Milk composition and flavor under different feeding systems: a survey of dairy farms.

Understanding the influence of regional dietary factors on the flavors of milk and dairy products will provide consumers with more options and promote the conservation of regional resources and the original terroir. The objective of this study was to evaluate the influence of regional differences in feeding systems on the composition, fatty acid content, and flavor of pasteurized milk at the farm level. Nine dairy farms using grass silage (GS), 6 farms using maize silage (MS), and 4 farms using by-products (BP) as the characteristic feed components were chosen for this survey. Fresh milk was sampled once per month from September 2008 to February 2009 at each dairy farm. The percentages of GS, MS, and BP (soybean curd residue or brewer's grain) in the feed were 32.4, 22.1, and 15.1%, respectively. The milk fat, protein, and lactose contents did not differ among the milks from farms with different feeding systems. Fatty acids with chain lengths of less than C16 and saturated fatty acids were present at higher concentrations in the milks from the GS and MS farms than in the milk from the BP farms; conversely, fatty acids with chain lengths greater than C18 and unsaturated fatty acids (UFA), including mono- (MUFA) and polyunsaturated fatty acids (PUFA), were present at higher concentrations in the milks from the BP farms than in the milks from the GS farms. No significant differences were detected in milk flavor, evaluated as sweetness, body, texture, aftertaste, and palatability, between the milks from the farms with different feeding systems. The proportion of BP in the cow's diet was positively correlated with the concentrations of fatty acids with chain lengths greater than C18 and with UFA, MUFA, and PUFA. In contrast, the proportion of GS in the diet was positively associated with the levels of milk fat, protein, fatty acids with chain lengths less than C16, and SFA. The MUFA, PUFA, UFA, and fatty acids with chain lengths greater than C18 were not associated with any of the milk flavors. These results suggest the regional differences in feeding systems contribute to the differences in the fatty acid compositions of milk at the farm level. However, these differences do not influence the flavor of pasteurized milk. Thus, more specific feed profiles will be required to provide a specific regional flavor to pasteurized milk.

Gene transfer and expression in progeny after intravenous DNA injection into pregnant mice.

Several methods that enable foreign genes to be transferred directly into germ cells and adult animals have been developed, which have stimulated great interest in manipulating genes in vivo. However, there have been no methods available for introducing genes into fetuses. We report here that a single intravenous injection of expression plasmid: lipopolyamine complexes into pregnant mice resulted in successful gene transfer into the embryos. The transgenes thus introduced were expressed in the fetuses and newborn progeny. This simple and new method of gene transfer into embryos will facilitate rapid analysis of transgene effects in the fetuses and will be useful for studying gene-deficient animal models to gain transgene functions at desired stages of embryogenesis.

The cooling mechanism of minuscule ribbed surfaces.

One reason human beings wear stockings is to warm their legs. Ordinary textile materials are thermally insulative, which prevents body's heat from dissipating. In contrary to this common sense, it was discovered that some knitted stockings made up of them permanently promote heat release and cool body. This non-intuitive phenomenon emerges when micro-size yarns are knitted to form wide spacing between neighboring yarns. However, the reason why they cool body was unclear because conventional principles of cooling garments cannot account for it. Here, in the basis of fluid-solid conjugate heat transfer analysis of natural convection, we have clarified the cooling mechanism originates from relative relationship between their geometric structure, a periodic alignment of minuscule ribs, and thermal boundary layer. Our novel finding revealed that sufficiently small ribs on the surface are exposed to steep temperature gradient within thermal boundary layer. Thereby, thermal conduction via ribs is enhanced complementarily as they are separated to guide cooler flow onto the surface. Our study provides a general insight into understanding permanent cooling mechanism on micro-size ribbed surfaces in contrast to conventional theory for heat sink, which is applicable not only to other clothes, but also to artificial devices or natural structures.

Hst-1 (FGF-4) antisense oligonucleotides block murine limb development.

