HeLa cell nucleus, a source of thymidine for Trichomonas vaginalis growing in vitro

Trichomonas vaginalis is a parasitic protist incapable of de novo purine and pyrimidine biosynthesis. The lack of these de novo syntheses of nucleotides is supplemented with purine and pyrimidine salvage pathways. Likewise, T. vaginalis is incapable of converting its ribonucleotides to deoxyribonucleotides. Therefore, the parasite must rely on the salvage of exogenous deoxyribonucleosides for DNA synthesis. It has been demonstrated that the parasite can incorporate external adenine and guanine in vitro, but no in vivo nucleotide source has been identified so far. Accordingly, we set out to determine if the parasite could incorporate 3H-thymidine from the nuclei of a cervical-derived cell line into its own DNA. By light and electron microscopy we found that the parasite was able to interact directly, both with mechanically isolated HeLa cell nuclei and with the nuclei released after the disruption of HeLa cell monolayers by the parasite. This study shows that T. vaginalis was capable of incorporating 3H-thymidine from labeled HeLa cells into its own DNA suggesting that the nuclei of this cervical cell line could be an in vivo source of nucleotides for T. vaginalis.     

Acute noise stress analgesia in relation to 5-HT2 and mu-opioid receptor changes in the frontal cortex of young mice

A number of studies have reported that exposure to stress provoked behavioural changes, including analgesia, in rodents. Differences have been observed in these responses to different types of stress and a link between hormones and neurotransmitters proposed. We studied the effect of acute noise stress on nociception and the possible changes in the serotonergic and opioidergic systems in young mice of both sexes. Naloxone pre-treatment was also investigated. Noise stress was produced by a sound source, nociception was measured by the hot-plate test and binding characteristics were evaluated by a radioligand binding technique using membrane preparation from the total frontal cortex. Acute noise stress provoked an antinociceptive effect, associated with an increase in plasma corticosterone levels, a decrease in the number of 5-HT2 receptors in stressed male and female mice and a decrease in the number of mu receptors in both sexes. The behavioural and biochemical effects were antagonized by 1 mg/kg of naloxone. Acute noise stress behaves like other types of stress on nociception. The opioidergic system seems to be involved in this behaviour but also the serotonergic system may play a role. Sex differences were detected in the number of 5-HT2 and mu receptors between male and female mice not subjected to stress, while the percentage decrease in 5-HT2 and mu receptors did not differ significantly between the two sexes.     

Mechanical Stress Downregulates MHC Class I Expression on Human Cancer Cell Membrane

In our body, cells are continuously exposed to physical forces that can regulate different cell functions such as cell proliferation, differentiation and death. In this work, we employed two different strategies to mechanically stress cancer cells. The cancer and healthy cell populations were treated either with mechanical stress delivered by a micropump (fabricated by deep X-ray nanolithography) or by ultrasound wave stimuli. A specific down-regulation of Major Histocompatibility Complex (MHC) class I molecules expression on cancer cell membrane compared to different kinds of healthy cells (fibroblasts, macrophages, dendritic and lymphocyte cells) was observed, stimulating the cells with forces in the range of nano-newton, and pressures between 1 and 10 bar (1 bar = 100.000 Pascal), depending on the devices used. Moreover, Raman spectroscopy analysis, after mechanical treatment, in the range between 700–1800 cm⁻¹, indicated a relative concentration variation of MHC class I. PCA analysis was also performed to distinguish control and stressed cells within different cell lines. These mechanical induced phenotypic changes increase the tumor immunogenicity, as revealed by the related increased susceptibility to Natural Killer (NK) cells cytotoxic recognition.     

Diffusion on Membrane Domes,Tubes, and Pearling Structures

Diffusion is a fundamental mechanism for protein distribution in cell membranes. These membranes often exhibit complex shapes, which range from shallow domes to elongated tubular or pearl-like structures. Shape complexity of the membrane influences the diffusive spreading of proteins and molecules. Despite the importance membrane geometry plays in these diffusive processes, it is challenging to establish the dependence between diffusion and membrane morphology. We solve the diffusion equation numerically on various static curved shapes representative for experimentally observed membrane shapes. Our results show that membrane necks become diffusion barriers. We determine the diffusive half-time, i.e., the time that is required to reduce the amount of protein in the budded region by one half, and find a quadratic relation between the diffusive half-time and the averaged mean curvature of the membrane shape, which we rationalize by a scaling law. Our findings thus help estimate the characteristic diffusive timescale based on the simple measure of membrane mean curvature.     

