Resorption of unemitted gametes in Lithognathus mormyrus (Sparidae,Teleostei): a possible synergic action of somatic and immune cells

The resorption of unemitted gametes during the post-spawning period of the male and female reproductive cycles in Lithognathus mormyrus was studied by histochemical, histological and cytological methods. The resorption of residual spermatozoa involved the phagocytotic activity of Sertoli cells bounding the seminiferous cysts of spermatozoa, and those associated with spermatogonia lining the lobular lumen. Spermatozoa remaining in the sperm duct were phagocytozed by the lining epithelial cells. Eosinophilic granulocytes and macrophages were identified in the vicinity of residual spermatozoa. The remnants of oocytes underwent an atretic phenomenon in which follicle cells were firstly involved, inducing a progressive fragmentation of the oocyte cytoplasm. Subsequently, eosinophilic granulocytes invaded oocyte degenerative areas and clung to the remaining vitelline inclusions ensuring their biotransformation into waste products (brown bodies). The analogy of the resorption processes of both male and female unemitted gametes during the post-spawning period of natural reproductive cycle, involving first the enveloping somatic cells and then immune cells, is emphasized.     

Raspberry ketone-supplemented diet has no effect on fitness parameters or lure responsiveness in male melon flies,Zeugodacus cucurbitae (Diptera: Tephritidae)

The male annihilation technique (MAT) and sterile insect technique (SIT) are often used to control pestiferous tephritid fruit flies (Diptera: Tephritidae). MAT involves the deployment of traps containing a male attractant and insecticide with the goal of drastically reducing male abundance and ultimately eliminating the entire population. SIT, which involves the mass production, sterilization, and release of the target species, may also be implemented to achieve final extirpation. Generally, simultaneous implementation of MAT and SIT is counterproductive, because the presence of large numbers of male-specific traps in the environment (MAT) would greatly reduce the number of sterile males available for copulating with wild females (SIT). However, studies on the Queensland fruit fly, Bactrocera tryoni (Froggatt), indicate that concurrent use of MAT and SIT may be feasible. Sexually mature males of B. tryoni are attracted to the raspberry ketone and its synthetic analogue cue-lure. Males of B. tryoni fed raspberry-ketone-supplemented diet when newly emerged showed lower attraction to cue-lure baited traps than control males. In addition, newly emerged males provided this diet displayed accelerated sexual maturation, which would allow the early release of sterile males and reduce pre-release holding costs. Here, we examined whether the addition of raspberry ketone to the adult diet of male melon flies, Zeugodacus cucurbitae (Coquillett), produced effects similar to those observed for B. tryoni. Despite using similar methods, no significant effect of raspberry ketone-supplemented diet on time to sexual maturity, survival, mating competitiveness, or attraction to cue-lure baited traps in mass-reared Z. cucurbitae males.     

Measuring the heart rate of the spider, Aphonopelma hentzi: a non-invasive technique

In this work we describe a non‐invasive and precise technique to record the heartbeats of a spider. A linear output Hall effect transducer in conjunction with a small magnet was used to monitor the micromovements on the dorsal surface of the abdomen of the tarantula Aphonopelma hentzi (Girard) (Theraphosidae). The exoskeleton in this region is in direct contact with suspensory ligaments connected to the heart, and the dorsal cuticle of the opisthosoma moves with each heartbeat. The technique allowed the discrimination of the different stages of the spider's cardiac cycle. The method can be also adapted for a smaller spider or other arthropods. We believe that the method proposed in this paper allows investigators to gain insights into a spider's natural heart rate by gathering unbiased data with a non‐invasive and very precise technique. We have found the resting heart rate of A. hentzi to be 5.6 ± 1.47 beats/min, which is lower than previously reported values.     

Absolute nannofossil abundance estimates: Quantifying the pros and cons of different techniques

A quick and inexpensive method to determine absolute nannofossil abundance in deep sea sediments – the “drop” technique (modified dilution method) – was compared to two other available methods – the filtration and random settling techniques. All techniques rely on the same basic principle, under which a volume of known concentration (bulk sediment weight/mL) is distributed evenly over a known total area (glass slide or filter) to then count particles within a set of (randomly) selected fields of view. The three preparation techniques were also calibrated by spiking the samples with microbeads to approach the “real values” as closely as possible. Significant offsets in abundance estimates between methods mainly reflect bias due to the uneven distribution and/or loss of particles. We show that the drop technique is most consistent and accurate in estimating “real values” and offers similar or better reproducibility than the other techniques. The drop method also allows detection of the same trends with or without calibration with microbeads. The filtration method holds the risk to drastically underestimate absolute abundances, while the settling technique is demanding in terms of time and may suffer from advection processes. The composition of nannofossil assemblages can be reliably determined by any of the three different techniques.     

