Comparison of Cholesterol Lowering Diets: Apple, Casein Cytochrom P450 protein and Cholesterol 7α Hydroxylase Activities in Hamsters

Lithogenic diet, casein and apple fiber diets were fed to hamsters for 3-5 weeks. For control group, animals were fed on normal Purina chow without any supplement. The cholesterol lowering effect of lithogenic diet, casein and apple diets were compared. After dietary regimen, animals were screened for any gall stone formation. The isolated liver microsomes were separated from animals and tested for the cholesterol-7α Hydroxylase (CH) enzyme activity measurement in all three groups. The control animals did not show any gall stone formation and their CH enzyme activities were normal. The lithogenic diet showed significantly enhanced CH enzyme activities while animals fed on casein and apple diet regimen showed moderate increase in microsomal CH enzyme activity indicated cholesterol lowering in liver. In conclusion, cholesterol 7α hydroxylase may be a biomarker of cholesterol status in the body and microsomal CH enzyme may be lowered down after treatment of casein and apple diets

21 Tesla MRI Microimaging of Rat Heart by Superparamagnetic Antimyoglobin Bound Nanoparticles

Superparamagnetic iron-oxide bound antimyoglobin (SPIOM) characteristics were analyzed at different pH. After injecting SPIOM in heart, ex vivo magnetic resonance microimaging (MRM) technique was used to visualize microvasculature of rat heart including cardiac arteries, veins, cordate tendons attached with valves. The measurement accuracy of ventricles, aorta, vasculature and dimensions of cordate tendons were close to coregistered histology measurements

The Hepatocellular Hypoxia Criteria:2’Nitroimidazole Effect on Hepatocyte Carbohydrate Metabolizing Enzymes

Aim: to understand the 2’-nitroimidazole induced hypoxia and liver cell interaction, we proposed a “Hapatocellular Hypoxia Criteria”. Hypothesis: The nitroimidazole induced metabolic energy loss and oxygen depletion (hypoxia) in liver cell mitochondria causes the phagocytosis. Based on it, ten control subjects with 2’-nitroimidazole therapy were studied for their carbohydrate metabolizing enzymes in serum and hepatocellular enzymes in liver biopsy tissues. Materials and Methods: Proven ten control subjects were studied for hypoxia by enzyme assays. The 2’nitroimidazole treated paired ten subjects were studied for hypoxia using enzyme assays and hepatocellular cytomorphology by electron microscopy. Results and Discussion: Out of ten subjects on 2’-nitroimidazole, nine showed elevated carbohydrate metabolizing and lysosomal enzyme levels in serum. The enzymes glucokinase (in 80% samples), aldolase (in 80% samples), phosphofructokinase (in 80% samples), malate dehydrogenase (in 75% samples), isocitrate dehydrogenase (ICDH) (in 60% patients) were elevated while succinate dehydrogenase and lactate dehydrogenase (LDH) levels remained unaltered. Lysosomal enzymes β-glucuronidase, alkaline phosphatase, acid phosphatase, showed enhanced levels in the serum samples. In control ten liver biopsies, the hepatocytes and Kupffer cell preparations showed altered enzyme levels. Hepatocytes showed lowered glucokinase (in 80%), LDH (in 80%), and higher content of aldolase (in 80%), pyruvate kinase (in 100%), malate dehydrogenase (in 80%), ICDH (in 80%), citrate dehydrogenase (in 70%), phosphogluconate dehydrogenase (in 80%). Kupffer cells showed higher enzyme levels of β-glucuroronidase (in 80%), leucine aminopeptidase (in 70%), acid phosphatase (in 80%) and aryl sulphatase (in 88%). In these 10 biopsy samples from subjects on 2’-nitronidazole clinical trial, the electron microscopy cytomorphology observations showed swollen bizarre mitochondria, proliferative endoplasmic reticulum, and anisonucleosis after 2’-Nitroimidazole effect in liver cell damage. Conclusion: The proposed “Hepatocellular Hypoxia Criteria” served to define origin of liver hypoxia and showed altered hepatic enzyme activities with active phagocytosis and cytotoxicity in subjects after 2’-nitroimidazole treatment. The study suggests the enzyme based evaluation of nitroimidazole induced hypoxia monitoring and treatment of hepatic tumors and infected liver

