Morphological and enzymatic analysis of the midgut of Anopheles darlingi during blood digestion

The midgut of adult female Anopheles darlingi is comprised of narrow anterior and dilated posterior regions, with a single layered epithelium composed by cuboidal digestive cells. Densely packed apical microvilli and an intricate basal labyrinth characterize each cell pole. Before blood feeding, apical cytoplasm contains numerous round granules and whorled profiles of rough endoplasmic reticulum. Engorgement causes a great distension of midgut. This provokes the flattening of digestive cells and their nuclei. Simultaneously, apical granules disappear, the whorls of endoplasmic reticulum disassemble and 3h post bloodmeal (PBM), nucleoli enlarge manyfold. An intense absorptive process takes place during the first 24 h PBM, with the formation of large glycogen inclusions, which persist after the end of the digestive process. Endoproteases activities are induced after bloodmeal and attain their maximum values between 10 and 36 h PBM. At least two different aminopeptidases seem to participate in the digestive process, with their maximum activity values at 36 and 48 h PBM, respectively. Coarse electrondense aggregates, possibly debris from digested erythrocytes, begin to appear on the luminal face of the peritrophic membrane from 18 h PBM and persist during all the digestive process, and are excreted at its end. We suggest that these aggregates could contain some kind of insoluble form of haem, in order of neutralize its toxicity.     

Effect of different soil and clove treatments in the control of white rot of garlic

Soil solarisation was consistently efficacious in reducing inoculum density to undetectable levels in a field naturally‐infested with Sclerotium cepivorum. This treatment delayed epidemic onset of white rot of garlic 2–3 months as compared with the untreated control or the inoculation of planting furrows with Glomus intraradices. Furthermore, significant reductions of disease incidence and of the standardised AUDPC were also observed in solarised plots, resulting in quantitative and qualitative yield improvement. Similar effects were observed in plots planted with tebuconazole‐treated cloves, confirming previous results, whereas Trichoderma harzianum was ineffective as a biocontrol agent, when applied to planting furrows. The inoculation of plots with G. intraradices before planting, in three consecutive years, was neither effective for disease control nor on the development of garlic, although the root systems of garlic plants from all the experimental treatments were heavily mycorrhizal at harvest, indicating the presence of native arbuscular mycorrhizal propagules in the soil and their survival after soil solarisation.     

Application of two machine learning algorithms to genetic association studies in the presence of covariates

Background

achaete-scute complexe (AS-C) has been widely studied at genetic, developmental and evolutional levels. Genes of this family encode proteins containing a highly conserved bHLH domain, which take part in the regulation of the development of central nervous system and peripheral nervous system. Many AS-C homologs have been isolated from various vertebrates and invertebrates. Also, AS-C genes are duplicated during the evolution of Diptera. Functions besides neural development controlling have also been found in Drosophila AS-C genes.

Results

We cloned four achaete-scute homologs (ASH) from the lepidopteran model organism Bombyx mori, including three proneural genes and one neural precursor gene. Proteins encoded by them contained the characteristic bHLH domain and the three proneural ones were also found to have the C-terminal conserved motif. These genes regulated promoter activity through the Class A E-boxes in vitro. Though both Bm-ASH and Drosophila AS-C have four members, they are not in one by one corresponding relationships. Results of RT-PCR and real-time PCR showed that Bm-ASH genes were expressed in different larval tissues, and had well-regulated expressional profiles during the development of embryo and wing/wing disc.

Conclusion

There are four achaete-scute homologs in Bombyx mori, the second insect having four AS-C genes so far, and these genes have multiple functions in silkworm life cycle. AS-C gene duplication in insects occurs after or parallel to, but not before the taxonomic order formation during evolution.     

