A cotton miRNA is involved in regulation of plant response to salt stress

The present study functionally identified a new microRNA (microRNA ovual line 5, miRNVL5) with its target gene GhCHR from cotton (Gossypium hirsutum). The sequence of miRNVL5 precursor is 104 nt long, with a well developed secondary structure. GhCHR contains two DC1 and three PHD Cys/His-rich domains, suggesting that GhCHR encodes a zinc-finger domain-containing transcription factor. miRNVL5 and GhCHR express at various developmental stages of cotton. Under salt stress (50–400 mM NaCl), miRNVL5 expression was repressed, with concomitant high expression of GhCHR in cotton seedlings. Ectopic expression of GhCHR in Arabidopsis conferred salt stress tolerance by reducing Na+ accumulation in plants and improving primary root growth and biomass. Interestingly, Arabidopsis constitutively expressing miRNVL5 showed hypersensitivity to salt stress. A GhCHR orthorlous gene At2g44380 from Arabidopsis that can be cleaved by miRNVL5 was identified by degradome sequencing, but no confidential miRNVL5 homologs in Arabidopsis have been identified. Microarray analysis of miRNVL5 transgenic Arabidopsis showed six downstream genes (CBF1, CBF2, CBF3, ERF4, AT3G22920, and AT3G49200), which were induced by salt stress in wild-type but repressed in miRNVL5-expressing Arabidopsis. These results indicate that miRNVL5 is involved in regulation of plant response to salt stress

Pathway-based expression profiling of benign prostatic hyperplasia and prostate cancer delineates an immunophilin molecule associated with cancer progression

Aberrant restoration of AR activity is linked with prostate tumor growth, therapeutic failures and development of castrate-resistant prostate cancer. Understanding the processes leading to ARreactivation should provide the foundation for novel avenues of drug discovery. A differential gene expression study was conducted using biopsies from CaP and BPH patients to identify the components putatively responsible for reinstating AR activity in CaP. From the set of genes upregulated in CaP, FKBP52, an AR co-chaperone, was selected for further analysis. Expression of FKBP52 was positively correlated with that of c-Myc. The functional cross-talk between c-Myc and FKBP52 was established using c-Myc specific-siRNA to LNCaP cells that resulted in reduction of FKBP52. A non-canonical E-box sequence housing a putative c-Myc binding site was detected on the FKBP4 promoter using in silico search. LNCaP cells transfected with the FKBP52 promoter cloned in pGL3 basic showed increased luciferase activity which declined considerably when the promoter-construct was co-transfected with c-Myc specific-siRNA. ChIP-PCR confirmed the binding of c-Myc with the conserved E-box located in the FKBP52 promoter. c-Myc downregulation concomitantly affected expression of FGF8. Since expression of FGF8 is controlled by AR, our study unveiled a novel functional axis between c-Myc, AR and FGF8 operating through FKBP52

Genome sequence of Mycobacterium yongonense RT 955-2015 isolate from a patient misdiagnosed with multidrug-resistant tuberculosis: First clinical detection in Tanzania

Background: Mycobacterium yongonense is a recently described novel species belonging to Mycobacterium avium complex, which is the most prevalent aetiology of non-tuberculous mycobacteria associated with pulmonary infections, and poses tuberculosis diagnostic challenges in high-burden, resource-constrained settings. Methods: Whole genome shotgun sequencing and comparative microbial genomic analyses were used to characterize the isolate from a patient diagnosed with multidrug-resistant tuberculosis (MDR-TB) after relapse. Results: The genome sequence of the first case of M. yongonense (M. yongonense RT 955-2015) in Tanzania is presented. Sequence analysis revealed that the RT 955-2015 strain had a high similarity to M. yongonense 05-1390(T) (98.74%) and Mycobacterium chimaera DSM 44623(T) (98%). Its 16S rRNA showed similarity to Mycobacterium paraintracellulare KCTC 290849(T) (100%), Mycobacterium intracellulare ATCC 13950(T) (100%), M. chimaera DSM 44623(T) (99.9%), and M. yongonense 05-1390(T) (98%). The strain exhibited a substantially different rpoB sequence to that of M. yongonense 05-1390 (95.16%), but closely related to that of M. chimaera DSM 44623(T) (99.86%), M. intracellulare ATCC 13950(T), (99.53%), and M. paraintracellulare KCTC 290849(T) (99.53%). Conclusions: In light of the OrthoANI algorithm and phylogenetic analysis, it was concluded that the isolate was M. yongonense Type II genotype, which is an indication that the patient was misdiagnosed with TB/MDR-TB and received inappropriate treatment. (C) 2018 The Author(s). Published by Elsevier Ltd on behalf of International Society for Infectious Diseases.

