Determination of bezafibrate, ciprofibrate and fenofibric acid in human plasma by high-performance liquid chromatography

A selective high-performance liquid chromatographic method to assess either bezafibrate, ciprofibrate or fenofibric acid plasma levels is described. Drugs are extracted with diethyl ether, after acidification with HCl. An isocratic acetonitrile-0.02 M H₃PO₄ (55:45) mobile phase, a C₁₈ (5 μm) column and UV detection are used. The LOQ found was 0.25 μg/ml for the three fibrates. Intra- and inter-assay accuracy ranges were 90–107% and 82–111%; 96–115% and 94–107%; 94–114% and 94–126% for bezafibrate, ciprofibrate and fenofibric acid, respectively. Intra- and inter-assay precision (C.V.% ranges) were 1.72–3.06% and 2.66–7.67%; 1.88–4.64% and 0.62–2.99%; 1.26–4.69% and 3.56–7.17% for the three fibrates studied. Its sensitivity, accuracy and precision make it a useful tool for monitoring plasma levels of these drugs in a clinical setting and for research purposes.     

Dry matter and nutrient inputs through litter fall in a dry tropical forest of India

The present paper elucidates the pattern of leaf and non-leaf fall and quantifies of the total annual input of litter in a dry tropical forest of India. In addition, concentration of selected nutrients in various litter species and their annual return to the forest floor are examined. Total annual input of litter measured in litter traps ranged between 488.0–671.0 g m^-2 of which 65–72% was leaf litter fall and 28–35% wood litter fall. 73–81% leaves fall during the winter season. Herbaceous litter fall ranged between 80.0–110.0 g m^-2 yr^-1. The annual nutrient return through litter fall amounted (kg ha^-1): 51.6–69.6 N, 3.1–4.3 P, 31.0–40.0 Ca, 14.0–19.0 K and 3.7–5.0 Na, of which 71–77% and 23–29% were contributed by leaf and wood litter fall, respectively for different nutrients. Input of nutrients through herbaceous litter was: 13.0–16.6 for N, 1.0–1.4 for P, 4.0–5.0 for Ca, 7.9–10.5 for K and 0.8–1.0 kg ha^-1 yr^-1 for Na.     

Studies on the effects of climate on livestock production in Iraq

Measurements were made of the roughage and water intake of young Friesian cross males and females throughout a 12-month period. Results showed that the roughage consumption fell by 10–16% between the temperature range 11°–20°C and 21°–30°C and by 20–43% between the temperature range 11°–20°C and 31°–40°C. Water intake increased from 30–44% between 11°–20°C and 21°–30°C and by 45–92% between 11°–20°C and 31°–40°C.

---

Zusammenfassung Rauhfutterverzehr und Wasserkonsum junger schwarzbunter Kreuzungsrinder wurden während eines Zeitraumes von zwölf Monaten gemessen. Die Ergebnisse zeigen, dass der Rauhfutterverzehr um 10–16% zurückging wenn die Lufttemperatur von 11°–20°C auf 21°–30°C stieg, und um 20–43% beim Anstieg von 11°–20°C auf 31°–40°C. Der Wasserkonsum nahm zwischen 11°–20°C und 21°–30°C um 30–44% und zwischen 11°–20°C und 31°–40°C um 45–92% zu.

---

Resume Pendant une période de douze mois, on a cherché à mesurer la quantité d'eau et de foin consommée par un jeune bétail de race Friesian en Irak. Les résultats furent les suivants: Lors d'une augmentation de température de 11°–20°C à 21°–30°C, la consommation de foin diminua de 10 à 16%, allant même jusqu'à diminuer de 20 à 43% lors de températures augmentant de 11°–20°C à 31°–40°C. La consommation d'eau, quant à elle, augmenta de 30 à 44% dans le premier cas et de 45 à 92% dans le second.

