Hexokinase (HXK) is the first key enzyme in the glycolytic pathway and plays an extremely important role in energy metabolism. By searching the microsporidian database, we found a sequence (NBO_27g0008) of Nosema bombycis Nägali, 1857 with high similarity to hexokinase-2, and named it as NbHXK2. The NbHXK2 gene has 894 bp and encodes 297 amino acids with 34.241 kD molecular weight and 5.26 isoelectric point. NbHXK2 contains 31 phosphorylation sites and 4 potential N-glycosylation sites with signal peptides and no transmembrane domain. Multiple sequence alignment showed that NbHXK2 shares more than 40% amino acid identity with that of other microsporidia, and the homology with hexokinase-2 of Nosema tyriae Canning, Curry, Cheney, Lafranchi-Tristem, Kawakami, Hatakeyama, Iwano et Ishihara, 1999, Nosema pyrausta (Paillot, 1927) and Nosema ceranae Fries, Feng, da Silva, Slemenda et Pieniazek, 1996 was 89.17%, 87.82% and 69.86%, respectively. Phylogenetic analysis based on the amino acid sequence of hexokinase showed that all microsporidia cluster together in the same clade, and are far away from animals, plants and fungi, and that N. bombycis is closely related to N. tyriae; N. pyrausta; N. ceranae and Nosema apis Zander, 1909. Immunolocalisation with the prepared polyclonal antibody showed that NbHXK2 was mainly distributed in the cytoplasm and plasmalemma in proliferative, sporulation stage and mature spore of N. bombycis. qRT-PCR assay showed that the NbHXK2 expressed at higher level during spore germination and at early stage of proliferation. These results indicate that N. bombycis may use its own glycolytic pathways to supply energy for infection and development, especially germination and in the early stage of proliferation, and acquire energy from the host through certain ways as well.
Tapinoma melanocephalum is a worldwide distributed, highly invasive ant species. It lives in close association with human societies and its distribution is human-mediated in large measure. The geographical origin of this ant species is unknown, but its introduction in areas previously devoided of its presence can represent a threat to the native biota, act as an agricultural pest or as a pathogen vector. To investigate the genetic structure and phylogeography of this species we identified 12 new polymorphic microsatellite markers, and in addition, we tested and selected 12 ant-universal microsatellites polymorphic in T. melanocephalum. We genotyped 30 individuals from several islands of Micronesia and Papua-New Guinea. All 24 loci exhibited strong homozygosity excess (45-100%, mean = 86%), while the number of alleles per locus reached usual values (2-18, mean = 6.5), resulting in levels of expected heterozygosity much higher than observed. Based on several robust tests, we were able to exclude artefacts such as null alleles and allelic dropout as a possible cause of the observed pattern. Homozygosity excess might be a consequence of founder effect, bottleneck and/or inbreeding. As our sample population was composed of individuals from several distinct localities, the Wahlund effect might have contributed to the increased homozygosity as well. Despite the provisionally observed deviation from the Hardy-Weinberg equilibrium, the newly developed microsatellites will provide an effective tool for future genetic investigations of population structure as well as for the phylogeographic study of T. melanocephalum., Jan Zima Jr., Ophélie Lebrasseur, Michaela Borovanská, Milan Janda., and Obsahuje bibliografii
Eggs of the migratory locust, Locusta migratoria (Orthoptera: Acrididae), hatch synchronously when in a pod, but only sporadically when kept separately. Here, we aimed to detect the vibrational stimuli emitted by eggs that initiate synchronous hatching. First, we recorded the vibrations emitted by an egg pod and single eggs. One bout of vibrations consisted of 2 to 46 vibrations. The total number and amplitude of vibrations in single eggs increased as the time to hatch decreased. Eggs kept separately were continuously subjected during the last 2 days before hatching to recordings of vibrations from a single egg. Recordings made during the last 2.5 h before hatching caused these eggs to hatch significantly earlier than those not subjected to this treatment, the control. In contrast, eggs subjected to recordings made 8 to 10 h before hatching significantly delayed their hatching relative to the controls, which indicates that synchronous hatching of eggs is induced by age-dependent changes in vibrations from neighbouring eggs. Exposure to one large bout of vibrations (consisting of 40 vibrations in 101 s) was sufficient to induce synchronous hatching in eggs kept separately when applied 5 h before hatching, but not 36 or 11.5 h before hatching. Visual inspection of the spectra indicated that the vibrations had two peaks at about 100 Hz and 1.5 kHz. Only exposure to the latter altered the hatching time of eggs. The embryo moved the posterior part of its abdomen when emitting the vibrations. These results indicate that the synchronous hatching of locust eggs is induced when the embryos emit particular vibrations.
