Abstract: A new sythetic gene coding for a Human
Elastin-Like Polypeptide was constructed and expressed. The
recombinant product was tested as coating agent to realize a
surface suitable for cell growth. Coatings showed peculiar
features and different human cell lines were seeded and
cultured. All cell lines tested showed to adhere and proliferate
on this substrate that has been shown also to exert a specific
effect on cells, depending on cell type.
Abstract: The effect of cross linking of the protein isolates of
three legumes with the microbial enzyme transglutaminase (EC
2.3.2.13) on the functional properties at different NaCl concentration
was studied. The reduction in the total free amino groups (OD340) of
the polymerized protein showed that TGase treatment cross-linking
the protein subunit of each legume. The solubility of the protein
polymer of each legume was greatly improved at high concentration
of NaCl. At 1.2 M NaCl the solubility of the native legumes protein
was significantly decreased but after polymerization slightly
improved. Cross linked proteins were less turbid on heating to higher
temperature as compared to native proteins and the temperature at
which the protein turns turbid also increased in the polymerized
proteins. The emulsifying and foaming properties of the protein
polymer were greatly improved at all concentrations of NaCl for all
legumes.
Abstract: The purpose of the present work was to study the
production and process parameters optimization for the synthesis of
cellulase from Trichoderma viride in solid state fermentation (SSF)
using an agricultural wheat straw as substrates; as fungal conversion
of lignocellulosic biomass for cellulase production is one among the
major increasing demand for various biotechnological applications.
An optimization of process parameters is a necessary step to get
higher yield of product. Several kinetic parameters like pretreatment,
extraction solvent, substrate concentration, initial moisture content,
pH, incubation temperature and inoculum size were optimized for
enhanced production of third most demanded industrially important
cellulase. The maximum cellulase enzyme activity 398.10±2.43
μM/mL/min was achieved when proximally analyzed lignocellulosic
substrate wheat straw inocubated at 2% HCl as pretreatment tool
along with distilled water as extraction solvent, 3% substrate
concentration 40% moisture content with optimum pH 5.5 at 45°C
incubation temperature and 10% inoculum size.
Abstract: Response Surface Methodology (RSM) is a powerful
and efficient mathematical approach widely applied in the
optimization of cultivation process. Cellulase enzyme production by
Trichoderma reesei RutC30 using agricultural waste rice straw and
banana fiber as carbon source were investigated. In this work,
sequential optimization strategy based statistical design was
employed to enhance the production of cellulase enzyme through
submerged cultivation. A fractional factorial design (26-2) was applied
to elucidate the process parameters that significantly affect cellulase
production. Temperature, Substrate concentration, Inducer
concentration, pH, inoculum age and agitation speed were identified
as important process parameters effecting cellulase enzyme synthesis.
The concentration of lignocelluloses and lactose (inducer) in the
cultivation medium were found to be most significant factors. The
steepest ascent method was used to locate the optimal domain and a
Central Composite Design (CCD) was used to estimate the quadratic
response surface from which the factor levels for maximum
production of cellulase were determined.
Abstract: Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth each of six Candida spp. was inhibited significantly by ~13 mM SHAM or 2 mM cyanide, albeit to differing extents. In C. dubliniensis, C. krusei and C. tropicalis the rate of O2 uptake was inhibited by 18-36% by 25 mM SHAM, but this had little or no effect on C. glabrata, C. guilliermondii or C. parapsilosis. Although SHAM substantially inhibited the growth of Candida spp., it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of Candida spp. infection.
Abstract: Enzymatic hydrolysis of starch from natural sources
finds potential application in commercial production of alcoholic
beverage and bioethanol. In this study the effect of starch
concentration, temperature, time and enzyme concentration were
studied and optimized for hydrolysis of Potato starch powder (of
mesh 80/120) into glucose syrup by immobilized (using Sodium
arginate) α-amylase using central composite design. The
experimental result on enzymatic hydrolysis of Potato starch was
subjected to multiple linear regression analysis using MINITAB 14
software. Positive linear effect of starch concentration, enzyme
concentration and time was observed on hydrolysis of Potato starch
by α-amylase. The statistical significance of the model was validated
by F-test for analysis of variance (p ≤ 0.01). The optimum value of
starch concentration, enzyme concentration, temperature, time and
were found to be 6% (w/v), 2% (w/v), 40°C and 80min respectively.
The maximum glucose yield at optimum condition was 2.34 mg/mL.
Abstract: The study was designed to evaluate the use of low
concentrations of separan flocculent (Less than 3 ppm) on
physicochemical properties of sugar cane juice. Colour, pH, purity,
turbidity, pol, brix, reducing sugars tannins and polyphenols of
crushed cane (green and burned) juice, mixed juice and clarified
juice were studied. The results showed that pol, brix, reducing sugar
and turbidity are higher in crushed cane juice. Clarified burned juice
had low turbidity, reducing sugars, pol and brix but had significantly
lower pH, purity and colour when compared to crushed juice.
