Abstract: Soybean Natto powder was added to the burger in order to enhance the oxidative stability as well as decreases the microbial spoilage. The soybean bioactives compound (soybean Natto) as antioxidant and antimicrobial were added at level of 1, 2 and 3%. Chemical analysis and physical properties were affected by soybean Natto addition. All the tested soybean Natto additives showed strong antioxidant properties. The microbiological indicators were significantly (P < 0.05) affected by the addition of the soybean Natto. Decreasing trends of different extent were also observed in samples of the treatments for total viable counts, Coliform, Staphylococcus aureus, yeast and molds. Storage period was significantly (P < 0.05) affected on microbial counts in all samples Staphylococcus aureus were the most sensitive microbe followed by Coliform group of the sample containing soybean Natto. Sensory attributes were also performed, added soybean Natto exhibits beany flavor which was clear about samples of 3% soybean Natto.
Abstract: Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.
Abstract: Protein hydrolysates prepared from a number of medicinal plants are promising sources of various bioactive peptides. In this work, proteins from dried whole plant of Euphorbia hirta Linn. were extracted and digested with pepsin for 12h. The hydrolysates of lesser than 3 KDa were fractionated by a cut-off membrane. The peptide hydrolysate was then purified by an anion-exchange chromatography on DEAE-Sephacel™ column and reverse-phase chromatography on Sep-pak C18 column, respectively. The cytotoxic effect of each peptide fraction against a gastric carcinoma cell line (KATO-III, ATCC No. HTB103) was investigated using colorimetric MTT viability assay. A human liver cell line (Chang Liver, CLS No. 300139) was used as a control normal cell line. Two purified peptide peaks, peak l and peak ll at 100µg peptides mL-1 affected cell viability of the gastric cancer cell lines to 63.85±4.94 and 66.92±6.46%, respectively. Our result showed for the first time that the peptide fractions derived from protein hydrolysate of Euphorbia hirta Linn. have anti-gastric cancer activity, which offers a potential novel and natural anti-gastric cancer remedy.
Abstract: We present a preliminary x-ray study on human-hair
microstructures for a health-state indicator, in particular a cancer
case. As an uncomplicated and low-cost method of x-ray technique,
the human-hair microstructure was analyzed by wide-angle x-ray
diffractions (XRD) and small-angle x-ray scattering (SAXS). The
XRD measurements exhibited the simply reflections at the d-spacing
of 28 Å, 9.4 Å and 4.4 Å representing to the periodic distance of the
protein matrix of the human-hair macrofibrous and the diameter and
the repeated spacing of the polypeptide alpha helixes of the
photofibrils of the human-hair microfibrous, respectively. When
compared to the normal cases, the unhealthy cases including to the
breast- and ovarian-cancer cases obtained higher normalized ratios of
the x-ray diffracting peaks of 9.4 Å and 4.4 Å. This likely resulted
from the varied distributions of microstructures by a molecular
alteration. As an elemental analysis by x-ray fluorescence (XRF), the
normalized quantitative ratios of zinc(Zn)/calcium(Ca) and
iron(Fe)/calcium(Ca) were determined. Analogously, both Zn/Ca and
Fe/Ca ratios of the unhealthy cases were obtained higher than both of
the normal cases were. Combining the structural analysis by XRD
measurements and the elemental analysis by XRF measurements
exhibited that the modified fibrous microstructures of hair samples
were in relation to their altered elemental compositions. Therefore,
these microstructural and elemental analyses of hair samples will be
benefit to associate with a diagnosis of cancer and genetic diseases.
This functional method would lower a risk of such diseases by the
early diagnosis. However, the high-intensity x-ray source, the highresolution
x-ray detector, and more hair samples are necessarily
desired to develop this x-ray technique and the efficiency would be
enhanced by including the skin and fingernail samples with the
human-hair analysis.
