Effects of a mannan-rich yeast cell wall-derived preparation on cecal concentrations and tissue prevalence of Salmonella Enteritidis in layer chickens.

Salmonella Enteritidis (SE) has been the most common Salmonella serotype associated with foodborne infections in the last several years. Dietary applications of yeast-based preparations in feed have shown to reduce Salmonella colonization in chickens augmenting SE control strategies. This study was conducted to evaluate the effects of a mannan-rich yeast cell wall-derived preparation (Actigen®) administered in feed at a rate of 400 g/ton on SE colonization in the cecum and internal organs of commercial layer chickens. Sixteen week-old layer pullets were orally challenged with a selected nalidixic acid resistant SE strain at a dose of 1.7×10^9 colony forming units (CFU) per bird. SE colonization was assessed by evaluating isolation rates from ovary and pooled liver/spleen samples as well as enumeration of SE in cecal pouches one week post-challenge. Recovery rates of SE from the ovaries of directly challenged birds receiving Actigen® were significantly lower (P <0.02) than those in directly challenged birds fed an unsupplemented control diet. Recovery rates of SE from pooled liver/spleen samples were not significantly different between Actigen®-treated pullets and controls (P = 0.22). Using direct plate count methods, cecal SE concentrations were 1 log10 lower (P <0.001) in challenged pullets in the Actigen®-supplemented group than in the challenged controls. The SE concentration distributions in the ceca were similar in groups testing positive and groups testing negative for SE in the ovaries and liver/spleens tissues. As a result, SE concentrations in the ceca could not be directly related to the occurrence or prevalence of SE in these tissues. In conclusion, Actigen® supplementation appears to decrease the prevalence of SE in ovarian tissue and concentrations of SE in cecal contents and may be useful as a tool for reducing the risk of eggshell contamination and transovarian transmission of SE in eggs.

Acute lion's mane jellyfish, Cyanea capillata (Cnideria: Scyphozoa), exposure to Atlantic salmon (Salmo salar L.).

Jellyfish-induced gill pathology relies upon occasional diagnostic observations yet the extent and impact of jellyfish blooms on aquaculture may be significant. Idiopathic gill lesions are often observed in apparently healthy fish. This study exposed Atlantic salmon (Salmo salar L.) smolts to macerated Cyanea capillata at 2.5 and 5 g/L for 2 hr under controlled laboratory conditions. Blood chemistry and gill histopathology were examined over a subsequent 4-week period. Fish showed an acute response to the presence of jellyfish, including characteristic external "whiplash" discoloration of the skin and acute increases in blood electrolytes and CO2 concentration; however, these were resolved within 4 days after exposure. Histopathologically, gills showed first an acute oedema with epithelial separation followed by focal haemorrhage and thrombus formation, and then progressive inflammatory epithelial hyperplasia that progressively resolved over the 4 weeks post-exposure. Results were consistent with the envenomation of gills with cytotoxic neurotoxins and haemolysins known to be produced by C. capillata. This study suggests that many focal hyperplastic lesions on gills, especially those involving focal thrombi, may be the result of jellyfish stings. Thus, the presence of jellyfish and their impact may be severe and understated in terms of marine fish aquaculture and fish welfare.

In vitro effect of garlic extract and metronidazole against Neoparamoeba pemaquidensis, page 1987 and isolated amoebae from Atlantic salmon.

Neoparamoeba pemaquidensis believed to be the most prevalent parasite of Atlantic salmon industry in Australia. In the present study, the in vitro effects of crude extract of garlic and metronidazole were investigated using a primary culture toxicity assay. Garlic extract appeared to be completely effective at killing a cultured strain (NP251002) of Neoparamoeba pemaquidensis in vitro at a dilution of 1:100 with in 24 h. The number of viable Amoebae after using garlic extract in lower dilutions (1:200, 1:400, 1:800, 1:1000) for 24 h, also were significantly lower than in the control group. Garlic extract was also efficacious at killing wild type Amoebae that isolated from the diseased fish showing clinical signs of AGD. Metronidazole had no clear effect against Neoparamoeba pemaquidensis (NP251002) even in a concentration of 50 mg L(-1) for 24 h. However some morphological changes have occurred in metronidazole-treated Amoebae after 5 days of exposure.

