Isolation of a Novel Auxin,Methyl 4-Chloroindoleacetate from Immature Seeds of Pisum sativum

Human casein was separated by gel filtration on a column of Sephadex G–200 with 0.1 m Tris buffer (pH 8.5) containing 1.0 m NaCl. The effluent which increased in turbidity at 25°C was centrifuged at 25,000 × g for 30 min and the precipitate was obtained as Fraction 6. After centrifugation, the effluent was separated into 5 elution fractions.

Disc gel electrophoretic patterns of each fraction showed occurrence of secondary bands other than major bands especially in Fractions 3, 4 and 5. The casein solutions unheated and heated at 100°C for 5 and 10 min were kept at 5°C for 5 days. No marked changes of electrophoretic pattern were observed among these casein solutions. However, when a casein solution heated at 100°C for 5 min was chroma to graphed under the same condition, secondary bands also appeared.     

The Occurrence of Ferredoxin-sulfite Reductase in Barley Roots

A simple method was established for determining 10 preservatives, butylhydroxyanisole and dibutylhydroxytoluene in food. Steam distillation was carried out, and the distillate was trapped with dichloromethane and distilled water. After acidification and addition of sodium chloride, food additives were extracted from aqueous phase with dichloromethane. The food additives were analyzed with a gas Chromatograph equipped with a dual column system of 10% FFAP and 5% DEGS + 1% H₃PO₄. Column temperature was increased from 140 to 210°C at the rate of 3°C/min, Fluorene was used as an internal standard.

Ethyl p-hydroxybenzoate and isopropyl p-hydroxybenzoate were not separated with the 10% FFAP column, but other food additives were simultaneously determined with this column. With the 5% DEGS + 1% H₃PO₄ column, isobutyl p-hydroxybenzoate and propyl p-hydroxybenzoate were not separated, but the others were simultaneously determined.

Added recovery tests were carried out on about 38 foods.     

Impact of In-Situ CO2 Nano-Bubbles Generation on Freezing Parameters of Selected Liquid Foods

The impact of in-situ CO₂ nano-bubbles generation on the freezing properties of soft serve, milk, and apple juice was investigated. Carbonated (0, 1000, and 2000 ppm) liquid foods contained in a tube were submerged and cooled for 90 min in a pre-set ethylene glycol bath (−15 °C). Before the enclosed liquid reached 0 °C, the vibration was discharged through ultrasound in the bath to create nano-bubbles within the carbonated food samples, and the changes in temperature for 90 min of each food were recorded as a freezing curve. The time for onset of nucleation of control soft serve mix was halved in samples with 2000-ppm CO₂ due to the presence of nano-bubbles. Likewise, the nucleation time for milk with and without nano-bubbles at the same CO₂ concentration of 2000 ppm was 7.9 ± 0.1 and 2.8 ± 0.8 min_, respectively. The generation of CO₂ nano-bubbles from 2000-ppm CO₂ level in 10 ^oBx apple juice displayed −9.3 ± 0.3 °C nucleation temperature while the control one had −11.7 ± 0.9 °C.

     

Application of Nitrate Reductase for the Determination of Nitrate in Meat and Fishery Products

A rapid and specific method is described for the determination of nitrate in meat and fishery products.

Nitrate separated from foods by extraction with 1/50Ν sodium hydroxide and ultrafiltration was readily reduced to nitrite by the use of respiratory nitrate reductase (NR) from Escherichia coli K-12. The nitrite so obtained can be determined by the specific diazotation-coupling reaction method.

The use of an enzymatic reaction resulted in quantitative reduction of nitrate, and the method was relatively free of interferences. Recoveries of 10 and 100 ppm of nitrate from 5 samples of meat and fishery products ranged from 92.8 to 97.8% for 10 ppm and 97.8 to 99.4% for 100 ppm with a detection limit of 0.5 ppm.     

Biochemical Properties of A Thermophilic Alkalophile

A thermophilic alkalophile (IC strain) which can grow well in an alkaline medium at over 55°C was isolated from soil samples, and identified as Bacillus licheniformis; its growth on a neutral medium was, however, very poor. This strain was able to grow at 37°C as well as at 55°C, but the specific growth rate at 55°C was about twice as high as that at 37°C under alkaliné conditions.

The intracellular pH remained below 9.5 when Na⁺ was present in the medium. Na ⁺ stimulated the alanine uptake by cells or membrane vesicles, but was not required ATP synthesis.

Intracellular enzymes were stable on heat treatment up to 60°C. The residual activity of enolase after heating at 60°C for 10 min was about 80%. Cytochrome oxidase in membrane vesicles was completely stable up to 58°C for 30 min. These enzymes were also resistant to SDS treatment, more than 50% of their activities remaining at 5% SDS.     

