The role of mast cells in the elicitation of experimental allergic encephalomyelitis

Experimental allergic encephalomyelitis (EAE), a T cell-mediated autoimmune disease can be transferred with lymphoid cells from actively immunized rats into naive recipients. In the mouse, previous studies have suggested a role for histamine/serotonin in the development of active EAE. We have found that myelin basic protein-reactive cells transfer a biphasic skin test response to naive rats analogous to what has been described in the mouse contact dermatitis system, where mast cell sensitization by Ag-specific T cell factors is required for the induction of skin test responses. Treatment of cell recipients with the serotonin receptor antagonists, cyproheptadine or methysergide, blocked or significantly reduced the development of EAE. Furthermore, it was found that treatment with cyproheptadine was effective in blocking clinical disease when administered day 3 to day 6 after cell transfer. In contrast, cyproheptadine treatments before induction of paralysis day 0 to 3, failed to alter the course of clinical disease. The inhibitor of mast cell degranulation, proxicromil, was also found to effectively block the elicitation of adoptively transferred EAE and was also found to be effective when administered just before the onset of clinical disease. Reserpine, a compound known to deplete mast cells of vasoactive amines by forcing granule contents into the cytoplasm where they are degraded by cell enzymes, was also effective in blocking both active and adoptively transferred EAE. Disease inhibition was found to be partially reversed with pargyline, an inhibitor of monoamine oxidase. In addition lymphocytes from treated animals were capable of transferring disease to naive recipients and appeared to have normal activity as assessed by Ag-or mitogen-driven proliferation in addition to IL-2 production.     

Growth response, reproductive activity, and serum growth hormone in fine-wool ewe lambs treated with triiodothyronine

Forty-four 6-mo-old, fine-wool ewe lambs were randomly allotted to one of three triiodothyronine (T(3)) treatment groups (14 or 15 ewe lambs/group). Treatments (0, 200, and 400 mug T(3)) were administered daily by subcutaneous injection during a 30-d treatment period in September. Following the treatments, ewe lambs were allowed to mate with Debouillet rams during a 34-d breeding season. Animal weights did not differ (P > 0.10) among treatment groups throughout the study. However, ewe lambs receiving 400 mug T(3)/d gained less (P < 0.05) during the treatment period than did those receiving 200 mug T(3)/d or controls. Grease fleece weights (6-mo clip) were similar (P > 0.10) among treatment groups. Serum growth hormone (GH) values in blood samples collected on Days 7 and 28 of the 30-d treatment were similar (P > 0.10) before treatments were administered and on Day 7 after treatments were administered. However, on Day 28 after treatments were administered, serum GH was lower (P < 0.01) in ewe lambs receiving 400 mug T(3)/d (1.5 ng/ml) than in those receiving 200 mug/d (2.1 ng/ml) or controls (2.2 ng/ml). Ovarian cyclicity was determined by monitoring serum progesterone. Forty percent of control ewe lambs were determined to have cycled during treatment compared with 20% of those receiving 200 mug T(3)/day and none of those receiving 400 mug T(3)/d (P < 0.10). Pregnancy rates were lower (P < 0.10) in ewe lambs receiving 400 mug T(3)/d (29%) compared with those receiving 200 mug T(3)/d (60%) or controls (66.7%). Day of conception, lambs born/ewe exposed, lambs born/ewe lambing, and lambs weaned/ewe lambing did not differ (P > 0.10) among treatments. Preweaning performance of offspring was not altered by maternal treatment with 200 or 400 mug T(3)/d (P > 0.10). These data indicate that 200 and 400 mug T(3)/d will not enhance GH production or improve reproductive performance of ewe lambs.     

Structural basis for a new tetracycline resistance mechanism relying on the TetX monooxygenase

The flavin-dependent monooxygenase TetX confers resistance to all clinically relevant tetracyclines, including the recently approved, broad-spectrum antibiotic tigecycline (Tygacil®) which is a critical last-ditch defense against multidrug-resistant pathogens. TetX represents the first resistance mechanism against tigecycline, which circumvents both the tet-gene encoded resistances, relying on active efflux of tetracyclines, and ribosomal protection proteins. The alternative enzyme-based mechanism of TetX depends on regioselective hydroxylation of tetracycline antibiotics to 11a-hydroxy-tetracyclines. Here, we report the X-ray crystallographic structure determinations at 2.1 Å resolution of native TetX from Bacteroides thetaiotaomicron and its complexes with tetracyclines. Our crystal structures explain the extremely versatile substrate diversity of the enzyme and provide a first step towards the rational design of novel tetracycline derivatives to counter TetX-based resistance prior to emerging clinical observations.     