Municipal waste compost as an alternative to cattle manure for supplying potassium to lowland rice.

The importance of the use of potassium in agriculture is increasing in South Asia for making most productive use of the nutrient in terms of economic returns. Nutrient supply traditionally by cattle manure is constrained by its insufficient availability. Municipal waste compost may be an alternative source of nutrient supplements. Field experiments were conducted at the Experimental Farm of Calcutta University, West Bengal, India during the wet seasons of 1997, 1998 and 1999 on flooded lowland rice. Potassium fractions in municipal waste compost and cattle manure were determined by sequential extraction and also the potassium uptake by rice to compare the effectiveness of municipal waste compost with traditional manure. Potassium was significantly bound to the organic matter in municipal waste compost. Potassium uptake by rice grain and straw increased significantly with the combined application of organics and fertilizers and it was higher in grain than in straw. Water-soluble and non-exchangeable potassium contents of municipal waste compost and cattle manure were highly correlated with the uptake of potassium by straw and grain. Exchangeable and residual potassium were also significantly correlated with the uptake of potassium by straw and grain of rice. Much higher uptake of K in rice straw and rain resulted from applying the manures in conjunction with fertilizers than when applied singly.

Trends in colon cancer screening procedures in the US Medicare and Tricare populations: 1999-2001.

BACKGROUND: This study examined trends in the numbers of double contrast barium enemas, flexible sigmoidoscopies, and colonoscopies and trends in the choices of colorectal cancer screening service providers. METHODS: Descriptive statistics were used to examine Medicare and Tricare data for the years 1999 to 2001. RESULTS: The total volume of procedures increased 5% and 14%, respectively, in Tricare and Medicare. Tricare and Medicare, respectively, saw 32% and 33% reductions in barium enemas and 28% and 41% reductions in flexible sigmoidoscopies. Colonoscopies increased by 45% and 34% in Tricare and Medicare, respectively. Gastroenterologists provided the majority of colonoscopies for both groups each year. CONCLUSIONS: The volume of colonoscopies increased from 1999 to 2001 for both groups while the volumes of barium enemas and flexible sigmoidoscopies decreased. Gastroenterologists appear to be the preferred providers of colonoscopies.

Use of traditional healers and modern medicine in Ghana.

PURPOSE: To gain understanding of the use of traditional and modern medicine among the people in Ghana, West Africa. METHODS: Data were collected from nine participants using a semi-structured questionnaire developed by the researchers based on review of the literature. FINDINGS: Data analysis was performed manually using reduction methodology to develop broad themes. Findings indicated that choices in healthcare modalities by literate Ghanaians included either traditional or modern medicine, or blending of both. Strong influences on these choices were the level of education and related themes, influence of family and friends, and spiritual/religious beliefs. IMPLICATIONS: Findings indicate that traditional and modern medicines will always be part of Ghanaian healthcare delivery and efforts should be made to integrate traditional practitioners into the national healthcare delivery system.

Hyperbaric oxygen pre-breathe modifies the outcome of decompression sickness.

Deep sea divers, aviators and astronauts are at risk of decompression sickness when the ambient pressure reductions exceed a critical threshold. Venous bubbles associated with decompression sickness have the potential to react with the vascular membrane and adjacent blood products, eliciting an inflammatory cascade. Preventive measures usually involve careful decompression procedures to avoid or reduce bubble formation. De-nitrogenation with 100% oxygen pre-breathing as a preventive measure has been well established at least in altitude decompression exposures. The objective of this study was to determine the physiological and biochemical effects of Hyperbaric Oxygen Pre-breathe (HBOP) upon decompression from a hyperbaric exposure. Male Sprague-Dawley rats were randomly assigned to one of eight groups. Two experimental groups received HBOP at 1 and 18 hours prior to decompression, as compared with ground level oxygen or non-treated groups that still experienced decompression stress, and the associated non-decompressed controls. The results showed decreased extravascular lung water (pulmonary edema), bronchoalveolar lavage and pleural protein and arterial, broncho-alveolar lavage, and urine leukotriene E4 (LKE4) levels in both the 1Hr and 18Hr HBOP decompressed rats compared to non-oxygenated decompressed rats, as well as a decreased overall expression of signs of decompression sickness. This study indicates that HBOP-treated rats exhibit fewer signs and complications of decompression sickness compared with non-treated or ground level oxygen treated rats.

