Chemical structures of oligosaccharides in milks of the American black bear (Ursus americanus americanus) and cheetah (Acinonyx jubatus)

The milk oligosaccharides were studied for two species of the Carnivora: the American black bear (Ursus americanus, family Ursidae, Caniformia), and the cheetah, (Acinonyx jubatus, family Felidae, Feliformia). Lactose was the most dominant saccharide in cheetah milk, while this was a minor saccharide and milk oligosaccharides predominated over lactose in American black bear milk. The structures of 8 neutral saccharides from American black bear milk were found to be Gal(β1–4)Glc (lactose), Fuc(α1–2)Gal(β1–4)Glc (2′-fucosyllactose), Gal(α1–3)Gal(β1–4)Glc (isoglobotriose), Gal(α1–3)[Fuc(α1–2)]Gal(β1–4)Glc (B-tetrasaccharide), Gal(α1–3)[Fuc(α1–2)]Gal(β1–4)[Fuc(α1–3)]Glc (B-pentasaccharide), Fuc(α1–2)Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–3)Gal(β1–4)Glc (difucosyl lacto-N-neotetraose), Gal(α1–3)Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–3)Gal(β1–4)Glc (monogalactosyl monofucosyl lacto-N-neotetraose) and Gal(α1–3)Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (Galili pentasaccharide). Structures of 5 acidic saccharides were also identified in black bear milk: Neu5Ac(α2–3)Gal(β1–4)Glc (3′-sialyllactose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)[Fuc(α1–2)Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (monosialyl monofucosyl lacto-N-neohexaose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)[Gal(α1–3)Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (monosialyl monogalactosyl lacto-N-neohexaose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3){Gal(α1–3)Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–6)}Gal(β1–4)Glc (monosialyl monogalactosyl monofucosyl lacto-N-neohexaose), and Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3){Gal(α1–3)[Fuc(α1–2)]Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–6)}Gal(β1–4)Glc (monosialyl monogalactosyl difucosyl lacto-N-neohexaose). A notable feature of some of these milk oligosaccharides is the presence of B-antigen (Gal(α1–3)[Fuc(α1–2)]Gal), α-Gal epitope (Gal(α1–3)Gal(β1–4)Glc(NAc)) and Lewis x (Gal(β1–4)[Fuc(α1–3)]GlcNAc) structures within oligosaccharides. By comparison to American black bear milk, cheetah milk had a much smaller array of oligosaccharides. Two cheetah milks contained Gal(α1–3)Gal(β1–4)Glc (isoglobotriose), while another cheetah milk did not, but contained Gal(β1–6)Gal(β1–4)Glc (6′-galactosyllactose) and Gal(β1–3)Gal(β1–4)Glc (3′-galactosyllactose). Two cheetah milks contained Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (lacto-N-neohexaose), and one cheetah milk contained Gal(β1–4)Glc-3’-O-sulfate. Neu5Ac(α2–8)Neu5Ac(α2–3)Gal(β1–4)Glc (disialyllactose) was the only sialyl oligosaccharide identified in cheetah milk. The heterogeneity of milk oligosaccharides was found between both species with respect of the presence/absence of B-antigen and Lewis x. The variety of milk oligosaccharides was much greater in the American black bear than in the cheetah. The ratio of milk oligosaccharides-to-lactose was lower in cheetah (1:1–1:2) than American black bear (21:1) which is likely a reflection of the requirement for a dietary supply of N-acetyl neuraminic acid (sialic acid), in altricial ursids compared to more precocial felids, given the role of these oligosaccharides in the synthesis of brain gangliosides and the polysialic chains on neural cell adhesion.

     

Novel stable analogues of the neurotensin C-terminal hexapeptide containing unnatural amino acids

Amino Acids - Neurotensin (NT) (pGlu–Leu–Tyr–Glu–Asn–Lys–Pro–Arg–Arg–Pro–Tyr–Ile–Leu) exerts a dual function as a...     

