Effects of muscular strength training and growth hormone (GH) supplementation on femoral bone tissue: analysis by Raman spectroscopy, dual-energy X-ray absorptiometry, and mechanical resistance.

The aim of the present study was to verify the effects of muscular strength training and growth hormone (GH) supplementation on femoral bone tissue by Raman spectroscopy (Raman), dual-energy X-ray absorptiometry (DXA), and mechanical resistance (F-max) analysis. A total of 40 male Wistar animals, 60 days old, were used. The animals were distributed into four groups: control (C), control with GH (GHC), muscular strength training (T), and muscular strength training with GH (GHT). Blood samples were collected for the quantification of creatine kinase (CK-MB) and the femurs were removed for analysis by Raman, DXA, and F-max. A more pronounced increase in the bone mineral components was verified in the T group, for all the variables obtained by the Raman (calcium, phosphate, amide, and collagen). In addition, for animals submitted to GH supplementation, there was a reduction in the variable bone mineral density (BMD) obtained by the DXA (p < 0.05). Finally, the animals that received GH supplementation presented a higher F-max, but without statistical significance (p > 0.05). It was concluded that animals that received GH supplementation demonstrated a decrease in BMD. In addition, T alone was able to promote increased calcium, phosphate, amide, and collagen compounds in bone tissue.

Effects of precipitation variability on carbon and water fluxes in the understorey of a nitrogen-limited montado ecosystem.

To date the implications of greater intra-annual variability and extremes in precipitation on ecosystem functioning have received little attention. This study presents results on soil and vegetation carbon and water fluxes in the understorey of a Mediterranean oak woodland in response to increasing precipitation variability, with an extension of the dry period between precipitation events from 3 to 6 weeks, without altering total annual precipitation inputs. With prolonged dry periods soil moisture did breach the stress thresholds for ecosystem processes, which led to short-term treatment differences in photosynthesis, but not in system carbon losses, with subsequent short-term decreases in net ecosystem exchange. Independent of treatment, irrigation events rapidly increased carbon and water fluxes. However, contradicting the predictions drawn from the 'bucket model', over the course of the growing season no all-over treatment differences were found in system assimilation and respiration, nor in evapotranspiration and ecosystem water use efficiency. This lack of responsiveness is attributed to the ecosystem's resilience to low soil moisture during the growing season of the herbaceous understorey, with temperature rather than soil moisture controlling key ecosystem processes. Moreover, severe nitrogen limitation of the studied ecosystem may explain the lack of moisture effects on net system carbon dynamics. Thus, although the bucket model predicts changes in soil water dynamics with increasing precipitation variability, ecosystem responses to more extreme precipitation regimes may be influenced by additional factors, such as inter-annual variability in nutrient availability.

The impact of drought on leaf physiology of Quercus suber L. trees: comparison of an extreme drought event with chronic rainfall reduction.

Understanding the responses of cork oak (Quercus suber L.) to actual and predicted summer conditions is essential to determine the future sustainability of cork oak woodlands in Iberia. Thermal imaging may provide a rapid method for monitoring the extent of stress. The ecophysiology of cork trees was studied over three years. Three treatments were applied by means of rainfall capture and irrigation, with plots receiving 120%, 100%, or 80% of natural precipitation. Despite stomatal closure, detected using both thermal imaging and porometry, leaf water potential fell during the summer, most drastically during the third year of accumulative stress. The quantum efficiency (ΦPSII) and the maximum efficiency Fv' /FM' of photosystem II also fell more intensely over the third summer, while non-photochemical quenching (NPQ) increased. The reduced precipitation treatment sporadically further reduced leaf water potential, stomatal conductance (gs), IG (an index of gs derived from thermal imaging), ΦPSII, and Fv' /FM', and increased leaf temperature and NPQ. It is concluded that these are very resilient trees since they were only severely affected in the third year of severe drought (the third year registering 45% less rainfall than average), and removing 20% of rainfall had a limited impact..

Leiomyomatosis peritonealis disseminata: a rare disease with a difficult diagnosis.

Leiomyomatosis peritonealis disseminata (LPD) is a rare condition, characterised by the proliferation of peritoneal smooth muscle nodules. LPD is a benign disease with a low rate of malignant degeneration. We describe the case of a 46-year-old, asymptomatic, woman presenting with a mass on the left renal hilum, identified by ultrasound. A CT scan showed three nodules near the left kidney, a mass anterior to the vena cava and bilateral iliac nodules. Biopsy revealed a mesenchymal low-grade tumour. The patient underwent a left nephrectomy and excision of the other masses. The histological diagnosis revealed smooth muscle nodular proliferation with no malignant features, compatible with LPD. The differential diagnosis between LPD and metastatic leiomyosarcoma is sometimes very difficult because they are clinically very similar and even on histology the diagnosis can be tricky. Treatment is conservative in most cases, with surgical excision reserved for high-risk patients.

Global genetic analyses reveal strong inter-ethnic variability in the loss of activity of the organic cation transporter OCT1.