The Jahamah Platform is a part of a structural depression called the Sirt basin, located in the northern central part of Libya. The Jahamah Platform spans latitude 〖29.95〗^° N to 〖30.55〗^° N and longitudes 〖19.32〗^° E to 〖19.77〗^° E with an estimated area of about 2,187 km2. Libyan Petroleum Institute provided the data of aeromagnetic that was used in this study. The data was used to study the structure beneath the Jahamah Platform by using Oasis montaj software. Various filters from the software have been applied to enhance determining the fault system within the study area. An RTP filter was applied to the magnetic data to construct a reduction to the pole anomaly map. The subsurface structural elements underneath the study area were identified using Total horizontal derivative (THD), CET analysis, and Euler deconvolution. 2-D forward modelling of the area was constructed based on gravity data, and then the basement depth was estimated to range from 2.2 km to 3.1 km based on the model. Based on the interpretation of the constructed maps, the area has a number of faults that trend in NE-SW, NNW-SSE, N-S and NW-SE and faults depth ranging between 790 m to 3102 m.
Studie je replikací experimentu, který přinesl podporu pro hypotézu, že lidé představující si konzumaci jídla sní následně méně jídla než lidé, kteří si konzumaci jídla nepředstavovali. Replikace byla provedena na vzorku 77 vysokoškolských studentů, kteří byli náhodně rozděleni do tří skupin podle různých druhů imaginace. V porovnání s replikovanou studií byl pro analýzy použit větší zkoumaný soubor, byla ověřována úspěšnost manipulace s nezávislou proměnnou a zkoumaným osobám za účast ve výzkumu nebyla nabízena žádná odměna. Na rozdíl od replikované studie nebyl zaznamenán statisticky významný vliv druhu imaginace konzumace jídla na jeho pozdější skutečnou konzumaci. Výsledky nepodporují výsledky původního experimentu a ukazují, že původní výsledky mohly být jen důsledkem náhody. Jsou potřeba další replikace, aby bylo zřejmé, zda je efekt imaginace konzumace na zkonzumované množství jídla skutečný. and This study is a replication of an experiment that has supported a hypothesis that people imagining a consumption of food before its real consumption eat less food than without the prior imagination. The sample consisted of 77 university students who were randomly assigned into three groups varying by amount of imagined objects being consumed. A larger sample has been collected, the level of predictor variable manipulation has been controlled and a no compensation has been offered to the participants in comparison to the replicated study. On the contrary to the primary study, no significant effect of the type of imagined consumption on the consecutive consumption has been found. Results do not support results of the original study and suggest the original study’s outcome might have been achieved at random. More replication is necessary to be able to assess whether the effect of imagination on food consumption is real.
Type 2 diabetes (T2D) is believed to be a non-autoimmune metabolic disorder. However, there are increasing reports that some T2D patients have immune abnormalities. In addition, it is known that there are sex differences in the onset of diabetes and immune responses in humans. Spontaneously Diabetic Torii (SDT) rats, a non-obese T2D model, also have sex differences in the onset of diabetes, but the involvement of immune abnormalities in diabetes is unknown. In this study, we investigated immune abnormalities in SDT rats. Immune cell subset analysis was performed in male and female SDT rats and control Sprague-Dawley (SD) rats at 5, 11, and 17 weeks of age. Male and female SDT rats had swelling of the spleen and lymph nodes and a higher number of T cells and B cells in the blood, spleen, and lymph nodes than SD rats. Only male SDT rats developed diabetes at 17 weeks of age, and the number of classical and non-classical monocytes in the blood and spleen of male SDT rats was higher than that in male SD rats and female SDT rats that did not develop diabetes. Most of these findings were observed before the onset of diabetes (~11 weeks of age), suggesting that classical and non-classical monocytes may contribute to the development of diabetes in male SDT rats. In conclusion, SDT rats may be a useful T2D model involved in immune abnormalities, and further research will help elucidate the pathophysiology of T2D with immune abnormalities and develop new therapeutic agents.
The elastase, which belongs to the serine protease family, hydrolyses various proteins and may be involved in the parasite invasion. In this study, complete sequence of elastase-1 (TsE) the nematode Trichinella spiralis (Owen, 1835) was cloned into the plasmid pcDNA3.1 as TsE DNA vaccine. After intramuscular vaccination, serum anti-Trichinella antibodies (IgG and subclass IgG1/IgG2a, and IgA), total and specific intestinal mucosal sIgA in mice vaccinated with pcDNA3.1/TsE were measured by ELISA. The results showed that vaccination with pcDNA3.1/TsE induced a systemic humoral immune response (high levels of serum IgG and subclass IgG1/IgG2a and IgA) and local intestinal mucosal immune responses (high levels of TsE-specific sIgA). Vaccination of mice with TsE DNA vaccine also triggered a systemic and local concomitant Th1/Th2 response, as demonstrated by significant elevation of Th1 (IFN-γ and IL-2) / Th2 (IL-4 and IL-10) cytokine levels after the spleen, mesenteric lymph node and Peyer's patch cells from vaccinated mice were stimulated with recombinant TsE (rTsE). The vaccination of mice with pcDNA3.1/TsE displayed a 17% reduction of intestinal adult worms and a 39% reduction of muscle larvae. Our results indicated that TsE DNA vaccine elicited a systemic concomitant Th1/Th2 response and an enteral local sIgA response, and produced a partial protection against infection with T. spiralis. The TsE may be regarded as a potential candidate vaccine target against Trichinella infection. The oral polyvalent vaccines should be developed to improve the protective efficacy of anti-Trichinella vaccines.