Ontogeny of the digestive system and the profile of proteases in larvae of cachara (Pseudoplatystoma reticulatum Siluriformes: Pimelodidae) and its hybrid (Pseudoplatystoma corruscans × Pseudoplatystoma reticulatum)

The development of the digestive system and digestive proteases was studied in cachara (Pseudoplatystoma reticulatum) and its hybrid (P. reticulatum × Pseudoplatystoma corruscans) during the first 25 days after fertilization. Both groups presented a fast and similar development of the digestive system, and the unspecific alkaline protease profile suggested that the digestive capacity of hybrid larvae is higher than that of cachara after the last half of the studied phase. The activities of trypsin and chymotrypsin were similar and observed since the oocyte. These observations suggested that cachara and its hybrid has similar digestive capacity and digestive system development.     

Effect of age, family history of prostate cancer, prostate enlargement and seasonality on PSA levels in a contemporary cohort of healthy Italian subjects

We assessed the joint effect of age at enrollment, age at follow-up, family history of prostate cancer, prostate enlargement and seasonality on prostate-specific antigen (PSA) estimated through log-normal mixed-effects modeling in an Italian cohort of healthy, 45- to 65-year-old subjects over a 4-year period. The median ratio was used as the measure of effect. Median and mean baseline PSA were 0.78 (interquartile range: 0.41-1.50) and 1.27 (95% CI: 1.19-1.35) ng/mL, respectively. A similar median annual increase of 5.7% (95% CI: 4.8%-6.5%) was found for age at enrollment and age at follow-up. Individuals with moderate to severe prostate enlargement had a median PSA ratio of 1.040 (95% CI: 0.919-1.176) and 1.318 (95% CI: 1.128-1.539), respectively. Median ratios of 1.200 (95% CI: 0.026-1.404) and 1.300 (95% CI: 0.915-1.845), respectively, were computed for subjects with only one or more than one prostate-cancer-affected relatives. Regarding seasonality, the highest value was shown in summertime, the lowest in wintertime, and the resulting median ratio was 1.280 (95% CI: 1.117-1.468). Irrespective of age, baseline PSA was in most cases about 1.00 ng/mL with a yearly median variation of about 5% over a 4-year period. Indeed, prostate enlargement, prostate cancer family history and seasonality showed a remarkable impact on PSA measurement. This should be considered when counseling patients with a PSA history.     

Supratidal foraminifera as ecological indicators in anthropically modified wetlands (Lagoon of Venice, Italy)

In transitional environments, the intertidal zones represent a peculiar case characterized by halophile vegetation and by a low diversity benthic community. On these areas just a few particular foraminiferal species, a class of Protoctista secreting a shell called test, can survive for a certain time out of water. They are distributed in well-defined vertical zonations with respect to mean sea level and they correspond to analogous marsh floral zonations. In particular, the Trochammina macrescens Brady + Trochammina inflata (Montagu) association characterizes the salt marsh zone above mean high water level. The potential of these taxa as bioindicators is tested, since their presence-absence-dominance differentiates the subtidal/supratidal environments. Over the last few centuries, various engineering works generated major physical changes in the Venetian Lagoon. These changes affected the natural evolution of the intertidal morphologies, the surface of which is decreasing. In an attempt to reverse this tendency, numerous artificial salt marshes have been constructed and more are under construction. In this study, the Mazzorbo artificial salt marsh, built during the second half of 1999, is considered. On its surface, 16 samples were collected along a transect line in May 2008 to verify the ecological role of this salting within the lagoon ecosystem. The sediment grain size distribution of the salt marsh reflects the dissipative role of the tide and the effect of sediment transport due to the wave and tidal action. However, the presence of only a few Trochammina individuals shows that the foraminiferal fauna did not recognise this morphology as a salt marsh. The lack of Trochammina colonisation can be related to the excessive elevation of the salt marsh surface. This hypothesis is confirmed by the lack of the salt-tolerant plant Spartina. The unsuccessful colonisation by the foraminifera seems to indicate that this artificial salting does not have the natural dynamism of the intertidal morphologies and it may only be classified as land recovery. The supratidal foraminiferal taxa can act as an ecological indicator: through their observation it is possible to verify whether an artificial salt marsh accomplishes its task of functioning as an ecological unit with the community of organisms.