Cloning characterization and expression of a novel metallothionein gene (mt-d) from triticum durum

Two different metallothionein genes, labelled as mt-d and mt-a were identified in wheat (Triticum durum and Triticum aestivum) genomic DNA sequences were characterized. mt-d and mta, were found to contain 416 and 399 nucleotides, respectively. Nucleic acid sequence alignment showed 95 % similarity between the two. Sequencing results showed that the difference resulted from two extra TTTTTA repeats in the intron regions. cDNAs encoding mt-genes were identifed by RT-PCR. Gene alignment algorithms strongly suggested that both of these cDNAs (mt-a and mt-d) encoded an open reading frame of 75 amino acids with two cysteine-rich domains featuring Cys-XCys motifs at the amino- and carboxy termininus. The deduced amino acid sequences of mt-a and mt-d genes show striking similarity to the MT-like proteins described within the Class II as Type 1 MTs and showed 100 % similarity with each other as deduced from cDNA sequencing results. These results indicate that mt-d from T.durum forms a “novel type 1” MT. For further studies of mt-d expression, localization of the durum metallothionein protein (dMT) and its interactions with other proteins mt-d gene was inserted into the 5’ MCS of pGFPuv vector. Verification was based on sequence data and restriction enzyme analysis. However, expression could not be validated by neither by visual detection of GFP expression nor by SDS-PAGE analysis. A more detailed sequence analysis indicated that the problem was due to a point mutation within the coding sequence of the GFPuv, resulting in a stop codon and premature termination of the fusion protein. Results presented here show the presence of metallothionein gene in the wheat Triticum durum. Although our attempts to express the gene as a fusion protein together with GFP to facilitate its localization in different systems was not successful it will be important in future studies to pursue this goal and achieve expression of labelled protein in plant systems to gain insights into its exact function in plants

Serum progranulin levels are elevated in infertile women with obesity

Introduction: The aim of the present study was to investigate the changes in serum progranulin and sex hormone levels in infertile women with obesity. Material and methods: A total of 171 infertile women who had fertility desire were included in this cross-sectional study. The initial assessment included measurements of weight, height, waist circumference, and hip circumference to calculate body mass index (BMI) and waist/hip ratio. All participants were categorised into two groups in accordance with BMI as a control group ( < 30 kg/m2, n = 135) and a study group (≥ 30 kg/m2, n = 36). After anthropometric measurements, venous blood samples were taken for analyses of oestradiol and follicle stimulating hormone (FSH), luteinising hormone, total testosterone (TT), 17-hydroxyprogesterone, dehydroepiandrosterone sulphate, anti-Müllerian hormone, and progranulin. Results: The present study demonstrated that the overweight women had higher FSH levels (p < 0.01). Elevated TT levels were detected in obese women (p < 0.05). Progranulin concentrations were higher in the study group than in the control group (p < 0.05). Regression analysis demonstrated that there was a relationship between the serum progranulin concentrations and BMI (p < 0.05). Conclusions: Our findings support that the elevated progranulin levels are associated with obesity in infertile women. Therefore, infertile and obese patients may benefit if their serum progranulin levels decrease. Further studies are needed to elucidate this issue

Effect of anticholinergic burden on the development of dementia in older adults with subjective cognitive decline

OBJECTIVE: Data on the effect of anticholinergic cognitive burden (ACB) in older adults with subjective cognitive decline (SCD) are limited. We aimed to study whether ACB increases the future risk of dementia in older adults with SCD. METHODS: The retrospective cohort analysis was carried out on 1496 older adults. Out of those, 109 older patients with SCD followed up over 36 months were studied. They were divided into two groups according to cognitive status at last visit: group I included the subjects with SCD who did not progress to dementia and group II included those who progressed to dementia. The drugs with anticholinergic effects that were received by subjects three months or more were identified from records. The drugs were categorized as having absent (ACB = 0), possible (ACB = 1), and definite (ACB = 2) anticholinergic properties based on an ACB scale. ACB was calculated for each subject by adding the score of each drug and classified as no or low ACB (ACB ≤ 2) and high ACB (ACB ≥ 3). RESULTS: The mean age of all subjects was 72.5 ± 6.3 years and 66.1% of the sample was female. The median follow-up time for all subjects was 75 months (range, 36–185). Fifteen (13.8%) of 109 participants with baseline SCD developed dementia. High ACB was present in 12 subjects (12.8%) in group I and 7 subjects (46.7%) in group II (p = .001). The 75–84 and 85+ age groups (hazard ratio (HR) = 3.595; CI: 1.117–11.574; p = .032 and HR = 12.203; CI: 2.889–51.537; p = .001, respectively), hypertension (HR = 7.835; CI: 1.020–60.189; p = .048), and high ACB (HR = 4.312; CI: 1.563–11.899; p = .005) were found to be possible risk factors for dementia among subjects with SCD in the univariate model. In the final multivariate Cox regression model, subjects with high ACB had a 4.2-fold the risk of the development of dementia. Metoprolol (28.6%), trazodone (21.4%), and trospium (12.9%) were leading used drugs with anticholinergic properties. Among subjects with a total ACB score ≥ 3, the majority were on trospium (29.0%), followed by metoprolol (16.2%), paroxetine (16.2%), and trazodone (16.2%). CONCLUSION: We found that high ACB increases 4.2-fold the risk of the development of dementia in older adults with SCD in long-term follow up. The results of our study are promising, however, the effect of ACB on cognitive status among subjects with SCD is still lacking. To clarify the association between ACB and the risk of dementia, large and longer prospective studies are needed in this population

Adjuvant systemic chemotherapy with or without bevacizumab in patients with resected pulmonary metastases from colorectal cancer

Introduction: We investigated the impact of modern chemotherapy regimens and bevacizumab following pulmonary metastasectomy (PM) from metastatic colorectal cancer (CRC)