<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Effect of dorsal hip loading,sedation, and general anesthesia on the dorsolateral subluxation score in dogs

OBJECTIVE: To determine whether dorsal loading of the pelvis and type of chemical restraint affected the dorsolateral subluxation (DLS) score of dog hips. STYDY DESIGN: In vivo testing of diagnostic method. ANIMALS: Labrador retrievers, Greyhounds, and crosses between both breeds (n = 119 dogs). METHODS: Dorsal load was applied to the hips through a strap that was placed over the hips, and the peak vertical and steady-state vertical, ground-reaction forces exerted on the stifles were measured in the DLS position. The DLS score was measured with their hips under dorsal load and compared with the DLS score without load. For 24 dogs, the DLS score was measured both under chemical restraint (medetomidine) and under general anesthesia. Wilcoxon-signed rank test and paired t test was used to compare effects of dorsal load and restraint method on the left and right DLS hip score, and P <.05 was considered significant. RESULTS: Both stifles together accepted median 15% body weight in the vertical plane in the unloaded steady state. Loading the hips increased the steady-state, vertical ground-reaction force to 34% body weight. At 8 months of age, loading significantly decreased the DLS score from 65% +/- 11% (mean +/- SD) to 60% +/- 12% for left hips and 66% +/- 11% to 63% +/- 13% for right hips. The left hip DLS score was affected by load more consistently than the right hip. In general, loading the hips decreased the DLS score in all breeds by 4% to 6% except for the greyhounds, in which the DLS score was unchanged by dorsal loading. When grouped by their DLS scores, load significantly decreased DLS scores in dogs with unloaded scores greater than 55% (nondysplastic), whereas DLS scores of dogs with unloaded scores between 55% and 45% (dysplastic) and less than 45% did not change significantly with load. Unloaded scores did not change significantly when compared under sedation versus general anesthesia. However, load significantly decreased the DLS score under general anesthesia for the left hip but not for the right hip. CONCLUSIONS: Less than half of the vertical ground-reaction force normally exerted by the hindlegs of a standing dog was sufficient to induce hip subluxation in the DLS position. Although dorsal loading approximately doubled the steady-state, vertical ground-reaction force, the decrease in the DLS score under load was never greater than 6%. Furthermore, the DLS scores of dogs most likely to be dysplastic (based on DLS scores <45%) were not affected by load, whereas load slightly decreased DLS cores of dogs with DLS scores greater than 55%. CLINICAL RELEVANCE: External influences can change the DLS score significantly, but the magnitude of change is unlikely to be of clinical importance, making the method useful as a field screening test for both dysplastic and nondysplastic hips in young dogs.     

Cytochrome P-based resistance mechanism and pyrethroid resistance in the field Anopheles albimanus resistance management trial

The relative rates of cytochrome P⁴⁵⁰ selection in southern Mexican Anopheles albimanus populations were investigated during a 3 years indoor residual house spraying intervention with a pyrethroid (PYR) or DDT, a mosaic of organophosphate (OP)-PYR, and the annual rotation of OP-PYR-carbamate (CARB). This insecticide resistance mechanism, initially evenly spread in the mosquito population, correlated with PYR resistance during the second treated year, when cytochrome P⁴⁵⁰ contents increased in most villages of the PYR, rotation and mosaic schemes. However, by the third year, mean cytochrome P⁴⁵⁰ contents declined to susceptible levels in mosquitoes of the rotation and one mosaic group but not in the PYR-treated villages. In DDT-treated villages, a continuous decrement of cytochrome P⁴⁵⁰ levels occurred since the first treatment year, and susceptible levels were observed at the end of the intervention. Most correlations of cytochrome P⁴⁵⁰ levels and PYR resistance were lost during the third year, indicating that another mechanism evolved in resistant mosquito populations.     

Evaluation of microbial culture techniques for the isolation of Pythium insidiosum from equine tissues.