     

A new species of amblycipitid catfish, <Emphasis Type="Italic">Xiurenbagrus gigas</Emphasis> (Teleostei: Siluriformes), from Guangxi,China

A new species of amblycipitid catfish is described based on five specimens (88.0–164.5mm in standard length: SL) collected from the vicinity of Duan, Guangxi Zhuangzu Autonomous Region, southern China. The new species, Xiurenbagrus gigas, is different from all other known amblycipitid catfish, but similar to X. xiurenensis by having two patches of vomerine teeth. The new species can be distinguished from X. xiurenensis by having a larger head (head length: 27.9–34.9% vs. 21.8–26.4% SL; head depth: 14.4–17.9% vs. 12.4–14.1% SL; head width: 22.4–29.4% vs. 18.1–20.3% SL), lower adipose fin (4.6–6.0% vs. 6.5–8.3% SL), shorter distance from dorsal origin to adipose origin (25.7–32.0% vs. 32.9–38.9% SL), longer dorsal fin (21.2–24.9% vs. 16.2–19.1% SL), longer pectoral fin (18.4–20.7% vs. 13.5–16.8% SL), longer pelvic fin (13.1–14.3% vs. 8.8–11.8% SL), and more gill rakers (7–9 vs. 5). Xiurenbagrus gigas, the largest known species in the family Amblycipitidae, is distributed in the Xijiang River, the longest tributary of the Zhujiang (Pearl) River basin.     

Growth and age structure of the swan mussel Anodonta cygnea (L.) at different depths in lake Mattsee (Salzburg, Austria)

Unionid clams were collected at 1–2 m, 3–4 m and 6–7 m depth in lake Mattsee, a moderately mesotrophic lake, to investigate the effect of depth on clam growth and age structure. No significant differences in age structure of Anodonta cygnea were found (p=0.65). Three and ten years old clams were present at all depths, but in different percentages. Whereas at 1–2 m 13.3% of the collected clams were <4 years old, this percentage was 4.4% at 6–7 m and 7.1% at 3–4 m. A greater percentage (6.7%) of older mussels (9, 10 years) were collected at 6–7 m than at 1–2 m (2.2%). Growth declined with depth. Total length at a given age of clams at 1–2 m and 3–4 m did not differ (p=0.54), whereas differences were significant between clams at 1–2 m and 6–7 m (p<0.05) as well as between 3–4 m and 6–7 m (p<0.05). The Growth constant k was highest at 1–2 m depth.     

Trophic structure and productivity of the lagoonal communities of Tikehau atoll (Tuamotu Archipelago,French Polynesia)

Carbon standing stocks and fluxes were studied in the lagoon of Tikehau atoll (Tuamotu archipelago, French Polynesia), from 1983 to 1988.The average POC concentration (0.7–2000 µm) was 203 mg C m^–3. The suspended living carbon (31.6 mg C m^–3) was made up of bacteria (53%), phytoplankton < 5 µm (14.2%), phytoplankton > 5 µm (14.2%), nanozooplankton 5–35 µm (5.7%), microzooplankton 35–200 µm (4.7%) and mesozooplankton 200–2000 µm (7.9%). The microphytobenthos biomass was 480 mg C m^–2.Suspended detritus (84.4% of the total POC) did not originate from the reef flat but from lagoonal primary productions. Their sedimentation exceeded phytobenthos production.It was estimated that 50% of bacterial biomass was adsorbed on particles. the bacterial biomass dominance was explained by the utilisation of 1) DOC excreted by phytoplankton (44–175 mg C m^–2 day ^–1) and zooplankton (50 mg Cm^–2 day^–1)2) organic compounds produced by solar-induced photochemical reactions 3) coral mucus.50% of the phytoplankton biomass belongs to the < 5 µm fraction. This production (440 mg C m^–2 day^–1) exceeded phytobenthos production (250 mg C m^–2 day^–1) when the whole lagoon was considered.The zooplankton > 35 µm ingested 315 mg C m^–2 day^–1, made up of phytoplankton, nanozooplankton and detritus. Its production was 132 mg C m^–2 day^–1.     

