Partial anomalous pulmonary venous connection in a dog

A 2-year-old male intact Belgian Malinois was presented for exercise intolerance. A grade III/VI left basilar systolic murmur was detected. Echocardiography revealed moderate right atrial and ventricular dilation and increased pulmonic outflow velocity. Thoracic radiographs showed right heart enlargement and a dilated caudal vena cava. In addition, on the left lateral projection, an enlarged aberrant right cranial pulmonary lobar vein was suspected to be diverging ventrally from the course of the right cranial lobar bronchus and inserting more ventrally than normal in the region of the right atrium. A left-to-right pulmonary vascular shunt was suspected, and the patient underwent further diagnostics under general anesthesia. An agitated saline study was positive, suggestive of a concurrent right to left shunt. A right heart catheterization was performed. Angiography was inconclusive. Oximetry testing revealed an increase in oxygen saturation within the right atrium at the level of the caudal cava supportive of a left-to-right shunt in this region. Computed tomography angiography revealed a large single pulmonary vein that anomalously entered into the caudolateral aspect of the right atrium (left-to-right shunt) and was suspicious for a small arteriovenous malformation between the right caudal pulmonary artery and the right pulmonary vein returning to the left atrium (right to left shunt). The patient was diagnosed with a partial anomalous pulmonary venous connection and a possible arteriovenous malformation.     

Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes

During fertilization in mammalian species, a sperm-induced intracellular Ca²⁺ signal ([Ca²⁺]_i) mediates both exit of meiosis and oocyte activation. Recently, we demonstrated in mouse oocytes that the phosphorylation levels of inositol 1,4,5 trisphosphate receptor type1 (IP₃R1), the channel responsible for Ca²⁺ release and oscillations during fertilization, changed during maturation and fertilization. Therefore, we examined the expression and phosphorylation of IP₃R1 during in vitro maturation of pig oocytes. Here, our present study shows that expression of IP₃R1 protein did not change during maturation, although the phosphorylation status of the receptor, specifically at an MPM-2 epitope, did. We found that while at the beginning of maturation IP₃R1 lacked MPM-2 immunoreactivity, it became MPM-2 reactive by 24 h and reached maximal reactivity by 36 h. Interestingly, the acquisition of MPM-2 reactivity coincided with the activation of p34^cdc2 kinase and mitogen-activated protein kinase (MAPK), which are involved in meiotic progression. Following completion of maturation, inactivation of MAPK by U0126 did not affect IP₃R1 phosphorylation, although inactivation of p34^cdc2 kinase by roscovitine dramatically reduced IP₃R1 phosphorylation. Neither inhibitor affected total expression of IP₃R1. Altogether, our results show that IP₃R1 undergoes dynamic phosphorylation during maturation and this might underlie the generation of oscillations at fertilization.     

Calibrating the liana crown occupancy index in Amazonian forests

Lianas hold an important, but understudied, role in forest dynamics, however they are difficult to measure and detailed liana measurements are time consuming. Many researchers have therefore used an ordinal scale index, the crown occupancy index (COI), to describe the liana load carried by trees. Here we assess the overall effectiveness, in terms of accuracy, precision, repeatability and efficiency, of the COI in tropical forests. We relate the COI to more detailed liana measurements at the individual tree-level and site-level, comparing sites with different levels of liana infestation. Our results show (1) that the COI accurately measures individual tree and plot level liana loads, indicated by the strong correlations between the COI and the number and basal area of lianas. However, (2) as expected, the COI is only weakly related to the basal area of lianas rooted close to the tree, which is a proxy for competition for below-ground resources. The COI is also (3) an efficient measure of liana loads, as the input time needed for a COI survey is considerably less than that of a detailed liana survey. We also (4) found a high degree of repeatability in COI classification between observers. Additionally (5), the COI can be used to differentiate between sites in terms of their overall liana canopy competition (precision), but (6) may not be a precise indicator of the site-level mean basal area of lianas in tree crowns.     

Cytotoxicity of benzyl isothiocyanate in normal renal proximal tubular cells and its modulation by glutathione

In the present study, we examined the toxicity of benzyl ITC (BITC) and its urinary mercapturic acid metabolite (BITC-NAC), using a normal renal proximal tubular cell line, pig LLC-PK1. BITC increased cell death with an IC(50) value of about 7 μM, whereas the cytotoxic effect of BITC-NAC was five times weaker than that of BITC. We observed a significant necrosis of the compounds on LLC-PK1 cells with oxidative stress. In the presence of 5 mM glutathione (GSH), comparable to physiological levels, the cytotoxicity of BITC-NAC as well as BITC was significantly reduced. Furthermore, the increase in intracellular GSH levels by pretreatment with NAC before the BITC treatment resulted in inhibition of the BITC-induced necrotic events as well as intracellular oxidative stress. These results suggest that GSH is a determinant of cellular resistance against the BITC-mediated and oxidative stress-dependent cytotoxicity in renal proximal tubular cells.     