The purpose of this study was to evaluate the effects of sample handling, storage, and culture techniques on the isolation of Pythium insidiosum from infected equine tissues. Tissue and kunker samples obtained immediately posteuthanasia from a horse with subcutaneous pythiosis were used to assess the effects of sample type (kunkers vs. tissues), media type (selective vs. nonselective), storage technique, and storage time on P. insidiosum isolation rate. Overall, isolation rates were higher from fresh kunkers (94.6%) and stored kunkers (76.4%) than from fresh tissues (8.3%) or stored tissues (4.6%). Isolation of P. insidiosum also occurred more often on antibiotic-containing media than on nonselective media for both fresh and stored samples. For samples that were stored for 1-3 days prior to culture, P. insidiosum isolation rates were highest for the following techniques: kunkers stored at room temperature and plated on selective media (100%), kunkers stored at 4 C and then plated on either nonselective (91.7%) or selective (95.8%) media, kunkers stored on cold packs and then plated on either nonselective (93.8%) or selective (100%) media, kunkers stored in ampicillin solution and plated on selective media (100%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). For samples stored for 4-5 days, P. insidiosum isolation rates were highest for kunkers stored at 4 C and then plated on either nonselective (81.3%) or selective (87.5%) media, kunkers stored in ampicillin solution and then plated on selective media (87.5%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). Results of this study suggest that optimal isolation rates of P. insidiosum from infected equine tissues are achieved by culturing fresh kunkers on selective media. For samples that cannot be processed immediately, acceptable handling techniques include storage at room temperature for up to 3 days, refrigeration for up to 5 days, shipping on cold packs, and storage in antibiotic solution, each combined with subsequent inoculation on selective media.     

Metallomics approach to trace element analysis in ustilago maydis using cellular fractionation, atomic absorption spectrometry, and size exclusion chromatography with ICP-MS detection

Huitlacoche is the ethnic name of the young fruiting bodies of Ustilago maydis, a common parasite of maize. In Mexico and other Latin American countries, this fungus has been traditionally appreciated as a local delicacy. In this work a metallomics approach was used with the determination of eight elements in huitlacoche by electrothermal atomic absorption spectrometry as one facet of this approach. The results obtained indicated relatively lower concentrations of commonly analyzed metals, as referred to the data reported for other mushroom types. This effect was ascribed to different accessibilities of elements, depending on fungus substrate (lower from plant than from soil). Subcellular fractionation was accomplished by centrifugation of cell homogenates suspended in Tris-HCl buffer. Recoveries of the fractionation procedure were in the range of 71-103%. For six elements (Cr, Cu, Fe, Mn, Ni, and Pb), the mean relative contributions in cytosol, cell walls, and mixed membrane fraction were 50.7, 48.2, and 1.1% respectively. To attain the molecular weight distribution of compounds containing target elements as an additional aspect of the metallomics approach, the fungus extract (1% sodium dodecyl sulfate in Tris-HCl, 30 mmol L(-)(1), pH 7.0) was analyzed by size exclusion chromatography with UV and ICP-MS detection. With spectrophotometric detection (280 nm), the elution of high molecular weight compounds was observed in the form of one peak (MW > 10 kDa), and several lower peaks appeared at higher retention times (MW < 10 kDa). On ICP-MS chromatograms, a coelution of (59)Co, (63)Cu, (57)Fe, (202)Hg, (60)Ni, and (80)Se with the first peak on the UV chromatogram was clearly observed, indicating that a fraction of each element incorporated with high molecular weight compounds (12.7, 19.8, 33.7, 100, 19.4, and 45.8%, respectively, based on the peak area measurements). From a comparison of (80)Se and (33)S chromatograms (for sulfur analysis, the extract was obtained in the absence of SDS), both elements coeluted with the first UV peak, but their lower molecular weight compounds were apparently different. These findings may contribute to a better understanding of the accumulation of elements in mushrooms.     