Detection of microbial biomass by intact polar membrane lipid analysis in the water column and surface sediments of the Black Sea

The stratified water column of the Black Sea produces a vertical succession of redox zones, stimulating microbial activity at the interfaces. Our study of intact polar membrane lipids (IPLs) in suspended particulate matter and sediments highlights their potential as biomarkers for assessing the taxonomic composition of live microbial biomass. Intact polar membrane lipids in oxic waters above the chemocline represent contributions of bacterial and eukaryotic photosynthetic algae, while anoxygenic phototrophic bacteria and sulfate-reducing bacteria comprise a substantial amount of microbial biomass in deeper suboxic and anoxic layers. Intact polar membrane lipids such as betaine lipids and glycosidic ceramides suggest unspecified anaerobic bacteria in the anoxic zone. Distributions of polar head groups and core lipids show planktonic archaea below the oxic zone; methanotrophic archaea are only a minor fraction of archaeal biomass in the anoxic zone, contrasting previous observations based on the apolar derivatives of archaeal lipids. Sediments contain algal and bacterial IPLs from the water column, but transport to the sediment is selective; bacterial and archaeal IPLs are also produced within the sediments. Intact polar membrane lipid distributions in the Black Sea are stratified in accordance with geochemical profiles and provide information on vertical successions of major microbial groups contributing to suspended biomass. This study vastly extends our knowledge of the distribution of complex microbial lipids in the ocean.     

Species Diversity and Floristic Characteristics of Vascular Plants in Nanji Island,Zhejiang Provience

A total of 393 species (sub type included) in 267 genera, 97 families were reported in Nanji Island, the main island of Nanji Islands National Nature Reserve. After eliminating all the cultivated and invasive species, there were still 304 species in 210 genera and 81 families belonging to wild vascular plants; To better understand the floristic characteristics of Nanji, we applied analysis in 3 levels: At family level, the largest families in pteridophyte were ranked as Pteridaceae、Thelypteridaceae、Lygodiaceae、Dryopteridaceae; In seed plants, almost the largest ones were Cosmopolitan, while the small ones dominented the most, quantitatively and proportionately; The characteristic families of seed flora were consisted of Liliaceae、Amaryllidaceae、Urticaceae、Verbenaceae, cause they got high ratio of VFICS/WFIW; At genus level, the Pantropical Distribution (3810%)、the North Temperate Distribution (1630%) and the East Asia distribution (1131%) formed the main part of seed flora; The proportion of the category of temperate distribution and tropical distribution was 6072% and 3928% respectively; Took 6 other sites as reference, through cluster analysis and R/T ratio comparisons, It was proved that Nanji Island were most similar to that of Wuyanling of Zhejiang Provience but showed more tropical affinities; At species level, wild seed plants can also be divided into 9 areal subtypes, The Yangtze River & Southern Proviences sub type dominented at 5390%, following by South China sub type at 2078%. The results showed that Nanji Island had a distinctive transitional characteristics of flora from north to south China with more tropical biological properties, thus the floristic division of Nanji should on the southern margin area of east China.     

Nonantibiotic Properties of Tetracyclines: Structural Basis for Inhibition of Secretory Phospholipase A2

Secretory phospholipase A₂ is involved in inflammatory processes and was previously shown to be inhibited by lipophilic tetracyclines such as minocycline (minoTc) and doxycycline. Lipophilic tetracyclines might be a new lead compound for the design of specific inhibitors of secretory phospholipase A₂, which play a crucial role in inflammatory processes. Our X-ray crystal structure analysis at 1.65 Å resolution of the minoTc complex of phospholipase A₂ (PLA₂) of the Indian cobra (Naja naja naja) is the first example of nonantibiotic tetracycline interactions with a protein. MinoTc interferes with the conformation of the active-site Ca²⁺-binding loop, preventing Ca²⁺ binding, and shields the active site from substrate entrance, resulting in inhibition of the enzyme. MinoTc binding to PLA₂ is dominated by hydrophobic interactions quite different from antibiotic recognition of tetracyclines by proteins or the ribosome. The affinity of minoTc for PLA₂ was determined by surface plasmon resonance, resulting in a dissociation constant K_d = 1.8 × 10^− 4 M.     