Pyrolysis of Mixed Date Stones and Pistachio Shells: Identification of Bio-Oil and Utilization of Bio-Char as Activated Carbon Precursor

Thermal pyrolysis of mixed date stones and pistachio shells in a semi-batch reactor was addressed in this study. The highest yield of liquids (51.20 %) was produced at 500 °C, 90 min, 20 °C/min heating rate, and 50 mesh particle size. Under these conditions, yield of liquid from date stones and pistachio shells separately was 49.12 % and 47.67 %, respectively. The FT-IR results confirmed the presence of multiple oxygen-containing compounds in the bio-oil. Results from GC–MS declared that it was predominately composed of acids (57.57 %), esters (21.35 %), phenols (4.63 5), and alcohols (3.49 5). The obtained biochar was transformed into activated carbon (AC) by the optimized ZnCl₂ activation method. The ideal AC was synthesized at 600 °C for 60 min using a 2 : 1 ZnCl₂: biochar impregnation ratio. FESEM and XRD measurements showed that the AC was amorphous. The prepared AC was effective in eliminating dibenzothiophene (DBT) from model fuel (200 ppm DBT/hexane) with a maximum performance 95.26 % at 40 °C for 1h using 0.35 g of the AC. The exhausted AC was regenerated and reutilized 4 times, and removal efficiency reached 88.23 % in the 4^th cycle under ideal working conditions.     

Modeling and optimization of lipase-catalyzed esterification of policosanols with conjugated linoleic acid by response surface methodology

Abstract

The aim of this study was to model the lipase-catalyzed esterification of policosanols with conjugated linoleic acid (CLA) in a solvent-free system to produce wax esters which had a lower melting point than that of their corresponding policosanol forms and to optimize the reaction conditions by response surface methodology (RSM). Novozym 435 was selected as a suitable biocatalyst for the reaction. The molar ratio of substrates (policosanols to CLA) was 1:2. A well-fitting quadratic polynomial regression model for the degree of esterification (DE) of policosanols with CLA was established with regard to temperature (35–65°C), enzyme loading (1–5% of weight of total substrates), and reaction time (10–50 min). Optimal reaction conditions were 61.3°C for temperature, 3.7% for enzyme loading, and 34.1 min for reaction time, and the DE was ? 95 mol% under these conditions. The policosanols and wax esters synthesized under optimal conditions had melting points of 79°C and 57°C, respectively.     

THE UPTAKE OF BIOGENIC MONOAMINES BY THE ISOLATED AURICLE OF THE SNAIL (HELIX POMATIA)

—The uptake of [³H]5HT, [³H]dopamine, [³H]noradrenaline and [³H]octopamine into the auricle of Helix pomatia was studied. When tissues were incubated at 25°C in media containing radioactive amines, tissue:medium ratios of about 49:1, 14:1 and 5:1 for 5-HT, dopamine, noradrenaline, and octopamine respectively were obtained after a 20–30 min incubation time. Tissues incubated at 25°C in media containing radioactive amines for 20–30 mins showed that almost all (96%) the radioactivity was present as unchanged [³H]5-HT, [³H]dopamine, [³H]octopamine or [³H]noradrenaline. The high tissue:medium ratios for 5-HT and dopamine, but not for noradrenaline and octopamine, showed saturation kinetics which were dependent upon temperature and sodium ions. From the Lineweaver–Burk plots, two uptake mechanisms for 5-HT at 25°C were resolved; the high affinity uptake process having a K_m1 value of 6.0 ± 10^?8m and a V_m1 value of 0.115 nmol/g/min while the lower affinity process had a K_m2 value of 1.04 ± 10^?6m and a V_m2 value of 0.66nmol/g/min. At 0°C a single uptake mechanism for 5-HT occurred which gave a K_m value of 5.02 ± 10^?8m and a V_m value of 0.0165 nmol/g/min. In the case of dopamine, the Lineweaver–Burk plot at 25°C showed a single uptake process with values for K_m and V_m of 1.55 ± 10^?7m and 0.086 nmol/g/min respectively. This process did not function at 0°C. The effect of various agents and ions upon the accumulation processes for all amines was also studied, and the data indicate that the same neurons probably accumulate more than one amine type. It is concluded that 5-HT and dopamine uptake in the auricle is a mechanism for inactivating these substances at 25°C and that an uptake mechanism for 5-HT also functions at 0°C. The results are discussed from the point of view of 5-HT's being the cardioexcitatory substance in the snail heart.     