Ascidiacyclamides containing oxazoline and thiazole motifs assume square conformations and show high cytotoxicity

Four cyclic octapeptides were designed from ascidiacyclamide [cyclo(–Ile–Oxz–D ‐Val– Thz–)₂] (ASC, 1 ) to investigate the effects of oxazoline (Oxz) and thiazole (Thz) rings on the structures and cytotoxicities of the peptides. cyclo(–Ile–Thz–D ‐Val–Oxz–)₂ ( 2 ) had the same number of Oxz and Thz rings as ASC, but the ring positions were switched. cyclo(–Ile–Oxz–D ‐Val–Thz–Ile–Thz–D ‐Val–Thz–) ( 3 ) and cyclo(–Ile–Thz–D ‐Val–Oxz–Ile–Thz–D ‐Val–Thz–) ( 4 ) contained one Oxz and three Thz rings within the molecule. All Oxz rings were substituted with Thz in cyclo(–Ile–Thz–D ‐Val–Thz–)₂ ( 5 ). These analogues had new Oxz and Thz blocks forming the 24‐membered ring. Based on CD spectra and X‐ray diffraction analyses, the structures of all four analogues were classified as square ASC forms. But the structures of 2 and 5 differed from the original square form of 1 , and they showed no cytotoxicity. The structure of 3 was very similar to that of 1 , and 3 showed 10 times greater cytotoxicity than 1 . Although no definite structure of 4 was obtained, it showed three times greater cytotoxicity than 1 . It appears that the position and number of Oxz residues are essential determinants in the structure‐cytotoxicity relationship of ASC analogues.     

Composition of the major elements and trace elements of 10 methanogenic bacteria determined by inductively coupled plasma emission spectrometry

The elemental composition of 10 methanogenic species was determined by inductively coupled plasma emission spectrometry and by a C-H-N-analyzer. The 10 species were representative of all three orders of the methanogens and were cultivated under defined conditions. Special emphasis was given toMethanosarcina barkeri, represented by 5 strains and cultivated on various substrates. The following elements with the lowest and highest values in parentheses were determined: C (37–44%, w/w), H (5.5–6.5%), N (9.5–12,8%); Na (0.3–4.0%), K (0.13–5.0%), S (0.56–1.2%), P (0.5–2.8%), Ca (order I: 85–550 ppm; order II: 1000–4500 ppm), Mg (0.09–0.53%), Fe (0.07–0.28%), Ni (65–180 ppm), Co (10–120 ppm). Mo (10–70 ppm), Zn (50–630 ppm), Cu (<10–160 ppm), Mn (<5–25 ppm). The biggest variations were found with respect to N and K, which both seem to have important physiological functions. Although it is unknown whether zinc and copper are essential trace elements for methanogens, all investigated species contained remarkably high zinc contents, whereas copper seemed to be present only in some species.     

Interaction of a dimeric distamycin analog with poly(dA)poly(dT), poly[d(A–T)]poly[d(A–T)], and duplex O<Subscript>23</Subscript> at the origin of replication of the herpes simplex virus