Control of the production of individual fusicoccins at different dissolved oxygen concentrations

The regulatory effect of different concentrations of dissolved oxygen on the production of fusicoccins by the fungus Fusicoccum amygdali Del. was studied. The maximum output of total fusicoccins was obtained by using a profiled dissolved oxygen tension (DOT) regime, in which the DOT was maintained at 15–20% during the biomass growth phase and at 5–8% during the fusicoccins production phase. In comparison with the profiled regime, the maintenance of DOT at 15–20% during the whole fermentation shortened the fusicoccins production phase. The fermentation performance at a low DOT (5–8%) inhibited both the accumulation of biomass and the production of fusicoccins. At high DOT (40–50%), an accelerated accumulation of the biomass with an expressed autolysis of mycelia took place, and the production of fusicoccins was lowered. The qualitative composition of individual fusicoccins varied substantially at different DOTs. Fusicoccins, A, C, D, J, H, 16-O-demethyl-J, detretpentenylfusicoccin and some minor fusicoccin metabolites were found in the fermentation broth using the method of liquid secondary ion mass spectrometry. It was established that the profiled DOT regime (15–20% to 5–8%) provided both the maximum concentration of fusicoccins and an enhanced accumulation of the main metabolite – fusicoccin A (FC A). The performance of the fermentation at a DOT of 15–20% decreased the content of FC A by 2–6% in comparison with the profiled DOT regime, and increased the content of fusicoccin C to 14–20% of the total fusicoccins. Fermentation at DOT of 5–8% was characterized by the highest content of the precursors of FC A, the less oxidized fusicoccins H and J, the contents of which were in range 7–12% and 16–17% of total fusicoccins, respectively.     

Experimental studies of rapid bioerosion of coral reefs in the Galápagos Islands

Experimental carbonate blocks of coral skeleton,Porites lobata (PL), and cathedral limestone (LS) were deployed for 14.8 months at shallow (5–6 m) and deep (11–13m) depths on a severely bioeroded coral reef, Champion Island, Galápagos Islands, Ecuador. Sea urchins (Eucidaris thouarsii) were significantly more abundant at shallow versus deep sites.Porites lobata blocks lost an average of 25.4 kg m^–2yr^–1 (23.71 m^–2yr^–1 or 60.5% decrease yr^–1). Losses did not vary significantly at depths tested. Internal bioeroders excavated an average of 2.6 kg m^–2 yr^–1 (2.41 m^–2 yr^–1 or 0.6% decrease yr^–1), while external bioeroders removed an average of 22.8 kg m^–2 yr^–1). (21.31 m^–2 yr^–1). or 59.9% decrease yr^–1). few encrusting organisms were observed on the PL blocks. Cathedral limestone blocks lost an average of 4.1 kg m^–2 yr^–1). (1.81 m^–2 yr^–1). or 4.6% decrease yr-'), also with no relation to depth. Internal bioeroders excavated an average of 0.6 kg m^–2 yr^–1). (0.31 m^–2 yr^–1). or 0.7% decrease yr^–1). and external bioeroders removed an average of 3.5 kg m^–2 yr^–1). (1.51 m^–2 yr^–1). or 3.9% decrease yr^–1). from the LS blocks. Most (57.6%) encrustation occurred on the bottom of LS blocks, and there was more accretion on block bottoms in deep (61.4 mg cm^–2 yr^–1). versus shallow (35.0 mg cm^–2 yr^–1) sites. External bioerosion reduced the average height of the reef framework by 0.2 cm yr^–1). for hard substrata (represented by LS) and 2.3 cm yr^–1). for soft substrata (represented by PL). The results of this study suggest that coral reef frameworks in the Galápagos Islands are in serious jeopardy. If rates of coral recruitment do not increase, and if rates of bioerosion do not decline, coral reefs in the Galápagos Islands could be eliminated entirely.     

