P-glycoprotein is localized in caveolae in resistant cells and in brain capillaries

A significant proportion of P-glycoprotein (P-gp) and caveolin was co-localized in caveolae isolated from resistant (CH(R)C5) cells overexpressing P-gp and from drug-sensitive Chinese hamster ovary cells (AuxB1). The proportion of P-gp and caveolin associated with caveolar microdomains was higher in CH(R)C5 cells grown in the presence of P-gp substrates (cyclosporin A or colchicine) than in untreated CH(R)C5 cells. Coimmunoprecipitation of P-gp and caveolin from CH(R)C5 lysates suggests that there is a physical interaction between them. Furthermore, co-localization of P-gp and caveolin was found in caveolae from brain capillaries, indicating that this association also takes place in vivo.     

Newcastle disease virus in double-crested cormorants in Alabama, Florida, and Mississippi.

In order to understand the epidemiology of Newcastle disease (ND) outbreaks in double-crested cormorants (Phalacrocorax auritus), a study was conducted on wintering migratory cormorants (P. a. auritus) in Alabama and Mississippi (USA) and non-migratory cormorants (P. a. floridanus) that breed in Florida (USA). Antibodies against ND virus were detected by the hemagglutination-inhibition method in sera from 86 of 183 (47%) migratory cormorants over-wintering in eight roosting sites in Alabama and Mississippi between November, 1997 and April, 1999. Titers ranged from 5 to 40. Antibody prevalences in sera collected from females in early winter (November and December) (26%) and late winter (February and March) (56%) were significantly different (P = 0.0007). None of 45 serum samples from 1- to 7-wk-old nestlings from 11 colonies in Florida during the 1997-98 and 1998-99 breeding seasons was positive. However, antibodies were detected in yolk samples from 98 of 126 (78%) eggs collected in these same colonies. Titers ranged from 4 to 256. The prevalence of antibodies in eggs collected from fresh-water colonies (63% prevalence, n = 30) and salt-water colonies (82% prevalence, n = 96) was significantly different (P = 0.041). ND virus was not isolated from tissues of 18 cormorants and cloacal and tracheal swabs from 202 cormorants collected in Alabama and Mississippi; virus was also not isolated from cloacal and tracheal swabs from 51 nestlings from Florida.     

Meningeal worm in free-ranging deer in Nebraska

The meningeal worm (Parelaphostrongylus tenuis) was found in 22 (7%) of 300 white-tailed deer (Odocoileus virginianus) (257 adults, 43 fawns) examined from Nebraska (USA) during November 1996. None of 53 mule deer (Odocoileus hemionus) (47 adults and 6 fawns) examined were infected. Twenty-two white-tailed deer from 18 counties in eastern Nebraska were infected with Parelaphostrongylus tenuis. This is the first record of P. tenuis in white-tailed deer from this state.     

Nuclear transfer in cattle using in vivo-derived vs. in vitro-produced donor embryos: Effect of developmental stage

To determine the best developmental stage of donor embryos for yielding the highest number of clones per embryo, we compared the efficiencies of nuclear transfer when using blastomeres from morulae or morulae at cavitation, or when using inner-cell-mass cells of blastocysts as nuclear donors. This comparison was done both on in vivo-derived and in vitro-produced donor embryos. In experiment 1, with in vivo-derived donor embryos, nuclei from morulae at cavitation supported the development of nuclear transfer embryos to the blastocyst stage (36%) at a rate similar to that of nuclei from morulae (27%), blastomeres from morulae at cavitation being superior (P < 0.05) to inner-cell-mass cells from blastocysts (21%). The number of blastocysts per donor embryo was significantly (P < 0.05) higher when using nuclei from morulae at cavitation (15.7 ± 4.1) rather than nuclei from morulae (9.8 ± 5.5) or blastocysts (6.3 ± 3.3). With in vitro-produced donor embryos (experiment 2), nuclei from morulae yielded slightly more blastocysts (32%) than nuclei from morulae at cavitation (29%), both stages being superior to nuclei from blastocysts (15% development to the blastocyst stage). Morulae at cavitation yielded a higher number of cloned blastocysts per donor embryo (11.5 ± 5.9) than did morulae (9.3 ± 3.2) and blastocysts (3.3 ± 1.4). Transfer of cloned embryos originating from in vivo-derived morulae, morulae at cavitation, and blastocysts resulted in four pregnancies (10%), three pregnancies (7%), and one (17%) pregnancy on day 45. The corresponding numbers of calves born were 3 (4%), 3 (7%), and 0, respectively. After transfer of blastocysts derived from in vitro nuclear donor morulae (n = 16) and morulae at cavitation (n = 7), two (20%) and two (50%) recipients, respectively, were pregnant on day 45. However, transfer of seven cloned embryos from in vitro donor blastocysts to three recipients did not result in a pregnancy. Using in vitro-produced donor embryos, calves were only obtained from morula-stage donors (13%). Our results indicate that the developmental stage of donor embryos affects the efficiency of nuclear transfer, with morulae at cavitation yielding a high number of cloned blastocysts. © 1996 Wiley-Liss, Inc.     