The binding of a dimeric distamycin analog (Pt–bis–Dst) to poly[d(A–T)]poly[d(A–T)], poly(dA)poly(dT), and duplex O₂₃ with the sequence 5’-GCCAATATATATATATTATTAGG-3’, which occurs at the origin of replication (OriS) of the herpes simplex virus, was studied via UV and CD spectroscopy. The synthetic polyamide differs from the natural antibiotic in having two distamycin moieties that are linked via a glycine cis-diamino platinum group. The Pt–bis–Dst binding to poly[d(A–T)]poly[d(A–T)] and poly(dA)poly(dT) reached saturation at approximately one ligand molecule per eight bp. As the ligand–base pair ratio further increased, the maximum wavelength band tended to shift toward longer wavelengths in the CD spectra of complexes with poly[d(A–T)]poly[d(A–T)] and a shoulder appeared in the 290–310 nm spectral region that was absent from the CD spectra of complexes with lower ligand coverages. At higher ligand–oligonucleotide molar ratios, Pt–bis–Dst could bind to poly[d(A–T)]poly[d(A–T)] in the form of hairpins or associations that result from interactions between the distamycin moieties of two neighbor Pt–bis–Dst molecules. The structures of the complexes were stabilized by interactions between the pirrolcarboxamide moieties of two Pt–bis–Dst molecules absorbed on adjacent overlapping binding sites. The interactions could also be responsible for the concentration-dependent spectral changes that were observed during the formation of a complex between Pt–bis–Dst and poly[d(A–T)]poly[d(A–T)]. Spectral changes were almost absent in the case of Pt–bis–Dst binding to poly(dA)poly(dT). The binding of Pt–bis–Dst to duplex O₂₃ reached saturation at two ligand molecules per duplex, which contained a cluster of 18 AT pairs. At higher molar-concentration ratios, duplex CD spectra underwent changes similar to those that were observed for Pt–bis–Dst binding to poly[d(A–T)]poly[d(A–T)]. Testing Pt–bis–Dst for antiviral activity identified 1.5 μg/mL as a concentration that halved the cytopathic effect of the herpes simplex virus on Vero E6 cells; the selectivity index of antiviral action was 65; cytotoxicity was relatively low. The Pt–bis–Dst concentration that caused the death of approximately half of the cells was estimated at 100 μg/mL.     

Chromatography and Atomic Absorption Spectrometry for the Assessment of Heavy Metal Distribution among Amino Acids Used in Parenteral Nutrition Formulations—Studies with Cadmium and Lead

The distribution of Cd (II) and Pb (II) among amino acids in parenteral nutrition formulations was investigated by coupling ion-chromatography (HPLC/IC) and electrothermal atomic absorption spectrometry. The methodology was based on ion-exchange separation and fluorimetric amino acid detection after post-column derivatization. Cd (II) and Pb (II) were assayed in 500-µL fractions of the column effluent. The distribution of Cd (II) and Pb (II) in alanine (Ala), aspartic acid (Asp), glutamic acid (Glu), glycine (Gly), histidine (His), methionine (Met), phenylalanine (Phe), serine (Ser), and threonine (Thr) were analyzed by monitoring changes in the concentration of free amino acids by HPLC/IC. The results indicated that Cd (II) and Pb (II) were distributed according to the following trend: Gly–Cd?>?Gly–Pb?>?Ala–Cd?>?Ala–Pb?>?His–Cd?～?His–Pb?>?Thr–Cd?>?Thr–Pb?>?Phe–Cd?～?Phe–Pb?～?Asp–Cd?～?Asp–Pb?～?Met–Cd?～?Met–Pb?～?Glu–Cd?～?Glu–Pb?>?Ser–Cd?～?Ser–Pb. The effects of amino acid concentration and stability constants of amino acid–metal complexes are discussed.     

ACE Inhibitory and Antioxidant Activities of Novel Peptides from <Emphasis Type="Italic">Scorpaena notata</Emphasis> By-product Protein Hydrolysate

This study describes the isolation of angiotensin I converting enzyme and antioxidative peptides from head protein hydrolysate of red scorpionfish (Scorpaena notata) prepared by treatment with a protease from the fungus Penicillium digitatum. After ultrafiltration, three peptides were isolated by a two-step procedure: size exclusion chromatography on a Toyopearl HW-40 followed by reversed-phase high performance liquid chromatography (RP-HPLC) with a high purification yield of 2.22 mg of peptide/g of initial protein. Active peptides were then identified by nanoscale liquid chromatography coupled to tandem mass spectrometry (nanoLC/MS–MS), corresponding to the following sequences: Gln–Gln–Pro–His–Ser–Arg–Ser–Lys–Gly–Phe–Pro–Gly–Pro (1424.724 Da), Gly–Gln–Lys–Ser–Val–Pro–Glu–Val–Arg (1000.565 Da) and Val–Glu–Gly–Lys–Ser–Pro–Asn–Val (830.448 Da). Peptides D-I, E-I and F-I showed high angiotensin-I converting enzyme inhibitory activity with an IC₅₀ values of 0.98, 1.69 and 1.44 µM, respectively as well as a synergistic antioxidant activity between the different fractions. Thus, we have demonstrated that underutilized wastes can be valorized by production of peptides that can be used as potential therapeutic compounds active against oxidative stress and hypertension.     

