Basic Fraction Research Articles

Prevention of osteoporosis by foods and dietary supplements. Milk basic protein (MBP) increases bone mineral density in young adult women and perimenopausal women

Milk has beneficial effects on bone health than other food sources. Recent in vitro and in vivo studies have shown that milk whey protein, especially its basic protein fraction (milk basic protein: MBP), contains components capable of promoting bone formation and inhibiting bone resorption. We tried to examine the effect of MBP on bone mineral density (BMD) and markers of bone metabolism in healthy young adult women and perimenopausal women. In the healthy young women study, we found that MBP increased BMD, by promoting bone formation and inhibiting bone resorption. In the healthy perimenopausal women study, we also found that MBP increased BMD, primarily by inhibiting bone resorption, while maintaining bone formation. Thus, bone remodeling balances become positive that leads to maintenance or increase of bone formation. MBP supplementation could be effective for bone health in a wide range of generation especially those who hate to drink milk.

Formas de fósforo no solo em sistemas de milho exclusivo e consorciado com feijão sob adubação orgânica e mineral

Os sistemas de manejo do solo modificam a dinâmica do fósforo alterando o conteúdo das diferentes formas de P. Objetivou-se avaliar o efeito de sistemas de cultivo em longo prazo (16 anos de plantio) sobre as diferentes formas de P no solo. Os tratamentos constaram de combinações entre dois sistemas de cultivo: milho exclusivo (M) e milho consorciado com feijão (MF), com duas doses (0 e 40 m³ ha-1 ano-1) de adubo orgânico (AO), e três doses (0, 250 e 500 kg ha-1) de N-P-K, 4-14-8 (AM). Solo sob um fragmento de Floresta Atlântica foi utilizado como referência de um estado em equilíbrio. Os valores de P orgânico total (Pot) variaram de 184,2 a 280,2 e de 147,9 a 282,9 mg kg-1, em amostras de solo das camadas de 0-10 e 10-20 cm, respectivamente, sendo os maiores valores observados para combinação 500 kg ha-1 + adubação orgânica, correspondendo, em média, a 26,4 % do P total no solo. Houve tendência da relação C/Pot manter-se constante, entre os tratamentos, constatando-se aumento dos valores de Pot com o aumento do teor de carbono orgânico total no solo. O adubo mineral promoveu incremento do P na biomassa microbiana (Pbm) apenas no sistema de milho exclusivo. Em média, o aumento do Pbm foi de 262 e 164 % para o sistema que recebeu o composto orgânico no sistema de milho exclusivo e consorciado com feijão, respectivamente. Em média, a fração de P orgânico solúvel em meio ácido correspondeu a 90 % do Pot predominando sobre a fração solúvel em base. Nos tratamentos com 500 kg ha-1 de 4-14-8 e 500 kg ha-1 + composto orgânico, no sistema de consórcio, foram obtidos aumentos nos valores de P total lábil de 53 e 157 %, respectivamente, comparados aos da testemunha. O P orgânico lábil (Pol) correspondeu, em média, a 3,7 % do Pot para os sistemas de cultivo, já para a Floresta Atlântica, esta relação foi de 10,7 %, nas duas profundidades. Os aumentos nos teores das formas mais lábeis de P, proporcionados pela adubação orgânica, evidenciam a importância deste sistema de manejo no favorecimento da ciclagem de P.

Antibacterial Studies of Essential oil and Different Extract Fractions from the Seeds of Nigella sativa L.

The antibacterial activity of the neutral, acidic, basic fractions and essential oil of Nigella sativa seeds were tested for their in vitro activity against eight species of bacteria, Staphylococcus aureus, Bacillus subtilis , Proteus mirablis , Salamonella typhi E. coli , Pseudomonas aeruginosa , Salamonella typhimurium , Shigella flexneri were investigated and compared with standard antibiotics Gentamycin and Amoxicillin. The result indicate that these essential oil, acidic and basic fractions have antibacterial activity against most of the tasted bacteria. We

