among the four groups. The length of diagnostic delay was positively correlated with the occurrence of bowel stenosis (odds ratio (OR) 1.76, P=0.011 for delay of ≥25 months) and intestinal surgery (OR 1.76, P=0.014 for delay of 10-24 months and OR 2.03, P=0.003 for delay of ≥25 months). Disease duration was positively associated and non-ileal disease location was negatively associated with bowel stenosis (OR 1.07, P<0.001, and OR 0.41, P=0.005, respectively) and intestinal surgery (OR 1.14, P<0.001, and OR 0.23, P<0.001, respectively). CONCLUSIONS: The length of diagnostic delay is correlated with an increased risk of bowel stenosis and CD-related intestinal surgery. Efforts should be undertaken to shorten the diagnostic delay.

Exploring the rate-limiting steps in visual phototransduction recovery by bottom-up kinetic modeling

Phototransduction in vertebrate photoreceptor cells represents a paradigm of signaling pathways mediated by G-protein-coupled receptors (GPCRs), which share common modules linking the initiation of the cascade to the final response of the cell. In this work, we focused on the recovery phase of the visual photoresponse, which is comprised of several interacting mechanisms. We employed current biochemical knowledge to investigate the response mechanisms of a comprehensive model of the visual phototransduction pathway. In particular, we have improved the model by implementing a more detailed representation of the recoverin (Rec)-mediated calcium feedback on rhodopsin kinase and including a dynamic arrestin (Arr) oligomerization mechanism. The model was successfully employed to investigate the rate limiting steps in the recovery of the rod photoreceptor cell after illumination. Simulation of experimental conditions in which the expression levels of rhodospin kinase (RK), of the regulator of the G-protein signaling (RGS), of Arr and of Rec were altered individually or in combination revealed severe kinetic constraints to the dynamics of the overall network. Our simulations confirm that RGS-mediated effector shutdown is the rate-limiting step in the recovery of the photoreceptor and show that the dynamic formation and dissociation of Arr homodimers and homotetramers at different light intensities significantly affect the timing of rhodopsin shutdown. The transition of Arr from its oligomeric storage forms to its monomeric form serves to temper its availability in the functional state. Our results may explain the puzzling evidence that overexpressing RK does not influence the saturation time of rod cells at bright light stimuli. The approach presented here could be extended to the study of other GPCR signaling pathways.

Coping with continuous human disturbance in the wild: insights from penguin heart rate response to various stressors.

ABSTRACT: BACKGROUND: A central question for ecologists is the extent to which anthropogenic disturbances (e.g. tourism) might impact wildlife and affect the systems under study. From a research perspective, identifying the effects of human disturbance caused by research-related activities is crucial in order to understand and account for potential biases and derive appropriate conclusions from the data. RESULTS: Here, we document a case of biological adjustment to chronic human disturbance in a colonial seabird, the king penguin (Aptenodytes patagonicus), breeding on remote and protected islands of the Southern ocean. Using heart rate (HR) as a measure of the stress response, we show that, in a colony with areas exposed to the continuous presence of humans (including scientists) for over 50 years, penguins have adjusted to human disturbance and habituated to certain, but not all, types of stressors. When compared to birds breeding in relatively undisturbed areas, birds in areas of high chronic human disturbance were found to exhibit attenuated HR responses to acute anthropogenic stressors of low-intensity (i.e. sounds or human approaches) to which they had been subjected intensely over the years. However, such attenuation was not apparent for high-intensity stressors (i.e. captures for scientific research) which only a few individuals experience each year. CONCLUSIONS: Habituation to anthropogenic sounds/approaches could be an adaptation to deal with chronic innocuous stressors, and beneficial from a research perspective. Alternately, whether penguins have actually habituated to anthropogenic disturbances over time or whether human presence has driven the directional selection of human-tolerant phenotypes, remains an open question with profound ecological and conservation implications, and emphasizes the need for more knowledge on the effects of human disturbance on long-term studied populations.

Diallel analysis for frogeye leaf spot resistance in soybean

Seven soybean cultivars (Bossier, Cristalina, Davis, Kent, Lincoln, Paraná and Uberaba), with different levels of resistance to Cercospora sojina, were crossed in a diallel design to determine the general (GCA) and specific (SCA) combining abilities relative to the inheritance of the resistance. Race 04 of the fungus was inoculated in the parents and in the 21 F1 hybrids in a greenhouse in a completely randomized design, with 12 replications. The reactions to the disease were evaluated 20 days after the inoculation, always on the most infected leaflet. Both GCA and SCA were significant for all the evaluated characters, being inferred that, for the expression of the characters, the additive, dominant and, possibly, epistatic genic actions were important. The largest values of estimated SCA effect (ij) were observed in the hybrid combinations where at least one parent presented high GCA. Cristalina, Davis and Uberaba cultivars showed the largest estimates for GCA effect (i), and from the analysis of ii, the contribution of these parents to heterosis of their hybrids will be towards the reduction of the disease symptoms. Therefore, these cultivars are indicated as parents in breeding programs that seek the development of soybean cultivars with resistance to frogeye leaf spot.