Polyphenols of the crushed juice (1.19%) decreased significantly in
the clarified juice to 0.006%. Addition of separan at a concentration
of 0.015 ppm reduced significantly colour, polyphenols and tannins
and reducing sugar compared to the control.
Abstract: Nowadays, biometrical characterizations of Artemia
cysts are used as one of the most important factors in the study of
Artemia populations and intraspecific particularity; meanwhile these
characters can be used as economical indices. For example, typically
high hatching efficiency is possible due to the small diameter of
cysts (high number per gram); therefore small diameter of cysts
show someway high quality of cysts. This study was performed
during a ten year period, including two different ecological
conditions: rainy and drought. It is important from two different
aspects because it covers alteration of A. urmiana during ten years
also its variation in the best and worst environmental situations in
which salinity increased from 173.8 ppt in 1994 to 280.8 ppt in
2003/4. In this study the biometrical raw data of Artemia urmiana
cysts at seven stations from the Urmia Lake in 1994 and their seven
identical locations at 26 studied stations in 2003/4 were reanalyzed
again and compared together. Biometrical comparison of untreated
and decapsulated cysts in each of the seven similar stations showed a
highly significant variation between 1994 and 2003/4. Based on this
study, in whole stations the untreated and decapsulated cysts from
1994 were larger than cysts of 2003/4 without any exception. But
there was no logical relationship between salinity and chorion
thickness in the Urmia Lake. With regard to PCA analyses the
stations of two different studied years certainly have been separated
with factor 1 from each other. In conclusion, the interaction between
genetic and environmental factors can determine and explain
variation in the range of cysts diameter in Artemia.
Abstract: To evaluate genetic variation of wheat (Triticum aestivum) affected by heat and drought stress on eight Australian wheat genotypes that are parents of Doubled Haploid (HD) mapping populations at the vegetative stage, the water stress experiment was conducted at 65% field capacity in growth room. Heat stress experiment was conducted in the research field under irrigation over summer. Result show that water stress decreased dry shoot weight and RWC but increased osmolarity and means of Fv/Fm values in all varieties except for Krichauff. Krichauff and Kukri had the maximum RWC under drought stress. Trident variety was shown maximum WUE, osmolarity (610 mM/Kg), dry mater, quantum yield and Fv/Fm 0.815 under water stress condition. However, the recovery of quantum yield was apparent between 4 to 7 days after stress in all varieties. Nevertheless, increase in water stress after that lead to strong decrease in quantum yield. There was a genetic variation for leaf pigments content among varieties under heat stress. Heat stress decreased significantly the total chlorophyll content that measured by SPAD. Krichauff had maximum value of Anthocyanin content (2.978 A/g FW), chlorophyll a+b (2.001 mg/g FW) and chlorophyll a (1.502 mg/g FW). Maximum value of chlorophyll b (0.515 mg/g FW) and Carotenoids (0.234 mg/g FW) content belonged to Kukri. The quantum yield of all varieties decreased significantly, when the weather temperature increased from 28 ÔùªC to 36 ÔùªC during the 6 days. However, the recovery of quantum yield was apparent after 8th day in all varieties. The maximum decrease and recovery in quantum yield was observed in Krichauff. Drought and heat tolerant and moderately tolerant wheat genotypes were included Trident, Krichauff, Kukri and RAC875. Molineux, Berkut and Excalibur were clustered into most sensitive and moderately sensitive genotypes. Finally, the results show that there was a significantly genetic variation among the eight varieties that were studied under heat and water stress.
Abstract: Gene expression profiling is rapidly evolving into a
powerful technique for investigating tumor malignancies. The
researchers are overwhelmed with the microarray-based platforms
and methods that confer them the freedom to conduct large-scale
gene expression profiling measurements. Simultaneously,
investigations into cross-platform integration methods have started
gaining momentum due to their underlying potential to help
comprehend a myriad of broad biological issues in tumor diagnosis,
prognosis, and therapy. However, comparing results from different
platforms remains to be a challenging task as various inherent
technical differences exist between the microarray platforms. In this
paper, we explain a simple ratio-transformation method, which can
provide some common ground for cDNA and Affymetrix platform
towards cross-platform integration. The method is based on the
characteristic data attributes of Affymetrix- and cDNA- platform. In
the work, we considered seven childhood leukemia patients and their
gene expression levels in either platform. With a dataset of 822
differentially expressed genes from both these platforms, we carried
out a specific ratio-treatment to Affymetrix data, which subsequently
showed an improvement in the relationship with the cDNA data.
