1H-NMR based metabolic study of MMTV-PyMT mice along with pathological progress to screen biomarkers for early diagnosis of breast cancer

1H NMR based metabonomic approach was applied to monitor the alterations of serum metabolic profiles in MMTV-PyMT transgenic mice to detect the dynamic changes related to the pathological process and...

Simultaneous Quantification of Propylthiouracil and Its N-β-d Glucuronide by HPLC-MS/MS: Application to a Metabolic Study

Propylthiouracil (PTU) is commonly prescribed for the management of hyperthyroidism and thyrotoxicosis. Although the exact mechanism of action is not fully understood, PTU is associated with hepatoxicity in pediatric population. Glucuronidation mediated by uridine 5′-diphospho-glucuronosyltransferases (UGTs), which possess age-dependent expression, has been proposed as an important metabolic pathway of PTU. To further examine the metabolism of PTU, a reliable HPLC-MS/MS method for the simultaneous quantification of PTU and its N-β-D glucuronide (PTU-GLU) was developed and validated. The chromatographic separation was achieved on a ZORBAX Extend-C18 column (2.1 × 50 mm, 1.8 μm) through gradient delivery of a mixture of formic acid, methanol and acetonitrile. The electrospray ionization (ESI) was operated in its negative ion mode while PTU and PTU-GLU were detected by multiple reaction monitoring (MRM). This analytical method displayed excellent linearity, sensitivity, accuracy, precision, recovery and stability while its matrix effect and carry-over were insignificant. Subsequently, the in vitro metabolism of PTU was assessed and UGT1A9 was identified as an important UGT isoform responsible for the glucuronidation of PTU. The information obtained from this study will facilitate future mechanistic investigation on the hepatoxicity of PTU and may optimize its clinical application.

Insight into the Metabolic Profiles of Pb(II) Removing Microorganisms

The objective of the study was to gather insight into the metabolism of lead-removing microorganisms, coupled with Pb(II) removal, biomass viability and nitrate concentrations for Pb(II) bioremoval using an industrially obtained microbial consortium. The consortium used for study has proven to be highly effective at removing aqueous Pb(II) from solution. Anaerobic batch experiments were conducted with Luria-Bertani broth as rich growth medium over a period of 33 h, comparing a lower concentration of Pb(II) with a higher concentration at two different nutrient concentrations. Metabolite profiling and quantification were conducted with the aid of both liquid chromatography coupled with tandem mass spectroscopy (UPLC-HDMS) in a “non-targeted” fashion and high-performance liquid chromatography (HPLC) in a “targeted” fashion. Four main compounds were identified, and a metabolic study was conducted on each to establish their possible significance for Pb(II) bioremoval. The study investigates the first metabolic profile to date for Pb(II) bioremoval, which in turn can result in a clarified understanding for development on an industrial and microbial level.

Evaluation of cassava foliage as a protein supplement for sheep

Three trials were conducted to evaluate the potential of cassava (Manihot esculenta) foliage (leaves and petioles) as a protein supplement for sheep. In the first trial, nylon bag degradability of cassava foliage was studied. Proximate analysis of cassava foliage was conducted before and after the incubation. Rumen Ammonia Concentration (RAC) and pH were determined at 0, 4 and 8 hours after feeding cassava foliage diet (30%). Proximate composition of sundried cassava foliage was (%) DM 91.25, CP 18.55, NDF, 31.41, ADF, 29.2, EE, 6.6, Ash, 12.95. Nylon-bag dry matter disappearance (DMD) increased significantly (P<0.05) from 15.75% to 75.5% RAC values increased significantly from 8.20mg NH3/100ml at zero hour to 11.41mg NH3/100ml at 8 hours post feeding. Rumen fluid pH was not significantly changed by the 30% cassava foliage diet. In the second trial lasting 56 days, twenty-eight Yankasa/ WAD yearling rams of initial average weight of 18kg were balanced for weight and randomly allotted to seven treatments. All rams except those on control diet were fed cassava foliage or groundnut haulms or both at 1.0% or 1.5% of body weight (BW) as supplements to a basal diet of Gamba hay. Feeding Gamba hay alone (control) resulted in weight loss (-30.5g/day) but with cassava foliage or groundnut haulms supplementation at 1.0% BW, significantly higher gains of 39.2 and 44.6g/day were achieved respectively. At 1.5%BW level of supplementation with cassava foliage or groundnut haulms higher weight gains of 41.2 and 51.7g/day were achieved while feeding the (50.50) combined supplements at 1.0 and 1.50% BW resulted in live weight gains of 65.1 and 69.3g/day respectively. The third trial consisted of a four week grazing trial followed by a metabolic study. Twenty-one Yankasa/WAD yearling rams were randomly allotted to seven treatments and were balanced for live weight. All rams were allowed to graze standing digitaria hay as basal diet, except the rams on control diet. Those on treatment were fed cassava foliage or groundnut haulms or a (50:50) combination of both at 0.5% and 1.0% (BW). The metabolic study consisted of a seven day adjustment period and a seven day total collection period. There was no significant difference (P>0.05) in the live weight gains of rams on the control diet compared with those supplemented except for those supplemented with cassava foliage and groundnut haulms combined at 1% level where the weight gain was significantly higher (P<0.05). Feed intake was not significantly (P>0.05) affected by feeding either supplement. Supplementing digitaria hay with cassava foliage at both 0.5% and 1.0% BW did not significantly increase the nitrogen retention values, but groundnut haulms supplemented at 1.0% BW significantly (P<0.05) increased the nitrogen retention values.