Feeding rates and energy deficits of juvenile and adult Japanese monkeys in a cool temperate area with snow coverage

Japanese monkeys, Macaca fuscata, living in a cool temperate forest experienced energy crises in winter. We measured feeding times and feeding rates (mass of foods eaten per unit time of feeding) in six different-sized, age–sex classes (1.2–12.6 kg body mass) in autumn and winter. One-, 2- and 3~4-year-olds spent 34–35% and 44–46% of the day feeding in autumn and winter, respectively. Monkeys less than 0 years old spent less time feeding (16–28%) than average in winter and autumn; adult females spent less (41%) in winter; and adult males spent less (25%) in autumn. All age–sex classes ate mainly fruits in autumn and the heavier classes fed more on tree bark than buds in winter. The feeding rate for fruits (2.3–53.5 g min^–1) was nine to 12 times faster than those for buds (1.0– 4.8 g min^–1) and bark (0.5–4.4 g min^–1), and energy content did not differ among fruits (22.1 kJ g^–1 dry mass), buds (19.9 kJ g^–1 dry mass) and bark (23.2 kJ g^–1 dry mass). Energy base feeding rates increased with body mass where the body mass exponent for buds (0.29) was smaller than those for bark (0.64) and fruits (0.63), which might be attributable to the unit size of food items and mass dependency of masticatory apparatus. Our monkeys obtained two to five times more energy in autumn (1567–1150 kJ day^–1) than in winter (604–3020 kJ day^–1). Adult females obtained 60% of expected energy expenditure and other classes obtained 77–88% of that in winter.     

Simultaneous determination of six penicillins in cows'' raw milk by a multiresidue high-performance liquid chromatographic method

A high-performance liquid chromatographic method based on C₁₈ solid-phase extraction and ultraviolet detection at 323 nm of analytes derivatized with benzoic anhydride and 1,2,4-triazole mercuric chloride solution was developed for the simultaneous determination of amoxicillin, penicillin G (benzylpenicillin), ampicillin, oxacillin, cloxacillin and dicloxacillin residues in raw milk. The detection limit of the method was 1.3 μg/l for penicillin G; 1.4 μg/l for amoxicillin, oxacillin and cloxacillin, 1.5 μg/l for ampicillin and 2.7 μg/l for dicloxacillin. The mean recovery was 95–102% for amoxicillin, penicillin G and ampicillin, 92–98% for oxacillin and cloxacillin and 87–94% for dicloxacillin, measured by using an internal standard. The relative repeatability standard deviation was 4–9% on level 4–15 μg/l, respectively, 2–7% on level 30–40 μg/l.     

Response surface optimization for the continuous glucose isomerization process

Production of fructose via a continuous glucose isomerization process was optimized using response surface methodology. Glucose isomerization was performed using immobilized glucose isomerase in a flow-through tubular reactor. Process factors eg pH (7.0–7.8), temperature (50–60°C), flow rate (5–17 ml min^–1) and glucose content (30–50% w/w) of the feedstock solution were simultaneously tested according to a central composite experimental design. Measured responses such as % isomerization, and fructose yield (gh^–1) has an excellent correlation with tested factors. The highest desirability,D, (geometric mean of % isomerization and fructose yield) was obtained when the feedstock (56–60°C) had 34–36% glucose, a pH of 7.4–7.8 and was pumped at 15 ml min^–1.     

Cultivation and carrageenan yield and quality ofKappaphycus alvarezii in the waters of Vietnam

The carrageenan-producing red algaKappaphycus alvarezii (Doty) Doty was brought to Vietnam from Japan in 1993. Branch fragments of this species were cultivated in a pond, lagoon, inlet and offshore in Vietnam for the first time. The best daily growth rate (DGR) of plants grown in the lagoon area attained 9–11 % day^–1 in May to June (cold season). The water temperature and salinity in this area ranged from 27.2–32.4 °C and 31.4–33.7 °C, respectively. DGR of plants grown in the inlet ranged from 7 to 9% day^–1 in June. Grazing by fish has been observed to occur in this area. The DGR of plants grown in the pond ranged from 5–6% in January–July, but decreased to less than 4% day^–1 in August (hot season). K. alvarezii in Vietnam showed a carrageenan yield of 18.8–24.6% and gel strength of 1566–1712 g cm^–2. These values are similar ones obtained fromK. alvarezii cultivated in the Philippines and Indonesia.     

