Determination of non-steroidal anti-inflammatory drug (NSAIDs) residues in water samples

Pharmacologically active substances used to treat human and animal illnesses can enter the aquatic environment via effluents from wastewater treatment plants or in the case of veterinary drugs directly through liquid manure discharge. Some of these substances enter the environment either as the parent compound or as active/inactive metabolites. Due to their pharmacological activity, their determination and understanding their behavior and fate in the environment are important. The scope of this paper was to develop an analytical procedure to determine common pharmaceutical residues in wastewaters. Pharmacologically active substances were chosen according to their wide spread application in Slovenia and Central Europe and are members of analgesics, e.g., non-steroidal anti-inflammatory drugs: ibuprofen, naproxen, ketoprofen and diclofenac. Selected compounds were isolated from synthetic water using a novel SPE sorbent Strata X. Due to the non-volatile nature of these compounds they were first silylised prior to gas chromatographic-mass spectrometric detection. The developed procedure was tested with synthetic wastewaters and their extraction efficiency (>84%) and method limits of detection (2-6 ng L(-1)) were determined. Our procedure has been adopted and optimised for "real" water samples and applied to eleven drinking and ten river water samples from Slovenia. The results showed no traces of NSAIDs in all potable water samples and low-range contamination (ng L(-1)) of Slovene rivers. These results show that NSAIDs contamination of Slovene waters is comparable with published results of water contamination in Central Europe.     

The effects of male and female genotype on variance in male fertilization success in the red flour beetle (<Emphasis Type="Italic">Tribolium castaneum</Emphasis>)

The number of eggs fertilized by a male at any given copulation (fertilization success) is affected by a large number of factors. Male insemination and sperm competition success and various female structures and/or processes that bias paternity in favor of some males over others (cryptic female choice) are all likely to affect fertilization success. We suggest that more comprehensive measures of male fertilization success can increase our understanding of postcopulatory sexual selection. To improve our understanding of the importance of various sources of variance in male fertilization success, we conducted a series of experiments using flour beetles. Different wild-type strains were used in reciprocal double mating experiments, against a phenotypic marker strain. We assessed the relative effects of female genotype, male genotype and mating order on independent and inclusive measures of male defense ability ( P ₁), male offense ability ( P ₂), and female remating behavior. Female genotype influenced both P ₁ and P ₂, and male genotype interacted strongly with female genotype in its effect on P ₂. We also documented an interaction between female and male genotypes in the effects of mating on female remating behavior, such that females tended to remate most rapidly when mated to males of their own genotype. It is clear from our experiments that cryptic female choice influences the pattern of fertilization success in flour beetles, and we suggest that cryptic female choice may often be an important component of postcopulatory sexual selection. Future investigations would benefit from studying the multiple components of variance in male fertilization success.     

Piercing the partner's skin influences sperm uptake in the earthworm Lumbricus terrestris

Sexual conflict between mating partners can give rise to strategies that are advantageous for one sex but harmful to the opposite sex. Usually, sperm donors develop (offensive) traits to enhance their chances in sperm competition, while sperm recipients evolve (defensive) traits that allow them to stay in control of who fathers their offspring. Here, we demonstrate that these processes are also at work in simultaneous hermaphrodites. The hermaphroditic earthworm Lumbricus terrestris uses 40 to 44 copulatory setae to pierce into its partner's skin, causing damage and injecting a substance from its setal glands. Experimental injection of the gland substance indicates that a refractory period may be induced. More importantly, removal of the copulatory setae shows that they influence the partner's sperm uptake. When the setae are present, more sperm are taken up and sperm are distributed more equally over the four spermathecae. We interpret this as a strategy that stacks the odds for the donor's sperm in fertilizing cocoons.     