Genetic variation in wholesale carcass cuts predicted from digital images in cattle

The objective of this study was to quantify the genetic variation in carcass cuts predicted using digital image analysis in commercial cross-bred cattle. The data set comprised 38,404 steers and 14,318 heifers from commercial Irish herds. The traits investigated included the weights of lower value cuts (LVC), medium value cuts (MVC), high value cuts (HVC), very high value cuts (VHVC) and total meat weight. In addition, the weights of total fat and total bones were available on the steers. Heritability of carcass cut weights, within gender, was estimated using an animal linear model, whereas genetic and phenotypic correlations among cuts were estimated using a sire linear model. Carcass weight was included as a covariate in all models. In the steers, heritability ranged from 0.13 (s.e. = 0.02) for VHVC to 0.49 (s.e. = 0.03) for total bone weight, and in the heifers heritability ranged from 0.15 (s.e. = 0.04) for MVC to 0.72 (s.e. = 0.06) for total meat weight. The coefficient of genetic variation for the different cuts varied from 1.4% to 3.6%. Genetic correlations between the different cut weights were all positive and ranged from 0.45 (s.e. = 0.08) to 0.89 (s.e. = 0.03) in the steers, and from 0.47 (s.e. = 0.14) to 0.82 (s.e. = 0.06) in the heifers. Genetic correlations between the wholesale cut weights and carcass conformation ranged from 0.32 (s.e. = 0.06) to 0.45 (s.e. = 0.07) in the steers, and from 0.10 (s.e. = 0.12) to 0.38 (s.e. = 0.09) in the heifers. Genetic correlations between the same wholesale cut traits in steers and heifers ranged from 0.54 (s.e. = 0.14) for MVC to 0.79 (s.e. = 0.06) for total meat weight; genetic correlations between carcass weight and carcass classification for conformation and fat score in both genders varied from 0.80 to 0.87. The existence of genetic variation in carcass cut traits, coupled with the routine availability of predicted cut weights from digital image analysis, clearly shows the potential to genetically improve carcass value.     

Differential alterations in antioxidant capacity in cells from Alzheimer patients

Oxidative stress occurs in brains of Alzheimer's disease (AD) patients. A major question in AD research is whether the oxidative stress is just secondary to neurodegeneration. To test whether oxidative stress is an inherent property of AD tissues, the ability of cultured fibroblasts bearing the AD Presenilin-1 246 Ala-->Glu mutation to handle reactive oxygen species (ROS) was compared to controls. Although ROS in cells from AD subjects were only slightly less than cells from controls under basal conditions (-10%) or after exposure to H(2)O(2) (-16%), treatment with antioxidants revealed clear differences. Pretreatment with DMSO, a hydroxyl radical scavenger, reduced basal and H(2)O(2)-induced ROS levels significantly more in cells from controls (-22%, -22%) than in those from AD subjects (-4%, +14%). On the other hand, pretreatment with Trolox diminished H(2)O(2)-induced ROS significantly more in cells from AD (-60%) than control subjects (-39%). In summary, cells from AD patients have greater Trolox sensitive ROS and less DMSO sensitive ROS than controls. The results demonstrate that fibroblasts bearing this PS-1 mutation have altered means of handling oxidative stress and appear useful for determining the mechanism underlying the altered redox metabolism.     