Quality of anaesthesia for insertion of tension-free vaginal tape using local analgesia and sedation.

BACKGROUND AND OBJECTIVE: The use of tension-free tape in the vagina is a relatively new surgical procedure for the treatment of urinary incontinence. Traditionally, the procedure is carried out using local anaesthesia and sedation. We aimed to assess the quality of anaesthesia obtained during insertion of tension-free vaginal tape using local anaesthetic infiltration and intravenous conscious sedation. METHODS: Twenty-four patients were studied using intravenous midazolam, fentanyl and infiltration with 0.5% prilocaine and epinephrine. Visual analogue scores before operation were used to assess anxiety. The digit symbol substitution test was used before and after surgery to assess psychomotor recovery, and amnesia for the procedure was assessed by means of picture recall. A nurse, anaesthetist and surgeon independently assessed quality of anaesthesia based on vocalization, facial expression, movement and co-operation with incontinence testing. RESULTS: The nurse, anaesthetist and surgeon reported good or excellent conditions in 18, 14 and 22 cases, respectively. There was no significant psychomotor impairment using the digit symbol substitution test, with mean (standard deviation) preoperative and postoperative scores of 22 (8) and 21 (7) correctly matched symbol digit pairs. There was a significant correlation between preoperative anxiety and intraoperative fentanyl requirement (r=0.48, P < 0.05). CONCLUSION: Satisfactory anaesthetic conditions can be achieved for the insertion of tension-free vaginal tape using local anaesthesia with sedation.

Describing individual incidents of sexual abuse: a review of research on the effects of multiple sources of information on children's reports.

OBJECTIVE: For successful prosecution of child sexual abuse, children are often required to provide reports about individual, alleged incidents. Although verbally or mentally rehearsing memory of an incident can strengthen memories, children's report of individual incidents can also be contaminated when they experience other events related to the individual incidents (e.g., informal interviews, dreams of the incident) and/or when they have similar, repeated experiences of an incident, as in cases of multiple abuse. METHOD: Research is reviewed on the positive and negative effects of these related experiences on the length, accuracy, and structure of children's reports of a particular incident. RESULTS: Children's memories of a particular incident can be strengthened when exposed to information that does not contradict what they have experienced, thus promoting accurate recall and resistance to false, suggestive influences. When the encountered information differs from children's experiences of the target incident, however, children can become confused between their experiences-they may remember the content but not the source of their experiences. CONCLUSIONS: We discuss the implications of this research for interviewing children in sexual abuse investigations and provide a set of research-based recommendations for investigative interviewers.

Naturally occurring dicistronic cricket paralysis virus RNA is regulated by two internal ribosome entry sites.

Cricket paralysis virus is a member of a group of insect picorna-like viruses. Cloning and sequencing of the single plus-strand RNA genome revealed the presence of two nonoverlapping open reading frames, ORF1 and ORF2, that encode the nonstructural and structural proteins, respectively. We show that each ORF is preceded by one internal ribosome entry site (IRES). The intergenic IRES is located 6,024 nucleotides from the 5' end of the viral RNA and is more active than the IRES located at the 5' end of the RNA, providing a mechanistic explanation for the increased abundance of structural proteins relative to nonstructural proteins in infected cells. Mutational analysis of this intergenic-region IRES revealed that ORF2 begins with a noncognate CCU triplet. Complementarity of this CCU triplet with sequences in the IRES is important for IRES function, pointing to an involvement of RNA-RNA interactions in translation initiation. Thus, the cricket paralysis virus genome is an example of a naturally occurring, functionally dicistronic eukaryotic mRNA whose translation is controlled by two IRES elements located at the 5' end and in the middle of the mRNA. This finding argues that eukaryotic mRNAs can express multiple proteins not only by polyprotein processing, reinitiation and frameshifting but also by using multiple IRES elements.