Abstract: Ethanol is generally used as a therapeutic reagent against Hepatocellular carcinoma (HCC or hepatoma) worldwide, as it can induce Hepatocellular carcinoma cell apoptosis at low concentration through a multifactorial process regulated by several unknown proteins. This paper provides a simple and available proteomic strategy for exploring differentially expressed proteins in the apoptotic pathway. The appropriate concentrations of ethanol required to induce HepG2 cell apoptosis were first assessed by MTT assay, Gisma and fluorescence staining. Next, the central proteins involved in the apoptosis pathway processs were determined using 2D-PAGE, SDS-PAGE, and bio-software analysis. Finally the downregulation of two proteins, AFP and survivin, were determined by immunocytochemistry and reverse transcriptase PCR (RT-PCR) technology. The simple, useful method demonstrated here provides a new approach to proteomic analysis in key bio-regulating process including proliferation, differentiation, apoptosis, immunity and metastasis.
Abstract: The habitat where the present study has been carried
out is productive in relation to nutrient quality and they may perform
several useful functions, but are also threatened for their existence.
Hence, the proposed work, will add much new information about
biodiversity of macrophytes in drains and their embankment. All the
species were identified with their different stages of growth which
encountered on the three selected sites (I, II and III). The number of
species occurring at each site is grouped seasonally, i.e. summer,
rainy and winter season and the species were further recorded for the
study of phytosociology. Phytosociological characters such as
frequency, density and abundance were influenced by the climatic,
anthropogenic and biotic stresses prevailing at the three study sites.
All the species present at the study sites have shown maximum
values of frequency, density and abundance in rainy season in
comparison to that of summer and winter seasons.
Abstract: β-Glucosidase is an important enzyme for production
of ethanol from lignocellulose. With hydrolytic activity on
cellooligosaccharides, especially cellobiose, β-glucosidase removes
product inhibitory effect on cellulases and forms fermentable sugars.
In this study, β-glucosidase encoding gene (BGL1) from traditional
starter yeast Saccharomycosis fibuligera BMQ908 was cloned and
expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared
98% nucleotide homology with the closest GenBank sequence
(M22475) but identity in amino-acid sequences of catalytic domains.
Recombinant plasmid pPICZαA/BGL1 containing the sequence
encoding BGL1 mature protein and α-factor secretion signal was
constructed and transformed into methylotrophic yeast P. pastoris by
electroporation. The recombinant strain produced single extracellular
protein with molecular weight of 120 kDa and cellobiase activity of
60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0
and the optimum temperature was 50°C.
Abstract: Sorghum flour was supplemented with 15 and 30%
chickpea flour. Sorghum flour and the supplement were fermented at
35 oC for 0, 8, 16, and 24 h. Changes in pH, titrable acidity, total
soluble solids, protein content, in vitro protein digestibility and
amino acid composition were investigated during fermentation and/or
after supplementation of sorghum flour with chickpea. The pH of the
fermenting material decreased sharply with a concomitant increase in
the titrable acidity. The total soluble solids remained unchanged with
progressive fermentation time. The protein content of sorghum
cultivar was found to be 9.27 and that of chickpea was 22.47%. The
protein content of sorghum cultivar after supplementation with15 and
30% chickpea was significantly (P ≤ 0.05) increased to 11.78 and
14.55%, respectively. The protein digestibility also increased after
fermentation from 13.35 to 30.59 and 40.56% for the supplements,
respectively. Further increment in protein content and digestibility
was observed when supplemented and unsupplemented samples were
fermented for different periods of time. Cooking of fermented
samples was found to increase the protein content slightly and
decreased digestibility for both supplements. Amino acid content of
fermented and fermented and cooked supplements was determined.
Supplementation was found to increase the lysine and therionine
content. Cooking following fermentation decreased lysine,
isoleucine, valine and sulfur containg amino acids.
Abstract: Lutein is a dietary oxycarotenoid which is found
to reduce the risks of Age-related Macular Degeneration
(AMD). Supercritical fluid extraction of lutein esters from
marigold petals was carried out and was found to be much
effective than conventional solvent extraction. The
saponification of pre-concentrated lutein esters to produce free
lutein was studied which showed a composition of about 88%
total carotenoids (UV-VIS spectrophotometry) and 90.7%
lutein (HPLC). The lipase catalyzed hydrolysis of lutein esters
in conventional medium was investigated. The optimal
temperature, pH, enzyme concentration and water activity
were found to be 50°C, 7, 15% and 0.33 respectively and the
activity loss of lipase was about 25% after 8 times re-use in at
50°C for 12 days. However, the lipase catalyzed hydrolysis of
lutein esters in conventional media resulted in poor
conversions (16.4%).