BACKGROUND: The organic cation transporter OCT1 (SLC22A1) mediates the uptake of vitamin B1, cationic drugs, and xenobiotics into hepatocytes. Nine percent of Caucasians lack or have very low OCT1 activity due to loss-of-function polymorphisms in OCT1 gene. Here we analyzed the global genetic variability in OCT1 to estimate the therapeutic relevance of OCT1 polymorphisms in populations beyond Caucasians and to identify evolutionary patterns of the common loss of OCT1 activity in humans. METHODS: We applied massively parallel sequencing to screen for coding polymorphisms in 1,079 unrelated individuals from 53 populations worldwide. The obtained data was combined with the existing 1000 Genomes data comprising an additional 1,092 individuals from 14 populations. The identified OCT1 variants were characterized in vitro regarding their cellular localization and their ability to transport 10 known OCT1 substrates. Both the population genetics data and transport data were used in tandem to generate a world map of loss of OCT1 activity. RESULTS: We identified 16 amino acid substitutions potentially causing loss of OCT1 function and analyzed them together with five amino acid substitutions that were not expected to affect OCT1 function. The variants constituted 16 major alleles and 14 sub-alleles. Six major alleles showed improper subcellular localization leading to substrate-wide loss in activity. Five major alleles showed correct subcellular localization, but substrate-specific loss of activity. Striking differences were observed in the frequency of loss of OCT1 activity worldwide. While most East Asian and Oceanian individuals had completely functional OCT1, 80 % of native South American Indians lacked functional OCT1 alleles. In East Asia and Oceania the average nucleotide diversity of the loss-of-function variants was much lower than that of the variants that do not affect OCT1 function (ratio of 0.03) and was significantly lower than the theoretically expected heterozygosity (Tajima's D = -1.64, P < 0.01). CONCLUSIONS: Comprehensive genetic analyses showed strong global variations in the frequency of loss of OCT1 activity with selection pressure for maintaining OCT1 activity in East Asia and Oceania. These results not only enable pharmacogenetically-based optimization of drug treatment worldwide, but may help elucidate the functional role of human OCT1.

The poorly membrane permeable antipsychotic drugs amisulpride and sulpiride are substrates of the organic cation transporters from the SLC22 family.

Variations in influx transport at the blood-brain barrier might affect the concentration of psychotropic drugs at their site of action and as a consequence might alter therapy response. Furthermore, influx transporters in organs such as the gut, liver and kidney may influence absorption, distribution, and elimination. Here, we analyzed 30 commonly used psychotropic drugs using a parallel artificial membrane permeability assay. Amisulpride and sulpiride showed the lowest membrane permeability (P e < 1.5 × 10(-6) cm/s) and will require influx transport to penetrate the blood-brain barrier and other physiological barriers. We then studied the uptake of amisulpride and sulpiride by the organic cation transporters of the SLC22 family OCT1, OCT2, OCT3, OCTN1, and OCTN2 Amisulpride was found to be transported by all five transporters studied. In contrast, sulpiride was only transported by OCT1 and OCT2. OCT1 showed the highest transport ability both for amisulpride (CLint = 1.9 ml/min/mg protein) and sulpiride (CLint = 4.2 ml/min/mg protein) and polymorphisms in OCT1 significantly reduced the uptake of both drugs. Furthermore, we observed carrier-mediated uptake that was inhibitable by known OCT inhibitors in the immortalized human brain microvascular endothelial cell line hCMEC/D3. In conclusion, this study demonstrates that amisulpride and sulpiride are substrates of organic cation transporters of the SLC22 family. SLC22 transporters may play an important role in the distribution of amisulpride and sulpiride, including their ability to penetrate the blood-brain barrier.

Morphine is a substrate of the organic cation transporter OCT1 and polymorphisms in OCT1 gene affect morphine pharmacokinetics after codeine administration.

We investigated whether morphine and its pro-drug codeine are substrates of the highly genetically polymorphic organic cation transporter OCT1 and whether OCT1 polymorphisms may affect morphine and codeine pharmacokinetics in humans. Morphine showed low transporter-independent membrane permeability (0.5 × 10⁻6 cm/s). Morphine uptake was increased up to 4-fold in HEK293 cells overexpressing human OCT1. The increase was concentration-dependent and followed Michaelis-Menten kinetics (KM = 3.4 µM, VMAX = 27 pmol/min/mg protein). OCT1-mediated morphine uptake was abolished by common loss-of-function polymorphisms in the OCT1 gene and was strongly inhibited by drug-drug interactions with irinotecan, verapamil and ondansetron. Morphine uptake in primary human hepatocytes was strongly reduced by MPP⁺, an inhibitor of organic cation transporters, and morphine was not a substrate of OCT3, the other organic cation transporter expressed in human hepatocytes. In concordance with the in vitro data, morphine plasma concentrations in healthy volunteers were significantly dependent on OCT1 polymorphisms. After codeine administration, the mean AUC of morphine was 56% higher in carriers of loss-of-function OCT1 polymorphisms compared to non-carriers (P = 0.005). The difference remained significant after adjustment for CYP2D6 genotype (P = 0.03). Codeine itself had high transporter-independent membrane permeability (8.2 × 10⁻6 cm/s). Codeine uptake in HEK293 cells was not affected by OCT1 overexpression and OCT1 polymorphisms did not affect codeine AUCs. In conclusion, OCT1 plays an important role in the hepatocellular uptake of morphine. Carriers of loss-of-function OCT1 polymorphisms may be at higher risk of adverse effects after codeine administration, especially if they are also ultra-rapid CYP2D6 metabolizers.