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: Among all microRNAs (miRNAs) in 12 plant species investigated in this study, only miR398 targeted the copper chaperone for superoxide dismutase (CCS). The nucleotide sequences of miRNA binding sites were located in the mRNA protein-coding sequence (CDS) and were highly homologous. These binding sites in CCS mRNA encoded a conservative GDLGTL hexapeptide. The binding sites for miR398 in the CDS of superoxide dismutase 1 mRNA encoded GDLGN pentapeptide. The conservative miR398 binding site located in the CDS of superoxide dismutase 2 mRNA encoded the GDLGNI hexapeptide. The miR398 binding site in the CDS of superoxide dismutase 3 mRNA encoded the GDLGNI or GDLGNV hexapeptide. Gene expression of the entire superoxide dismutase family in the studied plant species was regulated only by miR398. All members of the miR398 family, i.e. miR398a,b,c were connected to one site for each CuZnSOD and chaperone mRNA.
Abstract: This study examined the toxicological effects and
safety of polypeptide k isolated from the seeds of Momordica
charantia in laboratory rats. 30 male Sprague Dawley rats (12 weeks
old, bodyweight 180-200 g) were randomly divided into 3 groups
(1000 mg/kg, 500 mg and 0 mg/kg). Rats were acclimatized to
laboratory conditions for 7 days and at day 8 rats were dosed orally
with polypeptide k (in 2% DMSO/normal saline) and the controls
received the dosed vehicle only. Rats were then observed for 72
hours before sacrificed. Rats were anaesthetized by pentobarbital
(50 mg/kg ip) and 2-3.0 mL of blood was taken by cardiac puncture
and rats were scarified by anaesthetic overdose. Immediately, organs
(heart, lungs, liver, kidneys) were weigh and taken for histology.
Organ sections were then evaluated by a histopathologist. Serum
samples were assayed for liver functions (ALT and γ-GT) and kidney
functions (BUN and creatinine). All rats showed normal behavior
after the dosing and no statistical changes were observed in all blood
parameters and organ weight. Histological examinations revealed
normal organ structures. In conclusion, dosing of rats up to 1000
mg/kg did not have any effects on the rat behavior, liver or kidney
functions nor histology of the selected organs.
Abstract: The aim of the study was to determine how different
ripening processes (traditional vs. industrial) influenced the
proteolysis in traditional Serbian dry-fermented sausage Petrovská
klobása. The obtained results indicated more intensive pH decline
(0.7 units after 9 days) in industrially ripened products (I), what had a
positive impact on drying process and proteolytic changes in these
samples. Thus, moisture content in I sausages was lower at each
sampling time, amounting 24.7% at the end of production period
(90 days). Likewise, the process of proteolysis was more pronounced
in I samples, resulting in higher contents of non-protein nitrogen
(NPN) and free amino acids nitrogen (FAAN), as well as in faster
and more intensive degradation of myosin (≈220 kDa), actin (≈45
kDa) and other polypeptides during processing. Consequently, the
appearance and accumulation of several protein fragments were
registered.
Abstract: Recent research has shown that milk proteins can
yield bioactive peptides with opioid, mineral binding,
cytomodulatory, antihypertensive, immunostimulating, antimicrobial
and antioxidative activity in the human body. Bioactive peptides are
encrypted in milk proteins and are only released by enzymatic
hydrolysis in vivo during gastrointestinal digestion, food processing
or by microbial enzymes in fermented products. At present
significant research is being undertaken on the health effects of
bioactive peptides. A variety of naturally formed bioactive peptides
have been found in fermented dairy products, such as yoghurt, sour
milk and cheese. In particular, antihypertensive peptides have been
identified in fermented milks, whey and ripened cheese. Some of
these peptides have been commercialized in the form of fermented
milks. Bioactive peptides have the potential to be used in the
formulation of health-enhancing nutraceuticals, and as potent drugs
with well defined pharmacological effects.
Abstract: Bacterial molecular chaperone DnaK plays an essential role in protein folding, stress response and transmembrane targeting of proteins. DnaKs from many bacterial species, including Escherichia coli, Salmonella typhimurium and Haemophilus infleunzae are the molecular targets for the insect-derived antimicrobial peptide pyrrhocoricin. Pyrrhocoricin-like peptides bind in the substrate recognition tunnel. Despite the high degree of crossspecies sequence conservation in the substrate-binding tunnel, some bacteria are not sensitive to pyrrhocoricin. This work addresses the molecular mechanism of resistance of Helicobacter pylori DnaK to pyrrhocoricin. Homology modelling, structural and sequence analysis identify a single aminoacid substitution at the interface between the lid and the β-sandwich subdomains of the DnaK substrate-binding domain as the major determinant for its resistance.