Exchange rate predictability

The main goal of this article is to provide an answer to the question: "Does anything forecast exchange rates, and if so, which variables?". It is well known thatexchange rate fluctuations are very difficult to predict using economic models, andthat a random walk forecasts exchange rates better than any economic model (theMeese and Rogoff puzzle). However, the recent literature has identified a series of fundamentals/methodologies that claim to have resolved the puzzle. This article providesa critical review of the recent literature on exchange rate forecasting and illustratesthe new methodologies and fundamentals that have been recently proposed in an up-to-date, thorough empirical analysis. Overall, our analysis of the literature and thedata suggests that the answer to the question: "Are exchange rates predictable?" is,"It depends" -on the choice of predictor, forecast horizon, sample period, model, andforecast evaluation method. Predictability is most apparent when one or more of thefollowing hold: the predictors are Taylor rule or net foreign assets, the model is linear, and a small number of parameters are estimated. The toughest benchmark is therandom walk without drift.

Long diagnostic delay in Crohn's disease is associated with complicated disease course and increased operation rate

Background and Aims: The impact of d iagnostic delay ( a period from appearance of f irst s ymptoms t o diagnosis) o n the clinical c ourse o f Crohn's disease (CD) i s unknown. W e examined whether length of d iagnostic delay a ffects d isease outcome. Methods: Data from the Swiss IBD cohort study were analyzed. T he frequencies of o ccurrence of b owel s tenoses, internal fistulas, perianal f istulas, and CD-related surgery at distinct i ntervals a fter C D diagnosis (0 - < 2 , 2 - < 6,  6 years) were c ompared f or g roups o f patients w ith different length of d iagnostic delay. Results: T he data from a g roup o f 200 CD patients with long diagnostic delay (> 24 months, 76th - 100th p ercentile) were c ompared to t hose from a group of 4 61 patients with a short diagnostic delay ( within 9 months, 1st - 50th p ercentile). T reatment r egimens d id n ot d iffer between t he two groups. Two years following diagnosis, p atients with long diagnostic delay presented more frequently with bowel stenoses (25% vs. 13.1%, p = 0.044), internal fistulas (10% vs. 2%, p = 0.018), perianal f istulas ( 20% vs. 8 .1%, p = 0.023) a nd more frequently underwent intestinal surgery (15% vs. 5 .1%, p = 0.024) t han patients with short diagnostic delay. Intestinal surgery was a lso m ore frequently p erformed  6 y ears after diagnosis in t he group with long d iagnostic delay ( 56.2% vs. 42.3%, p = 0.005) w hen compared to t he g roup with short diagnostic delay. Conclusions: L ong diagnostic delay i s associated with worse o utcome c haracterized by t he development o f increased bowel damage, n ecessitating more frequently operations in t he years following CD d iagnosis. Efforts should be undertaken to shorten the diagnostic delay.

Infrared thermometry to schedule irrigation of common bean

The objective of this work was to determine the critical irrigation time for common bean (Phaseolus vulgaris L. cv. Carioca) using infrared thermometry. Five treatments were analyzed. Canopy temperature differences between plants and a well-watered control about 1, 2, 3, 4, and 5±0.5ºC were tested. Physiological variables and plant growth were analyzed to establish the best time to irrigate. There was a significant linear correlation between the index and stomatal resistance, transpiration rate, and leaf water potential. Although significant linear correlation between the index and mean values of total dry matter, absolute growth rate, and leaf area index was found, no correlation was found with other growth index like relative growth rate, net assimilation rate, and leaf area ratio. Plants irrigated when their canopy temperature was 3±0.5ºC above the control had their relative growth rate mean value increased up to 59.7%, yielding 2,260.2 kg ha-1, with a reduction of 38.0% in the amount of water used. Plants irrigated when their canopy temperature was 4±0.5ºC yielded 1,907.6 kg ha-1, although their relative growth rate mean value was 4.0% below the control. These results show that the best moment to irrigate common bean is when their canopy temperature is between 3ºC and 4±0.5ºC above the control.

Disturbed local auxin homeostasis enhances cellular anisotropy and reveals alternative wiring of auxin-ethylene crosstalk in Brachypodium distachyon seminal roots.