Effect of heat pretreatment on the yield and composition of oil extracted from corn fiber.

Previously, hexane extraction of corn fiber was reported to produce a unique and potentially valuable oil that contained high levels of several phytosterols (which have been noted for their cholesterol-lowering properties). Current studies revealed that heat treatment (over the range of 100-175 degrees C) of corn fiber in either a convection oven or a vacuum oven caused only a modest reduction in the levels of the phytosterol components. However, these same heat pretreatments caused a considerable increase (up to 10-fold) in the levels (increasing from 0.34 wt % to a maximum of 3.64 wt % gamma-tocopherol in the oil) and yields (increasing from 5.4 mg of gamma-tocopherol/100 g of corn fiber to a maximum of 52.1 mg of gamma-tocopherol/100 g of corn fiber) of gamma-tocopherol in corn fiber oil. The main differences between the convection oven and vacuum oven pretreatments were associated with the disappearance of free fatty acids and free phytosterols at the higher temperature pretreatments in the vacuum oven, probably due to the lower boiling points of these lipids. Microwave pretreatment was also effective but caused a much smaller increase in the levels of gamma-tocopherol.

The effects of repeated experience on children's suggestibility.

The effect of suggestive questions on 3- to 5-year-old and 6- to 8-year-old children's recall of the final occurrence of a repeated event was examined. The event included fixed (identical) items as well as variable items where a new instantiation represented the item in each occurrence of the series. Relative to reports of children who participated in a single occurrence, children's reports about fixed items of the repeated event were more accurate and less contaminated by false suggestions. For variable items, repeated experience led to a decline in memory of the specific occurrence; however, there was no increase in susceptibility to suggestions about details that had not occurred. Most errors after repeated experience were intrusions of details from nontarget occurrences. Although younger children and children who were interviewed a while after the event were more suggestible, respectively, than older children and those interviewed soon after the event, repeated experience attenuated these effects.

Detection of the hereditary hemochromatosis gene mutation by real-time fluorescence polymerase chain reaction and peptide nucleic acid clamping.

Hereditary hemochromatosis (HH), an iron overload disease, is the most common known inheritable disease. The most prevalent form of HH is believed to be the result of a single base-pair mutation. We describe a rapid homogeneous mutation analysis method that does not require post-polymerase chain reaction (PCR) manipulations. This method is a marriage of three emerging technologies: rapid cycling PCR thermal cyclers, peptide nucleic acid (PNA) probes, and a new double-stranded DNA-selective fluorescent dye, Sybr Green I. The LightCycler is a rapid thermal cycler that fluorometrically monitors real-time formation of amplicon with Sybr Green I. PNAs are DNA mimics that are more sensitive to mismatches than DNA probes, and will not serve as primers for DNA polymerases. PNA probes were designed to compete with PCR primers hybridizing to the HH mutation site. Fully complemented PNA probes at an 18:1 ratio over DNA primers with a mismatch result in suppression of amplicon formation. Conversely, PNA probes with a mismatch will not impair the binding of a complementary primer, culminating in amplicon formation. A LightCycler-based rapid genetic assay has been developed to distinguish HH patients from HH carriers and normal individuals using PNA clamping technology.

Immunoliposomes containing antibodies to costimulatory molecules as adjuvants for HIV subunit vaccines.