Two Deaths Attributed to the Use of 2,4-Dinitrophenol

We report the cases of two individuals, one in Tacoma, WA, and the second in San Diego, CA, whose deaths were attributed to ingestion of 2,4-dinitrophenol (2,4-DNP). 2,4-DNP has historically been used as a herbicide and fungicide. By uncoupling mitochondrial oxidative phosphorylation, the drug causes a marked increase in fat metabolism that has led to its use to aid weight loss. Both cases reported here involved its use for this purpose. Features common to both cases included markedly elevated body temperature, rapid pulse and respiration, yellow coloring of the viscera at autopsy, history of use of weight loss or body building supplements, and presence of a yellow powder at the decedent's residence. Because of its acidic nature, the drug is not detected in the basic drug fraction of most analytical protocols, but it is recovered in the acid/neutral fraction of biological extracts and can be measured by high-performance liquid chromatography or gas chromatography-mass spectrometry. The concentration of 2,4-DNP in the admission blood samples of the two deaths reported here were 36.1 and 28 mg/L, respectively. Death in both cases was attributed to 2,4-DNP toxicity. Review of information available on the internet suggests that, although banned, 2,4-DNP is still illicitly promoted for weight loss.

Chemical characterization of the ambient organic aerosol soluble in water: 2. Isolation of acid, neutral, and basic fractions by modified size‐exclusion chromatography

A method employing size‐exclusion chromatography (SEC) with Total Organic Carbon detection is developed to isolate and quantify the water‐soluble organic carbon (WSOC) component of ambient aerosol particles by acid, neutral, and basic functional groups. The method provides unique quantitative insights into the characteristics and possible sources of a large fraction of the organic aerosol. The SEC is combined with a XAD‐8 method that separates WSOC into hydrophilic and hydrophobic fractions. Calibrations show the hydrophilic fraction separates into short‐chain aliphatic acids (WSOCxp_a), neutrals (WSOCxp_n, e.g., saccharides, polyols, and short‐chain carbonyls), and organic bases. Recovered hydrophobic fractions are separated into acids (WSOCxrr_a, e.g., aromatic) and neutrals (WSOCxrr_n, e.g., phenols). 13Carbon–nuclear magnetic resonance on ambient samples support the calibration‐based conclusions; however, calibrations only provide a guide to the type of ambient organic compounds expected in each group. Comparisons are made between XAD‐8/SEC results from urban Atlanta summer and winter and biomass burning samples. The largest isolated fraction of Atlanta summer WSOC is WSOCxp_a (29% μg C/μg C), suggesting aliphatic acids of less than C4 or C5 are the dominant secondary organic aerosol product. Combined with WSOCxrr_a, these acid groups are a higher fraction of summer organic carbon (20%) than winter (14%). They are correlated with each other (R2 = 0.74), with WSOCxp_n (R2 = 0.61 and 0.52), and with gaseous 24‐hour averaged volatile organic compounds linked to mobile sources. In biomass burning samples, neutrals (WSOCxp_n and WSOCxrr_n) dominate the WSOC. Atlanta winter samples have characteristics that appear to be a combination of summer and biomass samples.

Antioxidant and antimicrobial activities of the methanol extracts from pummelo (Citrus grandis Osbeck) fruit albedo tissues

The present study was conducted to evaluate the antioxidant and antimicrobial properties of methanol (100 and 80% aqueous) extracts of pummelo fruits albedo (Citrus grandis Osbeck). The antioxidant and antibacterial activity for crude extracts and isolated compounds were evaluated using free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Paper disc diffusion method. A 100% Methanol (MeOH) extract was steeped in water at different pH levels and partitioned with ethyl acetate (EtOAc) to give basic, acidic, neutral, and phenolic fractions. The neutral extract was found to possess maximum antioxidant and antibacterial activity. Thereafter, neutral extract was carried out on a silica gel column and eluted with hexane:EtOAc:acetone and preparative TLC (PTLC) to give oil buntan compound, linoleic acid methyl ester, β-sitosterol, sigmasterol, limonin, nomilin and meranzin hydrate were isolated. While, 80% MeOH extract was fractionated also using a silica gel column and PTLC to give isomeranzin hydrate, p-coumaric acid and caffeic acid compound. The extract concentration providing 50% inhibition (IC50) was as follows; oil buntan compound 95 μg/mL, caffeic acid 45 μg/mL, p-coumaric acid 105 μg/mL, limonin + nomilin (mixture) 135 μg/mL was lower than that of synthetic antioxidant butylated hydroxyanisole (BHA) 40 μg/mL. The inhibitory zone (mm) of bacteria tested was 2.9–4.1 mm caffeic acid and 11.6–15.1 mm p-coumaric acid.