Prevalence of Yersinia enterocolitica in pigs slaughtered in Chinese abattoirs

The distribution of Yersinia enterocolitica in slaughtered pigs in China was studied. A total of 8,773 samples were collected and examined from different pig abattoirs in 11 provinces from 2009 to 2011. Of these, 4,495 were oral-pharyngeal swab (tonsils) samples from pigs, 1,239 were from intestinal contents, and 3,039 were feces samples from abattoirs or local pigpens. The data showed that 1,132 strains were obtained, from which the isolation rate for Yersinia enterocolitica was 19.53% (878/4,495) from the tonsil samples, 7.51% (93/1,239) from intestinal contents, and 5.30% (161/3,039) from feces. Of the 850 pathogenic Yersinia strains, except for three of bioserotype 2/O:9 and three of bioserotype 4/O:3, most (844/850) were of bioserotype 3/O:3. Interestingly, pathogenic Y. enterocolitica accounted for the majority of the isolated strains from most provinces (85.17% to 100%), whereas from Heilongjiang, 96.52% (111/115) were classified as nonpathogenic biotype 1A with various serotypes, and only 3.48% of the strains (4/115) were pathogenic 3/O:3. All of the pathogenic strains were analyzed using pulsed-field gel electrophoresis (PFGE), and 49 patterns were obtained for the O:3 pathogenic strains; most of them were K6GN11C30021 (53.13%: 450/847) and K6GN11C30012 (21.37%: 181/847). Several strains from diarrhea patient samples revealed PFGE patterns identical to that from samples of local pigs, suggesting a possible link between porcine isolates and human infection. The results above suggested that Yersinia enterocolitica in slaughtered pigs from Chinese abattoirs was characterized by region-specific PFGE patterns and confirmed that strains isolated from pigs are closely related to those from human infections.     

Antibodies to granulocytic ehrlichiae in white-footed and cotton mice in eastern United States

Serum samples, collected from Peromyscus leucopus (white-footed mouse) or Peromyscus gossypinus (cotton mouse) during 1987 through 1990 in Florida, Georgia, Maryland, Mississippi, and North Carolina (USA), and in 1997 in southern Connecticut were analyzed by indirect fluorescent antibody (IFA) staining methods or Western blot procedures for antibodies to granulocytic ehrlichiae. Of the 82 sera from white-footed mice in Connecticut tested by IFA methods with either the BDS or NCH-1 strain of the human granulocytic ehrlichiosis (HGE) agent, 45 (55%) and 42 (51%) of the samples contained antibodies to these strains, respectively, at concentrations ranging from 1:80 to 1:2560. One (2%) of 43 sera from P. leucopus captured in Assateague Island (Maryland) had a titer of 1:80, while three (20%) of 15 sera from P. gossypinus, captured in Sapelo Island (Georgia) and four (40%) of 10 sera from cotton mice caught in Amelia Island (Florida) had antibodies to the NCH-1 strain at titers of 1:160 to 1:1,280. Fifty-five sera from P. leucopus in Cape Hatteras (North Carolina) and 30 sera from P. gossypinus in Mississippi were negative. Western blot analyses confirmed seropositivity for 19 (95%) of 20 mouse sera positive by IFA staining methods, including samples from both mouse species captured in Connecticut, Maryland, or Florida. There were key banding patterns to proteins having molecular masses of about 44, 80, 105, 110, or 120 kDa. Both serologic assays can be used to determine if mice have been exposed to granulocytic ehrlichiae. These rodents also may be useful in surveillance programs to identify endemic sites for HGE and in performing laboratory studies on immune responses to the etiologic agent.     

Colicinogeny in Salmonella serovars isolated in Brazil

A study of colicinogeny was made in 748 strains of Salmonella (97 serovars) isolated from different sources: human (291), animal (119), environmental (141), food (102) and animal feed (95). Colicin production was detected in 64 strains (8.6%), particularly isolated from foods (30.4%). Col E1 (53) and Ia (44) were the most frequently observed, especially in S. agona for environment and food sources. Col V production was identified in 5 strains of S. typhimurium within 8 producer cultures isolated from humans. Its relationship with the sources and serovars of Salmonella are discussed.     