Replication and extension of association of choline acetyltransferase with nicotine dependence in European and African American smokers

Choline acetyltransferase is critical in the synthesis of acetylcholine and regulation of cholinergic neuron functions. We recently reported association of the encoding gene ChAT with both smoking cessation and nicotine dependence (ND) in two independent European American (EA) samples; however, in the replication sample, only limited SNPs partially covering the gene were examined. In this study, we examined the association of 14 SNPs, which cover the entire gene, with ND, assessed by smoking quantity (SQ), heaviness of smoking index (HSI), and Fagerström Test for ND (FTND), in 2,037 subjects from 602 families of African American (AA) or EA origin. Individual SNP-based association analysis revealed that five SNPs showed nominal association with at least one ND measure in one of the samples (P = 0.022–0.042); none remained significant after correction for multiple testing. Haplotype-based association analysis revealed that haplotypes G–G–A–C, formed by rs1880676–rs3810950–rs10082479–rs8178990 (P = 0.005–0.0178), and G–G–T–C–G–C, formed by rs1880676–rs3810950–rs10082479–rs8178990–rs3793790–rs12266458 (P = 0.00247–0.00468), displayed significant association with all three ND measures in the AA sample, as did haplotype T–C–G–A–T, formed by rs12266458–rs11101191–rs8178991–rs4838544–rs4838547 (P = 0.00741–0.0103), in the EA sample. All these detected haplotype-based associations remained significant after correction for all major haplotypes for a given SNP combination. Together, our findings, in conjunction with the previous report of the association, warrant further investigation of ChAT in ND.     

Metabolic stability of human parathyroid hormone peptide hPTH (1–34) in rat tissue homogenates: kinetics and products of proteolytic degradation

The present study aim to investigate the metabolic stability and degradation of cleavage sites of human parathyroid hormone peptide, hPTH (1–34), in rat tissue homogenate, and to identify the types of proteases involved in hPTH (1–34) processing degradation. The stability of hPTH (1–34) in rat kidney, lung and liver homogenates was evaluated by LC–ESI–MS, and the structures of the major degradation products were identified by MALDI–TOF MS and LC–ESI–MS/MS. The ability of protease inhibitors to inhibit hPTH (1–34) degradation was used to identify the class of proteases involved in the metabolism of hPTH (1–34). hPTH (1–34) peptide was readily degraded in rat kidney, liver, and lung homogenates, with half-lives of 5.7, 32.2, and 18.9 min, respectively. The degradation of hPTH (1–34) in each tissue can be inhibited by inhibitors of serine and metalloproteases. The major degradation products of hPTH (1–34) are similar in each tissue and suggest that hPTH (1–15) and hPTH (16–34) appear as the major degradation products. The degradation patterns of hPTH (1–34) incubated in rat kidney, liver and lung homogenates are largely overlapping, and a majority of the fragments are generated via cleavages at sites of Leu15–Asn16 peptide bond.     