A comparison between cervical dimensions of pregnant and nonpregnant Santa Gertrudis and Bos taurus cows

Cervical measurements were recorded on palpation per rectum for 603 Bos taurus , Santa Gertrudis, and crossbred cattle. Postmortem cervical measurements were recorded for 77 of these cows. Antemortem measurements were positively correlated with those taken at postmortem examination (cervical diameter: r=0.75, P<0.01 and cervical length: r=0.29, P<0.06). Cervical diameters from antemortem and postmortem specimens were greater in Santa Gertrudis cattle than in Bos taurus and crossbred cattle (P<0.05). Cervical shape was different between Santa Gertrudis cattle and Bos taurus or crossbred cattle (P<0.01), with Santa Gertrudis cows having a higher incidence of conical shaped cervices than other breeds. Within Santa Gertrudis cattle, the incidence of conical cervical shape increased with age (P<0.05). Conical cervical shape was associated with a lower pregnancy rate in all breeds (P<0.01). Within Santa Gertrudis cattle, conical cervical shape was associated with a lower pregnancy rate in 1- to 4-year-old cows (P<0.05) but not in cows more than 4 years of age (P>0.05).     

Treatments resulting in pregnancy in nonovulating, hormone-treated oocyte recipient mares

Synchronization of follicle growth between oocyte donor and recipient mares is difficult. To avoid this, recipient mares in a clinical program were used during a period of low follicular activity, and were treated with estrogen before transfer and progesterone after transfer. Five pregnancies were established after oocyte transfer to nonovulating, hormone-treated recipient mares. One pregnancy was lost before 30 d gestation, and the other 4 foals were carried to term. One foal died at birth. Establishment and maintenance of pregnancy in these mares indicates that nonovulating, hormone-treated mares may offer an alternative to cyclic recipients in oocyte transfer programs.     

Staphylococcus aureus helicase but not Escherichia coli helicase stimulates S. aureus primase activity and maintains initiation specificity

Bacterial primases are essential for DNA replication due to their role in polymerizing the formation of short RNA primers repeatedly on the lagging-strand template and at least once on the leading-strand template. The ability of recombinant Staphylococcus aureus DnaG primase to utilize different single-stranded DNA templates was tested using oligonucleotides of the sequence 5'-CAGA (CA)5 XYZ (CA)3-3', where XYZ represented the variable trinucleotide. These experiments demonstrated that S. aureus primase synthesized RNA primers predominately on templates containing 5'-d(CTA)-3' or TTA and to a much lesser degree on GTA-containing templates, in contrast to results seen with the Escherichia coli DnaG primase recognition sequence 5'-d(CTG)-3'. Primer synthesis was initiated complementarily to the middle nucleotide of the recognition sequence, while the third nucleotide, an adenosine, was required to support primer synthesis but was not copied into the RNA primer. The replicative helicases from both S. aureus and E. coli were tested for their ability to stimulate either S. aureus or E. coli primase. Results showed that each bacterial helicase could only stimulate the cognate bacterial primase. In addition, S. aureus helicase stimulated the production of full-length primers, whereas E. coli helicase increased the synthesis of only short RNA polymers. These studies identified important differences between E. coli and S. aureus related to DNA replication and suggest that each bacterial primase and helicase may have adapted unique properties optimized for replication.     

Analysis of reactive carbonyls in the expired air of transgenic mice.

Methods for the determination of trace levels of volatile carbonyl compounds in air expired from mice were developed and validated. Tumor bearing transgenic mice or nontransgenic control mice were placed into a glass chamber through which air was passed continuously at 90 ml/min for 1 h. The effluent gas stream was bubbled into an aqueous cysteamine solution or an aqueous methylhydrazine solution. Formaldehyde, acetaldehyde, and acetone in expired air were derivatized to thiazolidine with cysteamine and malonaldehyde was derivatized to 1-methyl-2-pyrazole with methylhydrazine. The derivatized compounds were analyzed by capillary gas chromatography with flame photometric or nitrogen-phosphorous-specific detection. The lowest level quantitated was 4 micrograms/ml thiazolidine, equivalent to 1.35 micrograms/ml formaldehyde. Formaldehyde was recovered at a level of 1356 +/- 234 nmol/kg0.75 (mean +/- SD) from mice with tumors and 898 +/- 97 nmol/kg0.75 from mice without tumors, suggesting that tumor bearing transgenic mice expired significantly more formaldehyde than did tumor free controls. Amounts of expired acetaldehyde and acetone were not different among mice. Malonaldehyde was not detected in either group of mice.