Evaluation and Validation of clinical 4.23 T sodium MRI in animals and human: Application of oblique multi-slice spin-echo pulse sequence

Objective: Application of high-field 4.23 T MRI clinical imager was demonstrated for sodium-magnetic resonance imaging (MRI) data acquisition. Primary hypothesis: Sodium [Na] in brain is MR visible. Secondary hypothesis was, if, application of multislice spin echo (MSSE) pulse sequence at selected scan parameters can sufficiently visualize the total sodium signal as indicator of sub-clinical activity. Material and Methods: MSSE pulse sequence technique was used to simulate sodium images of human brain. For validation purpose, inversion recovery pulse sequence was validated by optimization of scan inversion time (TI). Phantom of sodium and rat brain were imaged. Sodium images were validated and compared with proton MRI images. Results: MSSE pulse technique enabled to visualize the sodium signal at optimized scan parameters. Specifically, MSSE pulse technique enabled the identification of different sodium rich areas due to their subphysiological activity in the brain, comparable with proton MRI images. Reconstruction images of brain further enhanced the power to classify the brain tissue. Intracellular sodium images of agarose-saline solution filled-tube phantom were generated by use of inversion recovery pulse sequence. Conclusion: Using MSSE pulse sequence at 4.23 T, in vivo sodium images can be generated within acceptable scan time for routine clinical brain examination for achieving better sub-physiological information as obtained from proton MRI

Emerging Trends of Nanotechnology towards Picotechnology: Energy and Biomolecules

In nature, metal oxide particles display their existence at the level of picomolecules in solution state and bioactive states in the body. We present evidence of picomolar behavior of molecules different than nanomolar behavior of particles. These particles can be encapsulated in polymers and can be functionalized with protein, nucleotides, and drugs to develop as smart intracellular targeting pico-devices. The preparation technique and physiological conditions decide the size and functionality of these pico-carrier devices. Their usable success rate, feasibility and potentials are yet to be proven or we do not know. The major difference between nanodevices and pico-devices is their intermolecular and intramolecular thermodynamics in medium and their molecular conformational interaction with molecular assembly in cytoarchitecture of the neurofilament, actin-myosin, microtubule proteins. Pico-carrier devices can be presumed as potential spears without interacting with host signal transduction and immunoprotection. In conclusion, the ultrafine size of newer picotechnology products may be better suited and easier to functionalize for design of particle based picodrugs, picochemicals, and pico-targeting molecules

Detection of Metabolites by Proton Ex Vivo NMR, in Vivo MR Spectroscopy Peaks and Tissue Content Analysis: Biochemical-Magnetic Resonance Correlation: Preliminary Results

*Aim*: Metabolite concentrations by in vivo magnetic resonance spectroscopy and ex vivo NMR spectroscopy were compared with excised normal human tissue relaxation times and tissue homogenate contents.

*Hypothesis*: Biochemical analysis combined with NMR and MR spectroscopy defines better tissue analysis.

*Materials and Methods*: Metabolites were measured using peak area, amplitude and molecular weights of metabolites in the reference solutions. In normal brain and heart autopsy, muscle and liver biopsy tissue ex vivo NMR peaks and spin-lattice (T1) and spin-spin (T2) relaxation times, were compared with diseased tissue NMR data in meningioma brain, myocardial infarct heart, duchene-muscular-dystrophy muscle and diffused-liver-injury liver after respective in vivo proton MR spectroscopy was done. NMR data was compared with tissue homogenate contents and serum levels of biochemical parameters.

*Results*: The quantitation of smaller NMR visible metabolites was feasible for both ex vivo NMR and in vivo MR spectroscopy. Ex vivo H-1 NMR and in vivo MRS metabolite characteristic peaks (disease/normal data represented as fold change), T1 and T2, and metabolites in tissue homogenate and serum indicated muscle fibrosis in DMD, cardiac energy depletion in MI heart, neuronal dysfunction in meningioma brain and carbohydrate-lipid metabolic crisis in DLI liver tissues.

*Conclusion*: This preliminary report highlights the biochemical-magnetic resonance correlation as basis of magnetic resonance spectroscopic imaging data interpretation of disease