Iron-induced oxidative DNA damage in rat sperm cells in vivo and in vitro

We investigated whether acute iron intoxication causes oxidative DNA damage, measured in terms of 7-hydro-8-oxo-2'-deoxyguanosine, 8-oxodG, in nuclear DNA in testes and epididymal sperm cells in vivo and in vitro in rats. In addition, we investigated levels of the modified nucleoside in liver and kidney and measured its urinary excretion. Sperm cells were isolated from the epididymides and the testes cells were isolated after homogenisation. In vitro, the sperm and testes cells were incubated with increasing concentrations of FeCl2 ranging from 0 to 600 microM. The median (range) levels of 8-oxodG/10(5) dG in the epididymal sperm cells increased from 0.48 (0.42-0.90) to 15.1 (11.4-17.6) (p < 0.05), whereas the level rose from 0.63 (0.22-0.81) to 8.8 (4.5-11.6) (p < 0.05) at 0 and 600 microM, respectively, in the testicular cells. In vivo groups of 7-8 rats received 0, 200 or 400 mg iron/kg as dextran i.p. After 24 h, epididymal sperm cells, testes, kidneys and liver were collected for analysis. Kidney and sperm DNA showed a significant increase in 8-oxodG in the iron-treated animals. The median (range) values of the 8-oxodG/10(5) dG in the epididymal sperm cells rose from 0.66 (0.38-1.09) to 1.12 (0.84-5.88) (p < 0.05) at 0 and 400 mg iron/kg, respectively, whereas the values in the testes and liver showed no significant change. In the kidneys the 8-oxodG/10(5) dG median (range) values were 0.98 (0.73-1.24), 1.21 (1.13-1.69) and 1.34 (1.12-1.66) after 0, 200 and 400 mg iron/kg, respectively (p < 0.05). The 8-oxodG-excretion rate was measured in 24h urine before and after iron treatment. The rate of urinary 8-oxodG excretion increased from 129 (104-179) pmol/24 h before treatment to 147 (110-239) pmol/24 h after treatment in the group receiving 400 mg iron/kg (p < 0.05). The results indicate that acute iron intoxication may increase oxidative damage to sperm and kidney DNA.     

Changes in thick filament length in Limulus striated muscle

Here we describe the change in thick filament length in striated muscle of Limulus, the horseshoe crab. Long thick filaments (4.0 microns) are isolated from living, unstimulated Limulus striated muscle while those isolated from either electrically or K+-stimulated fibers are significantly shorter (3.1 microns) (P less than 0.001). Filaments isolated from muscle glycerinated at long sarcomere lengths are long (4.4 microns) while those isolated from muscle glycerinated at short sarcomere lengths are short (2.9 microns) and the difference is significant (P less than 0.001). Thin filaments are 2.4 microns in length. The shortening of thick filaments is related to the wide range of sarcomere lengths exhibited by Limulus telson striated muscle.     

Electromyogram and mechanomyogram changes in fresh and fatigued muscle during sustained contraction in men

In surface electromyogram (EMG) and mechanomyogram (MMG) the electrical and mechanical activities of recruited motor units (MU) are summated. Muscle fatigue influences the electrical and mechanical properties of the active MU. The aim of this study was to evaluate fatigue-induced changes in the electrical and mechanical properties of MU after a short recovery period, using an analysis of force, surface EMG and MMG. In seven subjects the EMG and MMG were recorded from the biceps brachii muscle during sustained isometric effort at 80% of the maximal voluntary contraction (MVC), before (test 1) and 10 min after (test 2) a fatiguing exercise. From the time and frequency domain analysis of the signals, the root mean square (rms) and the mean frequency (fˉ) of the power spectrum were calculated. The results were that the mean MVC was 412 (SEM 90) N and 304 (SEM 85) N in fresh and fatigued muscle, respectively; during tests 1 and 2 the mean EMG rms increased from 0.403 (SEM 0.07) mV to 0.566 (SEM 0.09) mV and from 0.476 (SEM 0.07) mV to 0.63 (SEM 0.09) mV, respectively; during test 1 the mean MMG rms decreased from 9.4 (SEM 0.8) mV to 5.7 (SEM 0.9) mV; in contrast, during test 2 constantly lower values were observed throughout contraction; during tests 1 and 2 the EMG fˉ declined from 122 (SEM 7) Hz to 74 (SEM 7) Hz and from 106 (SEM 8) Hz to 60 (SEM 7) Hz, respectively; during test 1 the MMG fˉ increased in the first 6 s from 19.3 (SEM 1.4) Hz to 23.9 (SEM 2.9) Hz, falling to 13.9 (SEM 1.3) Hz at the end of contraction; in contrast, during test 2 the MMG fˉ declined continuously from 18.7 (SEM 1) Hz to 12.4 (SEM 0.8) Hz. The lower MVC after the fatiguing exercise and the changes in the EMG parameters confirmed that 10 min after the fatiguing exercise, the mechanical and electrical activities of MU were altered. In addition, the MMG results suggested that after a 10-min recovery, some highly fatigable MU might not be recruitable. Accepted: 9 June 1998     