Asthma symptoms, severity, and control with and without a clinical diagnosis of asthma in early adolescence in sub-Saharan Africa: a multi-country, school-based, cross-sectional study.

BACKGROUND: Rapid urbanisation and population growth in sub-Saharan Africa has increased the incidence of asthma in children and adolescents. One major barrier to achieving good asthma control in these adolescents is obtaining a clinical diagnosis. To date, there are scant data on prevalence and severity of asthma in undiagnosed yet symptomatic adolescents. We therefore aimed to assess symptom prevalence and severity, the effect of symptoms on daily life, and objective evidence of asthma in young adolescents from sub-Saharan Africa with and without a clinical diagnosis of asthma by spirometry and fractional exhaled nitric oxide (FeNO). METHODS: We designed a two-phase, multi-country, school-based, cross-sectional study to assess symptom prevalence and severity in sub-Saharan African adolescents. In phase 1 we surveyed young adolescents aged 12-14 years who were attending selected primary and secondary schools in Blantyre in Malawi, Durban in South Africa, Harare in Zimbabwe, Kampala in Uganda, Kumasi in Ghana, and Lagos in Nigeria. The adolescents were screened for asthma symptoms using the International Study of Asthma and Allergies in Children (ISAAC) questionnaire. Then, after opt-in consent, symptomatic adolescents were invited to complete a detailed survey on asthma severity, treatment, and exposure to environmental risk factors for phase 2. Adolescents performed the European Respiratory Society's diagnostic tests for childhood asthma. A positive asthma test was classified as a forced expiratory volume in 1 sec (FEV1) predicted under 80%, a FEV1 under the lower limits of normal, or FEV1 divided by forced vital capacity (FEV1/FVC) under the lower limits of normal; positive bronchodilator responsiveness or reversibility was defined as either an increase in absolute FEV1 of 12% or more, or an increase of 200 mL or more, or both, after 400 μg of salbutamol (shortacting β2 agonist) administered via a metered-dose inhaler and spacer, or FeNO of 25 parts per billion or higher, or any combination of these. The study was registered with ClinicalTrials.gov (NCT03990402) and is complete. FINDINGS: Between Nov 1, 2018, and Nov 1, 2021, we recruited 149 schools from six regions in six sub-Saharan countries to participate in the study. We administered phase 1 asthma questionnaires from Jan 20, 2019 to Nov 11, 2021, and from 27 407 adolescents who were screened, we obtained data for 27 272 (99·5%). Overall, 14 918 (54·7%) adolescents were female and 12 354 (45·3%) adolescents were male, and the mean age was 13 years (IQR 12-13); nearly all recruited adolescents were of black African ethnicity (26 821 [98·3%] of 27 272). In phase 1, a total of 3236 (11·9% [95% CI 11·5-12·3]) reported wheeze in the past 12 months, and 644 (19·9%) of 3236 had a formal clinical diagnosis of asthma. The prevalence of adolescents with asthma symptoms ranged from 23·8% in Durban, South Africa to 4·2% Blantyre, Malawi. Using ISAAC criteria, severe asthma symptoms were reported by 2146 (66·3%) of 3236 adolescents, the majority of whom (1672 [77·9%] of 2146) had no diagnosis of asthma by a clinician. Between July 16, 2019, and Nov 26, 2021, we administered the phase 2 questionnaire to the 1654 adolescents who had asthma symptoms in phase 1 and consented to proceed to the second phase. In the phase 2 cohort, 959 (58·0%) were female and 695 (42·0%) were male, and the mean age was 13 years (IQR 12-14). One or more diagnostic tests for asthma were obtained in 1546 (93·5%) of 1654 participants. One or more positive asthma tests were found in 374 (48·8%) of 767 undiagnosed adolescents with severe symptoms, and 176 (42·4%) of 415 of undiagnosed adolescents with mild-to-moderate symptoms. Of the 392 adolescents in phase 2 with clinician-diagnosed asthma, 294 (75·0%) reported severe asthma symptoms, with 94 (32·0%) of those with severe symptoms not using any asthma medication. In general, findings in both phases 1 and 2 were consistent across sub-Saharan African countries. INTERPRETATION: A large proportion of adolescents in sub-Saharan Africa with symptoms of severe asthma do not have a formal diagnosis of asthma and are therefore not receiving appropriate asthma therapy. To improve the poor state of asthma control in sub-Saharan Africa, potential solutions such as educational programmes, better diagnosis, and treatment and screening in schools should be considered. FUNDING: UK National Institute for Health and Care Research and UK Medical Research Council