Immunoliposomes containing monoclonal antibodies (MAbs) to the costimulatory molecules CD28 and CTLA4 and their counterreceptors B7-1 (CD80) and B7-2 (CD86) were evaluated for the ability to increase the immune response to recombinant envelope protein rgp120 of the MN strain of human immunodeficiency virus type 1 (HIV-1) during vaccination. MAbs were attached to rgp120-containing liposomes via a biotin-avidin-biotin bridge. Mice vaccinated with immunoliposomes were found to have a strong delayed-type hypersensitivity (DTH) response to the weakly immunogenic gp120 that was dependent on the presence of the MAbs. However, this vaccination protocol did not induce humoral immunity. The DTH response was not accompanied by increased production of interferon gamma (IFN-gamma) or interleukin 4 (IL-4), implying that the primary cellular interaction was between the immunoliposomes and cells of the reticuloendothelial system and not helper T (Th) cells. This strategy of incorporating antibodies to costimulatory molecules on the surface of antigen-containing particulates, such as liposomes or microspheres, can be used to increase DTH immune responses to protein or peptide vaccines.

Methscopolamine bromide blocks hypothalamic-stimulated release of growth hormone in ewes.

Twenty-five nonlactating ewes were used to test the hypothesis that methscopolamine bromide (MB) blocks secretion of growth hormone (GH) by affecting hypothalamic rather than pituitary mechanisms. Ewes were randomly assigned to receive a s.c. injection of 96 mg of MB or 2 mL of saline at min = 0. Saline-treated ewes were assigned to receive a subsequent (at +60 min) i.v. injection of 10 microg of growth hormone-releasing hormone (GHRH) to test pituitary responsiveness or .3 mg of clonidine to test hypothalamic responsiveness. Methscopolamine bromide-treated ewes were assigned to receive a subsequent (at +60 min) i.v. injection of 10 microg of GHRH, .3 mg of clonidine, or 2 mL of saline. Jugular blood samples were collected at 10-min intervals from -120 min to +240 min, and serum concentrations of GH were quantified with a RIA. No difference was detected in serum concentrations of GH with respect to MB vs saline treatment (P = .20). Concentrations of GH increased in saline-pretreated ewes following injections of clonidine or GHRH (P < .01). Treatment of ewes with MB, however, limited the ability of clonidine-induced mechanisms to increase concentrations of GH, but did not affect pituitary responsiveness to GHRH (P < .01). These data support the hypothesis that MB inhibits hypothalamic and not pituitary mediated mechanisms that regulate the secretion of GH.

Carbogen and nicotinamide in the treatment of bladder cancer with radical radiotherapy.

Carbogen and nicotinamide have been evaluated in a phase II study as hypoxia-modifying agents during radical radiotherapy for bladder cancer using a standard daily 20-fraction schedule. Three groups of patients have received (a) nicotinamide alone, given orally in a dose of 80 mg kg(-1) daily with 52.5 Gy in 20 fractions over 4 weeks, (b) carbogen alone, with 50 Gy in 20 fractions over 4 weeks, and (c) carbogen and nicotinamide, with 50-52.5 Gy in 20 fractions over 4 weeks. Ten patients were treated in each group. All patients completed carbogen and radiotherapy as prescribed, but only 45% completed daily nicotinamide over the 4-week treatment period. The end points of this study were acute bowel and bladder morbidity and local control at cystoscopy 6 months after treatment. An expected level of acute bowel and bladder morbidity was seen that reverted to normal in most patients by 12 weeks with no difference between the three treatment groups. Complete response rates at 6 months were seven out of ten (100%) in the nicotinamide alone group, nine out of ten (90%) in the carbogen alone group and seven out of ten (70%) in the carbogen and nicotinamide group. It is concluded that carbogen and nicotinamide may improve the results of daily fractionated radiotherapy in bladder cancer and that further evaluation is required.

Expression of rat neuronal nitric oxide synthase in Saccharomyces cerevisiae.

The rat neuronal nitric oxide synthase (nNOS) cDNA was expressed in Saccharomyces cerevisiae under the control of a hybrid PGK/GAL promoter, PAL. Galactose induction resulted in the production of a soluble 160 kDa protein that was recognised by anti-neuronal NOS antibody. NOS activity was detected by the conversion of [3H]arginine to [3H]citrulline and required the addition of the cofactors, calmodulin and tetrahydrobiopterin. The activity was inhibited by the arginine analogues NG-nitro-L-arginine and NG-nitro-L-arginine methyl ester.