The initiation of limb development depends on the site specific proliferation of the mesenchyme by the signals from the apical ectodermal ridge (AER) in embryonic mouse. We have previously reported that the local expression of Hst-1/Fgf-4 transcripts in AER of the mouse limb bud is developmentally regulated, expressed at 11 and 12 days post coitus (p.c.) embryo. In an effort to further understand the role of Hst-1/FGF-4 in mouse limb development, an antisense oligodeoxynucleotides (ODNs) study was performed. We first established a novel organ culture system to study mouse limb development in vitro. This system allows mouse limb bud at 9.5-10-d p.c. embryo, when placed on a sheet of extracellular matrix in a defined medium, to differentiate into a limb at 12.5-d p.c. embryo within 4.5 d. Using this organ culture system, we have shown that exposure of 9.5-10-d p.c. embryonal limb bud explants to antisense ODNs of Hst-1/FGF-4 blocks limb development. In contrast, sense and scrambled ODNs have no inhibitory effect on limb outgrowth, suggesting that Hst-1/FGF-4 may work as a potent inducing factor for mouse limb development.

Effects of a high-fat diet on superoxide anion generation and membrane fluidity in liver mitochondria in rats.

BACKGROUND: Obesity is a primary factor of lifestyle-related diseases, and the age of its onset has decreased. The reactive oxygen species (ROS), the superoxide anion, is generated in the mitochondrial electron transport chain and the damage it induces in cells may be a contributing factor to obesity-related lifestyle diseases. In the present study, the influence of the ingestion of a high-fat diet (HFD) on superoxide anion generation in rat liver mitochondria (Mt) and membrane fluidity was investigated. METHODS: Male Wistar rats were fed a normal diet (ND, n = 6) or HFD (n = 6). Liver Mt were isolated and oxygen consumption, superoxide anion production (the adrenaline method), and membrane fluidity (the spin label method) were measured. RESULTS: After 11 weeks, body weights and abdominal circumferences were higher in the HFD group than in the ND group. Mt oxygen consumption was higher in the HFD group than in the ND group. Superoxide anion production was significantly lower in the HFD group than in the ND group, while no significant changes were observed in membrane fluidity. CONCLUSION: Although rats developed diet-induced obesity, it did not reach the level of disease development. The promotion of lipid metabolism appeared to reduce superoxide anion production, but did not influence membrane fluidity. While superoxide anion damages cells as an oxidative stress, ROS and superoxide dismutase are essential signaling molecules in the body. The present results suggest that the continuous ingestion of a HFD impairs Mt and induces disease development.

Effective prevention of thrombocytopenia in mice using adenovirus-mediated transfer of HST-1 (FGF-4) gene.

HST-1 (FGF-4) gene product is a member of the fibroblast growth factor family with a signal peptide and plays a crucial role in limb development. We showed previously that an intraperitoneal injection of replication-deficient adenovirus containing the HST-1 gene (Adex1HST-1) into normal mice caused a twofold increase in peripheral platelet count. To investigate whether Adex1HST-1 could effectively prevent experimentally induced thrombocytopenia in mice, we injected Adex1HST-1 intraperitoneally into thrombocytopenic mice induced by administration of a chemotherapeutic agent and/or by irradiation. A single Adex1HST-1 injection caused continuously increased levels of serum HST-1 protein for at least 30 d and increased the count of large megakaryocytes in bone marrow, which specifically recovered platelet counts and more efficiently diminished the extent and duration of thrombocytopenia than any other reported cytokine or any combination of cytokines so far. In the other peripheral hematological parameters, no discernible differences were detected. No other apparent side effects were observed. Therefore, this method could be useful for treatment and/or prevention of thrombocytopenia induced by chemotherapy and/or irradiation for cancer treatment.

Analysis of Donor Factors for Non-Heart-Beating Donors With Regard to Cadaveric Kidney Transplantation in the Western Region of Japan.