Detection of MicroRNA processing intermediates through RNA ligation approaches

MicroRNAs (miRNA) are small RNAs of 20–22 nt that regulate diverse biological pathways through the modulation of gene expression. miRNAs recognize target RNAs by base complementarity and guide them to degradation or translational arrest. They are transcribed as longer precursors with extensive secondary structures. In plants, these precursors are processed by a complex harboring DICER-LIKE1 (DCL1), which cuts on the precursor stem region to release the mature miRNA together with the miRNA*. In both plants and animals, the miRNA precursors contain spatial clues that determine the position of the miRNA along their sequences. DCL1 is assisted by several proteins, such as the double-stranded RNA binding protein, HYPONASTIC LEAVES1 (HYL1), and the zinc finger protein SERRATE (SE). The precise biogenesis of miRNAs is of utter importance since it determines the exact nucleotide sequence of the mature small RNAs and therefore the identity of the target genes. miRNA processing itself can be regulated and therefore can determine the final small RNA levels and activity. Here, we describe methods to analyze miRNA processing intermediates in plants. These approaches can be used in wild-type or mutant plants, as well as in plants grown under different conditions, allowing a molecular characterization of the miRNA biogenesis from the RNA precursor perspective.Fil: Moro, Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Rojas, Arantxa Maria Larisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Palatnik, Javier Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Universidad Nacional de Rosario. Centro de Estudios Interdisciplinarios; Argentin

Paclobutrazol treatment as a potential strategy for higher seed and oil yield in field-grown camelina sativa L. Crantz

<p>Abstract</p> <p>Background</p> <p><it>Camelina (Camelina sativa </it>L. Crantz) is a non-food oilseed crop which holds promise as an alternative biofuel energy resource. Its ability to grow in a variety of climatic and soil conditions and minimal requirements of agronomical inputs than other oilseed crops makes it economically viable for advanced biofuel production. We designed a study to investigate the effect of paclobutrazol [2RS, 3RS)-1-(4-Chlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)pentan-3-ol] (PBZ), a popular plant growth regulator, on the seed and oil yield of <it>Camelina sativa </it>(cv. Celine).</p> <p>Results</p> <p>A field-based micro-trial setup was established in a randomized block design and the study was performed twice within a span of five months (October 2010 to February 2011) and five different PBZ treatments (Control: T₀; 25 mg l^-1: T₁; 50 mg l^-1: T₂; 75 mg l^-1: T₃; 100 mg l^-1: T₄; 125 mg l^-1: T₅) were applied (soil application) at the time of initiation of flowering. PBZ at 100 mg l^-1concentration (T₄) resulted in highest seed and oil yield by 80% and 15%, respectively. The seed yield increment was mainly due to enhanced number of siliques per plant when compared to control. The PBZ - treated plants displayed better photosynthetic leaf gas exchange characteristics, higher chlorophyll contents and possessed dark green leaves which were photosynthetically active for a longer period and facilitated higher photoassimilation.</p> <p>Conclusion</p> <p>We report for the first time that application of optimized PBZ dose can be a potential strategy to achieve higher seed and oil yield from <it>Camelina sativa </it>that holds great promise as a biofuel crop in future.</p

Use of photosensitising diuretics and risk of skin cancer: a population-based case–control study

Diuretics have photosensitising properties. However, little is known about how these diuretics affect the risk of skin cancers. In North Jutland County, Denmark, we investigated whether the use of photosensitising diuretics was associated with an increased risk for developing basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and malignant melanoma (MM). From the cancer registry, we identified primary cases of BCC, SCC and MM during the period of 1989–2003. We selected four population controls for each case from the Danish Civil Registration System, matched on age and gender. Prescriptions for photosensitising diuretics before cancer diagnosis were ascertained in the county's Prescription Database. We used conditional logistic regression to compute incidence rate ratio (IRR), controlling for the chronic medical conditions and for the previous use of oral glucocorticoids. We found an increased risk of SCC (IRR of 1.79 (95% confidence interval (CI): 1.45–2.21)) and MM (IRR of 1.43 (95% CI: 1.09–1.88)) among users of combined amiloride and hydrochlorothiazide therapy. An increased risk of MM (IRR of 3.30 (95% CI: 1.34–8.10)) was found among users of indapamide. We found little associations with risk of BCC. Our findings provide evidence that the use of some photosensitising diuretics is associated with an increased risk for SCC and MM