Formation of Antibacterial Factor by Heating System Containing Nitrite and Tryptone

A Perigo-type antibacterial factor (PTF) was produced when tryptone (a pancreatic digest of casein) medium was heated with nitrite at 121°C for 20 min. This PTF was inhibitory against Staphylococcus aureus, Bacillus subtilis and Clostridium botulinum, but was not against Escherichia coli and Salmonella typhimurium. The inhibitory activity varied with the concentration of nitrite (5 ~ 100 ppm) and tryptone (1, 2, 4%), and with pH (4, 5, 6, 7). The maximum inhibitory activity was observed when the medium containing 4% tryptone and 0.2% thioglycolate was heated with more than 50 ppm nitrite at pH 6. The tryptone was separated into three fractions by gel filtration and PTF was produced in every fraction, although the inhibitory activity was different in each. Our PTF might be unstable towards oxygen because its activity was lost completely by shaking for more than 16 hr.     

Inactivation of Pseudomonas aeruginosa biofilms formed under high shear stress on various hydrophilic and hydrophobic surfaces by a continuous flow of ozonated water

Abstract

The inactivation of Pseudomonas aeruginosa biofilms grown on glass under high shear stress and exposed to a range of dissolved ozone concentrations (2, 5 and 7?ppm) at 10 and 20?min was investigated. The regression equation, log reduction (biofilm)?=?0.64?+?0.59×(C – 2)?+?0.33×(T – 10), described the dependence of biofilm inactivation on the dissolved ozone concentration (C, ppm) and contact time (T, min). The predicted D-values were 11.1, 5.7 and 2.2?min at 2, 5 and 7?ppm, respectively. Inactivation of biofilms grown on various surfaces was tested at a single dissolved ozone concentration of 5?ppm and a single exposure time of 20?min. Biofilms grown on plastic materials showed inactivation results similar to that of biofilms on glass, while biofilms grown on ceramics were statistically significantly more difficult to inactivate, suggesting the importance of utilizing non-porous materials in industrial and clinical settings.     

Reaction Products from Various Tocopherols with Trimethylamine Oxide and Their Antioxidative Activities

The structures of many reaction products obtained when various tocopherols (Toc’s) and trimethylamine oxide (TMAO) were treated in liquid paraffin under a nitrogen stream at 180°C, were determined and their antioxidative activities were investigated.

The reaction products (Toc dimers) isolated were as follows: α-tocopheryl ethane from α-Toc; 5-(γ-tocopheryloxy)-γ-Toc, 5-(γ-tocopheryl)-γ-Toc (two kinds) and α-tocopheryl ethane from γ-Toc; 5-(δ-tocopheryloxy)-δ-Toc from δ-Toc.

The two 5-(γ-tocopheryl)-γ-Toc’s are atropisomers of each other (TLC (Rf): 0.75, 0.45—benzene) and isomerization occurred within 20 min when they were treated under nitrogen at 180°C.

All Toc dimers, in particular 5-(γ-tocopheryloxy)-γ-Toc, have antioxidative activities and excellent synergism with TMAO in inhibiting the oxidation of lard kept in the dark at 60°C.     

Multinuclear magnetic resonance spectroscopy and crystal structures of iodo-bridged dinuclear Pt(II) complexes with amines

Complexes of the types cis-Pt(amine)₂I₂ were transformed into the iodo-bridged dimers, which were characterized mainly by multinuclear (¹⁹⁵Pt, ¹H and ¹³C) magnetic resonance spectroscopy. For bulby amines, the dinuclear species were synthesized directly from K₂[PtI₄]. Compounds with several primary aliphatic and cyclic amines and two secondary amines were studied. In ¹⁹⁵Pt NMR, two signals were observed between −3899 and −4080 ppm in acetone. These species were assigned to the cis and trans dinuclear compounds I(amine)Pt(μ-I)₂PtI(amine). We suggest that the most shielded compound is the trans isomer. The difference between the two isomers is 12-13 ppm for the primary amine system and 26-27 ppm for the two secondary amines. There seems to be a slight dependence of the proton affinity in the gas phase of the amine (linear amines) with the δ(Pt) chemical shifts of the dinuclear Pt(II) compounds. The ²J(¹⁹⁵Pt-¹HN) coupling constants are slightly larger for the trans isomers (average 67 Hz, vs. 56 Hz). The ³J(¹⁹⁵Pt-¹H) coupling constants were detected only for the dimethylamine compounds, 46 Hz (trans) and 44 Hz (cis). In ¹³C NMR, the values of ²J(¹⁹⁵Pt-¹³C) and ³J(¹⁹⁵Pt-¹³C) were also found to be very slightly larger for the trans complexes (average 19 and 25 Hz vs. 15 and 18 Hz). The structures were confirmed by X-ray diffraction studies of the n-butylamine and diethylamine compounds. The two crystals were those of the trans dinuclear complexes.     

