Summary
Brassinosteroids (BRs) are a group of plant steroid hormones involved in regulating growth, development, and stress responses. Many components of the BR pathway have previously been ...identified and characterized. However, BR phenotyping experiments are typically performed in a low‐throughput manner, such as on Petri plates. Additionally, the BR pathway affects drought responses, but drought experiments are time consuming and difficult to control. To mitigate these issues and increase throughput, we developed the Robotic Assay for Drought (RoAD) system to perform BR and drought response experiments in soil‐grown Arabidopsis plants. RoAD is equipped with a robotic arm, a rover, a bench scale, a precisely controlled watering system, an RGB camera, and a laser profilometer. It performs daily weighing, watering, and imaging tasks and is capable of administering BR response assays by watering plants with Propiconazole (PCZ), a BR biosynthesis inhibitor. We developed image processing algorithms for both plant segmentation and phenotypic trait extraction to accurately measure traits including plant area, plant volume, leaf length, and leaf width. We then applied machine learning algorithms that utilize the extracted phenotypic parameters to identify image‐derived traits that can distinguish control, drought‐treated, and PCZ‐treated plants. We carried out PCZ and drought experiments on a set of BR mutants and Arabidopsis accessions with altered BR responses. Finally, we extended the RoAD assays to perform BR response assays using PCZ in Zea mays (maize) plants. This study establishes an automated and non‐invasive robotic imaging system as a tool to accurately measure morphological and growth‐related traits of Arabidopsis and maize plants in 3D, providing insights into the BR‐mediated control of plant growth and stress responses.
Significance Statement
RoAD is an automated phenotyping system for assessing brassinosteroid and drought responses in soil‐grown Arabidopsis plants. The system performs daily weighing, watering, and imaging tasks and is capable of accurately measuring morphological and growth‐related traits of Arabidopsis in 3D.
Summary
The aims of this study were to determine concentrations of total homocysteine, cysteine, cysteinylglycine and glutathione in spermatozoa, seminal fluid and blood plasma and to analyse their ...relationships with sperm parameters. For this reason, a new highly effective method of spermatozoa lysis was developed, using methanol, freezing and subsequent thawing in ultrasonic bath. An HPLC‐FD assay was conducted on thiols concentrations in lysed spermatozoa, seminal fluid and blood plasma. Concentrations of thiols in spermatozoa were significantly lower in men with normozoospermia than in samples with pathological semen parameters. Statistical analysis found significant correlations between thiol concentrations in spermatozoa and semen parameters, while the same analysis with thiol concentrations in seminal fluid was substantially less powerful. Only cysteinylglycine concentrations in seminal fluid significantly correlated with pathological semen parameters. No significant differences or correlations were found with blood plasma concentrations.
Raf‐1 protein kinase has been identified as an integral component of the Ras/Raf/MEK/ERK signalling pathway in mammals. Activation of Raf‐1 is achieved by Ras.GTP binding and other events at the ...plasma membrane including tyrosine phosphorylation at residues 340/341. We have used gene targeting to generate a ‘knockout’ of the raf‐1 gene in mice as well as a rafFF mutant version of endogenous Raf‐1 with Y340FY341F mutations. Raf‐1−/− mice die in embryogenesis and show vascular defects in the yolk sac and placenta as well as increased apoptosis of embryonic tissues. Cell proliferation is not affected. Raf‐1 from cells derived from raf‐1FF/FF mice has no detectable activity towards MEK in vitro, and yet raf‐1FF/FF mice survive to adulthood, are fertile and have an apparently normal phenotype. In cells derived from both the raf‐1−/− and raf‐1FF/FF mice, ERK activation is normal. These results strongly argue that MEK kinase activity of Raf‐1 is not essential for normal mouse development and that Raf‐1 plays a key role in preventing apoptosis.