Survey onYersinia Enterocolitica and related bacteria in shellfish

Results from examining shellfish collected from ‘safe’ (2–34Escherichia coli/100 mL) and ‘unsafe’ (> 34E. coli/100 mL) waters for the presence ofYersinia enterocolitica and related bacteria are reported. The recovery of these microorganisms was low in bivalve mollusks destined for food consumption, high in those derived from prohibited areas. No human pathogenic bio-serogroup f Yersinia was found during the entire survey.     

Noradrenergic stimulation of cyclic adenosine monophosphate in rat Purkinje neurons: an immunocytochemical study

A specific immunofluorescent histochemical method for cyclic adenosine monophosphate was used to study rat cerebellum. After topical treatment with norepinephrine or stimulation of norepinephrine-containing afferents from locus coeruleus, there was a striking increase in the number of Purkinje cells with strong cyclic adenosine monophosphate reactivity. Other putative inhibitory transmitters had no significant effect on staining of Purkinje cells. The results provide the first histochemical support for the hypothesis that cyclic adenosine monophosphate can be generated postsynaptically in central neurons in response to noradrenergic stimuli.     

Aerobic bacterial flora of the nasal cavity in Gulf of California sea lion (Zalophus californianus) pups

Nasal swab samples from clinically healthy California sea lions pups (Zalophus californianus) from six different reproductive rookeries in the Gulf of California were collected to determine the type and frequency of the representative aerobic bacterial microflora of their nasal mucosa. A total of 114 samples were examined and 100 bacterial isolates were identified and typified by microbiological and biochemical standard tests. Fifty four isolates corresponded to Gram positive bacteria (54%) and 46 isolates to Gram negative bacteria (46%). Fifteen bacterial genera were identified, including Micrococcus, Arcanobacterium, Corynebacterium, Moraxella, Neisseria, Escherichia, Kurthia, Acinetobacter, Staphylococcus, Brevibacillus, Bacillus, Klebsiella, Stenotrophomonas, Pseudomonas and Aeromonas. The most frequently isolated genera were Moraxella (24%), Micrococcus (18%), and Corynebacterium (15%). These results show the presence in the nasal cavity of sea lions of several microorganisms. Although considered part of the normal microflora, they may also be opportunistic pathogens for their hosts and may act as a potential natural sentinel of environmental changes.     

East Coast fever immunisation field trial in crossbred dairy cattle in Hanang and Handeni districts in northern Tanzania

East Coast fever (ECF) causes considerable mortality and production losses in the Tanzania smallholder dairy sector and limits the introduction of improved dairy breeds in areas where the disease is present. The infection and treatment method (ITM) was adopted by smallholder dairy farms for ECF immunisation in Hanang and Handeni districts of Tanzania. This study recorded incidence rates for ECF and other tick-borne diseases (TBDs) for ECF-immunised and non-immunised cattle between 1997 and 2000. Approximately 80% of smallholder households from both sites (n?=?167) participated in this longitudinal study, with immunisations carried out at the request of the livestock owners. Efficacy of ITM for preventing ECF cases in these crossbred dairy cattle was estimated at 97.6%, while that for preventing ECF deaths was 97.9%. One percent of the cattle developed clinical ECF as a result of immunisation. Since ECF immunisation permits a reduction in acaricide use, an increase in other TBDs is a potential concern. Sixty-three percent of farmers continued to use the same acaricide after immunisation, with 80% of these reducing the frequency of applications. Overall, 78% of farmers increased the acaricide application interval after immunisation beyond that recommended by the manufacturer, resulting in annual savings in the region of USD 4.77 per animal. No statistical difference was observed between the immunised and non-immunised animals in the incidence of non-ECF TBDs. However, immunised animals that succumbed to these diseases showed fewer case fatalities. ITM would therefore appear to be a suitable method for ECF control in Tanzania's smallholder dairy sector.