Title: The Production of Genomic DNA Library of Aureobasidiumpullulansby Lambda Bacteriophage Method.
Author:Steve Andersen

Abstract:

The production of a genomic DNA library of Aureobasidiumpullulansusing

Promega’sPackagene System was undertaken but not completed.A. pullulans

genomic DNA was isolated, and analyzed by spectrophotometry to determine DNA

concentration and purity.The genomic DNA was digested by a serial

dilution of Sau A31 restriction endonuclease to determine the proper concentration of Sau A31 to yield genomic DNA fragments of 15 – 23 KB. Partial fill-in of the 4 base over-hang of the Sau 3A1 digested A. pullulans genomic DNA was performed using Klenow fragment and dGTP and dATP. Ligation of BamH1 arms of lambda bacteriophage DNA to a positive control insert by T4 DNA ligase was successfully performed.Subsequently, the ligated BamH1 arms containing the positive control insert were packaged into lambda phage. The phage titer was determined on LB plates (E. coli / top agar method).

Title: Use of acid whey powder in probiotic yogurt making.
Author: Rajesh Parmar
Abstract
A study was undertaken to explore the possibilities of partial replacement of skim milk powder (SMP) with acid whey powder (AWP) in yogurt formulation. Probiotic yogurts were made using commercially available two different brands of AWP (suppliers : Main Street Ingredients(MS) and Century Food (CF) ) . The yogurt samples were analyzed for change in titratable acidity, pH and lactic count during manufacturing. Also the fresh products were analyzed for viscosity, firmnessandsyneresis. Changes in acidity, pH and lactic count were recorded over a period of 10, 20, 30 and 40 days of storage of the product at 4 o C. The products were given for organoleptic evaluation to a panel of expert judges on day 1 and day 30.
Data obtained revealed that partial replacement of SMP with AWP substantially reduced the incubation time of yogurt with a culture used for long set method (culture ABT-1).In case of a fast growing culture (ABT-4) there was not a significant difference between incubation periods of control and treated yogurts. When AWPs were used to have 0.5 % protein fortification, viscosity andfirmness ofyogurt samples were more or less same as control samples but increasing the level of AWPs to have 1 % protein fortification led to weak body. However this has a positive effect on the water binding capacity of yogurts and hence addition of AWPs to have 1 % protein fortification reduced syneresis significantly. As far as lactic counts were concerned, addition of AWP (MS) helped one of the probiotic bacteria Lactobacillus acidophilus to maintain viable population well above 10 6cfu /gm for as long as 20 days. There was not a significant difference in the viable counts of Bifidobacteria and Streptococci between treated and control yogurt samples. However this bacterial population remained well above 10 7cfu /gm throughout the storage period of 40 days at 4 o C. There was not a significant difference between the sensory scores of yogurts made using AWP ( MS) and control samples. However yogurts made using AWP (CF) were unacceptable based on sensory scores obtained.


Title: Intestinal Regulation of Neonatal Development of B Lymphocyte Subsets
Author: Mike Graybill
Abstract:
The period immediate after birth is the most challenging period for the developing immune system.This is particularly true in domestic animals, which develop in utero in the absence of exogenous antigen and are immediately bombarded with a diverse array of environmental microflora upon birth.During this early period, very few circulating B cells are present and the majority of neonatal immunity is obtained passively from the mother via the milk and colostrums.Immediately post-birth, there is an explosion in the production of new B cells which eventually rise from minimal 5-10% of peripheral blood lymphocytes
(PBLs) at birth to a stable level of roughly 50% of PBLs at 6 months of age.Recently, it has become clear that colostrum contains not only maternal antibody, but also cells and soluble factors which may stimulate B cell production.Furthermore, a unique subset of non-circulating B cells develop beginning at 6-8 weeks of age, to reach levels of roughly 50% of PBLs.Coincidentally, the appearance of this subset develops alongside the development of reactivity to a number if intestinal microflora.This research project is aimed at investigating two distinct questions:(a) the role of colostrums, and specifically soluble CD14 and macrophages, at promoting the development and release of ilealPeyer’s patch B cells in the neonatal period.(b)the role of intestinal microflora in the development of B cell subsets and immune competence.Identification of unique stages of B cell development have relied and continue to rely upon 2, 3, and 4-color phenotypic analysis to define functional and developmental subsets.The experiments to define the function and development of these subsets will necessarily rely upon the use of fluorescent tracking dyes (PKH, CellTracker dyes, 5, 6-Carboxyfluorescein succinmydl ester) in conjunction with multicolor flow cytometry.Typically, mature B cell subsets can be defined based upon the combined expression of surface immunoglobulin, CD21, and either CD11b or CD11c.The addition of either a red or a green fluorescent tracking label to monitor the phenotypic changes and the physiological growth of the B cell pools will require 4-color cytometry to obtain statistically meaningful results.These data will allow a greater understanding of the unique development of the ruminant immune system, and enhance the utility and design of neonatal agricultural vaccines.