High developmental rates of mouse oocytes cryopreserved by an optimized vitrification protocol: the effects of cryoprotectants, calcium and cumulus cells

Unfertilized oocytes are one of the most desired germ cell stages for cryopreservation because these cryopreserved oocytes can be used for assisted reproductive technologies, including in vitro fertilization (IVF) and intracytoplasmic sperm injection. However, in general, the fertility and developmental ability of cryopreserved oocytes are still low. The aim of the present study was to improve vitrification of mouse oocytes. First, the effects of calcium and cryoprotectants, dimethyl sulfoxide and ethylene glycol (EG), in vitrification medium on survival and developmental ability of vitrified oocytes were evaluated. Oocytes were vitrified by a minimal volume cooling procedure using different cryoprotectants. Most of the vitrified oocytes were morphologically normal after warming, but their fertility and development were low independently of calcium and cryoprotectants. Second, the effect of cumulus cells on ability of oocytes to be fertilized and develop in vitro was examined. The fertility and developmental ability of denuded oocytes (DOs) after IVF were reduced compared with cumulus-oocyte complexes (COCs) both in fresh and cryopreserved groups. Vitrified COCs showed significantly (P<0.05) higher fertility and ability to develop to the 2-cell and blastocyst stages than those of vitrified DOs with cumulus cells and vitrified DOs alone. The vitrified COCs developed to term at a high success rate equivalent to the rate obtained with IVF using fresh COCs. Taken together, the current results clearly demonstrate that, in the presence of surrounding cumulus cells, matured mouse oocytes vitrified using calcium-free media and EG retain their developmental competence. These findings will contribute to improve oocyte vitrification in not only experimental animals but also clinical application for human infertility.     

Vaccine development for the prevention of staphylococcal mastitis in dairy cows

Staphylococci are the most common and costly mammary disease of dairy cattle worldwide. Target of RNAIII Activating Protein (TRAP), a membrane associated 167AA protein, is highly conserved among staphylococci and was shown in Staphylococcus aureus to be involved in bacterial stress response. The aims of this study were to test the safety and efficacy of recombinant TRAP (rTRAP) vaccine in dairy animals. The vaccine was safe as 2-3 subcutaneous injections of rTRAP (54-100 μg) with adjuvant ISA 206 to cows and goats did not lead to any abnormal symptoms of sensitivity to the vaccine. The rTRAP vaccine was immunogenic and caused the induction of a humoral immune response that remained high for at least 160 d post second immunization. The rTRAP vaccine was efficacious; at parturition only 13.5% (5/37) heifers in the immunized group were infected with Staphylococcus chromogenes as compared to 42.9% (18/42) in the non-immunized group. Additionally, when cows were immunized in mid-lactation, the difference between somatic cell count (SCC) in immunized and control animals was profound (45 ± 7 vs. 470 ± 194, respectively). At the same time, the difference in milk yield was also evident (48.3 ± 1.4 L d(-1)vs. 44.3 ± 0.9 L d(-1), respectively). Put together, these studies indicate the value of the rTRAP vaccine in preventing new udder infections by staphylococci, which significantly lead to lowered SCC and some increase in milk yield. TRAP is conserved among all strains and species and is constitutively expressed in any strain of S. aureus or CNS tested so far, including those isolated from cows. TRAP may thus serve as a universal anti-staphylococcus vaccine.     

Triterpenoids isolated from Zizyphus jujuba inhibit foam cell formation in macrophages

Because foam cell formation in macrophages is believed to play an essential role in the progression of early atherosclerotic lesions in vivo, prevention of foam cell formation is considered to be one of the major targets for the treatment of atherosclerosis. The present study examined the inhibitory effect of 50 crude plant extracts on foam cell formation. Among those crude extracts, Zizyphi Fructus (ZF) and Zizyphi Semen (ZS) extracts significantly inhibited the foam cell formation induced by acetylated LDL. Furthermore, triterpenoids such as oleanonic acid, pomolic acid, and pomonic acid were the major active compounds, and triterpenoids containing a carboxylic acid at C-28 play an important role in the inhibitory effect on foam cell formation in human macrophages. These data suggest that triterpenoids in Zizyphus jujuba , the plant source of ZF and ZS, may therefore be useful for the prevention of atherosclerosis.     