BACKGROUND: Kidneys from non-heart-beating donors are thought to be marginal, and careful evaluation is required. Mass analyzed data are limited, and each transplant surgeon must evaluate these organs on the basis of their own experience. METHODS: We analyzed the data of 589 kidneys used for kidney transplantation from 304 non-heart-beating donors from January 2002 through December 2013 at the Japan Organ Transplant Network West Japan Division. The age of the donors, cause of death, and total ischemic time of more than 24 hours were factors that influenced the graft survival of the organs. RESULTS: On the other hand, the final serum creatinine level before donation (maximum, 12.4 mg/dL), the presence and duration of anuria (maximum, 92 hours), and the presence of cannulation did not influence the graft survival rate. CONCLUSIONS: In multivariate analysis of Cox proportional hazards regression analysis, graft survival was significantly related to the age of the donor (over 70 years of age), cause of death (atherosclerotic disease), and total ischemic time of more than 24 hours.

EXAFS study of U(VI) uptake by calcium silicate hydrates.

Among the different cement minerals, calcium silicate hydrates (C-S-H) are the prime candidates for heavy metal binding because of their abundance and appropriate structure. Immobilization processes of heavy metals by cementitious materials, and in particular C-S-H phases, thus play an important role in multibarrier concepts developed worldwide for the safe disposal of hazardous and radioactive wastes. In this study, the uptake of U(VI) by C-S-H has been investigated using X-ray absorption fine structure (XAFS) spectroscopy. C-S-H phases were synthesized using two different procedures: One is based on the mixing of CaO and SiO2 solids ("direct reaction" method); for the other one starting solutions of Ca and Si are used ("solution reaction" method). XAFS investigations were carried out on samples doped with U(VI). U(VI) was either sorbed onto previously precipitated C-S-H phases (sorption samples) or added during C-S-H synthesis (coprecipitation samples). The coordination environment of U(VI) in the sorption samples was found to be independent of the procedure used for C-S-H synthesis. A split equatorial oxygen shell (Oeq1: R=2.23-2.27 A; Oeq2: R=2.36-2.45 A), neighboring silicon atoms at short (R=3.07-3.11 A) and long (R=3.71-3.77 A) distances, and neighboring Ca atoms (R=3.77-3.81 and 4.15-4.29 A) were observed for all the samples. The structural parameters resemble those reported for uranophane. The coordination environment of U(VI) in the coprecipitation samples depends on the method used for C-S-H synthesis, and further, the spectra differ from those determined for the sorption samples. UU backscattering contributions were observed in the samples prepared using the direct reaction method, whereas no split equatorial shell appeared in the samples prepared using the solution reaction method.

Cyclic hydroxamic-acid-containing peptide 31, a potent synthetic histone deacetylase inhibitor with antitumor activity.

Cyclic hydroxamic-acid-containing peptide 1 (CHAP1), designed as a hybrid of trichostatin A and trapoxin, is a lead compound for the development of potent inhibitors of histone deacetylase (HDAC). In this study, we synthesized a series of CHAP derivatives and evaluated their biological activities by monitoring the potency of their inhibition of HDAC activity, their ability to augment the expression of MHC class-I molecules in B16/BL6 cells, and their effect on cell proliferation. A structure-activity relationship study using these three assay systems revealed several requirements of their structure for the strong inhibition of HDAC not only in the cell-free situation, but also in cells. When the structures of CHAP derivatives are represented as cyclo(-Asu(NHOH)-AA(2)-AA(3)-Pro or Pip-)(n), where Asu(NHOH) and Pip are zeta-hydroxamide-alpha-aminosuberic acid and pipecolic acid, respectively, (a) the tetrapeptide structure (n = 1) was better than the octapeptide one (n = 2); (b) AA(2) and AA(3) should be hydrophobic; and (c) the combination of amino acid chirality should be LDLD for the strongest inhibition of HDAC in cells (LDLD > LLLD, LDLL > LLDL). cyclo(-L-Asu(NHOH)-D-Tyr(Me)-L-Ile-D-Pro-) or CHAP31 was selected as one of the strongest CHAPs, and its biological activity was characterized further. CHAP31 was much more stable in the presence of cultured cells (t(1/2) > 3000 h) than trichostatin A (t(1/2) = 14.7 h) or trapoxin A (t(1/2) = 2.10 h). CHAP31 exhibited antitumor activity in C57BL x DBA/2 F(1) (BD2F(1)) mice bearing B16/BL6 tumor cells. Furthermore, CHAP31 inhibited the growth in four of five human tumor lines implanted into nude mice. These results suggest CHAP31 to be promising as a novel therapeutic agent for cancer treatment.