An improved approach to evaluate car sharing options

We develop an improved approach to evaluate car sharing options under uncertain environments with the combination of Fuzzy Analytic Hierarchy Process (F-AHP) and Fuzzy Technique for Order Preference by Similarity to Ideal Solution (F-TOPSIS), which consists of three steps. In the first step, we propose a SCUMN (Specific, Comprehensive, Understandable, Measurable, and Neutral) methodology to identify appropriate indicators and obtain a final list of 24 indicators according to their relevance to car sharing options. In the second step, we determine the weight of each indicator with F-AHP and conduct consistency check of the comparison matrix of selected indicators. In the third step, comparison of different options is performed with selected indicators and F-TOPSIS. A case study is provided to validate the proposed approach. Twenty-four indicators are identified to evaluate five different car sharing options and rank them according to their closeness coefficients in decreasing order. And thirty-one sensitivity analysis experiments are conducted to figure out the influence of indicators on decision making. The experimental results show that the proposed approach is capable of evaluating car sharing options with uncertainty and vagueness. F-AHP is able to determine the weight for each selected indicator and F-TOPSIS demonstrates its advantage in comparing potential options.     

Meloidogyne incognita Surface Antigen Epitopes in Infected Arabidopsis Roots

Surface-coat epitopes of Meloidogyne incognita were detected in root tissues of Arabidopsis thaliana during migration and feeding site formation. A whole-mount root technique was used for immunolocalization of surface coat epitopes in A. thaliana, with the aid of a monoclonal antibody raised specifically against the outer surface of infective juveniles of M. incognita. The antibody, which was Meloidogyne-specific, recognized a fucosyl-bearing glycoprotein in the surface coat. During migration in host tissues the surface coat was shed, initially accumulating in the intercellular spaces next to the juvenile and later at cell junctions farther from the nematode. Upon induction of giant cell formation, the antibody bound to proximally located companion cells and sieve elements of the phloem.     

Immunocytochemical localization of tubulin with colloidal gold in cilia of rabbit tracheal epithelial cultures

Ciliated tracheal epithelia cell cultures were investigated immunocytochemically with anti-tubulin and colloidal gold. When rabbit tracheal cultures were fixed in paraformaldehyde, treated with acetone, anti-tubulin and a second antibody coupled to FITC, fluorescence was associated with cytoskeletal and axonemal microtubules. Cilia covering the apical surface of the ciliated tracheal cells fluoresced very brightly thus facilitating identification of this cell type. Electron microscopy of tracheal cultures fixed as above, treated with Triton-X 100 and incubated in anti-tubulin and protein A coupled to colloidal gold resulted in the highly specific localization of tubulin in ciliary axonemes and basal bodies. Omission of primary or secondary antibody resulted in extremely low levels of fluorescence while no colloidal gold particles could be detected in cultures at the electron microscopy level when rabbit anti-tubulin was omitted.     

Complex regulation of nucleoside transporter expression in epithelial and immune system cells

Nucleoside transporters have a variety of functions in the cell, such as the provision of substrates for nucleic acid synthesis and the modulation of purine receptors by determining agonist availability. They also transport a wide range of nucleoside-derived antiviral and anticancer drugs. Most mammalian cells coexpress several nucleoside transporter isoforms at the plasma membrane, which are differentially regulated. This paper reviews studies on nucleoside transporter regulation, which has been extensively characterized in the laboratory in several model systems: the hepatocyte, an epithelial cell type, and immune system cells, in particular B cells, which are non-polarized and highly specialized. The hepatocyte co-expresses at least two Na+-dependent nucleoside transporters, CNT1 and CNT2, which are up-regulated during cell proliferation but may undergo selective loss in certain experimental models of hepatocarcinomas. This feature is consistent with evidence that CNT expression also depends on the differentiation status of the hepatocyte. Moreover, substrate availability also modulates CNT expression in epithelial cells, as reported for hepatocytes and jejunum epithelia from rats fed nucleotide-deprived diets. In human B cell lines, CNT and ENT transporters are co-expressed but differentially regulated after B cell activation triggered by cytokines or phorbol esters, as described for murine bone marrow macrophages induced either to activate or to proliferate. The complex regulation of the expression and activity of nucleoside transporters hints at their relevance in cell physiology.     

The need and means to characterize sediment structure and behaviour prior to the selection and implementation of remediation plans

Remediation of contaminated sediments requires detailed characterizations of the speciation of the toxic substances and their transformations with regard to time and spatial distribution. While many approaches exist to address dissolved species of toxicants, there is a need to characterize sediments per se in terms of materials or particles which bind toxicants and modulate their bioavailability and rate of burial. Such specific information can be achieved through the correlative use of analytical microscopies, applied directly to native aquatic materials and used in conjunction with novel particle isolation methods and standard techniques of analytical chemistry. Such sedimentary `materials' are dominated by clays and other colloidal minerals, microorganisms, humic substances, organic debris, iron and manganese oxide coatings and extracellular polymeric substances. By using new technology to (1) identify particles and their relative abundances, (2) examine specific toxicant/particle associations at the scale of individual abundant particles, and (3) follow transformations over time, we produce information more insightful than was obtainable previously. Such knowledge will assist in determining which remediation technologies would be best for a given contaminated sediment (i.e. `intrinsic remediation' or dredging/disposal).