Bovine mammary gland UDP-GalNAc:GlcNAcbeta-R beta1-->4-N- acetylgalactosaminyltransferase is glycoprotein hormone nonspecific and shows interaction with alpha-lactalbumin

We have identified a novel N -acetylgalactosaminyltransferase activity in lactating bovine mammary gland membranes. Acceptor specificity studies and analysis of products obtained in vitro by 400 MHz1H-NMR spectroscopy revealed that the enzyme catalyses the transfer of N - acetylgalactosamine (GalNAc) from UDP-GalNAc to acceptor substrates carrying a terminal, beta-linked N -acetylglucosamine (GlcNAc) residue and establishes a beta1-->4-linkage forming a GalNAcbeta1-->4GlcNAc ( N, N '-diacetyllactosediamine, lacdiNAc) unit. Therefore, the enzyme can be identified as a UDP-GalNAc:GlcNAcbeta-R beta1-->4-N- acetylgalactosaminyltransferase (beta4-GalNAcT). This enzyme resembles invertebrate beta4-GalNAcT as well as mammalian beta4- galactosyltransferase (beta4-GalT) in acceptor specificity. It can, however, be clearly distinguished from the pituitary hormone-specific beta4-GalNAcT by its incapability of acting with an elevated activity on a glycoprotein substrate carrying a hormone-specific peptide motif. Furthermore, the GalNAcT activity appeared not to be due to a promiscuous action of a beta4-GalT as could be demonstrated by comparing the beta4-GalNAcT and beta4-GalT activities of the mammary gland, bovine colostrum, and purified beta4-GalT, by competition studies with UDP-GalNAc and UDP-Gal, and by use of an anti-beta4-GalT polyclonal inhibiting antibody. Interestingly, under conditions where mammalian beta4-GalT forms with alpha-lactalbumin (alpha-LA) the lactose synthase complex, the mammary gland beta4-GalNAcT was similarly induced by alpha-LA to act on Glc with an increased efficiency yielding the lactose analog GalNAcbeta1-->4Glc. This enzyme thus forms the second example of a mammalian glycosyltransferase the specificity of which can be modified by this milk protein. It is proposed that the mammary gland beta4-GalNAcT functions in the synthesis of lacdiNAc- based, complex-type glycans frequently occurring on bovine milk glycoproteins. The action of this enzyme is to be considered when aiming at the production of properly glycosylated protein biopharmaceuticals in the milk of transgenic dairy animals.      

Aquatic diversity in a former floodplain: Remediation in an urban context

The Lobau, a former floodplain area of the Danube River situated within the city limits of Vienna (Austria), was strongly affected by the river regulation in 1875. The reduced hydrological connectivity led to an increasing loss of aquatic habitats. A water enhancement scheme with a maximum water input of 0.5 m³ s^?1 was initiated in 2001. The present study assesses the effect of this scheme on biodiversity using three target species groups – aquatic molluscs, dragonflies and fish – following a common Before–After Control-Impact design (BACI). Dragonflies and molluscs were positively affected, reflecting the habitat alterations in the system. For fish, no significant impact was observed. The aim of the scheme has been achieved: increased habitat diversity and improved habitat conditions for the system's initial community and further rheophilic/rheotolerant species. Water enhancement schemes can be effective remediation measures and deserve further attention in the management of urban wetlands.     

High dispersal in a frog species suggests that it is vulnerable to habitat fragmentation

Global losses of amphibian populations are a major conservation concern and their causes have generated substantial debate. Habitat fragmentation is considered one important cause of amphibian decline. However, if fragmentation is to be invoked as a mechanism of amphibian decline, it must first be established that dispersal is prevalent among contiguous amphibian populations using formal movement estimators. In contrast, if dispersal is naturally low in amphibians, fragmentation can be disregarded as a cause of amphibian declines and conservation efforts can be focused elsewhere. We examined dispersal rates in Columbia spotted frogs (Rana luteiventris) using capture-recapture analysis of over 10,000 frogs in combination with genetic analysis of microsatellite loci in replicate basins. We found that frogs had exceptionally high juvenile dispersal rates (up to 62% annually) over long distances (>5km), large elevation gains (>750m) and steep inclines (36 degrees incline over 2km) that were corroborated by genetic data showing high gene flow. These findings show that dispersal is an important life-history feature of some amphibians and suggest that habitat fragmentation is a serious threat to amphibian persistence.     

Transgenic smooth muscle expression of the human CysLT1 receptor induces enhanced responsiveness of murine airways to leukotriene D4

Cysteinyl leukotrienes (CysLTs) exert potent proinflammatory actions and contribute to many of the symptoms of asthma. Using a model of allergic sensitization and airway challenge with Aspergillus fumigatus (Af), we have found that Th2-type inflammation and airway hyperresponsiveness (AHR) to methacholine (MCh) were associated with increased LTD(4) responsiveness in mice. To explore the importance of increased CysLT signaling in airway smooth muscle function, we generated transgenic mice that overexpress the human CysLT1 receptor (hCysLT(1)R) via the alpha-actin promoter. These receptors were expressed abundantly and induced intracellular calcium mobilization in airway smooth muscle cells from transgenic mice. Force generation in tracheal ring preparations ex vivo and airway reactivity in vivo in response to LTD(4) were greatly amplified in hCysLT(1)R-overexpressing mice, indicating that the enhanced signaling induces coordinated functional changes of the intact airway smooth muscle. The increase of AHR imposed by overexpression of the hCysLT(1)R was greater in transgenic BALB/c mice than in transgenic B6 x SJL mice. In addition, sensitization- and challenge-induced increases in airway responsiveness were significantly greater in transgenic mice than that of nontransgenic mice compared with their respective nonsensitized controls. The amplified AHR in sensitized transgenic mice was not due to an enhanced airway inflammation and was not associated with similar enhancement in MCh responsiveness. These results indicate that a selective hCysLT(1)R-induced contractile mechanism synergizes with allergic AHR. We speculate that hCysLT(1)R signaling contributes to a hypercontractile state of the airway smooth muscle.     