The relationship of endophytic fungi to the gametophyte of the fern Schizaea pusilla.

Schizaea pusilla is a rare and threatened fern restricted in North America to acidic bogs of Nova Scotia, Newfoundland, and New Jersey. The gametophyte lives in close association with two endophytic fungi. To characterize the nature of this fern's relationship with these fungi, we introduced axenic gametophytes to bog soil for colonization. Following colonization, the endophytic fungi were isolated and reintroduced to axenic gametophytes. The gametophytes introduced to bog soil were colonized by an aseptate fungus that formed vesicles and arbuscules within the gametophyte. However, culture of colonized gametophytes produced two fungal isolates: an aseptate fungus (fungus B) and a septate fungus (fungus A). Upon reintroduction of fungal isolates to axenically grown gametophytes, the aseptate fungus demonstrated a positive growth response to the presence of the gametophytes and colonized the gametophytes without harm to the host. The septate fungus did not exhibit any specific recognition but contacted the gametophytes randomly, leaving a large percentage of the host nonviable. We propose that the relationship of the septate fungus to the gametophyte of S. pusilla is nonmycorrhizal while the relationship of the aseptate fungus to the gametophyte is mycorrhizal. Furthermore, based on lack of nutrient availability in local soils, formation of specialized structures in the gametophyte for harboring fungi, and dependence of the fern on fungal presence for completion of its life cycle, we propose that S. pusilla maintains an obligatory relationship with the aseptate mycorrhizal fungus.

Structure of the saponin adjuvant QS-21 and its base-catalyzed isomerization product by 1H and natural abundance 13C NMR spectroscopy.

The saponin QS-21, derived from the bark of the Quillaja saponaria Molina tree, has shown great potential as an adjuvant with a number of vaccines. Kinetic studies carried out to establish the stability of vaccine formulations show that commercially supplied QS-21 (primarily QS-21A) is converted slowly at pH 5.5, and rapidly at higher pH, to an equilibrium mixture of two regioisomers, QS-21A and QS-21B, in a ratio of 20:1. NMR studies show that QS-21A and QS-21B differ only in the point of attachment of the fatty acyl moiety to the fucose sugar ring. The major isomer, QS-21A, has the fatty acyl portion attached at the 4-hydroxyl group whereas the minor isomer, QS-21B, has the fatty acyl portion attached at the 3-hydroxyl group. The isomerization most likely involves ionization of the 3-hydroxy group and intramolecular acyl transfer from the 4-hydroxy group. The relative stereochemistry of the triterpene and the sugar anomeric centers is also established by NMR methods.

Instrumental comparison for the determination of cadmium and lead in calcium supplements and other calcium-rich matrices.

Three brands of Ca supplement, a laboratory-reagent grade CaCO3 and a certified reference material (International Atomic Energy Agency H-5 Animal Bone) wee analysed for Cd and Pb by four different analytical techniques, viz., anodic stripping voltammetry inductively coupled plasma mass spectrometry, flame atomic absorption spectrometry and electrothermal atomic absorption spectrometry. The Pb levels measured by the four techniques in the bone powder were within the certified Pb level in this certified reference material. Similarly, no significant differences [p less than 0.05; analysis of variance (ANOVA)] were observed in samples with Pb concentrations greater than 1 microgram g-1. However, the Pb levels in the laboratory-reagent grade CaCO3 obtained by flame atomic absorption spectrometry (0.79 micrograms g-1) averaged about three times higher than those measured by the other three techniques (i.e., 0.25 micrograms g-1). Although no significant differences (p less than 0.05; ANOVA) in Cd levels were observed within any of the samples (intra-sample variability), the Cd concentration measured in the different Ca supplements (inter-sample variability) varied by three orders of magnitude (ranging from 0.07 to 3.59 micrograms g-1).