Vitamin D levels in the pre- and post-COVID-19 pandemic periods in pediatric patients with chronic kidney disease.

Introduction: Vitamin D (VD) deficiency is common in children with chronic kidney disease (CKD) because of multiple factors. During the coronavirus disease 2019 (COVID-19) pandemic, it increased because of medicine shortage and no enough medical service for patients with non-COVID-19 diseases. Objective: To analyze the effects of the COVID-19 pandemic-related lockdown on the serum levels and status of 25-hydroxyvitamin D3 (25-[OH]D) in children with CKD. Materials and methods: This retrospective study included patients (6-18 years old) who were diagnosed with CKD stage 2-5 and routinely measured for serum VD levels between May 2019 and December 2022. Serum 25-(OH)D levels were measured before, during, and after the pandemic (2019, 2020-2021, and 2022, respectively). The daily dose of cholecalciferol supplementation and the readjustment (if required) were recorded. Results: This study included 171 patients (median age: 12 years). Before the pandemic, the median serum VD level was 25.0 ng/mL (19.3% VD deficiency). Then, VD supplementation was adjusted to 400-1,200 UI daily in 98.8% (n = 169) of patients. During the pandemic, the median VD level decreased to 22.5 ng/mL (43.3% VD deficiency). Hence, the supplementation was readjusted, and after the pandemic, the level was 28.7 ng/mL (18.7% VD deficiency), indicating a statistically significant increase in serum VD levels from the prepandemic period (p = 0.007). Conclusion: Decreased serum VD levels and increased VD deficiency frequency were observed in patients with CKD during the COVID-19 but improved after readjustment of supplementation.

Neurotrophic fragments as therapeutic alternatives to ameliorate brain aging.

Aging is a global phenomenon and a complex biological process of all living beings that introduces various changes. During this physiological process, the brain is the most affected organ due to changes in its structural and chemical functions, such as changes in plasticity and decrease in the number, diameter, length, and branching of dendrites and dendritic spines. Likewise, it presents a great reduction in volume resulting from the contraction of the gray matter. Consequently, aging can affect not only cognitive functions, including learning and memory, but also the quality of life of older people. As a result of the phenomena, various molecules with notable neuroprotective capacity have been proposed, which provide a therapeutic alternative for people under conditions of aging or some neurodegenerative diseases. It is important to indicate that in recent years the use of molecules with neurotrophic activity has shown interesting results when evaluated in in vivo models. This review aims to describe the neurotrophic potential of molecules such as resveratrol (3,5,4'-trihydroxystilbene), neurotrophins (brain-derived neurotrophic factor), and neurotrophic-type compounds such as the terminal carboxyl domain of the heavy chain of tetanus toxin, cerebrolysin, neuropeptide-12, and rapamycin. Most of these molecules have been evaluated by our research group. Studies suggest that these molecules exert an important therapeutic potential, restoring brain function in aging conditions or models of neurodegenerative diseases. Hence, our interest is in describing the current scientific evidence that supports the therapeutic potential of these molecules with active neurotrophic.

Correction to: A protocol for custom CRISPR Cas9 donor vector construction to truncate genes in mammalian cells using pcDNA3 backbone.

The original article [1] contains three erroneous mentions of usage of a restriction enzyme-BstZ17I-in the Methods section as displayed in the following sentences.

A protocol for custom CRISPR Cas9 donor vector construction to truncate genes in mammalian cells using pcDNA3 backbone.

BACKGROUND: Clustered regularly interspaced short palindromic repeat (CRISPR) RNA-guided adaptive immune systems are found in prokaryotes to defend cells from foreign DNA. CRISPR Cas9 systems have been modified and employed as genome editing tools in wide ranging organisms. Here, we provide a detailed protocol to truncate genes in mammalian cells using CRISPR Cas9 editing. We describe custom donor vector construction using Gibson assembly with the commonly utilized pcDNA3 vector as the backbone. RESULTS: We describe a step-by-step method to truncate genes of interest in mammalian cell lines using custom-made donor vectors. Our method employs 2 guide RNAs, mutant Cas9D10A nickase (Cas9 = CRISPR associated sequence 9), and a custom-made donor vector for homologous recombination to precisely truncate a gene of interest with a selectable neomycin resistance cassette (NPTII: Neomycin Phosphotransferase II). We provide a detailed protocol on how to design and construct a custom donor vector using Gibson assembly (and the commonly utilized pcDNA3 vector as the backbone) allowing researchers to obtain specific gene modifications of interest (gene truncation, gene deletion, epitope tagging or knock-in mutation). Selection of mutants in mammalian cell lines with G418 (Geneticin) combined with several screening methods: western blot analysis, polymerase chain reaction, and Sanger sequencing resulted in streamlined mutant isolation. Proof of principle experiments were done in several mammalian cell lines. CONCLUSIONS: Here we describe a detailed protocol to employ CRISPR Cas9 genome editing to truncate genes of interest using the commonly employed expression vector pcDNA3 as the backbone for the donor vector. Providing a detailed protocol for custom donor vector design and construction will enable researchers to develop unique genome editing tools. To date, detailed protocols for CRISPR Cas9 custom donor vector construction are limited (Lee et al. in Sci Rep 5:8572, 2015; Ma et al. in Sci Rep 4:4489, 2014). Custom donor vectors are commercially available, but can be expensive. Our goal is to share this protocol to aid researchers in performing genetic investigations that require custom donor vectors for specialized applications (specific gene truncations, knock-in mutations, and epitope tagging applications).