Malolactomycin D, a potent inhibitor of transcription controlled by the Ras responsive element, inhibits Ras-mediated transformation activity with suppression of MMP-1 and MMP-9 in NIH3T3 cells.

To search for anti-cancer agents, a screening system for Ras signal inhibitors was developed using a NIH3T3 cell line with an introduced reporter gene which is controlled by the Ras-responsive element (RRE). With this screening system, malolactomycin D was identified as a selective inhibitor of transcription from the RRE. This compound was found to preferentially inhibit the anchorage-independent growth rather than the anchorage-dependent growth of Ras-transformed NIH3T3 cells. The expression of matrix metalloproteinases MMP-1 and MMP-9, which have RRE in their promoters, were reduced by treatment with malolactomycin D at the translational and transcriptional levels. Analysis of the activity of mitogen-activated protein (MAP) kinases, which play important roles in transduction of the Ras signal, showed that malolactomycin D inhibits the activation of p38 MAP kinase and Jun N-terminal-kinase (JNK) but not extracellular signal-regulated kinase 1 or 2 (ERK1 or 2). These findings suggest that by inhibiting the pathway that leads to the activation of p38 MAP kinase and JNK, malolactomycin D suppresses the expression of MMPs. Since MMPs play important roles in metastasis and maintenance of the microenvironment of tumor cells, both of which facilitate tumor growth, the inhibition of MMPs by malolactomycin D is believed to contribute to its ability to inhibit Ras-mediated tumorigenesis.

Effective prevention of thrombocytopenia using adenovirus-mediated transfer of HST-11FGF-4 gene: in vivo and in vitro studies.

A novel biological function of HST-1 protein (FGF-4) was investigated by constructing an adenovirus vector containing the HST-1 cDNA and applied for thrombocytopenia as a gene therapy. A single intraperitoneal injection of the replication-deficient adenovirus containing the HST-1 gene (Adex1HST-1) into mice caused a two-fold increase in peripheral platelet count for 30 days, and effectively prevented experimentally induced thrombocytopenia. Studies of Adex1HST-1-infected or HST-1 protein-treated megakaryocytic Dami cells suggested that HST-1 protein promotes megakaryocyte maturation, and increases cytokine secretion from megakaryocyte and adhesive interactions between megakaryocyte and endothelial cells. Colony assay revealed that HST-1 protein stimulated CFU-MK (colony-forming unit of megakaryocyte) not alone but synergistically with early-acting cytokines such as IL-3 or Tpo (c-mpl ligand) as a megakaryocyte potentiating factor. These results have important implications for clinical application of the Adex1HST-1 for thrombocytopenia.

Superoxide production from paraquat evoked by exogenous NADPH in pulmonary endothelial cells.

Superoxide production from paraquat in a pulmonary microvascular endothelial cell (PMEC) suspension was demonstrated using 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-alpha]pyraz in-3-one (MCLA), a chemiluminescence probe, to detect superoxide anions. Increased rates of superoxide production from paraquat, which were sensitive to superoxide dismutase (SOD), required the presence of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in the reaction medium, and occurred instantaneously after the addition of NADPH, which is impermeable to cell membranes. NADH as an electron donor was not as effective, and xanthine or succinate had no influence. Paraquat was anaerobically reduced in the presence of NADPH and PMECs to yield a one-electron reduced radical, and the reduction was inhibited by NADP+. Diphenyleneiodonium, an inhibitor of flavoprotein reductases, also markedly inhibited both paraquat reduction and superoxide production. These results indicate that NADPH-dependent superoxide production from paraquat probably occurs by a flavoprotein with NADPH-dependent reductase activity in cell membranes. NADPH-dependent superoxide production from paraquat was also reproduced using adherent PMECs on wells. Under these conditions, superoxide production was enhanced with agonists, including interleukin-1beta, A23187, and phorbol 12-myristate 13-acetate. The effect of the former two was blocked with staurosporine, while the latter's effect was suppressed with calyculin A.