Abstract: In order to characterize the soy protein hydrolysate obtained in this study, gel chromatography on Sephadex G-25 was used to perform the separation of the peptide mixture and electrophoresis in SDS-polyacrylamide gel has been employed. Protein hydrolysate gave high antioxidant activities, but didn't give any antimicrobial activities. The antioxidant activities of protein hydrolysate was in the same trend of peptide content which gave high antioxidant activities and high peptide content between fractions 15 to 50. With increasing peptide concentrations, the scavenging effect on DPPH radical increased until about 70%, thereafter reaching a plateau. In compare to different concentrations of BHA, which exhibited higher activity (90%), soybean protein hydrolysate exhibited high antioxidant activities (70%) at a concentration of 1.45 mg/ml at fraction 25. Electrophoresis analysis indicated that, low- MW hydrolysate fractions (F1) appeared, on average, to have higher DPPH scavenging activities than high-MW fractions. These results revealed that soybean peptides probably contain substances that were proton donors and could react with free radicals to convert them to stable diamagnetic molecules.
Abstract: Yogurt is a coagulated milk product obtained from
the lactic acid fermentation by the action of Lactobacillus
bulgaricus and Streptococcus thermophilus. The additions of fruits
into milk may enhance the taste and the therapeutical values of milk
products. However fruits also may change the fermentation
behaviour. In this present study, the changes in physicochemical, the
peptide concentration, total phenolics content and the antioxidant
potential of yogurt upon the addition of Hylocereus polyrhizus and
Hylocereus undatus (white and red dragon fruit) were investigated.
Fruits enriched yogurt (10%, 20%, 30% w/w) were prepared and the
pH, TTA, syneresis measurement, peptide concentration, total
phenolics content and DPPH antioxidant inhibition percentage were
determined. Milk fermentation rate was enhanced in red dragon fruit
yogurt for all doses (-0.3606 - -0.4126 pH/h) while only white
dragon fruit yogurt with 20% and 30% (w/w) composition showed
increment in fermentation rate (-0.3471 - -0.3609 pH/h) compared to
plain yogurt (-0.3369pH/h). All dragon fruit enriched yogurts
generally showed lower pH readings (pH 3.95 - 4.03) compared to
plain yogurt (pH 4.05). Both fruit yogurts showed a higher lactic
acid percentage (1.14-1.23%) compared to plain yogurt (1.08%).
Significantly higher syneresis percentage (57.19 - 70.32%)
compared to plain yogurt (52.93%) were seen in all fruit enriched
yogurts. The antioxidant activity of plain yogurt (19.16%) was
enhanced by the presence of white and red dragon fruit (24.97-
45.74%). All fruit enriched yogurt showed an increment in total
phenolic content (36.44 - 64.43mg/ml) compared to plain yogurt
(20.25mg/ml). However, the addition of white and red dragon fruit
did not enhance the proteolysis of milk during fermentation.
Therefore, it could be concluded that the addition of white and red
dragon fruit into yogurt enhanced the milk fermentation rate, lactic
acid content, syneresis percentage, antioxidant activity, and total
phenolics content in yogurt.
Abstract: Two commercial proteases from Bacillus
licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were
screened for the production of Z-Ala-Phe-NH2 in batch reaction.
Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal
amidation of Z-Ala-Phe-OMe using ammonium carbamate as
ammonium source. Immobilization of protease has been achieved by
the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and
tetramethoxysilane (TMOS) as precursors (unpublished results). In
batch production, about 95% of Z-Ala-Phe-NH2 was obtained at
30°C after 24 hours of incubation. Reproducibility of different
batches of commercial Alcalase 2.4 L FG preparations was also
investigated by evaluating the amidation activity and the entrapment
yields in the case of immobilization. A packed-bed reactor (0.68 cm
ID, 15.0 cm long) was operated successfully for the continuous
synthesis of peptide amides. The immobilized enzyme retained the
initial activity over 10 cycles of repeated use in continuous reactor at
ambient temperature. At 0.75 mL/min flow rate of the substrate
mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5
hours of substrate recycling. The product contained about 90%
peptide amide and 10% hydrolysis byproduct.
