Restricted Cubic Splines

Restricted cubic splines are a flexible regression technique for modeling nonlinear relationships between a continuous predictor and an outcome, while avoiding the instability of high-degree polynomials. They represent the predictor with separate cubic polynomial segments joined at selected knots, enforcing continuity of the fitted curve and its first and second derivatives, then constrain the boundary segments to be linear beyond the outer knots. In medicine, restricted cubic splines can characterize dose-response patterns, disease risk across age or exposure levels, and treatment effects when a straight line is inadequate. They support adjusted regression analyses and visualization of threshold-like or plateau behavior without imposing an arbitrary functional form.

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JoVE Science Education - Basic Biology

Restriction Enzyme Digests

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2023

Restriction enzymes or endonucleases recognize and cut DNA at a specific sequence. These enzymes occur naturally in bacteria as a defense against bacteriophages - viruses that infect bacteria. Bacterial restriction enzymes cut the invading bacteriophage DNA while leaving the bacterial genomic DNA unharmed due to addition of methyl groups. This video explains the basic principles of restriction enzymes including: how restriction enzymes are named and the types of recognition sites and...

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JoVE Journal - Medicine

Whole-Kidney Three-Dimensional Staining with CUBIC

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Cited by 5 •

2022

The present protocol describes a tissue clearing method and whole-mount immunofluorescent staining for three-dimensional (3D) kidney imaging. This technique can offer macroscopic perspectives in kidney pathology, leading to new biological discoveries.

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JoVE Lab Manual - Biology
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DNA Isolation and Restriction Enzyme Analysis - Student Protocol

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2019

DNA Isolation ExpandNOTE: In this experiment you will perform DNA isolation under two experimental conditions: one using a buffer containing the detergent SDS and one without detergent. Hypotheses: The alternate hypothesis for this experiment might be that the sample prepared without SDS, a strong anionic detergent that breaks apart cell membranes, will yield less DNA than the sample prepared with SDS. The null hypothesis for this experiment might be that both samples will yield an equal amount...

Imaging Serotonergic Fibers in the Mouse Spinal Cord Using the CLARITY/CUBIC Technique

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Cited by 13 •

2016

Supraspinal projections are important for pain perception and other behaviors, and serotonergic fibers are one of these fiber systems. The present study focused on the application of the combined CLARITY/CUBIC protocol to the mouse spinal cord in order to investigate the termination of these serotonergic fibers.

Research

JoVE Journal - Biochemistry
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Preparation and Delivery of Protein Microcrystals in Lipidic Cubic Phase for Serial Femtosecond Crystallography

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Cited by 20 •

2016

We describe procedures for the preparation and delivery of membrane protein microcrystals in lipidic cubic phase for serial crystallography at X-ray free-electron lasers and synchrotron sources. These protocols can also be applied for incorporation and delivery of soluble protein microcrystals, leading to substantially reduced sample consumption compared to liquid injection.

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