Abstract: Endemic Artemia franciscana populations can be found throughout the American continent and also as an introduced specie in several country all over the world, such as in the Mediterranean region where Artemia franciscana was identified as an invasive specie replacing native Artemia parthenogenetica and Artemia salina. In the present study, the characterization of the new invasive Artemia franciscana reported from Sabkhet Halk El-Menzel (Tunisia) was done based on the cysts biometry, nauplii instar-I length, Adult sexual dimorphism and fatty acid profile. The mean value of the diameter of non-decapsulated and decapsulated cysts, chorion thickness and naupliar length is 235.8, 226.3, 4.75 and 426.8 μm, respectively. Sexual dimorphism for adults specimen showed that maximal distance between compound eyes, diameter for compound eyes, length of first antenna and the abdomen length compared to the total body length ratio, are the most important variables for males and females discrimination with a total contribution of 62.39 %. The analysis of fatty acid methyl esters profile of decapsulated cysts resulted in low levels of linolenic acid (LLA, C18:3n-3) and high levels of eicosapentaenoic acid (EPA, C20:5n-3) with 3.11 and 11.10 %, respectively. Low quantity of docosahexaenoic acid (DHA, 22:6n-3) was also observed with 0.17 mg.g-1 dry weight.
Abstract: Artemia is one of the most conspicuous invertebrates
associated with aquaculture. It can be considered as a model
organism, offering numerous advantages for comprehensive and
multidisciplinary studies using morphologic or molecular methods.
Since DNA extraction is an important step of any molecular
experiment, a new and a rapid method of DNA extraction from adult
Artemia was described in this study. Besides, the efficiency of this
technique was compared with two widely used alternative techniques,
namely Chelex® 100 resin and SDS-chloroform methods. Data
analysis revealed that the new method is the easiest and the most cost
effective method among the other methods which allows a quick and
efficient extraction of DNA from the adult animal.
Abstract: Lectins have a good scope in current clinical
microbiology research. In the present study evaluated the
antimicrobial activities of a D-galactose binding lectin (PnL) was
purified from the annelid, Perinereis nuntia (polychaeta) by affinity
chromatography. The molecular mass of the lectin was determined to
be 32 kDa as a single polypeptide by SDS-PAGE under both reducing
and non-reducing conditions. The hemagglutinating activity of the
PnL showed against trypsinized and glutaraldehyde-fixed human
erythrocytes was specifically inhibited by D-Gal, GalNAc,
Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro
antibacterial screening studies against 11 gram-positive and
gram-negative microorganisms. From the screening results, it was
revealed that PnL exhibited significant antibacterial activity against
gram-positive bacteria. Bacillus megaterium showed the highest
growth inhibition by the lectin (250 μg/disc). However, PnL did not
inhibit the growth of gram-negative bacteria such as Vibrio cholerae
and Pseudomonas sp. PnL was also examined for in vitro antifungal
activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited
the mycelial growth of Alternaria alternata (24.4%). These results
indicate that future findings of lectin applications obtained from
annelids may be of importance to life sciences.
Abstract: The present study has been taken to explore the
screening of in vitro antimicrobial activities of D-galactose-binding
sponge lectin (HOL-30). HOL-30 was purified from the marine
demosponge Halichondria okadai by affinity chromatography. The
molecular mass of the lectin was determined to be 30 kDa with a
single polypeptide by SDS-PAGE under non-reducing and reducing
conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed
rabbit and human erythrocytes with preference for type O
erythrocytes. The lectin was subjected to evaluation for inhibition of
microbial growth by the disc diffusion method against eleven human
pathogenic gram-positive and gram-negative bacteria. The lectin
exhibited strong antibacterial activity against gram-positive bacteria,
such as Bacillus megaterium and Bacillus subtilis. However, it did
not affect against gram-negative bacteria such as Salmonella typhi
and Escherichia coli. The largest zone of inhibition was recorded of
Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in
diameter) at a concentration of the lectin (250 μg/disc). On the other
hand, the antifungal activity of the lectin was investigated against six
phytopathogenic fungi based on food poisoning technique. The lectin
has shown maximum inhibition (22.83%) of mycelial growth of
Botrydiplodia theobromae at a concentration of 100 μg/mL media.
These findings indicate that the lectin may be of importance to
clinical microbiology and have therapeutic applications.
Abstract: Candida albicans ATCC 10231 had low endogenous activity of the alternative oxidase compared with that of C. albicans ATCC 10261. In C. albicans ATCC 10231 the endogenous activity declined as the cultures aged. Alternative oxidase activity could be induced in C. albicans ATCC 10231 by treatment with cyanide, but the induction of this activity required the presence of oxygen which could be replaced, at least in part, with high concentrations of potassium ferricyanide. We infer from this that the expression of the gene encoding the alternative oxidase is under the control of a redoxsensitive transcription factor.