The Human SLC25A33 and SLC25A36 Genes of Solute Carrier Family 25 Encode Two Mitochondrial Pyrimidine Nucleotide Transporters

The human genome encodes 53 members of the solute carrier family 25 (SLC25), also called the mitochondrial carrier family, many of which have been shown to transport inorganic anions, amino acids, carboxylates, nucleotides, and coenzymes across the inner mitochondrial membrane, thereby connecting cytosolic and matrix functions. Here two members of this family, SLC25A33 and SLC25A36, have been thoroughly characterized biochemically. These proteins were overexpressed in bacteria and reconstituted in phospholipid vesicles. Their transport properties and kinetic parameters demonstrate that SLC25A33 transports uracil, thymine, and cytosine (deoxy)nucleoside di- and triphosphates by an antiport mechanism and SLC25A36 cytosine and uracil (deoxy)nucleoside mono-, di-, and triphosphates by uniport and antiport. Both carriers also transported guanine but not adenine (deoxy)nucleotides. Transport catalyzed by both carriers was saturable and inhibited by mercurial compounds and other inhibitors of mitochondrial carriers to various degrees. In confirmation of their identity (i) SLC25A33 and SLC25A36 were found to be targeted to mitochondria and (ii) the phenotypes of Saccharomyces cerevisiae cells lacking RIM2, the gene encoding the well characterized yeast mitochondrial pyrimidine nucleotide carrier, were overcome by expressing SLC25A33 or SLC25A36 in these cells. The main physiological role of SLC25A33 and SLC25A36 is to import/export pyrimidine nucleotides into and from mitochondria, i.e. to accomplish transport steps essential for mitochondrial DNA and RNA synthesis and breakdown

Eco-physiological response of two marine bivalves to acute exposition to commercial Bt-based pesticide

Microbial products based on the entomopathogenic bacterium Bacillus thuringiensis (Bt) are among the most common biopesticides used worldwide to suppress insect pests in forests, horticulture and agricultural crops. Some of the effects of commercial Bt have been recorded for terrestrial and freshwater non-target organisms but little research is available on marine fauna. Nevertheless, due to the contiguity of agro-ecosystems and coastal habitats, marine fauna may be highly influenced by this control method. We studied the effect of a commercial Bt product on the physiological and ecological responses and the energy budget of two of the most frequent marine intertidal bivalves in the Mediterranean, the native Mytilaster minimus and the invasive Brachidontes pharaonis. To test the effects experimentally, we simulated the worst scenarios possible using the average dose applied to fields and a hypothetical accumulation dose. The results showed the feeding rates of both species were affected detrimentally by the different experimental conditions; higher concentrations led to higher respiration rates, however neither species showed any significant difference in excretion rates. The biopesticide had a significant effect on the energy budget, the values decreasing with doses. In addition, it led to high mortality for the worst treatments and, in both species, induced significantly higher cardiac activity than in the controls. These results indicate a measurable effect of Bt commercial products on marine organisms, and great attention should be paid to biopesticides composed by entomopathogenic bacteria and addictive compounds. In addition, the results highlight the urgent need to study not only the effects of anthropogenic pressures on target organisms but also to extend our view to other ecosystems not expected to be influenced. Gaining data at the organismal level should help increase the sustainability of pest control and reduce the consequences of side-effects

A multi-objective DIRECT algorithm for ship hull optimization

The paper is concerned with black-box nonlinear constrained multi-objective optimization problems. Our interest is the definition of a multi-objective deterministic partition-based algorithm. The main target of the proposed algorithm is the solution of a real ship hull optimization problem. To this purpose and in pursuit of an efficient method, we develop an hybrid algorithm by coupling a multi-objective DIRECT-type algorithm with an efficient derivative-free local algorithm. The results obtained on a set of “hard” nonlinear constrained multi-objective test problems show viability of the proposed approach. Results on a hull-form optimization of a high-speed catamaran (sailing in head waves in the North Pacific Ocean) are also presented. In order to consider a real ocean environment, stochastic sea state and speed are taken into account. The problem is formulated as a multi-objective optimization aimed at (i) the reduction of the expected value of the mean total resistance in irregular head waves, at variable speed and (ii) the increase of the ship operability, with respect to a set of motion-related constraints. We show that the hybrid method performs well also on this industrial problem

Cytometric analysis, genetic manipulation and antibiotic selection of the snail embryonic cell line Bge from Biomphalaria glabrata, the intermediate host of Schistosoma mansoni.

