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Airborne system in Type 2 diabetes studies : research focused on Langerhans islets

Kavoosi, Negar LU (2012) KEMR13 20111
Department of Chemistry
Abstract
The explosive increase in expansion of diabetes mellitus, particularly type 2 diabetes (T2D) is becoming a threat to public health which is associated with modern lifestyle, abundant nutrient supply, reduced physical activity, and obesity. It is realized that T2D only develops in insulin-resistance subjects which can start with β-cell dysfunction. In order to clarify the relationship between hormone release and metabolism in isolated pancreatic islets, the secretory effects of a non-fuel stimulator like acetylcholine was studied.

The miniaturized "airborne system" (acoustic levitation and shooter) was combined with MALDI-MS to acquire data from single Langerhans islets and further β-cell metabolism. Levitation technique is a method... (More)
The explosive increase in expansion of diabetes mellitus, particularly type 2 diabetes (T2D) is becoming a threat to public health which is associated with modern lifestyle, abundant nutrient supply, reduced physical activity, and obesity. It is realized that T2D only develops in insulin-resistance subjects which can start with β-cell dysfunction. In order to clarify the relationship between hormone release and metabolism in isolated pancreatic islets, the secretory effects of a non-fuel stimulator like acetylcholine was studied.

The miniaturized "airborne system" (acoustic levitation and shooter) was combined with MALDI-MS to acquire data from single Langerhans islets and further β-cell metabolism. Levitation technique is a method which has been developed to study intra-and extra- cellular reactions at single or few cell levels. With this technique it is possible to handle 50 nl to 1500 nl sample volumes without contamination from solid walls. In combination with this method a CCD camera was used to check the islet in droplet during the experiments for surveillance of the droplet, for example it was possible to take pictures in different periods of time to calculate droplet size after evaporation to understand change in concentration by time. With usage of piezoelectric dispenser it was possible to keep size of droplet constant by time.

MALDI-TOF mass spectrometry is a sensitive analytical technique with good limit of detection. In this work it was combined with airborne system (levitation technique) to analyze proteins/peptides that are released from Langerhans islets. α-cyano-4-hydroxy Cinnamic acid (CHCA) was used as matrix to crystallize the sample on MALDI plate. Samples were analyzed with MALDI-MS spectrometry. Spectra observed from mass spectrometry gave information about molecular mass, intensity and charge of the detected proteins and peptides. (Less)
Abstract
Popular summary
Using Airborne System in Type 2 Diabetes
The explosive increase in expansion of diabetes mellitus, particularly type 2 diabetes (T2D) is becoming a threat to public health which is associated with modern lifestyle, abundant nutrient supply, reduced physical activity, and obesity. It is realized that T2D only develops in insulin-resistance subjects which can start with β-cell dysfunction. In order to clarify the relationship between hormone release and metabolism in isolated pancreatic islets, the secretory effects of a non-fuel stimulator like acetylcholine was studied.

The miniaturized "airborne system" (acoustic levitation and shooter) was combined with MALDI-MS to acquire data from single Langerhans islets and... (More)
Popular summary
Using Airborne System in Type 2 Diabetes
The explosive increase in expansion of diabetes mellitus, particularly type 2 diabetes (T2D) is becoming a threat to public health which is associated with modern lifestyle, abundant nutrient supply, reduced physical activity, and obesity. It is realized that T2D only develops in insulin-resistance subjects which can start with β-cell dysfunction. In order to clarify the relationship between hormone release and metabolism in isolated pancreatic islets, the secretory effects of a non-fuel stimulator like acetylcholine was studied.

The miniaturized "airborne system" (acoustic levitation and shooter) was combined with MALDI-MS to acquire data from single Langerhans islets and further β-cell metabolism. Levitation technique is a method which has been developed to study intra-and extra- cellular reactions at single or few cell levels. With this technique it is possible to handle 50 nl to 1500 nl sample volumes without contamination from solid walls. In combination with this method a CCD camera was used to check the islet in droplet during the experiments for surveillance of the droplet, for example it was possible to take pictures in different periods of time to calculate droplet size after evaporation to understand change in concentration by time. With usage of piezoelectric dispenser it was possible to keep size of droplet constant by time.