A METHOD FOR DETERMINATION OF ACETYLCHOLINE BY SLOW PYROLYSIS COMBINED WITH MASS FRAGMENTOGRAPHY ON A PACKED CAPILLARY COLUMN

A new procedure is described for the estimation of ACh by pyrolysis-gas chromatography/ mass spectrometry. ACh iodide and the iodides of ACh-d₁₆ or propionylcholine are slowly demethylated at 250°C by pyrolysis on the tip of a glass probe after insertion into the heated entrance of a glass tube leading to a packed capillary column. The volatile tertiary amines are then carried by helium to the column and trapped in its initial part which is kept at about 60°C. After 2–3 min the chromatography is started when the amines are released by heating this part to the ambient temperature in the oven (165°C). Peaks due to demethylated ACh and propionylcholine are well separated. The limit of detection is about 0.3pmol. After pyrolysis of mixtures of ACh and either ACh-d₁₆ or propionylcholine the peak amplitudes are linearly related to the doses.     

Studies on Tannin Acyl Hydrolase of Microorganisms

Tannin acyl hydrolase (Tannase) from Asp. oryzae No. 7 was purified. The purified enzyme was homogenous on column chromatography (DEAE-Sephadex A50, Sephadex G100), ultra centrifugation and electrophoresis.

The molecular weight of the enzyme estimated by gel filtration method was about 200,000.

The enzyme was stable in the range of pH 3 to 7.5 for 12 hr at 5°C, and for 25 hr at the same temperature in the range of pH 4.5 to 6. The optimum pH for the reaction was 5.5. It was stable under 30°C (over one day, in 0.05 M-citrate buffer of pH 5.5), and the optimum temperature was 30~40°C (reaction for 20min). The activity was lost completely at 55°C in 20 min at pH 5.5, or at 85°C in 10 min at the same pH.

Any metal salt tested did not activate the enzyme, Zink chloride and cupric chloride inhibited the activity or denatured the enzyme. The activity was lost completely by dialysis against EDTA-solution at pH 7.25, although it was not affected by dialysis against deionized water.     

Studies with dextran 40 in cryopreservation of blood

Whole blood from healthy donors was washed twice with phosphate-buffered saline (PBS) and then resuspended in sufficient PBS to give a final concentration of 2 × 10⁹/cells/ml. Aliquots were combined with equal volumes of the required diluents to give final dextran 40 concentrations of 0, 5, 10, 15, and 20% in PBS. Fifty-lambda samples in 50-lambda Micropets (Clay Adams) were frozen in alcohol baths at temperatures ranging from ?10 to ?80 °C. The specimens were frozen either for 1 min or 16 min, rapidly thawed, and resuspended in PBS or PBS plus dextran. Percentage of hemolysis was determined colorimetrically. Results indicate that concentraitons as low as 5% dextran exert a cryoprotective effect. Increased dextran concentration increases cryoprotection at high subzero bath temperatures (?10 ° and ?20 °C). Dextran concentrations beyond 12% have a damaging effect at low subzero bath temperatures (below ?30 °C). Based on this a two-factor hypothesis for cryopreservation is proposed. Apparent partial recovery of red blood cells without dextran or with 5% dextran during subzero storage was demonstrated.     

Isolation and characterization of a strain of <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> with citrinin-degrading activity

Monascus-fermented products have been widely used in Taiwan and other Asian countries as health foods. Unfortunately, many Monascus strains concurrently produce trace amounts of toxic citrinin. This study isolated a strain NPUST-B11 with the ability to degrade citrinin as the only carbon source. The isolated strain NPUST-B11 was characterised and identified as Klebsiella pneumoniae by 16S rRNA gene analysis using UNI-F and UNI-R primers. The isolated strain was then incubated in the mineral broth containing 10 ppm of citrinin, 1.2% of glucose, 0.3% of peptone and 100 ppm of vitamin C under optimal conditions, including pH 7, 200 rpm and 37°C. Citrinin was rapidly degraded with incubation from 97.9% at 1 h to 8.67% at 5 h and completely depleted at 10 h. Overall, this strain could be useful for the degradation of citrinin in food products and other medical applications.     