Abstract Purpose Reliable18 F-fluorodeoxyglucose (FDG) uptake quantification is crucial for cancer treatment monitoring. While interobserver variability has been found to be lower for a maximum ...standard uptake value (SUV)max than for an averaged SUV (SUVmean ), the repeatability has not been investigated yet. In this study, we determined the repeatability of SUV values in two sequential measurements 5 min apart. Methods Positron emission tomography data of malignant chest tumors were acquired dynamically during 45 min in 20 patients. SUV values were derived from the hottest (SUVmax ), the mean of the 5 (SUV5 ) and 10 (SUV10 ) hottest voxels and the mean of a volume of interest (SUVmean ). The repeatability of the SUV measurements was determined as the standard deviation of the difference between the values at 40 and 45 min and represented as Bland–Altman graphs. Results The standard deviation of the difference between the two sequential scans for SUVmax , SUV5 , SUV10 and SUVmean was 1.01, 0.53, 0.37 and 0.28. Conclusion The repeatability of SUV is markedly increased by deriving the value from multiple voxels. Compared to SUVmax , the variability in SUV measurements is reduced by a factor of 2.7 (2.7=1.01/0.37) if 10 voxels are pooled.
Purpose
Aim of this prospective observational study was to analyze fertility status of Hodgkin lymphoma (HL) patients treated with different types of chemotherapy while receiving GnRH analogues to ...preserve ovarian function.
Methods
Fertility status was assessed among 108 females in reproductive age treated by curative chemotherapy for freshly diagnosed HL between 2005 and 2010 in university-based tertiary fertility and oncology center. All patients received GnRH analogues during chemotherapy to preserve their ovarian function. Their reproductive functions were assessed by follicle-stimulating hormone (FSH) measurement and pregnancy achievement. Ovarian function was determined separately in three groups with increasing gonadotoxicity of chemotherapy.
Results
One year following the treatment, normal ovarian function was found in 89 (82.4 %) of patients. Two years after chemotherapy, 98 (90.7 %) of patients retained their ovarian function, and 23 (21.3 %) achieved clinical pregnancy during the follow-up period. Average FSH after chemotherapy was 11.6 ± 17.9 IU/l 1 year after the treatment resp. 9.0 ± 13.8 at the 2 years interval. There were significantly more patients with chemotherapy induced diminished ovarian reserve (chDOR) among the group receiving escalated BEACOPP chemotherapy in comparison with the other types of treatment (58.1 % vs. 87.9 % resp. 95.5 %).
Conclusion
The rate of chDOR is significantly higher after EB poly-chemotherapy and there is no tendency for improvement in time. The 2 + 2 chemotherapy with GnRH-a required for more advanced HL retained ovarian function significantly better after 2 years. Another important advantage of GnRH-a co-treatment is the excellent control of patient’s menstrual cycle.
BACKGROUND Frequent negative consequence of chemotherapy (CHT) is ovarian damage and premature ovarian failure (POF). Aim of this prospective case–control study is evaluation of GnRH analogue ...(GnRH-a) administration to patients with Hodgkin lymphoma (HL) during CHT and prevention of ovarian damage depending upon CHT regimen. METHODS Study group consists of 72 patients in fertile age (18–35 years) with HL diagnosis treated in 2004–2005 by curative CHT together with GnRH analogue (Triptorelin) administration according to a standardized protocol. Patients were divided into three groups according to the stage of disease and treated by three types of CHT regimens (A,B,C) with increased cytotoxicity. Ovarian function of all patients was assessed by gonadotrophin levels (FSH, LH) analysis from peripheral blood before treatment and also 6 and 12 month after it. The number of women with POF after CHT in study group was compared with control group (n = 45, age 18–35 years) of patients treated in 2002–2003 according to the same protocol but without protective GnRH analogue application. RESULTS In study group with GnRH analogue administration during CHT, there was significantly (P < 0.001) fewer cases with POF 6 and 12 month after the end of CHT (37.5% and 20.8%, respectively) than in control group (73.3% and 71.1%, respectively). Comparative analysis depending on cytotoxicity of CHT regimen used showed significant differences in percentage of patient with acquired POF between study and control group only in less aggressive CHT protocols. CONCLUSIONS Study showed a significant reduction of ovarian failure risk in women with HL treated with less aggressive CHT regimens plus a GnRH analogue.
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