Local formation of angiotensin peptides with paracrine activity by adipocytes

A local paracrine angiotensin (ANG) system influences the insulin sensitivity and cell differentiation of adipose tissue. The limited view of a merely systemic renin‐angiotensin‐aldosterone‐system with ANG II (1–8) as the main mediator of ANG‐related effects may oversimplify the situation. The aim was to analyze the degradation of ANG by using capillary electrophoresis (CE) techniques. The supernatant of cultured 3T3‐L1 adipocytes was used directly, and some data on degraded peptides were combined with a biological effect. The formation of several peptides such as ANG II (1–8), —III (2–8), —IV (3–8), and ANG (1–7) as degradation products is demonstrated; in addition low levels of ANG (3–7) are identified. The concentrations of the peptides ANG III (2–8) and ANG IV (3–8) (both are AT₄ receptor agonists) are modified in the vicinity of adipose tissue cells by amino‐terminal degradation which resulted in ANG (3–8), —(4–8) and —(5–8). ANG IV (3–8) and ANG II (1–8) were biologically highly effective in inhibiting IRAP (insulin regulated aminopeptidase, part of the AT₄ receptor). It is observed that ANG (1–7) is the main degradation product derived from ANG I via ANG (1–9) and that ANG III (2–8) is one important regulated peptide for IRAP. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd.     

Studies on the Chemical Constituents from Root Bark of Aralia armata (Wall.) Seem.

Eleven constituents were isolated from root bark of Aralia armata (Wall.) Seem. Their structures were elucidated by spectra(IR, MS, 1H-NMR and 13C-NMR) and chemical analyses. They were deglucose chikusetsusaponin Ⅳ (AT–Ⅰ), chikusetsusaponin Na (AT–Ⅱ ), zingibroside R1 (AT–Ⅲ), ginsenin R0 (AT–IV), decaisneanaside (AT–Ⅴ), armatoside (AT-Ⅶ), araloside A (AT–Ⅶ), octacosanoic acid (AT–Ⅷ), β-sitosterol (ATIX ), mixture of β-sitosterol and stigmastterol (AT–×), oleanolic acid (AT–Xl ). They were for the first time found in this plant, AT–Ⅵ is a new saponin.     

The impact of pharmacological and non‐pharmacological interventions to improve physical health outcomes in people with schizophrenia: a meta‐review of meta‐analyses of randomized controlled trials

We summarized and compared meta‐analyses of pharmacological and non‐pharmacological interventions targeting physical health outcomes among people with schizophrenia spectrum disorders. Major databases were searched until June 1, 2018. Of 3,709 search engine hits, 27 meta‐analyses were included, representing 128 meta‐analyzed trials and 47,231 study participants. While meta‐analyses were generally of adequate or high quality, meta‐analyzed studies were less so. The most effective weight reduction interventions were individual lifestyle counseling (standardized mean difference, SMD=–0.98) and exercise interventions (SMD=–0.96), followed by psychoeducation (SMD=–0.77), aripiprazole augmentation (SMD=–0.73), topiramate (SMD=–0.72), d‐fenfluramine (SMD=–0.54) and metformin (SMD=–0.53). Regarding waist circumference reduction, aripiprazole augmentation (SMD=–1.10) and topiramate (SMD=–0.69) demonstrated the best evidence, followed by dietary interventions (SMD=–0.39). Dietary interventions were the only to significantly improve (diastolic) blood pressure (SMD=–0.39). Switching from olanzapine to quetiapine or aripiprazole (SMD=–0.71) and metformin (SMD=–0.65) demonstrated best efficacy for reducing glucose levels, followed by glucagon‐like peptide‐1 receptor agonists (SMD=–0.39), dietary interventions (SMD=–0.37) and aripiprazole augmentation (SMD=–0.34), whereas insulin resistance improved the most with metformin (SMD=–0.75) and rosiglitazone (SMD=–0.44). Topiramate had the greatest efficacy for triglycerides (SMD=–0.68) and low‐density lipoprotein (LDL)‐cholesterol (SMD=–0.80), whereas metformin had the greatest beneficial effects on total cholesterol (SMD=–0.51) and high‐density lipoprotein (HDL)‐cholesterol (SMD=0.45). Lifestyle interventions yielded small effects for triglycerides, total cholesterol and LDL‐cholesterol (SMD=–0.35 to –0.37). Only exercise interventions increased exercise capacity (SMD=1.81). Despite frequent physical comorbidities and premature mortality mainly due to these increased physical health risks, the current evidence for pharmacological and non‐pharmacological interventions in people with schizophrenia to prevent and treat these conditions is still limited and more larger trials are urgently needed.     