Uterine tubes (UTs) are essential during physiological reproduction. The most intriguing part of its wall is the mucosa. Apart from the epithelial cells vital for its normal function, the connective tissue lamina propria contains wide spaces whose function, morphology and structure are yet to be elucidated. The present study used bioptic samples from 25 premenopausal (mean age 48.3 years, σ=3.56) and 25 postmenopausal women (mean age 57.8 years, σ=7.79). In both study groups, samples were obtained from two anatomically distinct parts of the UT – ampulla and infundibulum with fimbriae. The specimens were processed for scanning electron microscopy (SEM) and immunohistochemical detection of podoplanin (clone D2-40) and VEGFR-3 – two markers of lymphatic endothelial cells. The results showed that specimens from premenopausal and postmenopausal women contain wide lymphatic spaces, also known as lymphatic lacunae. The most probable function of the lacunae in the fimbriae is oocyte pick-up upon ovulation thanks to their ability to get engorged with lymph, thus serving as an erectile-like tissue. The ampullary lacunae are probably responsible for tubal fluid maintenance and recirculation. These results indicate that they are vital for normal reproduction because tubal fluid dynamics are as important as fluid composition. Further research on this topic is highly warranted because more detailed insights into UT function have a great potential to refine the methods of reproductive medicine, e.g. in vitro fertilization (IVF), which are still far from optimal regarding fertility outcomes.
MEHMO syndrome is a rare X-linked syndrome characterized by Mental retardation, Epilepsy, Hypogenitalism, Microcephaly, and Obesity associated with the defect of protein synthesis caused by the EIF2S3 gene mutations. We hypothesized that the defect in protein synthesis could have an impact on the immune system. We describe immunologic phenotype and possible treatment outcomes in patient with MEHMO syndrome carrying a frameshift mutation (I465fs) in the EIF2S3 gene. The proband (currently 9-year-old boy) had normal IgG and IgM levels, but had frequent respiratory and urinary tract infections. On subcutaneous immunoglobulin therapy achieving supraphysiological IgG levels the frequency of infections significantly decreased in Poisson regression by 54.5 % (CI 33.2-89.7, p=0.017). The MEHMO patient had had frequent acute infections despite normal IgG and IgM serum levels and responded well to the immunoglobulin treatment., Ivana Trochanová, Daniela Staníková, Martina Škopková, Klaudia Haštová, Daniela Gašperíková, Juraj Staník, Peter Čižnár., and Obsahuje bibliografii
Mitochondria are considered central regulator of the aging process; however, majority of studies dealing with the impact of age on mitochondrial oxygen consumption focused on skeletal muscle concluding (although not uniformly) a general declining trend with advancing age. In addition, gender related differences in mitochondrial respiration have not been satisfactorily described yet. The aim of the present study was to evaluate mitochondrial oxygen consumption in various organs of aging male and female Fischer 344 rats at the ages of 6, 12 and 24 months. Mitochondrial respiration of homogenized (skeletal muscle, left and right heart ventricle, hippocampus, cerebellum, kidney cortex), gently mechanically permeabilized (liver) tissue or intact cells (platelets) was determined using high-resolution respirometry (oxygraphs O2k, Oroboros, Austria). The pattern of age-related changes differed in each tissue: in the skeletal muscle and kidney cortex of both sexes and in female heart, parameters of mitochondrial respiration significantly declined with age. Resting respiration of intact platelets displayed an increasing trend and it did not correlate with skeletal muscle respiratory states. In the heart of male rats and brain tissues of both sexes, respiratory states remained relatively stable over analyzed age categories with few exceptions of lower mitochondrial oxygen consumption at the age of 24 months. In the liver, OXPHOS capacity was higher in females than in males with either no difference between the ages of 6 and 24 months or even significant increase at the age of 24 months in the male rats. In conclusion, the results of our study indicate that the concept of general pattern of age-dependent decline in mitochondrial oxygen consumption across different organs and tissues could be misleading. Also, the statement of higher mitochondrial respiration in females seems to be conflicting, since the genderrelated differences may vary with the tissue studied, combination of substrates used and might be better detectable at younger ages than in old animals.