Effects of bifidobacterial cytoplasm peptide and protein fractions on mouse lymphocyte proliferation and cytokine production

The ability of peptides and proteins from Bifidobacterium lactis Bb12 cytoplasm to elicit immune responses was measured in terms of murine splenocyte proliferation and cytokine secretion. Peptides and proteins from B. lactis Bb12 cytoplasm were separated into acidic and basic fractions by liquid-phase electrofocalization based on isoelectric pH. Both fractions were further fractionated by RP-HPLC based on hydrophobicity. Major peaks (of low hydrophobicity) selected in each fraction were found by mass spectrometry to contain an abundant peptide or protein with contaminants. The acidic fraction gave a stimulation index of 1.61±0.08 compared to 1.33±0.02 for the basic fraction and 2.96±0.35 for crude extract. The basic fraction stimulated IFN-γ secretion more than the acidic fraction (579±195 vs. 191±7 pg ml−1). The results demonstrate that peptides and proteins from B. lactis Bb12 exhibit immunostimulating activity, suggesting the use of cytoplasm as a nutraceutical product to enhance immune responses in children or elderly persons with reduced immune function.

A comparison between acidic and basic protein fractions from whey or milk for reduction of bone loss in the ovariectomised rat

The ability of whey acidic protein fractions to protect against bone loss due to ovariectomy (OVX) in the mature female rat was investigated. The bone bioactivity of these acidic protein fractions, isolated from both mineral acid whey protein concentrate (WPC) and rennet WPC, was compared with that of basic protein fractions isolated from both milk and rennet WPC. Fifty 6-month old rats that had been ovariectomised at 5.5 months were randomised into five groups of ten each. One group remained the OVX control while four groups were each fed one of the acidic or basic protein fractions as 0.3% (w/w) of the diet, for 4 months. Ten sham-operated rats served as a second control group. Sequential measurements of bone mineral density of the spine and femur indicated that the acidic protein fraction prepared from mineral acid WPC reduced bone loss due to OVX, maintaining bone density above OVX levels at week 16 of feeding. Biomechanical data indicated that both acidic fractions tended to increase bone stiffness, and hence resistance against breaking.

Ribosome-inactivating proteins in edible plants and purification and characterization of a new ribosome-inactivating protein from Cucurbita moschata

The basic protein fraction of tissue extracts from 40 edible plants inhibited cell-free protein synthesis and released adenine from herring sperm DNA, thus having adenine glycosylase activity. This suggested the presence of ribosome-inactivating proteins (RIPs) in the plant extracts. This indication was further strengthened by the presence of the two activities after a partial chromatographic purification of three extracts, including that from Lycopersicon esculentum (tomato), which had very low activity. From the extract of Cucurbita moschata (pumpkin), the most active one, a glycoprotein of 30,665 Da was purified which had the properties of a RIP, in that (i) it inhibited protein synthesis by a rabbit reticulocyte lysate with IC 50 (concentration giving 50% inhibition) 0.035 nM (1.08 ng ml −1) and by HeLa, HT29 and JM cells with IC 50 in the 100 nM range, (ii) deadenylated hsDNA and other polynucleotidic substrates, and (iii) depurinated yeast rRNA at a concentration of 0.1 ng ml −1, all values being comparable to those of other RIPs. The C. moschata RIP gave a weak cross-reaction only with an antiserum against dianthin 32, but not with antisera against other RIPs, and had superoxide dismutase, antifungal and antibacterial activities.