Title: Bioavailability of Vitamin D3 from Fortified Process Cheese
Author: Jana Johnson
Abstract:
Due to changes in the consumption patterns of milk and Process cheese, and to the availability of new methods for the fortification of Process cheese, the objective of this study was to determine the effect of 2 months of daily vitamin D fortified Process cheese consumption, delivering 600 IU/ vitamin D per day, on changes in serum vitamin D concentrations or bone markers.The three serum markers that were observed were 25-OHD, PTH, and OC.The population consisted of men and women over age 60 years old and healthy, which were split into three groups: one that consumed Process cheese fortified with vitamin D (200IU/serving), one group that consumed Process cheese with no additional vitamin D, and a control group that consumed no Process cheese.The hypotheses were that people who received vitamin D fortified Process cheese would show higher serum 25-OHD, lower PTH, and lower OC values.Preliminary results have shown that the group that consumed the fortified Process cheese had a drop in 25-OHD, an increase in PTH, and a drop in OC.The group that consumed regular non-fortified Process cheese exhibited an increase in 25-OHD, a drop in PTH, and a drop in OC.The control group showed 25-OHD levels that remained constant, a slight drop in PTH, and a drop in OC.These results are deviant from what was hypothesized, and thus, a continuing study is in the works to determine the bioavailability of vitamin D2 from Process cheese versus from a distilled water dilution given to various age groups.


Title: Utilization Of Corn Distillers Dried Grains In Dry Extruded Pet Foods
Author:ChiragShukla

Abstract:

The demand of protein has always been on the rise in any food product. There 

have been consistent efforts to find out different utilizable sources of 

protein that can be used successfully in human food and animal food/feed. 

Distillers dried grains (DDG), a rich source of protein and a co-product of 

the distillery industry, has been used to feed pigs and poultry. Our 

intention in this research is to manufacture pet food having DDG as one of 

the major component, using a single screw extruder. Different blends were 

made and extruded keeping processing parameters constant. We found that DDG 

can be successfully utilized to make pet food using extrusion technology. Up 

to 50% of DDG can be incorporated in a formulation to make pet food. A 

moisture level below 10% will ensure a desirable product.

