Placement report
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Transcript of Placement report
Work placement Presentation
43 weeks in a biochemistry laboratory in Lyon, France
Connor Downing N0293299
The placement
• Placement situated in Lyon, France• A laboratory that deals with rare errors of
inborn metabolism• Given the title “stagiaire” roughly translated as
“intern”.• Duration of one year August 29th 2011-12• Large cultural differences, language used was
French
The organisation
• Service Maladies Héréditaires du Métabolisme• Part of Hospices Civil de lyon• Sectors on the floor comprised of biochemical,
radiological, enzymatic and biomolecular laboratories• Full time staff consists of a “cadre” of trained doctors
and management and also trained technicians.• Public sector service with samples received from all
over mid-region France.
My responsibilities
• Performing technician duties such as:– Carrying out the manipulations– Responding rapidly to occasional emergency
samples– Storage and cleaning of equipment– Basic maintenance of machinery – Removal of Hazardous waste– Basic calculation and quantification – Filing(paper) work
Classical organic acid disorders
• Manipulation consisted of dilution adjustment, urea removal, adding of known concentration standards and silylation.
• Diagnosed by gas chromatography coupled with mass spectrometry (enabling identification and quantification)
• Emergency cases in patients do occur (Hyperammonemia) so some samples are prioritised for expendenicy.
• Detected by marker molecules that are the substrates (and their toxic by-products) from a missing enzyme
• Examples of analytes included homogenisitic acid, methylcitrate and methymalonic acid.
• Amino acid, carnitine and acyl-carnitine profiles commonly ordered alongside.
Brand chained amino acid metabolism
Niemann-Pick C-1 Assay• Cholesterol transportation failure in cells• We made an attempt to functionalise a
biochemical assay for this disease from an article– Involved saponification of sample with KOH
(removal of FAs from cholesterol derivatives) – A step of solid phase extraction in addition to
the normal liquid/liquid extraction– Analysed by GC/MS
• Currently Niemann-Pick disorders (A,B,C and D) are screened by cholesterol studies on cultured fibroblasts using immunocytofluorescence
Article and Procedure
Thin Layer chromatography• Various Lysosomal storage disorders screened by TLC
• Oligosaccridosises (family of disorders)• Sialic acid pathologies• Adenylosuccinyl lyase deficiency
• Extremely old technique (60+ years)• Enables identification and crude quantisation• Consists of
– Ultra filtration of the sample– Deposition of sample– Migration of plate– Revelation of bands
• Rf values are calculated from length of bands against a control used
• Intensity of bands and depot judged by eye and recorded
• Finally Rf values are compared with a reference table to identify the bands.
Skills learned
• Laboratory conduct and expertise• Practical professional knowledge of biochemistry• Professional behaviour in a work-place• French language learned• Initiative when dealing with problems• Improved team skills • Perseverance
END