Simultaneous determination of seven penicillins in muscle, liver and kidney tissues from cattle and pigs by a multiresidue high-performance liquid chromatographic method

A high-performance liquid chromatographic (HPLC) method based on solid-phase extraction (SPE) was developed for determination of amoxicillin, penicillin G (benzylpenicillin), ampicillin, oxacillin, cloxacillin, nafcillin and dicloxacillin in muscle, liver and kidney tissues of pigs and cattle. The compounds were extracted in aqueous solution by precipitation of organic materials with a mixture of sulphuric acid and sodium tungstate. The extract was cleaned up by SPE on a divinylbenzene-co-N-vinylpyrrolidone polymeric sorbent. Further clean-up was performed by liquid–liquid partition with diethyl ether. The extract was derivatised with benzoic anhydride and 1,2,4-triazole mercury (II) reagent. Chromatography was performed by reversed-phase gradient HPLC on a C₁₈ column with ultraviolet detection at 323 nm. The limits of detection estimated by a conservative model were in the range 8.9–11.1 μg/kg for amoxicillin, penicillin G, ampicillin, oxacillin, cloxacillin and nafcillin and 18.3–20.9 μg/kg for dicloxacillin. The mean recovery range was 66–77% for amoxicillin, 73–75% for penicillin G, 81–82% for ampicillin, 73–76% for oxacillin, 74–75% for cloxacillin, 66–72% for nafcillin and 58–65% for dicloxacillin.     

Biochemical properties of pigeon milk and its effect on growth

Summary The avian juvenile food pigeon milk was studied for its chemical composition and effect on growth in vivo and in vitro. Pigeon milk on a wet weight basis consisted of 9–13% protein, 9–11% fat, 0.9–1.5% carbohydrate, 0.8–1.1% ash, 0.10–0.12% non-protein nitrogen, energy content 5.6–6.8 kcal·g^-1. Except for proteins there was little or no decrease in pigeon milk constitutents during the first week of secretion. Pigeon milk proteins consisted of trichloroacetic acid (precipitable), trichloroacetic acid (soluble), and free amino acid components in the ranges 8.4–12.1%, 0.5–0.7% and 1.4–2.5%, respectively; whereas the level of trichloroacetic acid (precipitable) and trichloroacetic acid (soluble) components decreased by about 30%, that of the free amino acids increased by 9% in the first week. About 0.6–1.0% of pigeon milk sugar was found in the trichloroacetic acid (soluble) fraction and increased by 67% in the first week. The remainder was found in the trichloroacetic acid (precipitable) fraction and did not change during this period. Major lipids of pigeon milk were the neutral lipids (7.8–8.4%); the minor lipids were glycolipids (0.9–1.6%), phospholipids (0.5–1.4%) and cholesterol (0.5–0.6%). Squabs fed pigeon milk increased their body weight by 22-fold in the first 3 weeks after hatching, and crude extracts of pigeon milk stimulated the growth of cultured hamster ovary cells. These results reflect the ability of pigeon milk to stimulate growth both in vivo and in vitro.Abbreviations AOAC association of official analytical chemists - BRIT board of radiation and isotope technology - CHO chinese hamster ovary - DNA deoxyribonucleic acid - EDTA ethylenediaminetetra-acetic acid - FCS foetal calf serum - GF growth factor - GS goat serum - MEM minimum essential medium - NPN nonprotein nitrogen - PBS phosphate-buffered saline - PM pigeon milk - TCA(P) trichloroacetic acid precipitable fraction - TCA(S) trichloroacetic acid soluble fraction     

Winter fluxes of methane from Minnesota peatlands

Winter fluxes of methane were investigated in northern Minnesota during 1988–89 and 1989–90. Two bogs and a fen emitted methane throughout the snow-covered season (November through March). Fluxes decreased to a low level of 3–16 mg CH₄ m^–2 d^–1 in late March, reflecting decreasing peat temperatures and (in 1989–90) increasing depth of frost in the peat. Winter fluxes calculated by integration for an open poor fen, an open bog, a forested bog hollow, and a hummock site in the forested bog averaged 49, 12, 13, and 5 mg m^–2 d^–1, respectively, in 1989–1990 (the year most measurements were made). These comprised 11%, 4%, 15%, and 21% of total annual flux.     