Observations gained from model organisms are essential, yet it remains unclear to which degree they are applicable to distant relatives. For example, in the dicotyledon Arabidopsis thaliana (Arabidopsis), auxin biosynthesis via indole-3-pyruvic acid (IPA) is essential for root development and requires redundant TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (TAA1) and TAA1-RELATED (TAR) genes. A promoter T-DNA insertion in the monocotyledon Brachypodium distachyon (Brachypodium) TAR2-LIKE gene (BdTAR2L) severely down-regulates expression, suggesting reduced tryptophan aminotransferase activity in this mutant, which thus represents a hypomorphic Bdtar2l allele (Bdtar2l(hypo) ). Counterintuitive however, Bdtar2l(hypo) mutants display dramatically elongated seminal roots because of enhanced cell elongation. This phenotype is also observed in another, stronger Bdtar2l allele and can be mimicked by treating wild type with L-kynerunine, a specific TAA1/TAR inhibitor. Surprisingly, L-kynerunine-treated as well as Bdtar2l roots display elevated rather than reduced auxin levels. This does not appear to result from compensation by alternative auxin biosynthesis pathways. Rather, expression of YUCCA genes, which are rate-limiting for conversion of IPA to auxin, is increased in Bdtar2l mutants. Consistent with suppression of Bdtar2l(hypo) root phenotypes upon application of the ethylene precursor 1-aminocyclopropane-1-carboxylic-acid (ACC), BdYUCCA genes are down-regulated upon ACC treatment. Moreover, they are up-regulated in a downstream ethylene-signaling component homolog mutant, Bd ethylene insensitive 2-like 1, which also displays a Bdtar2l root phenotype. In summary, Bdtar2l phenotypes contrast with gradually reduced root growth and auxin levels described for Arabidopsis taa1/tar mutants. This could be explained if in Brachypodium, ethylene inhibits the rate-limiting step of auxin biosynthesis in an IPA-dependent manner to confer auxin levels that are sub-optimal for root cell elongation, as suggested by our observations. Thus, our results reveal a delicate homeostasis of local auxin and ethylene activity to control cell elongation in Brachypodium roots and suggest alternative wiring of auxin-ethylene crosstalk as compared to Arabidopsis.

Understanding the gains from wage flexibility: The exchange rate connection

We study the gains from increased wage flexibility and their dependence on exchange rate policy, using a small open economy model with staggered price andwage setting. Two results stand out: (i) the impact of wage adjustments on employment is smaller the more the central bank seeks to stabilize the exchange rate,and (ii) an increase in wage flexibility often reduces welfare, and more likely so ineconomies under an exchange rate peg or an exchange rate-focused monetary policy.Our findings call into question the common view that wage flexibility is particularlydesirable in a currency union.

Effect of neem extract on the cotton aphid

The aphid Aphis gossypii Glover (Hemiptera: Aphididae), a harmful pest on cotton, causes direct damage, reducing plant vigor, and indirect damage by honeydew secretion and transmission of several viruses. Due to the problem of pesticide resistance, alternative techniques for chemical control, such as the use of natural insecticides, have been object of research. The effects of aqueous extracts of neem (Azadirachta indica A. Juss) seed powder on the development, survival and fecundity of A. gossypii were evaluated. Treatments consisted of neem seed powder in the concentrations of 23.8, 122.0, 410.0 and 1,410.0 mg/100 mL of distilled water. Mortality rate during the nymphal development for aphids maintained on cotton leaf discs treated with the two highest concentrations were, respectively, 60.0% and 100.0%. With the exception of the highest concentration (1,410.0 mg/100 mL), neem concentrations did not extend the aphids' development period. The net reproductive rate (R0) was of 35.0 nymphs/female for control aphids and of 0.0 nymph/female when the group of females was exposed to neem seed powder at 1,410.0 mg/100 mL since birth. The aqueous extract of neem seeds is efficient against the aphid A. gossypii, causing nymph mortality and reducing their survival period and fecundity.

Is it appropriate to index glomerular filtration rate for body surface area

Purpose: Obesity is an established independent risk factor for chronic kidney disease. Thus, measurement of glomerular filtration rate (GFR) is important in this population. Traditionally, GFR has been indexed for body surface area (BSA), but this indexation may not be appropriate in obese individuals. Therefore, the objective of the study was to compare absolute GFR with GFR indexed for BSA and with GFR indexed for height. Methods and materials: The study was conducted in 66 families from the Seychelles islands that included several members with hypertension. GFR and effective renal plasma flow (ERPF) were measured using inulin and PAH clearances, respectively. Antihypertensive treatment, if used, was withheld 2 weeks before conducting the clearances. Participants with diabetes mellitus were excluded from the analysis. BSA was calculated using the Dubois formula. We assessed trend across BMI categories using a non parametric test. Results: Participants included 174 women and 127 men. The prevalence of hypertension was 61%, of which 68% were treated. The table shows that absolute GFR, GFR indexed for height, ERPF, filtration fraction were significantly higher across BMI categories. When GFR was indexed for BSA, the association between GFR and BMI categories was lost. Conclusion: Indexing GFR for BSA in overweight and obese individuals leads to a substantial underestimation of GFR. Filtration fraction, which does not depend on BSA, is higher in obese individuals, which suggests glomerular hyperfiltration. Indexing GFR for BSA therefore would mask the underlying glomerular hyperfiltration. As the number of nephrons does not increase with weight gain, absolute GFR represents a better marker of single nephron GFR and is more appropriate.