The antioxidant action of 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-alpha]pyra z in-3-one (MCLA), a chemiluminescence probe to detect superoxide anions.

The antioxidant effect of 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-alpha]pyraz in-3-one (MCLA), a Cypridina luciferin analog that acts as a chemiluminescence probe to detect O2.-, was investigated. MCLA produced a lag in oxygen consumption induced by cumene hydroperoxide in microsomes or by 2,2'-azobis (2-amidinopropane) dihydrochloride in liposomes and disappeared during the duration of the lag. MCLA profoundly inhibited the propagation reaction in Fe2+-dependent lipid peroxidation in liposomes, and MCLA disappearance accompanied by suppression of oxygen consumption markedly occurred in liposomes susceptible to peroxidation. Thiobarbituric acid-reactive substances in all systems used were also suppressed by MCLA dose dependently. These results indicate that MCLA has an antioxidant property through scavenging free radicals.

High prevalence of anti-cardiolipin antibody, C1q-, C3d-, and mRF-IgG immune complexes, and anti-nuclear antibody in hemophiliacs irrespective of infection with human immunodeficiency virus type 1.

We investigated the prevalence of various autoantibodies [anti-cardiolipin antibody (aCL), lupus anticoagulant (LA), immune complexes (ICs), anti-nuclear antibody (ANA), and anti-deoxyribonucleic acid antibody (aDNA)] in hemophiliac individuals with (n = 50) and without (n = 42) infection by human immunodeficiency virus type 1 (HIV-1). The positivity rate for ANA was similar in both groups, and none of the patients was positive for LA and aDNA. aCL was positive in 35 of 50 (70%) HIV-1-positive hemophiliac individuals and 33 of 42 (79%) HIV-1-negative hemophiliac individuals. However, the majority of the aCL was revealed to be beta 2-glycoprotein I independent, thus corresponding to a syphilis type aCL that does not cause the so-called antiphospholipid syndrome. A total of 39 of the 45 HIV-1 positive hemophiliac individuals (87%) and 34 of 41 HIV-1-negative hemophiliac individuals (83%) had at least one type of IC [C1q-, C3d-, and/or murine monoclonal rheumatoid factor (mRF)- IgG]. The mechanism producing various autoantibodies in hemophiliac persons irrespective of their HIV-1 status is still unclear, but pathogens (e.g., HIV-1, hepatitis B, and hepatitis C) and alloantigens in the blood products that these patients require may be possible candidates. The clinical significance of the presence of these autoantibodies and the underlying mechanisms involved both need to be clarified further.

Cytochalasin-E-Resistant Mutants of Coprinus cinereus: Isolation and Genetic, Biochemical, and Cytological Analyses

We first obtained a cytochalasin-E supersensitive strain (CES14) of the basidiomycete Coprinus cinereus and then isolated 1000 revertants of CES14, expecting that at least some of the revertants have defects in the functions of actin filaments. Microscopic examination revealed that three of the revertants lack septa in their hyphae. Of the 1000 revertants, 18 including the 3 septumless strains were genetically analyzed. In 11 of the 18 revertants, reversion was due to extragenic suppressor mutations of ces14. Ten of the 11 mutations were in the same locus designated cer1, while the locus of the remaining mutation, which inhibits septum formation, could not be determined because of its failure to mate with any other cer1 strains. In one (CES14R42) of the cer1 mutants, actin appeared to be altered in the affinity to actin antibody in Western blotting after isoelectric focusing, suggesting that cer1 is a gene encoding actin. Phenotypic examination revealed that the 10 cer1 mutations all confer resistance to cytochalasin-E and that two of the cer1 mutations, both of which block septation, inhibit the formation of the actin ring.