Stability of Soybean Ribosomal RNA in Various Conditions

The stability of RNA preparations which were prepared from soybean cotyledons was examined by incubating the RNA solutions of high salt at 20°C in the presence or absence of PVS. Sedimentation profiles of incubated RNA were given by ultracenirifugal analysis and compared with that of original RNA. RNA retained its original size after incubating for 4 hr in the presence of PVS and 200 mM KCl, while RNA was completely degraded into small fragments in the absence of PVS after the same treatment.

The purified rRNA which was prepared from 3 day-old hypocotyls was treated with heating, EDTA or urea in the presence of PVS. L-rRNA component was obviously disappeared by heating at 50°C for 5 min. Partial disruption of L-rRNA component occurred by dialyzing against urea solution. L-rRNA separated by zonal ultracentrifugation was decomposed into components by heat treatment or leaving at 4°C for 20 hr in the buffer from which KC1 was omitted. No back-conversion of heated RNA to original L-rRNA occurred by gradual cooling at room temperature for 40 min. S-rRNA, however, seemed to be stable in these treatments compared with L-rRNA.     

Aerobic degradation of a hydrocarbon mixture in natural uncontaminated potting soil by indigenous microorganisms at 20 °C and 6 °C

A hydrocarbon mixture containing p-xylene, naphthalene, Br-naphthalene and straight aliphatic hydrocarbons (C₁₄ to C₁₇) was aerobically degraded without lag phase by a natural uncontaminated potting soil at 20 °C and 6 °C. Starting concentrations were approximately 46 ppm for the aromatic and 13 ppm for the aliphatic compounds. All aliphatic hydrocarbons were degraded within 5 days at 20 °C, to levels below detection (ppb levels) but only down to 10% of initial concentration at 6 °C. Naphthalene was degraded within 12 days at 20 °C and unaffected at 6 °C. At 20 °C p-xylene was degraded within 20 days, but no degradation occurred at 6 °C. Br-naphthalene was only removed down to 30% of initial concentration at 20 °C, with no significant effect at 6 °C. The biodegradation was monitored with head space solid-phase microextraction and gas chromatography–mass spectrometry. Received: 5 October 1998 / Received revision: 4 December 1998 / Accepted: 5 December 1998     

Water Crystallisation of Model Sugar Solutions with Nanobubbles Produced from Dissolved Carbon Dioxide

This study was conducted to examine the influence of CO₂ nanobubbles on crystallisation behaviour of water during freezing of model sugar (2–5%w/v) solutions. CO₂ gas was dissolved at 0, 1000, and 2000-ppm concentrations before freezing. Carbonated sugar solutions in 50 mL plastic tubes were immersed in a pre-cooled (−15 °C) ethylene glycol bath and left to freeze at −15 °C for 90 min. When the temperature of the solutions reached 0 °C, ultrasound (US; 20 kHz) was emitted in the bath for 20 s duration through a metal horn transducer. The US wave applied in the ethylene glycol bath was expected to propagate to the sugar solutions in the tube and promote gas bubble formation from dissolved CO₂, which will trigger the ice nucleation. Obtained freezing curves were analysed for nucleation time and temperature, supercooling degree, and time taken for phase change. In general, the CO₂ gas promoted freezing of water, causing a noticeable shift in nucleation parameters. For example, nucleation time of 2000-ppm carbonated water coupled with sonication emission for 20 s (7.8 min) was much shorter than that of controls (pure water without any treatment = 19.1 min and US only = 14.3 min). The former initiated ice nucleation just below sub-zero temperature (−0.2 °C) whereas the onset temperature of controls (pure water without any treatment = −11.3 °C and the US only treatment = −10.3 °C). A similar effect was observed with different model sugar solutions. The current findings can be applied to refine the manufacturing process of ice-cream and frozen desserts by the food industries.

     

Pathogenesis and midgut histopathology of Bacillus thuringiensis in Simulium vittatum (Diptera: Simuliidae)

The pathogenesis and midgut histopathology which resulted when larvae of the blackfly, Simulium vittatum, were exposed to Bacillus thuringiensis at various temperatures and periods of exposure were investigated. The onset of mortality was studied at 10°, 15°, 19°, and 24°C. For each 4–5°C increase in temperature above 15°C, the onset of mortality was shortened by 24 hr. Exposures as brief as 15 min to 10 ppm of a whole spore preparation resulted in an average mortality of 29% in late-instar larvae. Mortality increased sharply for exposures up to 3 hr, approaching a maximum of 80%.The gross signs of disease included cessation of feeding and tetany with brachytosis. The tissue most affected was the midgut epithelium in the regions of the gastric caeca and posterior stomach. The formation of cytoplasmic vacuoles followed by cell lysis and/or sloughing were very apparent in moribund larvae. Death resulted without bacteremia.