Powassan virus in Ixodes cookei and Mustelidae in New England.

Powassan virus was recovered from a pool of 3 nymphal and 1 adult female Ixodes cookei removed from a striped skunk (Mephitis mephitis) trapped in Massachusetts during 1967 and from a pool of 9 nymphal I. cookei from a long-tailed weasel (Mustela frenata) captured in Connecticut during 1978. Virus was detected in the blood of both mammals. Hemagglutination-inhibiting (HI) antibody to Powassan virus was demonstrated in 16.0% of the skunks sampled in Connecticut, and neutralizing antibody was detected in 83.3% of the skunks tested from Massachusetts. HI antibody was found in 1 of 6 long-tailed weasels from Connecticut and 1 of 6 short-tailed weasels (Mustela erminea) from Maine.     

Occurrence of macrolide-lincosamide-streptogramin B resistant isolates of Staphylococcus aureus in clinical specimens in 2003-2004

The aim of the study was the analysis of frequency of macrolide-lincosamide-streptogramin B (MLS(B)) resistance among MSSA (n=1682) and MRSA (n=272) strains which were isolated in 2002-2004 from various clinical materials from patients hospitalized in the University Hospital at the L. Rydygier Collegium Medicum in Bydgoszcz, University of Nicolaus Copernicus in Toruń. Susceptibility testing and examination of methicillin-resistant strains were performed by the disc diffusion techniques according to recommendation of NCCLS. Resistance to the MLS(B) antimicrobials agents was higher among MRSA compared to MSSA isolates. The MLS, constitutive phenotype was more prevalent than the MLS(B) inducible phenotype among investigated MRSA (65.4%) and MSSA (7.6%) isolates. Inducible resistance had only 2.5% of the MSSA and 2.6% of the MRSA strains. Moreover in 2004 there were found increasing frequency of inducible MLS(B) resistance from 1.1% to 5.7% and decreasing frequency of constitutive MLS(B) resistance from 9.2% to 4.7% among MSSA strains, in comparison to 2003. The investigated MSSA MLS(B)-, MLS(B)- and MRSAMLS(B)+, MLS(B)- strains were the most frequently isolated from pus (adequately 5.2%, 28.8% and 30.5%, 10.7%) and also from nosopharynx swabs (1.7% MSSA MLS(B)+ and 22.9% MSSA MLS(B)-) and biomaterials (15,1% MRSA MLS(B)+ and 9.6% MRSA MLS(B)-). They mainly came from patients of the outpatient clinic (2,4% MSSA MLS(B)+ and 19.9% MSSA MLS(B)-) and patients treated at the neurosurgical ward (20.6% MRSA MLS(B)+ and 12.1% MRSA MLS(B)-).     

广州市十种森林生态系统的碳循环

为了探讨南亚热带森林生态系统碳循环的规律,在广泛收集资料和试验数据的基础上,对广州10种森林生态系统的碳循环进行研究.结果表明：10种森林生态系统的碳密度在108.35～151.85 t C·hm^-2,其中乔木层碳密度在10.85～48.86 t C·hm^-2,0～60 cm土壤层在87.74～99.01 t C·hm^-2,均低于全国平均水平;从大气流向植被层的碳流量为4.41～9.15 t C·hm^-2·a^-1,植被层流向土壤层的碳流量为0.74～2.06 t C·hm^-2·a^-1,土壤层流向大气层的碳流量为3.94～5.42 t C·hm^-2·a^-1,即系统从大气净吸收碳在0.47～4.97 t C·hm^-2·a^-1之间.各种林分的净系统生产力不同,阔叶林大于针叶林,混交林大于纯林,天然次生林大于人工林.     