The importance of avian-contributed nitrogen (N) and phosphorus (P) to Lake Grand-Lieu,France

The largest natural lake in France, Grand-Lieu, has suffered eutrophication. The objective of the study was to estimate the annual input of nutrients (N, P) resulting from avian excrement, deposited by birds feeding out of the lake and returning to its waters for breeding or roosting, as compared to the input by the rivers that enter in the lake. Two years are compared: 1981–82 and 1990–91. About 1600–2000 breeding herons and cormorants, 20 000–33 000 wintering ducks, gulls and cormorants and 1–2.4 million starlings deposited about 5800 kg total N in 1981–82 and 7640 kg in 1990–91. Respectively, 2000 and 2530 kg total P were deposited over the same time periods. These represent 0.7% and 0.4% of the total N input of the lake and 2.4 and 6.6% of the total P input in 1981–82 and 1990–91. Starlings account for 74% of the N and mallards most of the rest. P input by starlings (36% in 1981–82, 41% in 1990–91), and by mallards and herons (35% and 27% in 1981–82 and 22% and 24% in 1990–91 respectively) plays an appreciable role among birds. During the plant growing period (April–September), the contribution by birds can increase to 37% of total P input of the lake. Piscivorous bird colonies concentrate Phosphorus 42 times more within the colony than outside the colony. Overall, the role birds play in total N and P input is relatively small due to very high inputs from human sewage and agriculture run off. The monthly mean concentration of the water of the two rivers reaches currently 10 mg l^–1 of N (to 23 mg during peak floods) and 394 mg m^–3 of P (to 468 mg during peak floods). Earlier, for example in the 1960's, water in Brittany only contained 0.1 to 1.1 mg 1^–1 of N and 1 to 5 mg m^–3 of P during the maximum flow period. At this time, birds could probably have represented annually up to 37% of the N input and up to 95% of the P input to the lake.     

Interannual and between species comparison of the lipids, fatty acids and sterols of Antarctic krill from the US AMLR Elephant Island survey area

Antarctic euphausiids, Euphausia superba, E. tricantha, E. frigida and Thysanoessa macrura were collected near Elephant Island ¦ during 1997 and 1998. Total lipid was highest in E. superba small juveniles (16 mg g⁻¹ wet mass), ranging from 12 to 15 mg in other euphausiids. Polar lipid (56–81% of total lipid) and triacylglycerol (12–38%) were the major lipids with wax esters (6%) only present in E. tricantha. Cholesterol was the major sterol (80–100% of total sterols) with desmosterol second in abundance (1–18%). 1997 T. macrura and E. superba contained a more diverse sterol profile, including 24-nordehydrocholesterol (0.1–1.7%), trans-dehydrocholesterol (1.1–1.5%), brassicasterol (0.5–1.7%), 24-methylenecholesterol (0.1–0.4%) and two stanols (0.1–0.2%). Monounsaturated fatty acids included primarily 18:1(n−9)c (7–21%), 18:1(n−7)c (3–13%) and 16:1(n−7)c (2–7%). The main saturated fatty acids in krill were 16:0 (18–29%), 14:0 (2–15%) and 18:0 (1–13%). Highest eicosapentaenoic acid [EPA, 20:5(n−3)] and docosahexaenoic acid [DHA, 22:6(n−3)] occurred in E. superba (EPA, 15–21%; DHA, 9–14%), and were less abundant in other krill. E. superba is a good source of EPA and DHA for consideration of direct or indirect use as a food item for human consumption. Lower levels of 18:4(n−3) in E. tricantha, E. frigida and T. macrura (0.4–0.7% of total fatty acids) are more consistent with a carnivorous or omnivorous diet as compared with herbivorous E. superba (3.7–9.4%). The polyunsaturated fatty acid (PUFA) 18:5(n−3) and the very-long chain (VLC-PUFA), C₂₆ and C₂₈ PUFA, were not present in 1997 samples, but were detected at low levels in most 1998 euphausiids. Interannual differences in these biomarkers suggest greater importance of dinoflagellates or some other phytoplankton group in the Elephant Island area during 1998. The data have enabled between year comparisons of trophodynamic interactions of krill collected in the Elephant Island region, and will be of use to groups using signature lipid methodology.     