Wing and leg amputation induced oviposition pattern of tropical tasar silk moth,Antheraea mylitta Drury (Lepidoptera: Saturniidae) in oviposition devices – Earthen cups and paper sheets

Antheraea mylitta Drury (Lepidoptera: Saturniidae) is a tropical semi-domesticated wild tasar silkworm reared by marginal tribal farmers of India. Due to improper egg laying frequency, oviposition is generally kept for three continuous days after amputating the gravid moth wings (W^–) in earthen cups (C). In order to systematize the oviposition frequency, wing cut with leg cut (W^–L^–) was performed in ecorace Daba trivoltine (DTV) completing three life-cycles in a year. Oviposition of (C)W^–L^– was observed in all three grainages during the year 2020 and compared with (C)W^– moths. Oviposition was also accomplished on plain paper sheets (S). It is observed that (C)W^– obtained 63–67% and (C)W^–L^– obtained 72–80% egg laying on first day. Similarly, (S)W^– and (S)W^–L^– obtained 68–71% and 74–83% on first day. Further, significant high oviposition was observed within first four hours in both earthen cups and on paper sheets by W^–L^– moths laying 49–69% and 55–65% eggs compared to W^– laying 25–45% and 39–44% eggs, respectively. A total of 44–51, 47–54, 44–48 and 46–52 eggs/g moth weight was obtained in (C)W^–, (C)W^–L^–, (S)W^– and (S)W^–L^–, respectively in three consecutive grainage. Oviposition of W^– moths in earthen cups and on paper sheets are not significantly different indicating earthen cups in contemporary tasar grainage could be replaced with paper sheets. Thus, the paper demonstrates for the first time a fast, efficient and scalable cellular oviposition of A. mylitta on paper sheets comparable to Bombyx mori.     

Concentrations of Selected Trace Elements in Marine Bivalves and the Dietary Risk to Residents of Dalian City,Northern China

To assess the potential health risks associated with selected trace elements due to consumption of bivalves, a screening-level risk assessment was conducted through consideration of internationally accepted dietary guidelines and the calculation of hazard quotients. Seventeen edible bivalve species were collected from a local market in Dalian City, China. The ranges obtained for the trace elements analyzed in mg/kg dry weight were as follows: Cu (1.8–133), Zn (56–693), Mn (5.7–150), Se (0.2–3.8), Ni (0.56–7.67), Cd (0.04–10.3), Cr (0.48–2.23), Pb (0.01–1.63), Hg (0.07–0.35), and As (0.22–4.00). The estimated daily intake (EDI) of bivalves was 15.0–255.1 times lower than the RfD guidelines for all the trace elements studied. Further, the calculated hazard quotients (HQs) of the trace elements analyzed were well less than 1. Therefore, there were no potential human health risks to Dalian City consumers of marine bivalves.     

Influence of poly(n‐isopropylacrylamide)–CNT–polyaniline three‐dimensional electrospun microfabric scaffolds on cell growth and viability