Title: Protective effects of Nicotinamide in Human Cortical Neurons
Author: Suraj Bhansali
Abstract:
   Neurodegenerative diseases are quite prevalent in United States, especially in older people and are the third highest natural cause of death in USA. These diseases include ischemia, Alzheimer’s disease and Parkinson’s disease among others.  Previous studies have shown that cell death (apoptosis) in neurodegenerative diseases occurs because of oxidative stress of free radicals in the brain.  The oxygen radical generating toxin tertiary butylhydroperoxide (t-BuOOH), which induces DNA fragmentation and apoptosis in neuronal cells, was used to mimic the oxidative injury that has been implicated in these diseases.  It was observed that t-BuOOH induces DNA fragmentation and apoptosis in all regions of mouse brain, and that it also causes significant cell death in human cortical neurons (HNC1-A and HNC2) in culture.  In addition, previous studies have also shown that the poly (ADP ribose) polymerase (PARP) inhibitor nicotinamide is able to prevent DNA fragmentation and apoptosis induced by t- BuOOH in mouse brain and in human neuronal cells HCN1-A and HCN2.  Evidence indicate that nicotinamide is able to prevent the up-regulation of the pro-apoptotic proteins p53 and p21/WAF-1, and the down regulation of the anti-apoptotic protein bcl-2 that is induced by t-BuOOH in HCN1-A and HCN2 cells, though exact molecular mechanism at cellular level is not known.  My research involves delineation of molecular mechanism(s) by which nicotinamide is able to protect HCN1-A and HCN2 cells in presence of free radical generating toxin.  This project applies the techniques of genomics to examine the molecular mechanism underlying neuronal protection. The central hypothesis of my project is that nicotinamide protects human brain cells from the toxic effects of free radical generating toxin by regulating the levels of various pro- and anti-apoptotic proteins and that this protection can be enhanced in combination with other neuro-protective agents.
    Significance of this project includes evaluation of two human cortical neuronal cell lines, HCN1-A and HCN2, which can be used as cell culture model systems to evaluate various potential neuro-protective agents.  This study will provide insight into the effect that PARP inhibitors can have in preventing neuronal death and hence can be potential therapeutic agents against neurodegenerative diseases.  It will also help in better understanding of the underlying molecular mechanism(s) involved in the protective effects of PARP inhibitors in the brain by studying the effects of PARP inhibitors on regulation of pro-and anti-apoptotic proteins.  Once a clear mechanism is established, therapeutic agents with different mechanism of action will be used in combination with   nicotinamide to observe synergistic effect in protecting human neurons.
 The experimental techniques involve cell culture of human cortical neuronal cells, extraction and purification of RNA from these cells, RNA gel electrophoresis, DNA microarray to compare up- and down-regulation of various genes.  Enzyme Linked Immono Sorbent Assay (ELISA) and Western immunoblot assays will be performed to determine the levels of various proteins.  The results from the DNA microarray will be corroborated with RT-PCR. Here at SDSU, I have successfully learned cell culture laboratory techniques and have been able to extract and purify RNA from human brain cells. I also have done RNA gel electrophoresis. In near future I will be doing all the aforementioned tests and assays.
 Following the completion of the project, I should be able to elicit the molecular mechanism(s) of protective effects of nicotinamide and identification of other potential drug candidates, which will show synergistic effect in combination with nicotinamide. It will open new horizons in the research field of   neurosciences.

Title: Application of biorenewable fibers in biocomposites.
Author: Subba Rao.
Abstract:
    Interest in the use of natural fibers has grown during the last decade due to their low costs and the search for renewable sources. Composites consisting of polypropylene (PP), or high-density polyethylene (HDPE), and biorenewable fibers from soybean hulls, wood, and big blue stem (BBS) were prepared by extrusion processing. The test samples were prepared by injection molding. Mechanical properties of the composites were evaluated. The effect of the fiber type and content on the mechanical properties of natural fibers/PP composites and natural fibers/HDPE composites were studied. The results showed that the fiber content influenced tensile and flexural properties. The wood and big blue stem fiber composites had higher Young’s modulus and tensile strength than soybean hull fiber composites.  The Young’s modulus of wood, big blue stem and soybean hull fiber composites compared with pure polypropylene and polyethylene were comparable or higher. Biorenewable fibers like soybean hulls, big blue stem can be used as excellent reinforcing materials for low cost composites. The melt flow index of these composites was reduced due to the fibers, restricting the flow of the polymer molecules.

Title: Incorporation of acid whey in making yogurt
Author: Gaurav Singhal
Abstract
Cottage cheese whey is a cheap and good source of pre-digested whey proteins. Our objective was to optimize the incorporation of acid whey solids and make them a functional yogurt ingredient. Cottage cheese whey (0.87% protein) and ultrafiltered cottage cheese whey (0.5% and 1.0% protein) were used to replace 1.9%, 5.5% and 11.1% respectively of the total milk protein (4.5%) in yogurt. For long set yogurt cultures, curd making time was reduced to 8hrs and 10hrs with 11.1% and 5.5% protein supplementation respectively compared to control and 1.9% protein supplementation that took 12hrs. There was, however, not much difference in curd set time for short set cultures. Gel strength of yogurt (measured as cone penetration) was 355mm, 389.5mm, 360.5mm and 357.5mm for control, 11.1%, 5.5% and 1.9% protein supplementation respectively. Syneresis in control and 11.1% protein supplementation was less (42.9ml and 43.6ml respectively) than the samples with 5.5% and 1.9% protein supplementation (46ml and 44.8ml respectively). Viscosity of control was the highest (11627cp) followed by 1.9% protein supplementation (11367cp), then was 5.5% protein supplementation (9483cp) and finally 11.1% protein supplementation (7923cp). There was no adverse effect on the yogurt and probiotic bacteria when supplemented with different ingredients.