The invertebrate cell line, Bge, from embryos of the snail Biomphalaria glabrata, remains to date the only established cell line from any species of the Phylum Mollusca. Since its establishment in 1976 by Eder Hansen, few studies have focused on profiling its cytometrics, growth characteristics or sensitivity to xenobiotics. Bge cells are reputed to be challenging to propagate and maintain. Therefore, even though this cell line is a noteworthy resource, it has not been studied widely. With growing interest in functional genomics, including genetic transformation, to elucidate molecular aspects of the snail intermediate hosts responsible for transmission of schistosomiasis, and aiming to enhance the convenience of maintenance of this molluscan cell line, we deployed the xCELLigene real time approach to study Bge cells. Doubling times for three isolates of Bge, termed CB, SL and UK, were longer than for mammalian cell lines - longer than 40 h in complete Bge medium supplemented with 7% fetal bovine serum at 25 °C, ranging from ∼42 h to ∼157 h when 40,000 cells were seeded. To assess the potential of the cells for genetic transformation, antibiotic selection was explored. Bge cells were sensitive to the aminonucleoside antibiotic puromycin (from Streptomyces alboniger) from 5 μg/ml to 200 ng/ml, displaying a half maximal inhibitory concentration (IC50) of ∼1.91 μg/ml. Sensitivity to puromycin, and a relatively quick kill time (<48 h in 5 μg/ml) facilitated use of this antibiotic, together with the cognate resistance gene (puromycin N-acetyl-transferase) for selection of Bge cells transformed with the PAC gene (puroR). Bge cells transfected with a plasmid encoding puroR were partially rescued when cultured in the presence of 5 μg/ml of puromycin. These findings pave the way for the development of functional genomic tools applied to the host-parasite interaction during schistosomiasis and neglected tropical trematodiases at large

Can the plasma PD-1 levels predict the presence and efficiency of tumor-infiltrating lymphocytes in patients with metastatic melanoma?

Background: The immune response in melanoma patients is locally affected by presence of tumor-infiltrating lymphocytes (TILs), generally divided into brisk, nonbrisk, and absent. Several studies have shown that a greater presence of TILs, especially brisk, in primary melanoma is associated with a better prognosis and higher survival rate. Patients and Methods: We investigated by enzyme-linked immunosorbent assay (ELISA) the correlation between PD-1 levels in plasma and the presence/absence of TILs in 28 patients with metastatic melanoma. Results: Low plasma PD-1 levels were correlated with brisk TILs in primary melanoma, whereas intermediate values correlated with the nonbrisk TILs, and high PD-1 levels with absent TILs. Although the low number of samples did not allow us to obtain a statistically significant correlation between the plasma PD-1 levels and the patients' overall survival depending on the absence/presence of TILs, the median survival of patients having brisk type TILs was 5 months higher than that of patients with absent and nonbrisk TILs. Conclusions: This work highlights the ability of measuring the plasma PD-1 levels in order to predict the prognosis of patients with untreated metastatic melanoma without a BRAF mutation at the time of diagnosis

Interleukin-13 (IL-13)in autoimmune rheumatic diseases: relationship with autoantibody profile