Nitrogen Uptake by Radish,Spinach and “Chingensai” from Composted Tea Leaves,Coffee Waste and Kitchen Garbage

A pot experiment was conducted to determine the effects of the application of composted tea leaves (TC), coffee waste (CC), and kitchen garbage (KC) on the nitrogen and nitrate accumulated in radish (Raphanus sativus L. cv. ‘radicula pers’), Chingensai (Brassica campestris L. cv. ‘Choyo No. 2’), and spinach (Spinacia oleracea L. cv. ‘Ban chu paruku’) as compared with the effect of inorganic ¹⁵N labeled fertilizer (IN) application. The compost was applied at the rate of 24 g kg^?1 soil, corresponding to about 250 to 300 kg N ha^?1; the A value method was used to estimate nitrogen uptake. Dry matter production was significantly higher in the IN and TC treatments than in the KC and CC treatments for all the species and tissue. Of the composts used, TC was most effective in increasing N uptake and N content in the vegetables. The composts derived N recovery as a percentage of total N uptake varied with plant species, 50.8%-62.9% in radish root, 35.3%-60.4% in radish leaf, 29.9%-48.2% in spinach leaf, and 31.3%-54.8% in Chingensai leaf. The N-use efficiencies of IN, TC, CC, and KC were 6.3%, 6.3%, 5.3%, and 6.6% in radish root; 13.6%, 9.7%, 8.4%, and 6.7% in radish leaf; 22.4%, 14.4%, 3.6%, and 5.8% in spinach leaf; and 61.2%, 39.5%, 25.5%, and 21.5% in Chingensai leaf, respectively. Nitrate accumulation in edible portions was highest in plants provided with IN as compared with those grown with composts, and nitrate content in radish root was markedly higher than that in the leaf. It is observed that the fate of compost derived N differed noticeably with vegetable species, plant part, and compost source.     

Measurement of Total Dietary Fiber Using AOAC Method 2009.01 (AACC International Approved Method 32‐45.01): Evaluation and Updates

The Codex Committee on Methods of Analysis and Sampling recently recommended 14 methods for measurement of dietary fiber, eight of these being type I methods. Of these type I methods, AACC International Approved Method 32‐45.01 (AOAC method 2009.01) is the only procedure that measures all of the dietary fiber components as defined by Codex Alimentarius. Other methods such as the Prosky method (AACCI Approved Method 32‐05.01) give similar analytical data for the high‐molecular‐weight dietary fiber contents of food and vegetable products low in resistant starch. In the current work, AACCI Approved Method 32‐45.01 has been modified to allow accurate measurement of samples high in particular fructooligosaccharides: for example, fructotriose, which, in the HPLC system used, chromatographs at the same point as disaccharides, meaning that it is currently not included in the measurement. Incubation of the resistant oligosaccharides fraction with sucrase/β‐galactosidase removes disaccharides that interfere with the quantitation of this fraction. The dietary fiber value for resistant starch type 4 (RS₄), varies significantly with different analytical methods, with much lower values being obtained with AACCI Approved Method 32‐45.01 than with 32‐05.01. This difference results from the greater susceptibility of RS₄ to hydrolysis by pancreatic α‐amylase than by bacterial α‐amylase, and also a greater susceptibility to hydrolysis at lower temperatures. On hydrolysis of samples high in starch in the assay format of AACCI Approved Method 32‐45.01 (AOAC method 2009.01), resistant maltodextrins are produced. The major component is a heptasaccharide that is highly resistant to hydrolysis by most of the starch‐degrading enzymes studied. However, it is hydrolyzed by the maltase/ amyloglucosidase/isomaltase enzyme complex present in the brush border lining of the small intestine. As a consequence, AOAC methods 2009.01 and 2011.25 (AACCI Approved Methods 32‐45.01 and 32‐50.01, respectively) must be updated to include an additional incubation with amyloglucosidase to remove these oligosaccharides.     

Heartwood extractives from the Amazonian trees <Emphasis Type="Italic">Dipteryx odorata,Hymenaea courbaril</Emphasis>, and <Emphasis Type="Italic">Astronium lecointei</Emphasis> and their antioxidant activities

Heartwood extracts from Amazonian trees cumaru-ferro (Dipteryx odorata), jatoba (Hymenaea courbaril), and guarita (Astronium lecointei) exhibit antioxidant activities comparable with that of α-tocopherol, a well-known antioxidant. This article reports the characterization of the antioxidant compounds in the extracts of the three heartwoods. Silica gel column chromatography of the cumaru-ferro EtOAc extract yielded (−)-(3R)-7,2′,3′-trihydroxy-4′-methoxyisoflavan and (+)-(3R)-8,2′,3′-trihydroxy-7,4′-dimethoxyisoflavan. Silica gel column chromatography followed by preparative high-performance liquid chromatography of the jatoba EtOAc extract yielded (−)-fisetinidol and (+)-trans-taxifolin. Chemical structures were assigned using electron-ionization mass spectrometry, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy including nuclear Overhauser effect spectroscopy (NOESY), as well as optical rotation and circular dichroism. Gas chromatography-mass spectrometry demonstrated that the isolated compounds were predominant in the EtOAc extracts. In the guarita EtOAc extract, catechin and gallic acid were identified by comparing their retention times and mass fragmentation patterns with those of authentic samples. Antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl assay demonstrated that all these compounds had activities comparable with that of α-tocopherol. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007