Identification of aroma compounds in Chinese ‘Yanghe Daqu’ liquor by normal phase chromatography fractionation followed by gas chromatography[sol ]olfactometry

Aroma compounds in Chinese ‘Yanghe Daqu’ liquor were investigated with normal phase chromatography fractionation followed by gas chromatography–olfactometry (GC–O). Yanghe Daqu liquor was extracted with Freon 11, and then separated into acidic and neutral[sol ]basic fractions. The neutral[sol ]basic fraction was further separated into four fractions, using silica gel normal phase chromatography. Aroma compounds in each fraction were analysed by GC–O and GC–MS. The results showed that Yanghe Daqu liquor aroma was mainly contributed by esters and fatty acids. Among them, ethyl hexanoate (Osme value =15), ethyl butanoate (Osme value ≥14) were probably the most important esters, based on their Osme values. In addition, ethyl octanoate, 3-methylbutyl hexanoate and ethyl pentanoate could be important (Osme values ≥10). Acids with high Osme value included hexanoic, butanoic, 3-methylbutanoic and pentanoic acids. They contributed to sour, acid, cheesy and sweaty aromas. 1-Hexanol and 3-methylbutanol also had high Osme values. In addition, 4-ethylguaiacol, 4-methylguaiacol and 4-ethylphenol were identified in Yanghe Daqu liquor and they contributed to clove, sweet, smoky and goaty aromas. 1,1-Diethoxy ethane, 1,1-diethoxy-2-methylpropane and 1,1-diethoxy-3-methylbutane were identified and they gave fruity and floral aromas. Copyright © 2005 John Wiley & Sons, Ltd.

A controlled trial of the effect of milk basic protein (MBP) supplementation on bone metabolism in healthy menopausal women

Milk has more beneficial effects on bone health than other food sources. Recent in vitro and in vivo studies have shown that milk whey protein, especially its basic protein fraction (milk basic protein, MBP), contains several components capable of promoting bone formation and inhibiting bone resorption. The object of this study was to examine the effect of MBP on the bone metabolism of healthy menopausal women. Thirty-two healthy menopausal women were randomly assigned to treatment with either placebo or MBP (40 mg per day) for 6 months. The bone mineral density (BMD) of the lumbar vertebrae L2-L4 of each subject was measured by dual-energy X-ray absorptiometry (DXA) at 0 and 6 months of treatment. Serum and urine indices of bone metabolism were measured at 0, 3 and 6 months. Twenty-seven subjects who completed the study in accordance with the protocol were included in the analysis. The mean rate of gain of lumbar BMD in the MBP group (1.21%) was significantly higher than in the placebo group (-0.66%, P=0.046). When compared with the placebo group, urinary cross-linked N-telopeptides of type-I collagen (NTx) were significantly decreased in the MBP group at 6 months, but no significant difference in serum osteocalcin was observed between the two groups. The urinary NTx excretion was found to be related to serum osteocalcin in the MBP group at 3 and 6 months, indicating that MBP maintained the balance of bone remodeling. These results suggested that MBP supplementation was effective in preventing bone loss in menopausal women and that this improvement in BMD may be primarily mediated through the inhibition of bone resorption while maintaining the balance of bone remodeling by MBP supplementation.

Biological and chemical transformation of atrazine in coastal aquatic sediments

The degradation and distribution potential of atrazine, a persistent triazine herbicide, into three chemical fractions were measured in coastal aquatic sediments in the laboratory over time. Sediments with varying organic carbon contents were extracted with an organic solvent followed by an alkali hydrolysis reaction, and atrazine, deethylatrazine (DEA) and deisopropylatrazine (DIA) were quantified in the aqueous, solvent, and basic fractions using gas chromatography–mass spectrometry. The total amount of atrazine and its metabolites recovered after 95 days varied by site and ranged from 5% to 30% in which 95% was atrazine found primarily in the solvent fraction. Sediment organic carbon was positively correlated with the distribution of atrazine into the basic fraction and the decline in the total amount recovered. No DIA was detected in laboratory spiked sediments and transformation to DEA was limited in all sediments and made up less than 1% of the mass balance. The production and persistence of DEA were inversely correlated to organic carbon; sediments with less carbon and limited binding sites had increased formation and persistence of DEA. A secondary metabolite, methylated atrazine (M-ATR) not previously documented to be derived from atrazine, was chemically produced, detected in all sediments and time points, and concentrations were an order of magnitude higher than DEA. Based on results from spiked estuarine sediments, atrazine and M-ATR may have the potential to persist in the environment while DEA and DIA may not be an ecological threat due to their limited formation.