This study investigates the effect on: (1) the bulk surface and (2) the three‐dimensional non‐woven microfabric scaffolds of poly(N‐isopropylacrylamide)–CNT–polyaniline on growth and viability of cells. The poly(N‐isopropylacrylamide)–CNT–polyaniline was prepared using coupling chemistry and electrospinning was then used for the fabrication of responsive, non‐woven microfabric scaffolds. The electrospun microfabrics were assembled in regular three‐dimensional scaffolds with OD: 400–500 μm; L: 6–20 cm. Mice fibroblast cells L929 were seeded on the both poly(N‐isopropylacrylamide)–CNT–polyaniline bulk surface as well as non‐woven microfabric scaffolds. Excellent cell proliferation and viability was observed on poly(N‐isopropylacrylamide)–CNT–polyaniline non‐woven microfabric matrices in compare to poly(N‐isopropylacrylamide)–CNT–polyaniline bulk and commercially available Matrigel? even with a range of cell lines up to 168 h. Temperature dependent cells detachment behavior was observed on the poly(N‐isopropylacrylamide)–CNT–polyaniline scaffolds by varying incubation at below lower critical solution temperature of poly(N‐isopropylacrylamide). The results suggest that poly(N‐isopropylacrylamide)–CNT–polyaniline non‐woven microfabrics could be used as a smart matrices for applications in tissue engineering. © 2012 Wiley Periodicals, Inc. Biopolymers 99: 334–341, 2013.     

VARIATION IN APPLE CYTOCHIMERAS

Diploid-tetraploid cytochimeras of apple were investigated by cytological examination of buds from branches selected by the characteristics of the fruits, flowers or pollen. Eight types of cytochimeras were identified on the basis of relative size of cells, nuclei and metaphase mitoses in the apical meristem (protomeristem) and mitoses in meristematic primary tissues. There appeared to be five apical layers which contributed to the stem and four which contributed to the leaves and flower parts. Buds of various cytochimeral patterns (designated by a formula giving the ploidy of the apical layers) were found on some bearing trees propagated from known cytochimeral sources. The most frequently associated types were (a) 2–4–2–2–2 and 2–4–4–4–4 (and 2–4–2–4–4 in one cultivar), or (b) 2–2–2–4–4 and 2x. Some sports were uniformly 2–4–4–4–4. The stability of apple cytochimeras under normal conditions appeared greater in some eultivars than in others. Sprouts from severely pruned 2–2–4–4–4 trees were more variable than unpruned branches. Buds of shoots which grew from radiation-damaged buds were more variable than those from non-irradiated buds and included types not yet found by branch selection. Cytochimeral variation was interpreted to be due to layer replacement resulting from infrequent periclinal divisions in apical or axillary meristems, or from wounding of meristems by ionizing radiation.     

Chemical structures of oligosaccharides in milk of the raccoon (<Emphasis Type="Italic">Procyon lotor</Emphasis>)

In this study on milk saccharides of the raccoon (Procyonidae: Carnivora), free lactose was found to be a minor constituent among a variety of neutral and acidic oligosaccharides, which predominated over lactose. The milk oligosaccharides were isolated from the carbohydrate fractions of each of four samples of raccoon milk and their chemical structures determined by ¹H-NMR and MALDI-TOF mass spectroscopies. The structures of the four neutral milk oligosaccharides were Fuc(α1–2)Gal(β1–4)Glc (2′-fucosyllactose), Fuc(α1–2)Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (lacto-N-fucopentaose IV), Fuc(α1–2)Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (fucosyl para lacto-N-neohexaose) and Fuc(α1–2)Gal(β1–4)GlcNAc(β1–3)[Fuc(α1–2)Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (difucosyl lacto-N-neohexaose). No type I oligosaccharides, which contain Gal(β1–3)GlcNAc units, were detected, but type 2 saccharides, which contain Gal(β1–4)GlcNAc units were present. The monosaccharide compositions of two of the acidic oligosaccharides were [Neu5Ac]₁[Hex]₆[HexNAc]₄[deoxy Hex]₂, while those of another two were [Neu5Ac]₁[Hex]₈[HexNAc]₆[deoxy Hex]_3. These acidic oligosaccharides contained α(2–3) or α(2–6) linked Neu5Ac, non reducing α(1–2) linked Fuc, poly N-acetyllactosamine (Gal(β1–4)GlcNAc) and reducing lactose.     

