The Effects of Isoflavones on Androgens and Glucocorticoids During Puberty on Male Wistar Rats

Isoflavones are the most common phytoestrogens found in human diets. However, it is still not clear whether isoflavones have effects on the reproductive and the endocrine systems under normal dietary intake and overdose. The aim of this study was to determine how the most important isoflavones, genistein and daidzein, affect androgen and glucorticoid levels on male prepuberal rats. A hundred and seventy‐five 30‐day‐old male Wistar rats were dosed orally by stomach tube every day for 35 days, with saline solution, low and high doses of genistein, daidzein and a mixture of both. Serum samples were analysed by an enzyme immunoassay for hormone determinations. In control group, there was a peak of testosterone (T) and dihydrotestosterone levels associated to the onset of puberty, at the third week. However, in low‐dose groups, the same peak was found at the fourth week (p < 0.05), indicating a delay in the onset of puberty in these groups. Moreover, high doses groups serum androgen levels were significantly lower (p < 0.05) than the control group from the first week until fifth week. This fact was supported by a epididymal histological analysis that indicate in low doses there were several content of spermatozoa at fourth week and in high doses there were few content of spermatozoa. Besides, corticosterone levels followed the same pattern of androgens in all groups. We can conclude that oral administration of isoflavones in male rats decreased the secretion of androgens and glucocorticoids causing a delay in the onset of puberty and may cause physiological and developmental problems.     

Force-clamp spectroscopy monitors the folding trajectory of a single protein

We used force-clamp atomic force microscopy to measure the end-to-end length of the small protein ubiquitin during its folding reaction at the single-molecule level. Ubiquitin was first unfolded and extended at a high force, then the stretching force was quenched and protein folding was observed. The folding trajectories were continuous and marked by several distinct stages. The time taken to fold was dependent on the contour length of the unfolded protein and the stretching force applied during folding. The folding collapse was marked by large fluctuations in the end-to-end length of the protein, but these fluctuations vanished upon the final folding contraction. These direct observations of the complete folding trajectory of a protein provide a benchmark to determine the physical basis of the folding reaction.     

Assessment of neutrophil function in canine cancer patients undergoing chemotherapy and correlation with neutrophil numbers

Decreased neutrophil function following administration of chemotherapy has been reported in dogs with lymphoma. The first objective of our study was to determine if neutrophil oxidative burst and phagocytic activity are affected by chemotherapy 7 to 10 days following initiation of treatment in dogs with lymphoma and non-lymphoma malignancies. The second objective was to determine if there is a correlation between neutrophil numbers and neutrophil function before or after initiation of chemotherapy. Flow cytometric assessment of neutrophil oxidative burst and phagocytosis following stimulation with Escherichia coli was performed in 9 dogs diagnosed with lymphoma and 17 non-lymphoma tumor-bearing dogs pre- and post-chemotherapy, as well as 14 tumor-free control dogs. Spearman rank correlation was performed to determine if blood neutrophil numbers and neutrophil function were significantly correlated. Lymphoma patients showed significantly reduced percentage neutrophil oxidative burst post-chemotherapy compared to healthy controls as well as compared to pre-chemotherapy values (P = 0.0022 and P = 0.0020, respectively). Lymphoma patients also exhibited significantly reduced neutrophil phagocytosis activity post-chemotherapy compared to controls and pre-chemotherapy values (P = 0.0016 and P = 0.014, respectively). Dogs with non-lymphoma malignancies also showed a significant decrease in both percentage oxidative burst and phagocytosis post-chemotherapy compared to pre-chemotherapy values (P = 0.00040 and P = 0.029, respectively). Neutrophil numbers and function were not significantly correlated. The results of the study suggest that chemotherapeutic treatment decreases neutrophil oxidative burst and phagocytic activity 7 to 10 days post-treatment in dogs with various malignancies. Furthermore, neutrophil numbers cannot be used to predict neutrophil function.     

Predicting likelihood of in vivo chemotherapy response in canine lymphoma using ex vivo drug sensitivity and immunophenotyping data in a machine learning model

We report a precision medicine platform that evaluates the probability of chemotherapy drug efficacy for canine lymphoma by combining ex vivo chemosensitivity and immunophenotyping assays with computational modelling. We isolated live cancer cells from fresh fine needle aspirates of affected lymph nodes and collected post‐treatment clinical responses in 261 canine lymphoma patients scheduled to receive at least 1 of 5 common chemotherapy agents (doxorubicin, vincristine, cyclophosphamide, lomustine and rabacfosadine). We used flow cytometry analysis for immunophenotyping and ex vivo chemosensitivity testing. For each drug, 70% of treated patients were randomly selected to train a random forest model to predict the probability of positive Veterinary Cooperative Oncology Group (VCOG) clinical response based on input variables including antigen expression profiles and treatment sensitivity readouts for each patient's cancer cells. The remaining 30% of patients were used to test model performance. Most models showed a test set ROC‐AUC > 0.65, and all models had overall ROC‐AUC > 0.95. Predicted response scores significantly distinguished (P < .001) positive responses from negative responses in B‐cell and T‐cell disease and newly diagnosed and relapsed patients. Patient groups with predicted response scores >50% showed a statistically significant reduction (log‐rank P < .05) in time to complete response when compared to the groups with scores <50%. The computational models developed in this study enabled the conversion of ex vivo cell‐based chemosensitivity assay results into a predicted probability of in vivo therapeutic efficacy, which may help improve treatment outcomes of individual canine lymphoma patients by providing predictive estimates of positive treatment response.     