Abstract: Dried soy protein hydrolysate powder was added to
the burger in order to enhance the oxidative stability as well as
decreases the microbial spoilage. The soybean bioactive compounds
(soy protein hydrolysate) as antioxidant and antimicrobial were added
at level of 1, 2 and 3 %.Chemical analysis and physical properties
were affected by protein hydrolysate addition. The TBA values were
significantly affected (P < 0.05) by the storage period and the level of
soy protein hydrolysate. All the tested soybean protein hydrolysate
additives showed strong antioxidant properties. Samples of soybean
protein hydrolysate showed the lowest (P < 0.05) TBA values at each
time of storage.
The counts of all determined microbiological indicators were
significantly (P < 0.05) affected by the addition of the soybean
protein hydrolysate. Decreasing trends of different extent were also
observed in samples of the treatments for total viable counts,
Coliform, Staphylococcus aureus, yeast and molds. Storage period
was being significantly (P < 0.05) affected on microbial counts in all
samples Staphylococcus aureus were the most sensitive microbe
followed by Coliform group of the sample containing protein
hydrolysate, while molds and yeast count showed a decreasing trend
but not significant (P < 0.05) until the end of the storage period
compared with control sample. Sensory attributes were also
performed, added protein hydrolysate exhibits beany flavor which
was clear about samples of 3% protein hydrolysate.
Abstract: The paper is included within the framework of a
complex research program, which was initiated from the hypothesis
arguing on the existence of a correlation between pineal indolic and
peptide hormones and the somatic development rhythm, including
thus the epithalamium-epiphysis complex involvement. At birds,
pineal gland contains a circadian oscillator, playing a main role in the
temporal organization of the cerebral functions. The secretion of
pineal indolic hormones is characterized by a high endogenous
rhythmic alternation, modulated by the light/darkness (L/D)
succession and by temperature as well. The research has been carried
out using 100 chicken broilers - “Ross" commercial hybrid,
randomly allocated in two experimental batches: Lc batch, reared
under a 12L/12D lighting schedule and Lexp batch, which was photic
pinealectomised through continuous exposition to light (150 lux, 24
hours, 56 days). Chemical and physical features of the meat issued
from breast fillet and thighs muscles have been studied, determining
the dry matter, proteins, fat, collagen, salt content and pH value, as
well. Besides the variations of meat chemical composition in relation
with lighting schedule, other parameters have been studied: live
weight dynamics, feed intake and somatic development degree. The
achieved results became significant since chickens have 7 days of
age, some variations of the studied parameters being registered,
revealing that the pineal gland physiologic activity, in relation with
the lighting schedule, could be interpreted through the monitoring of
the somatic development technological parameters, usually studied
within the chicken broilers rearing aviculture practice.
Abstract: Acute disseminated encephalomyelitis (ADEM) has
been reported to develop after a hymenoptera sting, but its
pathogenesis is not known in detail. Myelin basic protein (MBP)-
specific T cells have been detected in the blood of patients with
ADEM, and a proportion of these patients develop multiple sclerosis
(MS). In an attempt to understand the mechanisms underlying
ADEM, molecular mimicry between hymenoptera venom peptides
and the human immunodominant MBP peptide was scrutinized,
based on the sequence and structural similarities, whether it was the
root of the disease. The results suggest that the three wasp venom
peptides have low sequence homology with the human
immunodominant MBP residues 85-99. Structural similarity analysis
among the three venom peptides and the MS-related HLA-DR2b
(DRA, DRB1*1501)-associated immunodominant MHC
binding/TCR contact residues 88-93, VVHFFK showed that
hyaluronidase residues 7-12, phospholipase A1 residues 98-103, and
antigen 5 residues 109-114 showed a high degree of similarity
83.3%, 100%, and 83.3% respectively. In conclusion, some wasp
venom peptides, particularly phospholipase A1, may potentially act
as the molecular motifs of the human 3HLA-DR2b-associated
immunodominant MBP88-93, and possibly present a mechanism for
induction of wasp sting-associated ADEM.