Cromatografia de troca-iônica aplicada ao isolamento da fração ácida do óleo de copaíba (Copaifera multijuga) e da sacaca (Croton cajucara)

ION EXCHANGE CHROMATOGRAPHY APPLIED TO THE FRACTIONATION OF THE COPAIBA OIL (Copaifera multijuga) AND SACACA (Croton cajucara) EXTRACTS. Plant extracts are usually complex mixtures which contain several molecules of different sizes with varied functional groups. Such extracts are a challenge to the chemist of natural products. Ion exchange chromatography in non-aqueous medium, used for separation of basic or acidic fractions from plant extracts, is an important unit operation in preparative scale separations. Anionic macroporous resin in non-aqueous medium was used with success in this study for separation of the acid fraction of Copaifera multijuga (Copaiba oil), rich in labdanic diterpenes and for the methanolic extract of Croton cajucara (acetyl aleuritoric acid).

The effect of whey acidic protein fractions on bone loss in the ovariectomised rat

Bovine milk has been shown to contain bioactive components with bone-protective properties. Earlier studies on bovine milk whey protein showed that it suppressed bone resorption in the female ovariectomised rat. A new osteotropic component was subsequently identified in the whey basic protein fraction, but bone bioactivity may also be associated with other whey fractions. In the present study, we investigated whether acidic protein fractions isolated from bovine milk whey could prevent bone loss in mature ovariectomised female rats. Six-month-old female rats were ovariectomised (OVX) or left intact (sham). The OVX rats were randomised into four groups. One group remained the control (OVX), whereas three groups were fed various whey acidic protein fractions from milk whey as 3 g/kg diet for 4 months. Outcomes were bone mineral density, bone biomechanics and markers of bone turnover. Bone mineral density of the femurs indicated that one of the whey AF over time caused a recovery of bone lost from OVX. Plasma C-telopeptide of type I collagen decreased significantly in all groups except OVX control over time, indicating an anti-resorptive effect of whey acidic protein. Biomechanical data showed that the AF may affect bone architecture as elasticity was increased by one of the whey AF. The femurs of AF-supplemented rats all showed an increase in organic matter. This is the first report of an acidic whey protein fraction isolated from milk whey that may support the recovery of bone loss in vivo.

Extracellular xylanases from two pathogenic races of Fusarium oxysporum f. sp. ciceris: enzyme production in culture and purification and characterization of a major isoform as an alkaline endo-β-(1,4)-xylanase of low molecular weight

Fusarium oxysporum f. sp. ciceris, the causal agent of Fusarium wilt of chickpea, comprises eight pathogenic races and two pathotypes. Races 0 and 5, representative of the least virulent yellowing pathotype and the most virulent wilt pathotype, respectively, produced extracellular xylanases when grown on minimal medium supplemented with either 1% commercial birchwood xylan or 0.3% chickpea cell walls. The pattern of extracellular proteins analysed by denaturing polyacrylamide gel electrophoresis in the two media presented some minor but distinctive differences between fungal races. By preparative isoelectrofocusing, the xylanase activity in cell wall-culture filtrates could be resolved into basic and neutral fractions with pI values around to 10 and 8, respectively, whereas the xylan-culture filtrates contained an additional acidic fraction of pI around 4. A common major xylanase was purified 7-fold to homogeneity by cation-exchange chromatography and chromatofocusing. The purified xylanase has a molecular weight of 21.6 kDa, optimum pH and temperature of 5.5 and 55 degrees C, respectively, pI in the range of 8.2 to 9.0, and Km and Vmax values of 2.24 mg ml(-1) (birchwood xylan as substrate) and 1200 nkat mg(-1) protein (72 U mg(-1) protein), respectively. The enzyme has an endo mode of action, hydrolysing xylan to xylobiose and higher short-chain xylooligosaccharides without forming free xylose.

