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Cystic Fibrosis: An autosomal recessive genetic disease of the Exocrine glands. It is caused by mutations in the gene encoding the Cystic fibrosis transmembrane conductance regulator expressed in several organs including the Lung, the Pancreas, the Biliary system, and the Sweat glands. Cystic fibrosis is characterized by epithelial secretory dysfunction associated with ductal obstruction resulting in Airway obstruction; chronic Respiratory infections; Pancreatic insufficiency; maldigestion; salt depletion; and Heat prostration.

Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients

1Foundation Hubrecht Organoid Technology, 2Department of Pediatric Pulmonology, Regenerative Medicine Centre Utrecht, Wilhelmina Children's Hospital, University Medical Centre Utrecht, 3Department of Stem Cells and Cancer, Walter and Eliza Hall Institute of Medical Research, 4Hubrecht Institute for Developmental Biology and Stem Cell Research, University Medical Centre Utrecht

JoVE 55159

 Medicine

Punnett Squares

JoVE 10772

A Punnett square displays the possible genotypes offspring can inherit from two parental genotypes. If a trait’s inheritance pattern (e.g., dominant or recessive) is known, Punnett squares can also be used to determine the probability of inheriting a phenotype. Punnett squares are applicable in situations where trait inheritance is determined by a single gene locus and traits are independently inherited. However, they cannot predict trait probabilities for more complex genetic inheritance scenarios. Punnett squares are visual representations that display possible offspring genotypes resulting from a cross between two parental genotypes. They can depict inheritance of one or multiple phenotypes, or traits, although other tools are more appropriate for investigating the inheritance of more than two traits. Punnett squares can be used to determine the likelihood of offspring inheriting a specific genotype, or pair of alleles causing a particular characteristic (i.e., phenotype or trait), provided that the phenotype is caused by a single gene locus and is independently assorted during meiosis. In other words, Punnett squares are useful for determining inheritance probabilities in scenarios where the likelihood of inheriting one trait does not affect the probability of inheriting another. Although there are many exceptions to these assumptions (e.

 Core: Classical and Modern Genetics

Chromatography-based Biomolecule Purification Methods

JoVE 5683

In biochemistry, chromatography-based purification methods are employed to isolate compounds from a complex mixture. Two such methods used commonly by biochemists are size-exclusion chromatography and affinity chromatography. In size-exclusion chromatography, a column packed with porous beads separates components of a mixture based on size. On the other hand, affinity chromatography allows for …

 Biochemistry

Preparing and Administering Inhaled Medications

JoVE 10390

Source: Madeline Lassche, MSNEd, RN and Katie Baraki, MSN, RN, College of Nursing, University of Utah, UT


Inhaled medications are prescribed for conditions affecting the bronchi, which branch off of the trachea, and bronchioles, which are progressively smaller conducting airways spread throughout the lung tissue. These conditions can be…

 Nursing Skills

Mutations

JoVE 10793

Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations can be characterized in a number of different ways: whether and how they alter the amino acid sequence of the protein, whether they occur over a small or large area of DNA, and whether they occur in somatic cells or germline cells.

Mutations that occur at a single nucleotide are called point mutations. When point mutations occur within genes, the consequences can vary in severity depending on what happens to the encoded amino acid sequence. A silent mutation does not change the amino acid identity and will have no effect on an organism. A missense mutation changes a single amino acid, and the effects might be serious if the change alters the function of the protein. A nonsense mutation produces a stop codon that truncates the protein, likely rendering it nonfunctional. Frameshift mutations occur when one or more nucleotides are inserted into or deleted from a protein-coding DNA sequence, affecting all of the codons downstream of the location of the mutation. The most drastic type of mutation, chromosomal alteration, changes the physical structure of a chromosome. Chromosomal alterations can include deletion, duplication, or inversion of large stretches of DNA within a single chromosome, or integration o

 Core: DNA Structure and Function

Pedigree Analysis

JoVE 10775

A pedigree is a diagram displaying a family’s history of a trait. Analyzing pedigrees can reveal (1) whether a trait is dominant or recessive, (2) the type of chromosome, autosomal or sex, a trait is linked to, (3) genotypes of family members, and (4) probabilities of phenotypes in future generations. For families with a history of autosomal or sex-linked diseases, this information can be crucial to family planning. In various plant and animal species, scientists study the inheritance of phenotypes, or traits, using carefully controlled mating experiments called crosses. For example, monohybrid crosses can establish trait dominance or recessiveness, and test crosses can determine the genotype (homozygous or heterozygous) of an organism exhibiting a known dominant phenotype. Humans, however, cannot be ethically or feasibly crossed. Therefore, researchers analyze pedigrees, or family trees, to understand how human traits and diseases are inherited. Pedigrees display a family’s history of a trait across generations and family members. Using the same principles that apply to crosses to analyze reproductive events that have already occurred, information about trait heritability can be inferred. On a typical pedigree, squares represent males and circles represent females. Shaded squares or circles signify the presence of a trait of intere

 Core: Classical and Modern Genetics

Protein Folding

JoVE 10679

Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation which is critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.

Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing immunological defense, storage, transport, cellular communication, movement, and structural support. A protein’s function mostly depends on its ability to recognize and bind other molecules, analogous to a lock and key. Hence the specific activity of each protein depends on its unique three-dimensional architecture. For a protein to be functional, it must fold accurately. Most proteins go through several intermediate forms before folding into the most stable, biologically active structure. Misfolding of proteins has detrimental effects on the overall functioning of the cell. In humans, several diseases are due to the accumulation of misfolded or unfolded proteins. These include cystic fibrosis, Alzheimer’s, Parkinson’s, ALS, and Creutzfeldt-Jakob disease. Proteins are made up of one or more chains of amino acids, called polypeptides. A polypeptide is synthesized as a linear chain which rapid

 Core: Macromolecules

Evolutionary Relationships- Concept

JoVE 10561

Humans have been attempting to properly classify living things since Aristotle made the first attempt during the 4th century BC. Aristotle’s system was improved upon during the Renaissance and then, subsequently, by Carolus Linnaeus in the mid 1700’s. These more formal classification and organization systems grouped species by their physical similarity to one another. For example,…

 Lab Bio

Respiratory Exam I: Inspection and Palpation

JoVE 10028

Source: Suneel Dhand, MD, Attending Physician, Internal Medicine, Beth Israel Deaconess Medical Center


Disorders of the respiratory system with a chief complaint of shortness of breath are among the most common reasons for both outpatient and inpatient evaluation. The most obvious visible clue to a respiratory problem will be whether the…

 Physical Examinations I

SNP Genotyping

JoVE 5544

Single nucleotide polymorphisms, or SNPs, are the most common form of genetic variation in humans. These differences at individual bases in the DNA often do not directly affect gene expression, but in many cases can still be useful for locating disease-associated genes or for diagnosing patients. Numerous methodologies have been established to identify, or…

 Genetics

An Introduction to Saccharomyces cerevisiae

JoVE 5081

Saccharomyces cerevisiae (commonly known as baker’s yeast) is a single-celled eukaryote that is frequently used in scientific research. S. cerevisiae is an attractive model organism due to the fact that its genome has been sequenced, its genetics are easily manipulated, and it is very easy to maintain in the lab. Because many yeast proteins are similar in sequence and function…

 Biology I

Standardized Measurement of Nasal Membrane Transepithelial Potential Difference (NPD)

1Department of Medicine and the Gregory Fleming James Cystic Fibrosis Center, University of Alabama at Birmingham, 2Department of Pulmonology and Tuberculosis, University Medical Center Utrecht, 3Center for Experimental Medicine, Queens University, Northern Ireland, 4Hadassah Hebrew University Medical Center, Jerusalem, 5Centro Fibrosi Cistica, Azienda Ospedaliera Universitaria Integrata, 6Service de Pneumologie et Allergologie Pédiatriques and Center de Ressources et de Compétence de la Mucoviscidose, Hôpital Necker Enfants Malades, 7INSERM U 1151, Institut Necker Enfants Malades

JoVE 57006

 Medicine

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

1Molecular Virology and Gene Therapy, KU Leuven, 2Department of Woman and Child, KU Leuven, 3Neurobiology and Gene Therapy, KU Leuven, 4Division of Nuclear Medicine, KU Leuven, 5Biomedical NMR Unit/ MoSAIC, KU Leuven

JoVE 4219

 Medicine

Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants

1Department of Pediatrics, Mucosal Immunology and Biology Research Center, Massachusetts General Hospital, 2Department of Molecular Biology, Massachusetts General Hospital, 3Department of Genetics, Harvard Medical School, 4Department of Pediatrics, Harvard Medical School

JoVE 57298

 Immunology and Infection

Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos

1Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS, UMR 535, Université Montpellier, 2Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé, CNRS, FRE 3689, Université Montpellier, 3Unité de Formation et de Recherche des Sciences de la Santé, EA3647-EPIM, Université Versailles St Quentin

JoVE 53130

 Immunology and Infection

Genetic Engineering of Primary Mouse Intestinal Organoids Using Magnetic Nanoparticle Transduction Viral Vectors for Frozen Sectioning

1Department of Medicine, Division of Hematology, Johns Hopkins University School of Medicine, 2Department of Natural Sciences, School of Arts & Sciences, Lebanese American University, 3Department of Cell Biology, Johns Hopkins University School of Medicine, 4Department of Oncology, Johns Hopkins University School of Medicine, 5Department of Pathology, Johns Hopkins University School of Medicine, 6Institute for Cell Engineering, Johns Hopkins University School of Medicine

JoVE 57040

 Developmental Biology

Live Cell Analysis of Shear Stress on Pseudomonas aeruginosa Using an Automated Higher-Throughput Microfluidic System

1Department of Chemistry, Doane University, 2Department of Biology, Doane University, 3Department of Pathology and Microbiology, University of Nebraska Medical Center, 4Department of Physics and Engineering, Doane University

JoVE 58926

 Bioengineering

A Fluorescence-based Assay for Characterization and Quantification of Lipid Droplet Formation in Human Intestinal Organoids

1Division of Pediatrics, Department of Pediatric Gastroenterology, Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht University, 2Regenerative Medicine Center, University Medical Center Utrecht, Utrecht University, 3Science for Life Laboratory, Department of Medical Biochemistry and Microbiology, Uppsala University

JoVE 60150

 Medicine

Polysome Profiling in Leishmania, Human Cells and Mouse Testis

1Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, 2Department of Biological Sciences, Texas Tech University, 3CISER (Center for the Integration of STEM Education & Research), Texas Tech University

JoVE 57600

 Biochemistry

A Simple Approach to Perform TEER Measurements Using a Self-Made Volt-Amperemeter with Programmable Output Frequency

1Interfaculty Institute of Biochemistry, University of Tübingen, 2Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 3Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University

JoVE 60087

 Biology

Real-Time, Semi-Automated Fluorescent Measurement of the Airway Surface Liquid pH of Primary Human Airway Epithelial Cells

1Epithelial Research Group, Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, 2Respiratory Group, Institute of Cellular Medicine, Faculty of Medical Sciences, Newcastle University, 3Paediatric Respiratory Medicine, Great North Children's Hospital, Newcastle upon Tyne Hospitals NHS Foundation Trust, 4Boehringer Ingelheim Pharma GmbH & Co

JoVE 59815

 Biochemistry
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