Laser stimulation of the acupoint ‘Zusanli’ (ST.36) on the radiopharmaceutical biodistribution in <Emphasis Type="BoldItalic">Wistar</Emphasis> rats

Laser used to stimulate acupoints is called laser acupuncture (LA). It is generally believed that similar clinical responses to manual acupuncture can be achieved. Here we analysed the effects of the laser (904 nm) at the ‘Zusanli’ acupoint (ST.36) of the stomach meridian on the biodistribution of the radiopharmaceutical Na^99mTcO₄. Wistar rats were divided into control (CG) and experimental groups (EG). The EG were exposed daily to the laser (904 nm) at ST.36 with 1 joule/min (40 mW/cm²) for 1 min. The animals of the CG were not exposed to laser at all. On the 8th day after LA, the animals were sedated and Na^99mTcO₄ was administered. After 10 min, the animals were all sacrificed and the organs removed. The radioactivity was counted in each organ to calculate the percentage of radioactivity of the injected dose per gram (%ATI/g). Comparison of the %ATI/g in EG and CG was performed by Mann-Whitney test. The %ATI/g was significantly increased in the thyroid due to the stimulation of the ST.36 by laser. It is possible to conclude that the stimulation of ST.36 does lead to biological phenomena that interfere with the metabolism of the thyroid.     

Drought effects on the dynamics of leaf production and senescence in field-grown Medicago arborea and Medicago citrina

Forage shrub production in the Mediterranean region is frequently limited by soil water availability. To study plant responses to water deficit under such conditions is important for improving crop management and for selecting better yielding forage shrub species. Pre-dawn leaf water potential (Ψ_pd), plant leaf area (PLA), leaf area per stem (LA_s), leaf appearance rate (LAR₁;), leaf senescence rate (LSR), individual leaf area (LA) and maximal leaf elongation rate (LER) were studied throughout the year for Medicago arborea (MA) and Medicago citrina (MC) under irrigated (control) and low rainfall field conditions, at the experimental field site of the University of the Balearic Islands in Spain. With irrigation, the highest LA and LER were observed in autumn and spring and the lowest in winter and summer. LAR; was similar for both species in autumn and winter. Throughout the spring, LAR₁ was higher for MC compared to MA. PLA was similar for both species during the autumn, winter and spring seasons; however, during the summer PLA of MA was significantly reduced by 53%. This decline was attributed to higher leaf senescence during seed maturity. As a consequence, MC maintained higher leaf area (∼ 5 m² plant⁻¹) than MA (3 m² plant⁻¹). Under natural field conditions, soil water deficit increased from February to late August. The main effect of water stress was a marked reduction in LAR₁, LA and LER reflected in lower LA_s and PLA. Leaf area was severely reduced for both species during the summer, but much more intensively in MA, which developed full leaf senescence. Thus, MC maintained higher PLA than MA (0.5 m² compared to 0.0 m²). Throughout the year, but especially in the driest months, MC was superior to MA in leaf growth parameters and PLA.     

The function of TLR4 in interferon gamma or interleukin-13 exposed and lipopolysaccharide stimulated gingival epithelial cell cultures

Gingival epithelial cells are part of the first line of host defense against infection. Toll-like receptors (TLRs) serve important immune and nonimmune functions. We investigated how interferon gamma (INF-γ) and interleukin 13 (IL-13) are involved in the TLR4 ligand-induced regulation of interleukin-8 (IL-8) effects on gingival epithelial cells. We used immunohistochemistry to localize TLR4 in ten healthy and ten periodontitis tissue specimens. Gingival epithelial cells then were primed with Th1 cytokine (INF-γ) or Th2 cytokine (IL-13) before stimulation with Escherichia coli-derived lipopolysaccharide (LPS) and enzyme-linked immunosorbent assay (ELISA) was performed to detect the level of IL-8 secretion in cell culture supernatants. Although both healthy and periodontitis gingival tissue samples expressed TLR4, the periodontitis samples showed more intense expression on gingival epithelial cells. Gingival epithelial cell cultures were primed with either INF-γ or IL-13 before stimulation with TLR4 ligand. Supernatants from co-stimulated epithelial cells exhibited IL-8 production in opposite directions, i.e., as one stimulates the release, the other reduces the release. INF-γ significantly increased TLR4 function, whereas IL-13 significantly decreased TLR4 function, i.e., production of IL-8. Pathogen associated molecular pattern-LPS, shared by many different periodonto-pathogenic bacteria, activates the gingival epithelial cells in a TLR-dependent manner. Diminished or increased TLR function in gingival epithelial cells under the influence of different Th cell types may protect or be harmful due to the altered TLR signaling.