A Kinome-wide screen identifies a CDKL5-SOX9 regulatory axis in epithelial cell death and kidney injury

© 2020, The Author(s). Renal tubular epithelial cells (RTECs) perform the essential function of maintaining the constancy of body fluid composition and volume. Toxic, inflammatory, or hypoxic-insults to RTECs can cause systemic fluid imbalance, electrolyte abnormalities and metabolic waste accumulation- manifesting as acute kidney injury (AKI), a common disorder associated with adverse long-term sequelae and high mortality. Here we report the results of a kinome-wide RNAi screen for cellular pathways involved in AKI-associated RTEC-dysfunction and cell death. Our screen and validation studies reveal an essential role of Cdkl5-kinase in RTEC cell death. In mouse models, genetic or pharmacological Cdkl5 inhibition mitigates nephrotoxic and ischemia-associated AKI. We propose that Cdkl5 is a stress-responsive kinase that promotes renal injury in part through phosphorylation-dependent suppression of pro-survival transcription regulator Sox9. These findings reveal a surprising non-neuronal function of Cdkl5, identify a pathogenic Cdkl5-Sox9 axis in epithelial cell-death, and support CDKL5 antagonism as a therapeutic approach for AKI

Rare coding variants in ten genes confer substantial risk for schizophrenia

Rare coding variation has historically provided the most direct connections between gene function and disease pathogenesis. By meta-analysing the whole exomes of 24,248 schizophrenia cases and 97,322 controls, we implicate ultra-rare coding variants (URVs) in 10 genes as conferring substantial risk for schizophrenia (odds ratios of 3–50, P < 2.14 × 10−6) and 32 genes at a false discovery rate of <5%. These genes have the greatest expression in central nervous system neurons and have diverse molecular functions that include the formation, structure and function of the synapse. The associations of the NMDA (N-methyl-d-aspartate) receptor subunit GRIN2A and AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) receptor subunit GRIA3 provide support for dysfunction of the glutamatergic system as a mechanistic hypothesis in the pathogenesis of schizophrenia. We observe an overlap of rare variant risk among schizophrenia, autism spectrum disorders1, epilepsy and severe neurodevelopmental disorders2, although different mutation types are implicated in some shared genes. Most genes described here, however, are not implicated in neurodevelopment. We demonstrate that genes prioritized from common variant analyses of schizophrenia are enriched in rare variant risk3, suggesting that common and rare genetic risk factors converge at least partially on the same underlying pathogenic biological processes. Even after excluding significantly associated genes, schizophrenia cases still carry a substantial excess of URVs, which indicates that more risk genes await discovery using this approach

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Appraisal of biochemical and genetic diversity of mango cultivars using molecular markers

Mango (Mangifera indica L.) is one of the oldest fruit crops and is broadly cultivated worldwide. To determine the level of genetic diversity, a total of 13 mango genotypes have been collected from different farms of Fayoum oasis in Egypt and were analyzed using molecular (DNA) and biochemical (SDS-PAGE) markers along with the quantification of soluble carbohydrates, chlorophyll and carotenoids. These profiles were evaluated as characters to identify the taxonomic relationships of these genotypes. A total of 433 protein bands (ranged from 8 to 180 KDa) from all genotypes, were detected in SDS-PAGE. A total of 306 RAPD fragments were produced by 19 primers and among them 123 (40.2%) were polymorphic. The similarities between different taxa were estimated by Jaccard’s similarity index and clustered in neighbour joining clustering tree. Among the 13 tested mango samples, the total carbohydrate contents ranged between 31.9 and 40.8 µg/100 mg fresh weights, which represents Taymour cultivar and accession No. 7, respectively. Of the 13 mango cultivars and accessions studied, the highest chlorophyll content (386.9 µg/g) was found in accession No. 10; whereas, the lowest value was observed with accession No. 12 (202.5 µg/g). The amounts of carotenoids were wide-ranging and reached a maximum value of 106.2 µg/g with accession No. 9, however, accession No. 8 recorded the lowest concentration (19.9 µg/g). In conclusion, RAPD-PCR and SDS-PAGE were proved to be an efficient tool in assessing the genetic diversity of mango genotypes. It will also provide an important input to breeders for mango improvement program.Keywords: Mango, genetic diversity, chlorophyll, SDS-PAGE, RAPD.African Journal of Biotechnology Vol 13(28) 2796-280

Cigarette smoking among female students in five medical and nonmedical colleges

Hamza M Abdulghani,1 Norah A Alrowais,2 Ali I Alhaqwi,3 Ahmed Alrasheedi,2 Mohammed Al-Zahir,2 Ahmed Al-Madani,2 Abdulaziz Al-Eissa,2 Bader Al-Hakmi,2 Redwan Takroni,2 Farah Ahmad1 1Department of Medical Education, 2Department of Family and Community Medicine, King Saud University, 3Department of Family Medicine, King Saud bin Abdulaziz University for Health Sciences, Riyadh, Saudi Arabia Objectives: This study was conducted to determine the prevalence of smoking, knowledge about the ill effects of smoking on health, and the influence of family members&#39; smoking habits among Saudi female students. Methods: This is a type of cross-sectional study. A sample of 1,070 female students was selected by a nonrandom and convenient sampling method from five colleges (Medicine, Business and Administration, Computer Sciences, Education, and Languages and Translation) of King Saud University, Riyadh, Saudi Arabia. A self-administrated questionnaire was used to determine the personal, social, and educational characteristics of the respondents. In addition, questions about their smoking types, status, duration of smoking, knowledge about the ill effects of smoking, daily cigarette consumption, and reasons for quitting smoking were included. Results: The students&#39; response rate was 85%. The prevalence of current smoking was 4.3% and 5.6% for cigarettes and water-pipes, respectively, whereas 3.9% of the participants were ex-smokers. The prevalence of current smoking was highest in the College of Business and Administration (10.81%) and lowest in the College of Medicine (0.86%). The majority (77%) of the smokers&#39; parents (current and ex-smokers) were also smokers. More than half (54%) of the smokers started their smoking habit for entertainment, and 44.4% of the participants did not know that smoking causes serious health problems. The most common factors for quitting smoking were health concerns (54%), religious beliefs (29%), and parent&#39;s advice (17%). Conclusion: The study concludes that the prevalence of smoking varies in different subject streams and that family and friends have a great influence on individuals starting or stopping smoking. Extensive health education programs are needed to educate young women on the health hazards of smoking and help stop them from smoking. Keywords: smoking prevalence, quitting smoking, female students, cigarettes per da