Greening of intermittent-light-grown bean plants in continuous light: thylakoid components in relation to photosynthetic performance and capacity for photoprotection

Phaseolus vulgaris (cvv. Windsor longpod and snap bean) plants, etiolated during germination, were exposed to intermittent light (2 min light every 2 hr) for up to 68 hr and then transferred to continuous white light. On transfer of the plants to continuous light (100 photons mumol m-2 s-1, 24 degrees C), the quantum yield of oxygen evolution increased two-fold in about 30 hr. The chlorophyll content per unit leaf area or unit fresh weight increased dramatically, but the fresh weight per unit leaf area was relatively constant. The changes were expressed on the basis of fresh weight or leaf area. On this basis, the contents of photosystem (PS) I and II increased in continuous light, by a factor of 3 and 8, respectively. While the chlorophyll b content and the contents of apoproteins of light-harvesting chlorophyll-protein complexes (LHCIIb, CP29, CP26 and CP24) increased markedly, neither the total carotenoid content nor the de-epoxidation state of the xanthophylls [ratio of zeaxanthin(Z) + antheraxanthin(A) to (Z + A + violaxanthin) was about 0.4)] responded significantly on transfer to continuous light. The fast rise of the flash-induced electrochromic signal (delta A518) was well correlated with the increases in PS I and PS II reaction centres, and with chlorophyll b and total carotenoid contents. The increase in the quantum yield of oxygen evolution during greening in continuous light is attributed to a more balanced distribution of excitation energy between the two photosystems, facilitated by the increased number of PS II units, the increased antenna size of each unit and the enhancement of grana formation. The chloroplast in intermittent light was found to contain abundant xanthophyll cycle pigments and the psbS gene product, presumably adequate for photoprotection in continuous light as soon as chlorophyll a/b- protein complexes are synthesized. The results suggest that greening in continuous light is accompanied by adjustments that include enhanced quantum efficiency of photosynthesis and development of a capacity for harmless dissipation of excess excitation energy.     

The early conception factor (ECF) lateral flow assay for non-pregnancy determination in the mare

The horse early conception factor (ECF) test is designed for qualitative determination of the ECF glycoprotein in the mare that has conceived. The objectives of this study were to determine the performance of the horse ECF test for the detection of the non-pregnant mare, and to determine the agreement among subjects or "readers" regarding the interpretation of the test. Blood samples from 60 mares were collected on Days 0, 5, 8, 11 and 18 following ovulation. Pregnancy status diagnosed with the ECF test was compared (2 x 2 table) to pregnancy status diagnosed by palpation per rectum and ultrasound examination on Day 18 following ovulation. Three readers interpreted the ECF test outcome independently. Two laboratories independently interpreted the ECF test outcome from the same serum samples. Agreement was tested by kappa coefficient. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy ranged from 0.74 to 0.84, 0.14 to 0.33, 0.62 to 0.66, 0.33 to 0.44 and 0.57 to 0.60, respectively. Agreement between readers was substantial (0.60相似文献   

Contrasting B cell- and T cell-based protective vaccines

A substantial research effort is devoted to the development of vaccines based on T cells. Such a vaccine would provide a means to protect against infection with HIV and stop the current pandemic. Here we investigate the possibility to develop a protective T cell-based vaccine. We do this by means of a mathematical model which describes the dynamics of a pathogen and the immune system in the early stages of infection. We compare an immune response that is near immediate--as is the case for a humoral response--with that of a response in which the effector cells have to be formed from precursor cells--as occurs in T cell responses. The latter applies to a T cell-based vaccine. A near immediate response is associated with a threshold number of effector cells above which an infection cannot take hold. For a T cell-based vaccine this threshold increases with the amount of antigen the immune system is exposed to. For small initial doses, as one would naturally expect to occur, this gives rise to impractically large thresholds. Thus, although a T cell vaccine might work against a high dose exposure, it might fail when exposed against to a low-dose exposure. This limits, we argue, the efficacy of T cell-based vaccines.     

Discovering new insect viruses: whitefly iridovirus (Homoptera: Aleyrodidae: Bemisia tabaci)

Adult whiteflies, Bemisia tabaci (Gennadius), collected from the field were screened for viral pathogens using a cell line from the silverleaf whitefly, B. tabaci, B biotype (syn. B. argentifolii). Homogenates from the field-collected whiteflies were applied to cell cultures and checked for cytopathic effects (CPE). Cells were observed to develop cytoplasmic inclusions and to have a change in morphology. Cells displaying CPE were observed using a transmission electron microscope and found to be infected with a virus. The virus particles had an icosahedral shape and an approximate size of 120-130 nm. The virus was observed in defined areas of the cytoplasm adjacent to the cell nucleus. Analysis using polymerase chain reaction, Southern blot hybridization, and DNA sequencing confirmed that the virus discovered infecting the whitefly cell cultures was an iridovirus. Sequence analysis showed that the amplimer (893 bp) had a 95% homology to the invertebrate iridescent virus type 6 major capsid protein gene. Discovery of new viruses of whiteflies may provide renewed interest in using pathogens in the development of innovative management strategies. This is the first report of an iridescent virus isolated from whiteflies, B. tabaci, collected from the field.