Formation of extracellular and intracellular ice during warming of vitrified mouse morulae and its effect on embryo survival

In vitrified solutions, ice can form during warming if the concentration of the cryoprotectant is insufficient. For the cryopreservation of cells, ice is innocuous when it remains outside the cell, but intracellular ice (ICI) is lethal. We tried to estimate the conditions in which ICI forms in vitrified mouse morulae during warming. The solutions for the experiments (EFS10–EFS50) contained 10–50% ethylene glycol plus Ficoll plus sucrose. When vitrified EFS20, EFS30, and EFS40 were kept at −80 °C, they remained transparent after 3 min, but turned opaque after 60 min (EFS20, EFS30) or 24 h (EFS40). Morulae were vitrified with EFS solutions after exposure for 30–120 s at 25 °C. They were warmed by various methods and survival was assessed in culture. After rapid warming (control), survival was high with EFS30 (79–93%) and EFS40 (96–99%). After slow warming, survival decreased with both EFS30 (48–62%) and EFS40 (44–64%). This must be from the formation of ICI. To examine the temperature at which ICI formed during slow warming, vitrified embryos were kept at various sub-zero temperatures during warming. Survival with EFS30 and EFS40 decreased on keeping samples for 3 min at −80 (25–75%), −60 (7–49%), −40 (0–41%), or −20 °C (26–60%). When samples were kept at −80 °C for 24 h, the survival decreased to 0–14%. These results suggest that ICI forms at a wide range of temperatures including −80 and −20 °C, more likely between −60 and −40 °C, and the ice forms not only quickly but also slowly.     

Combined preparative enzymatic synthesis of dTDP-6-deoxy-4-keto-D-glucose from dTDP and sucrose

dTDP–6–deoxy–4–keto–D–glucose (1), the common intermediate in the biosyntheses of the mainfold deoxysugars, was synthesized on a gram–scale by the combination of sucrose synthase and dTDP–D–glucose 4,6–dehydratase in a fed batch, starting the reaction with dTDP. This process allowed a dTDP conversion with a 100% rate. An easy and efficient three–step purification with anion–exchange chromatography and gel filtration gave 1.1 g of 1 in an overall yield of 73%. This work realizes a first step for an economic access to activated deoxysugars.     

Effect of brassinolide on callus growth and regeneration in Spartina patens (Poaceae)

The effect of brassinolide (BL) on cultured calluses of Spartina patens (Ait.) Muhl. (Poaceae), a halophyte monocot was studied. BL at 0.03–0.04 mg l^–1 at fixed concentrations of IAA (0.2 mg l^–1) and BA (3.0 mg l^–1) added in MS medium increased the ratio for fresh weight (CIRFW) to dry weight (CIRDW) by 96–111% and 235–326%. Similarly, in callus regeneration capacity, BL at 0.03 mg l^–1 was most effective, increasing the shoot regeneration ratio (SRR) by 425%. BL at 0.04 mg l^–1 had not such an increasing effect as BL at 0.03 mg l^–1, which increased SRR by 79%. However, BL at 0.005 mg l^–1 promoted regenerated shoot growth most significantly, increasing the shoot height increasing ratio (SHIR) by 395% after a 40-day culture. BL at 0.05 mg l^–1 was least effective in the callus regeneration and regenerated shoot growth, decreasing SRR by 27% and SHIR by 52%. Present results suggest that BL at 0.03 mg l^–1 is suitable for the callus growth and shoot regeneration, while BL at 0.005 mg l^–1 effectively enhanced the regenerated shoot growth.