Characterization of Aroma Compounds Responsible for the Rosy/Floral Flavor in Cheddar Cheese

The aroma-active compounds that contribute to the rosy/floral flavor in Cheddar cheese were characterized using both instrumental and sensory techniques. Two cheeses (>12 months old) with rosy/floral flavor and two Cheddar cheeses of similar ages without rosy/floral flavors were selected. After direct solvent extraction/solvent-assisted flavor evaporation and separation into neutral/basic and acidic fractions, samples were analyzed by gas chromatography-olfactometry with aroma extract dilution analysis. Selected compounds were quantified using internal standard methodology. Some of the intense aroma-active compounds in the neutral basic fraction of the rosy/floral cheeses included 2-phenethanol (rosy), phenylethyl acetate (rosy), and phenylacetaldehyde (rosy/floral). Quantification, threshold analysis, and sensory analysis of model cheeses confirmed that increased concentrations of phenylacetaldehyde and phenylacetic acid caused rosy/floral flavor when spiked into Cheddar cheese.

Allergenicity of acidic peptides from bovine β-lactoglobulin is reduced by hydrolysis with Bifidobacterium lactis NCC362 enzymes

Abstract Cow milk proteins, particularly β-lactoglobulin (BLG) are a leading cause of food allergy in children. In vivo, gastrointestinal hydrolysis of milk proteins decreases their allergenicity, but the intestinal microbiota also contributes to their breakdown. This study investigated the effect of hydrolysis of BLG-derived tryptic-chymotryptic (TC) peptides by Bifidobacterium lactis NCC362 enzymes, on their allergenicity. B. lactis enzymes have been shown to hydrolyze both acidic and basic peptide fractions isolated from tryptic-chymotryptic hydrolysate, but not whole proteins. The IgE binding capacity of acidic TC peptides was reduced by B. lactis peptidases-driven hydrolysis. This is partly explained by the breakdown of known allergenic peptides. Moreover, the resulting peptide fragments significantly up-regulated IFN-γ and IL-10 production and down-regulated IL-4 secretion by murine splenocytes. These results indicate that B. lactis NCC362 could be a potential probiotic for preventing cow's milk allergy through hydrolysis of the allergenic portion of BLG and the release of peptides which down-regulate the allergic immune response, but further studies using living bacteria are needed to confirm these data.

Screening Antimicrobial Activities of Basic Protein Fractions from Dry and Germinated Wheat Seeds

Two small cationic peptide fractions (5 kDa) were isolated from dry and germinated seeds of wheat, named WAP and GWAP, respectively. The antifungal and antibacterial activities of the peptides were analyzed using disk diffusion and turbidity measurement assays. The peptides in vitro exhibited effective antifungal activity against four plant pathogenic fungi at minimum concentration of 15 μg(protein) cm−3. Their antimicrobial activity was negatively affected by the presence of 5 mM CaCl2. The peptides were less effective against Gram-negative bacterium Erwinia carotovora subsp. carotovora, but they demonstrated inhibitory activity against Gram-positive bacterium Clavibacter michiganensis subsp. sepedonicus. The antimicrobial activity of GWAP was more effective than WAP.

Conversion of Agricultural Residues to Fuel Products via Supercritical Fluid Extraction

In this study, various agricultural residues such as wheat straw, corncob, corn stover, tobacco stalk, tobacco leaf, olive waste, walnut shell, and almond shell were subjected to supercritical fluid extraction (SFE) using acetone, methanol, and water as solvent. The highest yield (59.3%) of SFE was obtained from OW with water solvent at 678 K. While carbon content of the selected samples varies from about 41.2% to 53.6%, carbon content of the extracts from SFEs varies from 52.8% to 67.8%. The hydrogen content of the extracts varies from 6.8% to 8.8%. The oxygen content of the extracts ranges from 21.7% to 38.1%, and nitrogen content ranges from 0.4% to 2.3%. Decreases of oxygen content in the supercritical fluid extracts range from 38.9% to 13.8%. The supercritical fluid extracts were fractionated into water soluble, acetone soluble, phenolic, fatty acid, polar, nonpolar, and basic fractions.

Preparation in and Use of the Nemeth Braille Code for Mathematics by Teachers of Students with Visual Impairments

This study examined the preparation in and use of the Nemeth braille code by 135 teachers of students with visual impairments. Almost all the teachers had taken at least one course in the Nemeth code as part of their university preparation. In their current jobs, they prepared a variety of materials, primarily basic operations, word problems, tactile graphics, and fractions.