MALDI-TOF mass spectrometry is a sensitive analytical technique with good limit of detection. In this work it was combined with airborne system (levitation technique) to analyze proteins/peptides that are released from Langerhans islets. α-cyano-4-hydroxy Cinnamic acid (CHCA) was used as matrix to crystallize the sample on MALDI plate. There are three different components in the MS 1.Ionizer, 2.Mass analyzer, and 3.Detector. An Ionizer is used to ionize the proteins and peptides from activated single Langerhans islet, then there is a Mass analyzer that differentiates the Ions with respect to the Mass/Charge (M/Z) finally a detector to measure the Ion beam current.

In this study, a single Langerhans islet picked for each experiment and it was positioned in the levitator with a fused silica capillary. Stimulation of insulin secretion from the islets of Langerhans was accomplished by adding acetylcholine to the levitated droplet and addition of stimulators were stopped after desired concentration. A piezoelectric dispenser was used to add all the reagents to the droplet. At the end, epinephrine was added to stop the reaction and a fused silica capillary was used to transfer the droplet to MALDI plate. Sample was crystallized with CHCA matrix and it was analyzed with MALDI-MS spectrometry.

Spectra observed from mass spectrometry gave information about molecular mass, intensity and charge of the detected proteins and peptides. (Less)
Please use this url to cite or link to this publication:
author
Kavoosi, Negar LU
supervisor
organization
course
KEMR13 20111
year
type
H2 - Master's Degree (Two Years)
subject
keywords
Insulin, acetylcholine chloride, MALDI-TOF mass spectrometry, MALDI LTQ Orbitrap XL, Epinephrine, biokemi
language
English
id
3458359
date added to LUP
2013-02-11 11:47:41
date last changed
2013-02-11 11:47:41
@misc{3458359,
  abstract     = {Popular summary
Using Airborne System in Type 2 Diabetes
The explosive increase in expansion of diabetes mellitus, particularly type 2 diabetes (T2D) is becoming a threat to public health which is associated with modern lifestyle, abundant nutrient supply, reduced physical activity, and obesity. It is realized that T2D only develops in insulin-resistance subjects which can start with β-cell dysfunction. In order to clarify the relationship between hormone release and metabolism in isolated pancreatic islets, the secretory effects of a non-fuel stimulator like acetylcholine was studied. 

The miniaturized "airborne system" (acoustic levitation and shooter) was combined with MALDI-MS to acquire data from single Langerhans islets and further β-cell metabolism. Levitation technique is a method which has been developed to study intra-and extra- cellular reactions at single or few cell levels. With this technique it is possible to handle 50 nl to 1500 nl sample volumes without contamination from solid walls. In combination with this method a CCD camera was used to check the islet in droplet during the experiments for surveillance of the droplet, for example it was possible to take pictures in different periods of time to calculate droplet size after evaporation to understand change in concentration by time. With usage of piezoelectric dispenser it was possible to keep size of droplet constant by time.

MALDI-TOF mass spectrometry is a sensitive analytical technique with good limit of detection. In this work it was combined with airborne system (levitation technique) to analyze proteins/peptides that are released from Langerhans islets. α-cyano-4-hydroxy Cinnamic acid (CHCA) was used as matrix to crystallize the sample on MALDI plate. There are three different components in the MS 1.Ionizer, 2.Mass analyzer, and 3.Detector. An Ionizer is used to ionize the proteins and peptides from activated single Langerhans islet, then there is a Mass analyzer that differentiates the Ions with respect to the Mass/Charge (M/Z) finally a detector to measure the Ion beam current. 

In this study, a single Langerhans islet picked for each experiment and it was positioned in the levitator with a fused silica capillary. Stimulation of insulin secretion from the islets of Langerhans was accomplished by adding acetylcholine to the levitated droplet and addition of stimulators were stopped after desired concentration. A piezoelectric dispenser was used to add all the reagents to the droplet. At the end, epinephrine was added to stop the reaction and a fused silica capillary was used to transfer the droplet to MALDI plate. Sample was crystallized with CHCA matrix and it was analyzed with MALDI-MS spectrometry.
 
Spectra observed from mass spectrometry gave information about molecular mass, intensity and charge of the detected proteins and peptides.},
  author       = {Kavoosi, Negar},
  keyword      = {Insulin,acetylcholine chloride,MALDI-TOF mass spectrometry,MALDI LTQ Orbitrap XL,Epinephrine,biokemi},
  language     = {eng},
  note         = {Student Paper},
  title        = {Airborne system in Type 2 diabetes studies : research focused on Langerhans islets},
  year         = {2012},
}