Abstract: Alpfa-fetoprotein and its fragments may be an important vehicle for targeted delivery of radionuclides to the tumor. We investigated the effect of conditions on the labeling of biologically active synthetic peptide based on the (F-afp) with technetium-99m. The influence of the nature of the buffer solution, pH, concentration of reductant, concentration of the peptide and the reaction temperature on the yield of labeling was examined. As a result, the following optimal conditions for labeling of (F-afp) are found: pH 8.5 (phosphate and bicarbonate buffers) and pH from 1.7 to 7.0 (citrate buffer). The reaction proceeds with sufficient yield at room temperature for 30 min at the concentration of SnCl2 and (Fafp) (F-afp) is to be less than 10 mkg/ml and 25 mkg/ml, respectively. Investigations of the test drug accumulation in the tumor cells of human breast cancer were carried out. Results can be assumed that the in vivo study of the (F-afp) in experimental tumor lesions will show concentrations sufficient for imaging these lesions by SPECT.
Abstract: Shadoo protein (Sho) was described in 2003 as the newest member of Prion protein superfamily [1]. Sho has similar structural motifs like prion protein (PrP) that is known for its central role in transmissible spongiform enchephalopathies. Although a great number of functions have been proposed, the exact physiological function of PrP is not known yet. Investigation of the function and localization of Sho may help us to understand the function of the Prion protein superfamily. Analyzing the subcellular localization of YFP-tagged forms of Sho, we detected the protein in the plasma membrane and in the nucleus of various cell lines. To reveal the localization of the endogenous protein we generated antibodies against Shadoo as well as employed commercially available anti-Shadoo antibodies: i) EG62 anti-mouse Shadoo antibody generated by Eurogentec Ltd.; ii) S-12 anti-human Shadoo antibody by Santa Cruz Biotechnology Inc.; iii) R-12 anti-mouse Shadoo antibody by Santa Cruz Biotechnology Inc.; iv) SPRN antibody against human Shadoo by Abgent Inc. We carried out immunocytochemistry on non-transfected HeLa, Zpl 2-1, Zw 3-5, GT1-1, GT1-7 and SHSY5Y cells as well as on YFP-Sho, Sho-YFP, and YFP-GPI transfected HeLa cells. Their specificity (in antibody-peptide competition assay) and co-localization (with the YFP signal) were assessed.
Abstract: Radiolabeled cyclic RGD peptides targeting integrin αvβ3 are reported as promising agents for the early diagnosis of metastatic tumors. With an aim to improve tumor uptake and retention of the peptide, cyclic RGD peptide dimer E[c (RGDfK)] 2 (E = Glutamic acid, f = phenyl alanine, K = lysine) coupled to the bifunctional chelator DOTA was custom synthesized and radiolabelled with 68Ga. Radiolabelling of cyclic RGD peptide dimer with 68Ga was carried out using HEPES buffer and biological evaluation of the complex was done in nude mice bearing HT29 tumors.
Abstract: 16-Mercaptohexadecanoic acid (MHDA) and tripeptide glutathione conjugated with gold nanoparticles (Au-NPs) are characterized by Fourier Transform InfaRared (FTIR) spectroscopy combined with Surface-enhanced Raman scattering (SERS) spectroscopy. Surface Plasmon Resonance (SPR) technique based on FTIR spectroscopy has become an important tool in biophysics, which is perspective for the study of organic compounds. FTIR-spectra of MHDA shows the line at 2500 cm-1 attributed to thiol group which is modified by presence of Au-NPs, suggesting the formation of bond between thiol group and gold. We also can observe the peaks originate from characteristic chemical group. A Raman spectrum of the same sample is also promising. Our preliminary experiments confirm that SERS-effect takes place for MHDA connected with Au-NPs and enable us to detected small number (less than 106 cm-2) of MHDA molecules. Combination of spectroscopy methods: FTIR and SERS – enable to study optical properties of Au- NPs and immobilized bio-molecules in context of a bio-nano-sensors.