90874 - Applied Biochemistry.

Learning outcomes

“Applied Biochemistry” has as objectives the study of the theoretical foundations, the instrumental implementations and the main techniques used in cosmetics, in the preparation and analysis of samples for biochemical investigations. The student: o knows the physical and chemical principles underlying the main techniques and instruments employed in biochemical investigations on cosmetic products with reference to the new in vitro tests; o knows the different parameters that may be modified to change biochemical analysis on cosmetic products; o includes a working protocol and the main variables on which to act to optimize it; o can describe the structural elements of the main tools of a biochemical laboratory/cosmetology; o knows the correct terminology used in biochemical methods in cosmetics; o can use the knowledge acquired to evaluate the results of an experiment, identify the problems and optimize it; o knows how to evaluate the possible impact of changes of the parameters in an experiment for the evaluation of corrosion and skin irritation, skin absorption measures and in the main imaging techniques to measure the phototoxicity; o can orientate between the main qualitative and quantitative methodologies, to select the one appropriate to the study of the problem with reference to the new in vitro tests available for the analysis of cosmetic products.

Course contents

 Histochemical assay discussion of results using microscope 

Front teaching (classroom lectures: 40 hours)

Basic principles: Biochemical and molecular biology studies, Quantitative biochemical measurements, Safety in the laboratory.

Cell culture techniques: The cell culture laboratory and equipment, Safety considerations in cell culture, Aseptic techniques and good cell culture practice, Types of animal cell, characteristics and maintenance in culture, Stem cell and bacterial cell culture, Potential use of cell cultures.

Centrifugation: Basic principles of sedimentation, Types, care and safety aspects of centrifuges, Preparative centrifugation, Analytical centrifugation.

Microscopy: The light microscope, Optical sectioning, Imaging living cells and tissues, Measuring cellular dynamics, The electron microscope (EM), Image archiving.

Immunochemical techniques: Making antibodies, Immunoassay formats, Immuno-microscopy, Lateral flow devices, Epitope mapping, Immunoblotting, Fluorescent activated cell sorting , Cell and tissue staining techniques, Immuno-capture polymerase chain reaction (PCR), Immuno-affinity chromatography, Antibody-based biosensors, Therapeutic antibodies.

In vitro analysis of the pro-sensitising potential of a cosmetic product.

Proteins: structure (Hair and Skin), purification, characterisation and function analysis. 

Electrophoretic techniques: general principles and Support media, Electrophoresis of proteins and of nucleic acids, Capillary electrophoresis.

 

Laboratory training

Protein extraction from from cell cultures and colorimetric quantification via spectrophotometer and discussion of results. Elisa Assay. Different Histochemical assay on skin tissues, fibrosis assessment colorimetric quantification via spectrophotometer and discussion of results

 

Readings/Bibliography

"Principles and Techniques of Biochemistry and Molecular Biology"

Seventh Edition

Edited by Keith Wilson and John Wolker

 

"Principles and Techniques of Practical  Biochemistry"

Fifth Edition

Edited by Keith Wilson and John Wolker

 

 

Teaching methods

Lectures supported by powerpoint presentations of each topic of the course will be shown

The course is based both on classroom lectures (40 hours) and on laboratory practice (15 hours).

Assessment methods

Learning will be assessed by a final oral examination on the course content.

The final mark will also take into account the attendance and active participation of students in laboratory work.

Teaching tools

PC, overhead projector, biological laboratory

Office hours

See the website of Valeria Righi