Vacuolar ATPase depletion contributes to dysregulation of endocytosis in bloodstream forms of Trypanosoma brucei

BACKGROUND Vacuolar H-ATPase (V-ATPase) is a highly conserved protein complex which hydrolyzes ATP and pumps protons to acidify vacuolar vesicles. Beyond its role in pH maintenance, the involvement of V-ATPase in endocytosis is well documented in mammals and plants but is less clear in Trypanosoma brucei. METHODS In this study, the subcellular localization of V-ATPase subunit B (TbVAB) of T. brucei was assessed via in situ N-terminal YFP-tagging and immunofluorescence assays. Transgenic bloodstream forms (BSF) of T. brucei were generated which comprised either a V-ATPase subunit B (TbVAB) conditional knockout or a V-ATPase subunit A (TbVAA) knockdown. Acridine orange and BCECF-AM were employed to assess the roles of V-ATPase in the pH regulation of BSF T. brucei. The endocytic activities of three markers were also characterized by flow cytometry analyses. Furthermore, trypanosomes were counted from trypanolysis treatment groups (either containing 1% or 5% NHS) and endocytosed trypanosome lytic factor (TLF) was also analyzed by an immunoblotting assay. RESULTS TbVAB was found to localize to acidocalcisomes, lysosomes and probably also to endosomes of BSF of T. brucei and was demonstrated to be essential for cell growth. TbVAB depletion neutralized acidic organelles at 24 hours post-tetracycline depletion (hpd), meanwhile the steady state intracellular pH increased from 7.016 ± 0.013 to 7.422 ± 0.058. Trypanosomes with TbVAB depletion at 24 hpd were found to take up more transferrin (2.068 ± 0.277 fold) but less tomato lectin (49.31 ± 22.57%) by endocytosis, while no significant change was detected in dextran uptake. Similar endocytic dysregulated phenotypes were also observed in TbVAA knockdown cells. In addition, TbVAB depleted trypanosomes showed a low uptake of TLF and exhibited less sensitive to lysis in both 1% and 5% NHS treatments. CONCLUSIONS TbVAB is a key component of V-ATPase and was found to play a key function in endocytosis as well as exhibiting different effects in a receptor/cargo dependent manner in BSF of T. brucei. Besides vacuolar alkalinization, the dysregulation of endocytosis in TbVAB depleted T. brucei is considered to contribute to the reduced sensitivity to lysis by normal human serum

Theory and application of Fermi pseudo-potential in one dimension

The theory of interaction at one point is developed for the one-dimensional Schrodinger equation. In analog with the three-dimensional case, the resulting interaction is referred to as the Fermi pseudo-potential. The dominant feature of this one-dimensional problem comes from the fact that the real line becomes disconnected when one point is removed. The general interaction at one point is found to be the sum of three terms, the well-known delta-function potential and two Fermi pseudo-potentials, one odd under space reflection and the other even. The odd one gives the proper interpretation for the delta'(x) potential, while the even one is unexpected and more interesting. Among the many applications of these Fermi pseudo-potentials, the simplest one is described. It consists of a superposition of the delta-function potential and the even pseudo-potential applied to two-channel scattering. This simplest application leads to a model of the quantum memory, an essential component of any quantum computer.Comment: RevTeX4, 32 pages, no figure

Human African trypanosomiasis : the current situation in endemic regions and the risks for non-endemic regions from imported cases

Human African trypanosomiasis (HAT) is caused by Trypanosoma brucei gambiense and T. b. rhodesiense and caused devastating epidemics during the 20th century. Due to effective control programs implemented in the last two decades, the number of reported cases has fallen to a historically low level. Although fewer than 977 cases were reported in 2018 in endemic countries, HAT is still a public health problem in endemic regions until it is completely eliminated. In addition, almost 150 confirmed HAT cases were reported in non-endemic countries in the last three decades. The majority of non-endemic HAT cases were reported in Europe, United States and South Africa, due to historical alliances, economic links or geographic proximity to disease endemic countries. Furthermore, with the implementation of the “Belt and Road” project, sporadic imported HAT cases have been reported in China as a warning sign of tropical diseases prevention. In this paper, we explore and interpret the data on HAT incidence and find no positive correlation between the number of HAT cases from endemic and non-endemic countries.This data will provide useful information for better understanding the imported cases of HAT globally in the post-elimination phase

1+1+2 Electromagnetic perturbations on general LRS space-times: Regge-Wheeler and Bardeen-Press equations

We use the, covariant and gauge-invariant, 1+1+2 formalism developed by Clarkson and Barrett, and develop new techniques, to decouple electromagnetic (EM) perturbations on arbitrary locally rotationally symmetric (LRS) space-times. Ultimately, we derive 3 decoupled complex equations governing 3 complex scalars. One of these is a new Regge-Wheeler (RW) equation generalized for LRS space-times, whereas the remaining two are new generalizations of the Bardeen-Press (BP) equations. This is achieved by first using linear algebra techniques to rewrite the first-order Maxwell equations in a new complex 1+1+2 form which is conducive to decoupling. This new complex system immediately yields the generalized RW equation, and furthermore, we also derive a decoupled equation governing a newly defined complex EM 2-vector. Subsequently, a further decomposition of the 1+1+2 formalism into a 1+1+1+1 formalism is developed, allowing us to decompose the complex EM 2-vector, and its governing equations, into spin-weighted scalars, giving rise to the generalized BP equations