Perceived enjoyment,concentration, intention,and speed violation behavior: Using flow theory and theory of planned behavior

Objective: Road accidents are an important public health concern, and speeding is a major contributor. Although flow theory (FLT) is a valid model for understanding behavior, currently the nature of the roles and interplay of FLT constructs within the theory of planned behavior (TPB) framework when attempting to explain the determinants of motivations for intention to speed and speeding behavior of car drivers is not yet known. The study aims to synthesize TPB and FLT in explaining drivers of advanced vehicles intentions to speed and speed violation behaviors and evaluate factors that are critical for explaining intention and behavior.

Method: The hypothesized model was validated using a sample collected from 354 fully licensed drivers of advanced vehicles, involving 278 males and 76 females on 2 occasions separated by a 3-month interval. During the first of the 2 occasions, participants completed questionnaire measures of TPB and FLT variables. Three months later, participants' speed violation behaviors were assessed.

Results: The study observed a significant positive relationship between the constructs. The proposed model accounted for 51 and 45% of the variance in intention to speed and speed violation behavior, respectively. The independent predictors of intention were enjoyment, attitude, and subjective norm. The independent predictors of speed violation behavior were enjoyment, concentration, intention, and perceived behavioral control.

Conclusions: The findings suggest that safety interventions for preventing speed violation behaviors should be aimed at underlying beliefs influencing the speeding behaviors of drivers of advanced vehicles. Furthermore, perceived enjoyment is of equal importance to driver's intention, influencing speed violation behavior.     

Sympatry and allopatry in the deeply divergent mitochondrial DNA clades of the estuarine pulmonate gastropod genus <Emphasis Type="Italic">Phallomedusa</Emphasis> (Mollusca,Gastropoda)

DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene were collected from estuarine snails in the genus Phallomedusa to examine the effects of estuarine isolation on population structure and gene flow. Three clades were recovered, one corresponding to Phallomedusa austrina and two others with the morphology of Phallomedusa solida. The haplotype diversity in all three clades indicated recent population expansion. Phallomedusa austrina was restricted to the west of a previous land bridge in the Bass Strait between mainland Australia and Tasmania, and P. solida to its east and to northern Tasmania. Phylogeographic analysis of P. austrina and P. solida shows strong geographic separation of species, but no local genetic structure indicative of regional or estuarine isolation. The clades of P. solida exhibit substantial genetic divergence and were sympatric across their entire distribution in eastern Tasmania and mainland Australia. Such a situation, which has not previously been observed in phylogeographic studies of southeast Australia, suggests that P. solida has had a complex refugial history during periods of environmental challenge.     

Cx43 regulates mechanotransduction mechanisms in human preterm amniotic membrane defects

Objective

The effects of mechanical stimulation in preterm amniotic membrane (AM) defects were explored.

Methods

Preterm AM was collected from women undergoing planned preterm caesarean section (CS) due to fetal growth restriction or emergency CS after spontaneous preterm prelabour rupture of the membranes (sPPROM). AM explants near the cervix or placenta were subjected to trauma and/or mechanical stimulation with the Cx43 antisense. Markers for nuclear morphology (DAPI), myofibroblasts (αSMA), migration (Cx43), inflammation (PGE₂) and repair (collagen, elastin and transforming growth factor β [TGFβ₁]) were examined by confocal microscopy, second harmonic generation, qPCR and biochemical assays.

Results

In preterm AM defects, myofibroblast nuclei were highly deformed and contractile and expressed αSMA and Cx43. Mechanical stimulation increased collagen fibre polarisation and the effects on matrix markers were dependent on tissue region, disease state, gestational age and the number of fetuses. PGE₂ levels were broadly similar but reduced after co-treatment with Cx43 antisense in late sPPROM AM defects. TGFβ₁ and Cx43 gene expression were significantly increased after trauma and mechanical stimulation but this response dependent on gestational age.

Conclusion

Mechanical stimulation affects Cx43 signalling and cell/collagen mechanics in preterm AM defects. Establishing how Cx43 regulates mechanosignalling could be an approach to repair tissue integrity after trauma.