Fate of airborne metal pollution in soils as related to agricultural management. 1. Zn and Pb distributions in soil profiles

The fate of airborne metal pollutants in soils is still relatively unknown. We studied the incorporation of such airborne metal pollution in two soils under long-term permanent pasture (PP) and conventional arable land (CA). Both soils were located at an almost equal distance from a former zinc smelter complex and developed under comparable pedogenetic conditions. Profiles of total concentrations of Zn, chosen as a mobile, and Pb as a little- or non-mobile element, were examined and compared with macro- and micromorphological soil characteristics (soil colour, biological activity). The two soils showed different profiles of total Zn and Pb concentrations, with a marked decrease of concentrations of both elements under the plough layer in CA, whereas the decrease was more progressive in PP. However, the stocks of Zn and Pb for the 1-m soil profiles of CA and PP were comparable. Correlation of Zn and Pb concentration at different depths with total Fe contents and comparison with estimated data for the local geochemical background (LGCB), suggests transport of Zn from the surface to depth in CA and PP, and Pb movement in PP. In CA, 53% of Zn and 92.5% of Pb stocks derived from airborne metal pollution were located at depths < 26 cm. In PP, only 40% of Zn and 82% of Pb, derived from airborne pollution, were found in the A11 and A12 horizons (< 26 cm), the remaining 18% of the Pb stock being incorporated until 50 cm depth; one-third of total Zn stock ascribed to airborne pollution was found at depths > 50 cm. Studies of the composition of gravitational water collected in soils from the same study area suggest two mechanisms for metal movement. First, mobile metal ions (Zn²⁺) move in the soil solution and are intercepted by iron-clay complexes in deeper soil horizons. Second, observed only in PP, simultaneous movement of Zn and Pb is ascribed to bioturbation by earthworms.     

Isolation of amoebae of the genera Naegleria and Acanthamoeba from public fountains in Galicia (N.W. Spain)

Tests for free-living amoebae in water from 11 public fountains in Galicia were conducted at two seasons of the year, winter and summer. Only one fountain gave a negative result for the presence of amoebae in both samples. Twenty five strains were isolated, 14 belonging to the genus Acanthamoeba and 10 to the genus Naegleria. The Acanthamoeba strains were made up of the species A. polyphaga, A. quina, A. castellanii and A. paradivionensis. No relation was found between temperature, presence of free chlorine in the water and whether the water was drinkable and the presence or absence of amoebae, nor with the number of strains present in the water samples. The pathogenic capacity of the strains isolated from the genera Naegleria and Acanthamoeba was tested in vivo. It was found that of the 22 strains that would grow at 37 °C, 4 (3 strains of Acanthamoeba and 1 of Naegleria) caused the death of a statistically significant number of mice that had been inoculated intracerebrally, and the presence of amoebae was confirmed in the brains of all the animals inoculated. Intranasal inoculation caused less mice deaths than intracerebral inoculation, and less organs were found containing amoebae.     

Analysis of host genotype x rhizobial strain interaction in N2 fixation

Summary A method of analyzing host genotype x rhizobial strain interaction for N₂ fixation potential, based on the principle of structural relationship analysis, is proposed. When this interaction is significant, selecting genotypes for high N₂ fixation potential is incorrect, because the N₂ fixation efficiency is dependent on the rhizobial strain used. Under such circumstances, grouping genotypes based on average fixing ability (AFA), a linear response to the effectiveness of rhizobial strain, and specific fixing ability (SFA), deviation from the linear response in terms of the magnitude of the error variance, is useful before initiating a breeding program for enhanced N₂ fixation. Host genotype x strain interaction effect is partitioned into two components, and two parameters are derived which estimate AFA and SFA. N₂ fixation data from 3 mungbean genotypes and 6 rhizobial strains are used to illustrate this method.     

Shifting undesirable correlations

Wheat functions as a complex well-integrated organism with adaptation to many environments. Traits of wheat may be correlated to each other in a positive or a negative manner. The direction of the correlation is independent of breeding objectives and may change from one production environment to another. Grain yield is correlated positively with time to maturity in environments without terminal stress. The negative correlation between grain yield and grain protein concentration exemplifies an undesirable relationship in a wheat quality type in which protein concentration is positively correlated with bread loaf volume. However, this same correlation has an advantage to those end-products that are favored by low protein concentration such as confectionary products. To shift the negative correlation between grain yield and protein concentration requires assembling a more photosynthetically efficient, nitrogen-use efficient and\or water-use efficient genotype in combination or separately. Simultaneous selection for quantitative and quality traits is a breeding strategy, which with the inclusion of marker assisted selection, can shift undesirable correlations.     

Are stock assessment methods too complicated?

This critical review argues that several methods for the estimation and prediction of numbers‐at‐age, fishing mortality coefficients F, and recruitment for a stock of fish are too hard to explain to customers (the fishing industry, managers, etc.) and do not pay enough attention to weaknesses in the supporting data, assumptions and theory. The review is linked to North Sea demersal stocks. First, weaknesses in the various types of data used in North Sea assessments are summarized, i.e. total landings, discards, commercial and research vessel abundance indices, age‐length keys and natural mortality (M). A list of features that an ideal assessment should have is put forward as a basis for comparing different methods. The importance of independence and weighting when combining different types of data in an assessment is stressed. Assessment methods considered are Virtual Population Analysis, ad hoc tuning, extended survivors analysis (XSA), year‐class curves, catch‐at‐age modelling, and state‐space models fitted by Kalman filter or Bayesian methods. Year‐class curves (not to be confused with ‘catch‐curves’) are the favoured method because of their applicability to data sets separately, their visual appeal, simple statistical basis, minimal assumptions, the availability of confidence limits, and the ease with which estimates can be combined from different data sets after separate analyses. They do not estimate absolute stock numbers or F but neither do other methods unless M is accurately known, as is seldom true.