Effect of stocking density on growth and yield of GIFT tilapia under rice-fish production system

The study was conducted at the Central Experimental Station, Philippine Rice Research Institute, Maligaya, Science City of Munoz, Nueva Ecija, Philippines during the wet season to determine the suitable stocking density(s) for better growth and yield of fish under rice-fish production systems. Recovery rate of GIFT tilapia in different stocking densities ranged from 75.74 to 83.47%. Among different treatments, rice +5,000 fingerlings/ha and rice +10,000 fingerlings/ha resulted in the highest recovery rate of 83.33% and 83.47%, respectively. The lowest recovery rate of 75.75% was obtained from rice +20,000 fingerlings/ha, but similar to that was obtained (78.56%) from rice +15,000 fingerlings/ha. Significantly higher rate of gain in body weight and that of specific growth rate were recorded in the treatment from rice +5,000 fingerlings/ha, while other treatments resulted in similar absolute and specific growth rate. Fish yield increased significantly with relatively higher stocking densities, but higher densities produced maximum number of smaller fishes and also lower recovery rate

Significance testing of rank cross-correlations between autocorrelated time series with short-range dependence

Statistical dependency measures such as Kendall’s Tau or Spearman’s Rho are frequently used to analyse the coherence between time series in environmental data analyses. Autocorrelation of the data can, however, result in spurious cross correlations if not accounted for. Here, we present the asymptotic distribution of the estimators of Spearman’s Rho and Kendall’s Tau, which can be used for statistical hypothesis testing of cross-correlations between autocorrelated observations. The results are derived using U-statistics under the assumption of absolutely regular (or β-mixing) processes. These comprise many short-range dependent processes, such as ARMA-, GARCH- and some copula-based models relevant in the environmental sciences. We show that while the assumption of absolute regularity is required, the specific type of model does not have to be specified for the hypothesis test. Simulations show the improved performance of the modified hypothesis test for some common stochastic models and small to moderate sample sizes under autocorrelation. The methodology is applied to observed climatological time series of flood discharges and temperatures in Europe. While the standard test results in spurious correlations between floods and temperatures, this is not the case for the proposed test, which is more consistent with the literature on flood regime changes in Europe

1+1+2 Electromagnetic perturbations on non-vacuum LRS class II space-times: Decoupling scalar and 2-vector harmonic amplitudes

We use the covariant and gauge-invariant 1+1+2 formalism of Clarkson and Barrett \cite{Clarkson2003} to analyze electromagnetic (EM) perturbations on non-vacuum {\it locally rotationally symmetric} (LRS) class II space-times. Ultimately, we show how to derive six real decoupled equations governing the total of six EM scalar and 2-vector harmonic amplitudes. Four of these are new, and result from expanding the complex EM 2-vector which we defined in \cite{Burston2007} in terms of EM 2-vector harmonic amplitudes. We are then able to show that there are four precise combinations of the amplitudes that decouple, two of these are polar perturbations whereas the remaining two are axial. The remaining two decoupled equations are the generalized Regge-Wheeler equations which were developed previously in \cite{Betschart2004}, and these govern the two EM scalar harmonic amplitudes. However, our analysis generalizes this by including a full description and classification of energy-momentum sources, such as charges and currents.Comment: 9 page

Growth performance, recovery rate and fish yield of GIFT strain at various water depths under rice-fish culture systems

Effect of water depth on recovery rate, growth performance and fish yield of GIFT in the rice-fish production systems was studies in experimental plots of 123 m2 with a pond refuge of I meter deep which covered 10% of the total land area. Mortality rate of fish was very low ranging from 0.81-1.63%. However, at harvest, recovery rate ranged from 76.69-82.93% with the highest recovery at 11-15 em of water depth. Significantly the highest absolute growth (99.97) and specific growth rate (2.42%) were found at 21-25 cm water depth. The same treatment also produced significantly higher fish yield (909.76 kg/ha) although statistically similar to the fish yield (862.60 kg/ha) obtained at ll-15 em of water depth. Results also suggested that higher water depth can produce bigger fish but no significant effects of water depth was found on fish yield in the treatments 11-15 cm and 21-25 cm water depths of this experiment

Systematic Theoretical Search for Dibaryons in a Relativistic Model

A relativistic quark potential model is used to do a systematic search for quasi-stable dibaryon states in the $u$, $d$, and $s$ three flavor world. Flavor symmetry breaking and channel coupling effects are included and an adiabatic method and fractional parentage expansion technique are used in the calculations. The relativistic model predicts dibaryon candidates completely consistent with the nonrelativistic model.Comment: 12 pages, latex, no figure