The adhesion molecules, l-selectin and sialyl lewis x, relate to the formation of the follicular dendritic cell-lymphocyte cluster in the mantle zone.

The follicular dendritic cell (FDC)-lymphocyte cluster is rich in the follicular light zone of the secondary lymphoid follicles (LFs). Although, the mantle zone (MZ) also has FDC-lymphocyte cluster, it has not known about what kind of adhesion molecules relates to cluster formation. In the present study, we investigated whether the adhesion molecules, L-selectin (CD62L) and sialyl Lewis x (CD15s) can mediate the formation of the cluster in human tonsillar LFs. The MZ only expressed both the adhesion molecules in the secondary LF. Isolated FDC-lymphocyte clusters were composed of CD62L(+) lymphocytes and CD15s(+) FDCs. Stamper-Woodruff binding assay revealed that the binding of IgD(+) lymphocytes was significantly inhibited by pretreatment with anti-CD62L antibody or with anti-CD15s antibody. These results indicate that CD62L on MZ lymphocytes and CD15s on FDCs may play a role of the cluster formation, unlike the clusters in the other parts of LFs.

Localization of blood coagulation factors in the germinal centers of human Peyer's patches.

The immunohistochemical distribution of 15 blood coagulation factors in the germinal centers (GCs) of human Peyer's patches (PPs) was studied. Although factor VIII, active alpha-thrombin, and fibrinogen were hardly evident in the GCs, the majority of coagulation factors, such as kallikrein, high-molecular-weight kininogen, factors XII, X, IX, VII, V, XIIIa and XIIIb, prothrombin, anti-thrombin III and inactive alpha-thrombin were found, showing a lace-like staining pattern similar to that obtained with a monoclonal antibody, R4/23, specific for follicular dendritic cells (FDCs) in the GCs. By immunoelectron microscopy, positive reactions for factor X and XIIIa were found on the surfaces of FDCs, GC cells, and/or in the intercellular spaces of GCs, being especially marked on the surface of the labyrinth-like structure of FDCs. It is concluded that a majority of coagulation factors are localized in the GCs of human PPs. Furthermore, it is suggested that some of these coagulation factors have a close topographical relationship with FDCs.

An evaluation of segmental gastrectomy for gastric ulcer: one to ten year follow-up.

UNLABELLED: Of 61 patients with gastric ulcer subjected to an improved technique of segmental gastrectomy, 26 were available for follow-up study for 1 to 10 years; evaluations were made mainly on the postoperative reduction of gastric acid secretion and motor function of the gastric remnant. RESULTS: (1) x-ray observation revealed the gastric remnant to be larger than usually seen following Billroth I or II gastrectomy, with no dilatation or remarkable deformity; sphincteric function of the pylorus and peristaltic activity of the gastric remnant were satisfactory; (2) gastric emptying was usually complete in 60 to 180 minutes in 73 percent of the patients, similar to control patients; (3) no instance of dumping syndrome was recognized; (4) average rates of postoperative acid reduction were 58.3 percent for maximal acid concentration, 67.1 percent for maximal acid output, and 67.6 percent for peak acid output, indicating the reduction of gastric acid secretion to be fairly satisfactory. In none of the patients available for follow-up was identified a recurrence of ulcer. These follow-up results suggest the use of segmental gastrectomy for gastric ulcer and for other benign lesions in the mid portion of the stomach.

Left ventricular pseudoaneurysm and intracardiac fistulas after replacement of mitral valve prosthesis.

Operation was performed on a 61-year-old woman with left ventricular pseudoaneurysm, left ventricular-right atrial fistula, and left ventricular-coronary sinus fistula after mitral valve replacement. The diagnostic and therapeutic approaches to these complications are described briefly, and the literature on intracardiac fistula after mitral valve replacement is reviewed.