Casein-derived tripeptide Ile-Pro-Pro improves angiotensin-(1-7)- and bradykinin-induced rat mesenteric artery relaxation

AimsMilk casein-derived bioactive tripeptides isoleucine–proline–proline (Ile–Pro–Pro) and valine–proline–proline (Val–Pro–Pro) lower blood pressure in animal models of hypertension and humans. In some studies, their angiotensin-converting enzyme (ACE)-inhibitory effect has been demonstrated. Besides classical ACE-angiotensin II-AT₁-receptor pathway (ACE-Ang II- AT₁), the significance of ACE2-angiotensin-(1–7)-Mas-receptor (ACE2-Ang-(1–7)-Mas) axis in the blood pressure regulation has now been acknowledged. The present study was aimed to further evaluate the renin–angiotensin system (RAS)-related vascular effects of Ile–Pro–Pro in vitro using rat mesenteric arteries.Main methodsSuperior mesenteric arteries of spontaneously hypertensive rat (SHR) were isolated, cut into rings and mounted in standard organ bath chambers. Endothelium-intact arterial rings were incubated in Krebs solution either with Ile–Pro–Pro, proline–proline (Pro–Pro), isoleucine (Ile), proline (Pro) or captopril for 6 h at + 37 °C and vascular reactivity was measured.Key findingsIn the presence of AT₁-antagonist valsartan, Ang II induced vasodilatation, which was more pronounced in the arteries incubated with Ile–Pro–Pro (P < 0.05) compared to the other compounds. Ang-(1–7)-induced vasodilatation was augmented by Ile–Pro–Pro or Pro (P < 0.001 vs. control). Mas-receptor antagonist A-779 did not alter the responses. Ile–Pro–Pro and Pro augmented also bradykinin-induced relaxations (P < 0.001 vs. control). Control arteries and arteries incubated with captopril showed only slight relaxations at higher bradykinin concentrations.SignificanceCasein-derived tripeptide Ile–Pro–Pro and amino acid Pro enhance the vasodilatory effect of Ang-(1–7) and bradykinin. The role of ACE2-Ang–(1–7)-Mas axis in the modulation of vascular tone by these compounds seems probable.     

Morphometric study of Ecklonia cava (Laminariales, Phaeophyta) sporophytes in two localities with different temperature conditions

Sporophytes of Ecklonia cava Kjellman (Laminariales, Phaeophyta) were collected seasonally from within 3–4 replicate, 1‐m² quadrates, haphazardly placed in dense assemblages at 6–9 m depth in Tei, Tosa Bay (southern Japan; water temperature 15–29°C) from 1995 to 1996, and in Nabeta Bay, Shimoda (central Japan; water temperature 13–25°C) from 1996 to 1997. Growth rings were checked for all samples and mean values of each biometric parameter of sporophytes 1‐year‐old and over were compared. The plant length (stipe length + primary blade length) was always shorter in Tei sporophytes (24–52 cm) than Nabeta sporophytes (70–100 cm), the difference being mainly owing to the shorter stipe length in Tei sporophytes (7–14 cm) than in Nabeta sporophytes (54–83 cm). However, the primary blade length was sometimes longer in Tei sporophytes (12–38 cm) than in Nabeta sporophytes (14–21 cm). Stipe diameter, longest bladelet length and primary blade width were mostly less in Tei sporophytes (8.6–12.4 mm, 5.5–7.0 cm and 23.4–38.0 cm, respectively) than Nabeta sporophytes (16.4–20.2 mm, 9.2–12.0 cm and 43.0–52.6 cm, respectively). Nevertheless, the number of bladelets of sporophytes from Tei (15–28) and Nabeta (18–29) were within the same range. At Tei, 32–43% of the sporophytes had wrinkled blades in summer and autumn, whereas wrinkled primary blades or bladelets were not observed in Nabeta sporophytes throughout the experimental period. These morphometric differences of E. cava between the two localities are suggested to be dependent on environmental factors, especially seawater temperature.