Arachidonic acid turnover in peritoneal macrophages is altered in endotoxin-tolerant rats

Peritoneal macrophages from endotoxin-tolerant rats have been found to exhibit depressed metabolism of arachidonic acid (AA) to prostaglandins and thromboxane in response to endotoxin. The effect of endotoxin tolerance on AA turnover in peritoneal macrophages was investigated by measuring [14C]AA incorporation and release from membrane phospholipids. Endotoxin tolerance did not affect the amount of [14C]AA incorporated into macrophages (30 min-24 h). However, the temporal incorporation of [14C]AA into individual phospholipid pools (15 min-24 h) was altered. In endotoxin-tolerant macrophages, [14C]AA incorporation into phosphatidylcholine (PC) (2, 4, 24 h) and phosphatidylethanolamine (PE) (8 h) was increased, while the incorporation into phosphatidylserine (PS) (2-24 h) was reduced (P less than 0.005) compared to control macrophages. There was no change in [14C]AA incorporation into phosphatidylinositol (PI). Following 2 or 24 h of incorporation of [14C]AA, macrophages were incubated (3 h) with endotoxin (50 micrograms/ml) or A23187 (1 microM), and [14C]AA release was measured. Endotoxin-tolerant macrophages released decreased (P less than 0.05) amounts of [14C]AA in response to both endotoxin and the calcium ionophore A23187 compared to controls. Control macrophages in response to endotoxin released [14C]AA from PC, PI and PE. In contrast, tolerant cells released [14C]AA only from PC (P less than 0.05). A23187 released [14C]AA from all four pools in the control cells, but only from PC and PE in the tolerant cells. These data demonstrate that endotoxin tolerance alters the uptake and release of AA from specific macrophage phospholipid pools. These results suggest that changes in AA turnover and/or storage are associated with endotoxin tolerance.     

Age-related changes in the mevalonate metabolism in vivo in chick kidneys

The mevalonate incorporation in vivo into total nonsaponifiable lipids by chick kidneys drastically increased after hatching, reaching similar levels to those previously observed in liver. Cholesterol was the major sterol formed from mevalonate from 11 days onward, while a fraction of polar nonsaponifiable lipid(s) was observed as the major compound(s) synthesized at 5-8 days. Relative percentages of squalene, squalene oxide(s) and lanosterol synthesized from mevalonate also increased between 11-18 days after hatching. Results in this paper demonstrate for the first time the accumulation of a fraction of nonsaponifiable lipid(s) identified as lanosterol derivatives and cholesterol precursors formed by kidneys from [5-14C]mevalonate in experiments carried out in vivo, as well as their evolution during postnatal period.     

Tuberculosis in Tanzanian wildlife

Bovine tuberculosis, caused by Mycobacterium bovis, is a pathogen of growing concern in free-ranging wildlife in Africa, but little is known about the disease in Tanzanian wildlife. Here, we report the infection status of Mycobacterium bovis in a range of wildlife species sampled from protected areas in northern Tanzania. M. bovis was isolated from 11.1% (2/18) migratory wildebeest (Connochaetes taurinus) and 11.1% (1/9) topi (Damaliscus lunatus) sampled systematically in 2000 during a meat cropping program in the Serengeti ecosystem, and from one wildebeest and one lesser kudu (Tragelaphus imberbis) killed by sport hunters adjacent to Tarangire National Park. A tuberculosis antibody enzyme immunoassay (EIA) was used to screen serum samples collected from 184 Serengeti lions (Panthera leo) and 19 lions from Ngorongoro Crater sampled between 1985 and 2000. Samples from 212 ungulates collected throughout the protected area network between 1998 and 2001 also were tested by EIA. Serological assays detected antibodies to M. bovis in 4% of Serengeti lions; one positive lion was sampled in 1984. Antibodies were detected in one of 17 (6%) buffalo (Syncerus caffer) in Tarangire and one of 41 (2%) wildebeest in the Serengeti. This study confirms for the first time the presence of bovine tuberculosis in wildlife of northern Tanzania, but further investigation is required to assess the impact on wildlife populations and the role of different wildlife species in maintenance and transmission.