Objective: Several cytokines play a role in the production of autoantibodies such as RF and ANA by B-lymphocytes; the role of IL- 13 in this process has not been previously studied. We investigated the relationship between the serum concentration of this cytokine and circulating autoantibodies. Methods: IL-13 serum levels, as well as RF and ANA, were evaluated in 282 patients with autoimmune rheumatic diseases including RA (n=84), SLE (n = 114), SS (n = 52) and Scl (n =32). Results: Serum levels of IL-13 (pg/ml) were significantly higher in patients with RA (p < 0.00003), SLE (p < 0.03), SS (p < 0.0007), or Scl (p < 0.025) compared to controls. IL-13 serian levels correlated with those of RF in RA (p < 0.00001), SLE (p < 0.003) and Scl (p < 0.03). IL- 13 levels were higher in RA (p<0.0003), SLE (p<0.005) and Scl (p<0.05) patients with RF than in patients,without RE SS patients with anti-SSA/Ro antibodies had significantly higher IL-13 levels than SS patients without this autoantibody (p < 0.04). No statistically significant correlation was found between IL-13 levels and any other antinuclear autoantibody, total immunoglobulin levels or the main clinical features of each disease. Conclusion: The evidence of higher IL- 13 levels in our RA, SLE, SS and Scl patients confirms that this cytokine is involved in the pathogenesis of autoimmune rheumatic diseases. The relationship of this cytokine with RF in RA, SLE and Scl, as well as with antiSSA/ Ro antibody in SS, strengthens the hypothesis that it plays a role in autoantibody production. However the different autoantibody synthesis by Bcells recognises different pathways depending on the underlying autoimmune disease

First studies on Giardia duodenalis in the water buffalo

A cross-sectional survey of Giardia duodenalis infection in the water buffalo was carried out in Central Italy. The survey was conducted on a sample of 90 farms, selected using a grid approach within a Geographical Information System, followed by proportional allocation. On each farm, faecal samples were collected from three to five asymptomatic buffalo calves, aged from 1 to 9 weeks (total number = 347). Each faecal sample was tested for the presence of copro-antigens of G. duodenalis using a commercially available ELISA. Out of the 90 farms, 27 (30.0%) resulted positive. With respect to animals, out of the 347 faecal samples, 63 (18.1%) were found to have antigens of G. duodenalis. The results of the logistic regression model showed a positive association between the positivity to G. duodenalis and the presence of sheep on farm

Colorimetric paper-based device for hazardous compounds detection in air and water: A proof of concept

In the last decades, the increase in global industrialization and the consequent technological progress have damaged the quality of the environment. As a consequence, the high levels of hazardous compounds such as metals and gases released in the atmosphere and water, have raised several concerns about the health of living organisms. Today, many analytical techniques are available with the aim to detect pollutant chemical species. However, a lot of them are not affordable due to the expensive instrumentations, time-consuming processes and high reagents volumes. Last but not least, their use is exclusive to trained operators. Contrarily, colorimetric sensing devices, including paper-based devices, are easy to use, providing results in a short time, without particular specializations to interpret the results. In addition, the colorimetric response is suitable for fast detection, especially in resource-limited environments or underdeveloped countries. Among different chemical species, transition and heavy metals such as iron Fe(II) and copper Cu(II) as well as volatile compounds, such as ammonia (NH3) and acetaldehyde (C2 H4 O) are widespread mainly in industrialized geographical areas. In this work, we developed a colorimetric paper-based analytical device (PAD) to detect different contaminants, including Fe2+ and Cu2+ ions in water, and NH3 and C2 H4 O in air at low concentrations. This study is a “proof of concept” of a new paper sensor in which the intensity of the colorimetric response is proportional to the concentration of a detected pollutant species. The sensor model could be further implemented in other technologies, such as drones, individual protection devices or wearable apparatus to monitor the exposure to toxic species in both indoor and outdoor environments

Cystic echinococcosis in water buffaloes (Bubalus bubalis)

An epidemiological and molecular survey of cystic echinococcosis (CE) caused by Echinococcus granulosus in the water buffalo (Bubalus bubalis) of the Italian Mediterranean breed was carried out in the Campania region of southern Italy. Out of a total of 799 water buffaloes examined at slaughterhouses, 80 (10.0%) were found infected. The molecular study was performed on 58 hydatid cysts in order to determine the E. granulosus strain(s) present in this host. A region of cytocrome c oxydase 1 gene (CO1) was amplified by polymerase chain reaction and the PCR products were then purified and sequenced. DNA amplification of the partial CO1 gene gave a 446 bp fragment for all isolates examined. After sequencing, a region of 419 bp was identified for each sample. Thirty-two isolates were identified as the common sheep strain G1, 15 as the buffalo strain G3, 3 as the Tasmanian sheep strain G2, and 3 as the G1 c genotype (GenBank AF458873). In addition, 5 isolates presented 99% identity with the G2 genotype (Tasmanian sheep strain)