CHM683M/D Seminar

Critique Paper
Theme: Advanced Techniques in Biochemistry

Name: KARINA L. DAMO Term/AY: 2ND Term 2018-2019

ID No: 11784601 Date Submitted: April 5, 2019
Section: GO1 Score:

Title of Seminar: “Shwshshshswshshs Sepsis!”

Urine metabolomics: An Approach in finding Biomarkers and Pathways for Sepsis

Critique:
1. Sypnosis of the study presented in the seminar
According to World Health Organization, of the 30 million estimated cases of sepsis
yearly, 6 million leads to death (which is mostly children). Sepsis is due to a massive
immune system response to bacterial infection that enters the bloodstream and it often
causes organ failure or injury. In new born babies, early onset sepsis takes place in the first
72 hours. Late onset sepsis takes place after 72 hours. Sepsis mostly affects older adults,
pregnant women, people with chronic conditions and weakened immune systems.
The study aims to identify potential metabolic profile and biomarker related to sepsis.
The report also aims to evaluate on the capability of metabolomics in identifying a potential
metabolic profile.
The three studies made use of urine samples from ICU patients, and neonates.
Metabolic profiling made use of 1H NMR, and were further analyzed with either GC-MS or LC-
MS.
Results of the studies showed that acetone, lactose and glucose were the metabolites
found in neonates. Adult sepsis includes glucose, hippurate and ethanol as the metabolites.
Energy-producing biosynthetic pathways have the mainly relevant metabolic alterations in
sepsis.
In conclusion, metabolomics is a powerful tool in metabolite profiling that leads to a
more efficient treatment of diseases.

2. Concept map

have
 Summary:
Sepsis was investigated in new born babies (early onset and late onset sepsis) and adult ICU
patients. Metabolomic profiling were done using 1H NMR and further investigated with either
GC-MS or LC-MS. Results showed that the metabolites that may have a great contribution in the
metabolic pathway of sepsis include lactose, ethanol, hippurate, acetone and glucose.

3. Molecular/biochemical technique highlighted in the seminar

4.
The biochemical technique highlighted in the seminar was proton nuclear magnetic
resonance (1H NMR). The principle of 1H NMR is based on the spins of atomic nuclei. The
magnetic measurements depend upon the spin of unpaired electron whereas nuclear
magnetic resonance measures magnetic effect caused by the spin of protons and
neutrons. Both these nucleons have intrinsic angular momenta or spins and hence act as
elementary magnet.
The existence of nuclear magnetism was revealed in the hyper fine structure of
spectral lines. If the nucleus with a certain magnetic moment is placed in the magnetic
field, the phenomenon of space quantization can be observed and for each allowed
direction there will be a slightly different energy level.
GC-MS and LC-MS/MS were also used in the study. GC is used to vaporize the sample
to prepare pure compounds from a mixture. LC was used to separate the mixture into its
components before further analysis in MS.

5. Purpose the technique used in the study

1
H-NMR spectroscopy was used for the analysis of the urine samples from neonates
and adults in an attempt to obtain an overall view of the metabolic profile.

6. Main findings of the study upon the use of the technique

Table 1 shows the dominant metabolites in the urine sample identified from control
and septic newborns. The 1H-NMR signals corresponding to 3.4, 3.36, and 1.36 ppm
showed indicates that septic samples contained relatively higher contents of glucose and
lactate with respect to the control group (Fig. 1b). In contrast, control samples appeared
to be richer in citrate and creatinine, as shown at 2.56, 4.08 ppm.
In the study of urine metabolomics of neonates with late-onset sepsis, researchers
were able to document a clear separation between neonates fulfilling the diagnostic
criteria for LOS and non-septic controls. Non-targeted metabolomics investigation
revealed significant changes in 10 urine metabolites (Table 2). On day 3 and day 10, there
were no significant alterations between the groups.
With the targeted metabolomics approach (LC-MS/MS) that was applied next for the
quantification of additional important metabolites, researchers found statistically
significant alternation in 17 metabolites in the urine, three of which were also found to be
statistically significant by 1H-NMR spectroscopy.
Table 1. 1H NMR chemical shift of metabolites found in urine samples of control and septic
groups (1)

Fig. 1. 1H-NMR spectra of urine samples from (a) control and (b) septic subjects with main
assignments (1).

Table 2. Metabolites found by 1H NMR to have significantly changed in septic neonates and
on the onset of the disease (2)
7. Evaluation of the study’s overall success in achieving the objectives
The objectives of the study were met. The urine metabolic profile that causes sepsis
was properly investigated. The samples were properly prepared and separated prior to
chemical analysis with the use of gas chromatography and liquid chromatography.
Further analysis with mass spectroscopy highly supported the identity of the distinct
metabolites obtained from septic research participants. The statistical tools that were
used showed a clearer research result. The principal component analysis, PLS-DA and
multivariate statistical model facilitated the identification of groupings of the metabolites
that serves as biochemical markers for the detection and timely treatment of sepsis.

References:
[1] Sarafides, et al. 2017. Urine metabolomics in neonates with late-onset sepsis in a case-control
study. Scientific Reports | 7:45506 | DOI: 10.1038/srep45506

[2] V. Fanos et al. 2014. Urinary 1H-NMR and GC-MS metabolomics predicts early and late onset
neonatal Sepsis. Early Human Development 90S1, S78–S83