Difficulties in the molecular diagnosis of Helicobacter rodent infections

Molecular diagnostic methods using the polymerase chain reaction (PCR) are the gold standard in Helicobacter diagnostics. Most rely on the amplification of parts of the 16S rRNA gene sequence. Therefore, the validity and accuracy of results depends heavily on the PCR design and the time of its publication because new sequences are continually being submitted to databases. Here we report the presence of helicobacter in commercially bred mice supposedly free of this infection. Furthermore, three out of six different commercial laboratories performing helicobacter testing on the same spiked faecal samples failed to detect and identify H. hepaticus. We designed a simple generic PCR assay that amplifies a 261bp amplicon spanning two of the seven variable regions in the 16S rRNA of helicobacter. Using this assay together with an established generic assay designed by Bohr [Bohr, U.R., Primus, A., Zagoura, A., Glasbrenner, B., Wex, T., Malfertheiner, P., 2002. A group-specific PCR assay for the detection of Helicobacteraceae in human gut. Helicobacter 7, 378-383] and then cloning and sequencing their products, we detected the H. hepaticus used in the study that three commercial laboratories failed to detect. We think these assays together could detect all the currently known species of helicobacter and hopefully the new ones as well. In addition, we have been able to identify different species of helicobacter and their relative proportions infecting a single animal. This information has also shown that some helicobacters may have a much broader host range than originally reported.     

Effect of pre-medication on gastroduodenoscopy in isoflurane-anesthetized cats

The pre‐medicant chosen may influence the ease with which gastroduodenoscopy (GD) is performed. The purpose of this study was to evaluate the relative ease of GD in cats under ketamine and isoflurane anesthesia after IM injection of hydromorphone (H, 0.1 mg kg^?1), hydromorphone plus glycopyrrolate (HG, 0.1 mg kg^?1 (H), 0.01 mg kg^?1 (G)), medetomidine (M, 0.03 mg kg^?1), or butorphanol (B, 0.4 mg kg^?1). Eight cats were assigned randomly to receive each treatment in a cross‐over design with at least 7 days between treatments. Twenty minutes after pre‐medication, medetomidine produced greater (p = 0.001) sedation than the other treatments when assessed, using a subjective ordinal scale. The cats were injected with ketamine (10 mg kg^?1 IM), orotracheally intubated, connected to a pediatric circle breathing system, and allowed to spontaneously breathe isoflurane in oxygen. Once end‐tidal isoflurane concentration was stable at 1.4% for 15 minutes, endoscopy was started. A single endoscopist (REG), who was unaware of the treatment used, performed all endoscopies. The endoscopist scored the difficulty of endoscopy subjectively (0–3). The significance of differences between treatments was evaluated using Friedman's test. Time for entering the stomach was 9.4 (4.7–15.9) (median (minimum–maximum)), 6.6 (5.2–11.7), 8.4 (6.3–16.5), and 7.7 (5.1–14.7) seconds and for entering the duodenum from the stomach was 20.5 (13.8–40.9), 18.2 (10.3–39.8), 20.2 (16.2–119.5), and 22.2 (11.8–83.8) seconds for H, HG, M, and B treatments, respectively. There were no significant differences in the time for, or difficulty of, endoscopy. We conclude that any of these drugs can be used satisfactorily at the doses and combinations tested to pre‐medicate cats prior to general anesthesia for GD.     

In vitro Utilization of Gold and Green Kiwifruit Oligosaccharides by Human Gut Microbial Populations

We examined the effects of whole kiwifruit on gut microbiota using an in vitro batch model of gastric-ileal digestion and colonic fermentation. Faecal fermentations of gold and green kiwifruit, inulin and water (control) digests were performed for up to 48?h. As compared to the control, gold and green kiwifruit increased Bifidobacterium spp. by 0.9 and 0.8 log(10) cfu/ml, respectively (P?相似文献   

Imaging electron wave functions of quantized energy levels in carbon nanotubes

Carbon nanotubes provide a unique system for studying one-dimensional quantization phenomena. Scanning tunneling microscopy was used to observe the electronic wave functions that correspond to quantized energy levels in short metallic carbon nanotubes. Discrete electron waves were apparent from periodic oscillations in the differential conductance as a function of the position along the tube axis, with a period that differed from that of the atomic lattice. Wave functions could be observed for several electron states at adjacent discrete energies. The measured wavelengths are in good agreement with the calculated Fermi wavelength for armchair nanotubes.     

An Improved Technique for Counting Leucocytes in Milk by the Direct Microscopic Method

An improved microscopic method for counting leucocytes in milk has been developed, which results in acceptable levels of reproducibility. To overcome the lack of uniform cell distribution within a film a stratified sampling of the total film area is examined, which prevents the counting of any one area twice. The mean between-film coefficient of variation (S.D./x X 100) for all milk samples examined which contained more than 350,000 leucocytes/ml. was less than 10%.     

Metabolic adaptations associated with irreversible glucose loss are different to those observed during under-nutrition

In this study the hypothesis that irreversible glucose loss results in an 'uncoupling' of the somatotrophic axis (increasing plasma GH levels and decreasing plasma IGF-I) was tested. During periods of negative energy balance the somatotrophic axis respond by increasing plasma GH and decreasing plasma IGF-I levels. In turn, elevated GH repartitions nutrient by increasing lipolysis and protein synthesis, and decreases protein degradation. Irreversible glucose loss was induced using sub-cutaneous injections of phloridizin. Seven non-lactating cows were treated with 8g/day phloridizin (PHZ) and seven control animals (CTRL, 0g/day), while being restricted to a diet of 80% maintenance. PHZ treatment increased urinary glucose excretion (P<0.001), resulting in hypoglycemia (P<0.001). As a response to this glucose loss, the PHZ treated animals had elevated plasma NEFA (P<0.005) and BHBA (P<0.001) levels. Average plasma insulin concentrations were not altered with PHZ treatment (P=0.059). Plasma GH was not different between the two groups (P>0.1), whereas plasma IGF-I levels decreased significantly (P<0.001) with PHZ treatment. The decline in plasma IGF-I concentrations was mirrored by a decrease in the abundance of hepatic IGF-I mRNA (P=0.005), in addition the abundance of hepatic mRNA for both growth hormone receptors (GHR(tot) and GHR(1A)) was also decreased (P<0.05). Therefore, the irreversible glucose loss resulted in a partial 'uncoupling' of the somatotrophic axis, as no increase in plasma GH levels occurred although plasma IGF-I levels, hepatic IGF-I mRNA declined, and the abundance of liver GH receptor mRNA declined.