Adolescent transport and unintentional injuries: a systematic analysis using the Global Burden of Disease Study 2019

Background: Globally, transport and unintentional injuries persist as leading preventable causes of mortality and morbidity for adolescents. We sought to report comprehensive trends in injury-related mortality and morbidity for adolescents aged 10–24 years during the past three decades. Methods: Using the Global Burden of Disease, Injuries, and Risk Factors 2019 Study, we analysed mortality and disability-adjusted life-years (DALYs) attributed to transport and unintentional injuries for adolescents in 204 countries. Burden is reported in absolute numbers and age-standardised rates per 100 000 population by sex, age group (10–14, 15–19, and 20–24 years), and sociodemographic index (SDI) with 95% uncertainty intervals (UIs). We report percentage changes in deaths and DALYs between 1990 and 2019. Findings: In 2019, 369 061 deaths (of which 214 337 [58%] were transport related) and 31·1 million DALYs (of which 16·2 million [52%] were transport related) among adolescents aged 10–24 years were caused by transport and unintentional injuries combined. If compared with other causes, transport and unintentional injuries combined accounted for 25% of deaths and 14% of DALYs in 2019, and showed little improvement from 1990 when such injuries accounted for 26% of adolescent deaths and 17% of adolescent DALYs. Throughout adolescence, transport and unintentional injury fatality rates increased by age group. The unintentional injury burden was higher among males than females for all injury types, except for injuries related to fire, heat, and hot substances, or to adverse effects of medical treatment. From 1990 to 2019, global mortality rates declined by 34·4% (from 17·5 to 11·5 per 100 000) for transport injuries, and by 47·7% (from 15·9 to 8·3 per 100 000) for unintentional injuries. However, in low-SDI nations the absolute number of deaths increased (by 80·5% to 42 774 for transport injuries and by 39·4% to 31 961 for unintentional injuries). In the high-SDI quintile in 2010–19, the rate per 100 000 of transport injury DALYs was reduced by 16·7%, from 838 in 2010 to 699 in 2019. This was a substantially slower pace of reduction compared with the 48·5% reduction between 1990 and 2010, from 1626 per 100 000 in 1990 to 838 per 100 000 in 2010. Between 2010 and 2019, the rate of unintentional injury DALYs per 100 000 also remained largely unchanged in high-SDI countries (555 in 2010 vs 554 in 2019; 0·2% reduction). The number and rate of adolescent deaths and DALYs owing to environmental heat and cold exposure increased for the high-SDI quintile during 2010–19. Interpretation: As other causes of mortality are addressed, inadequate progress in reducing transport and unintentional injury mortality as a proportion of adolescent deaths becomes apparent. The relative shift in the burden of injury from high-SDI countries to low and low–middle-SDI countries necessitates focused action, including global donor, government, and industry investment in injury prevention. The persisting burden of DALYs related to transport and unintentional injuries indicates a need to prioritise innovative measures for the primary prevention of adolescent injury. Funding: Bill & Melinda Gates Foundation

Phenotyping male infertility in the mouse: how to get the most out of a ‘non-performer’

BACKGROUND: Functional male gametes are produced through complex processes that take place within the testis, epididymis and female reproductive tract. A breakdown at any of these phases can result in male infertility. The production of mutant mouse models often yields an unexpected male infertility phenotype. It is with this in mind that the current review has been written. The review aims to act as a guide to the 'non-reproductive biologist' to facilitate a systematic analysis of sterile or subfertile mice and to assist in extracting the maximum amount of information from each model. METHODS: This is a review of the original literature on defects in the processes that take a mouse spermatogonial stem cell through to a fully functional spermatozoon, which result in male infertility. Based on literature searches and personal experience, we have outlined a step-by-step strategy for the analysis of an infertile male mouse line. RESULTS: A wide range of methods can be used to define the phenotype of an infertile male mouse. These methods range from histological methods such as electron microscopy and immunohistochemistry, to hormone analyses and methods to assess sperm maturation status and functional competence. CONCLUSION: With the increased rate of genetically modified mouse production, the generation of mouse models with unexpected male infertility is increasing. This manuscript will help to ensure that the maximum amount of information is obtained from each mouse model and, by extension, will facilitate the knowledge of both normal fertility processes and the causes of human infertility