Cluster analysis of Anopheles Stephensi fourth instar larvae behavior to explore sequential organisation of larvae movement in a microcosm

Experiments were conducted on 20 fourth instar Anopheles stephensi larvae to explore behavior organisation. Twenty fourth instar larvae were placed in a glass aquarium and filmed using a handy cam video recorder and the recordings analysed in a laptop computer. Data of transitions from one behavior to another for all observations were collated into a matrix of preceding and succeeding behaviors to study sequential organisation and relationship among behaviors. Significant testing for first-order transition was done using G test at P< 0.005 and a kinematic graph constructed from significant transitions. A time budget and transition frequency data constructed for each behavior were subjected to cluster analysis to explore relationship between the behaviors. Result of the analysis showed that fourth instar Anopheles stephensi larvae behaviors occur in clusters in specific locations in their aquatic environment. Furthermore, the sequential organisation of behaviors is influenced by behavior frequency and the amount time a larva spent doing that behavior. When food is kept constant, other factors such as gas exchange requirements, behavior variation due to day-night cycle, presence of a predator, interlarval competition for food and the size of the aquarium (depth and width) maybe determining behavior organization.

A suppressive oligodeoxynucleotide inhibits ocular inflammation.

Synthetic oligodeoxynucleotides (ODN) expressing 'suppressive' TTAGGG motifs down-regulate a variety of proinflammatory and T helper type 1 (Th1)-mediated pathological immune responses. The ability of the archetypal suppressive ODN A151 to inhibit ocular inflammation was examined in two murine models: experimental autoimmune uveitis, induced by immunization with a retinal antigen, interphotoreceptor retinoid-binding protein (IRBP) and adoptively transferred ocular inflammation, induced by transferring Th1 cells specific to hen egg lysozyme (HEL) into recipient mice that express HEL in their eyes. A151 treatment suppressed the inflammation in both models. In addition, A151 inhibited IRBP-specific cytokine production and lymphocyte proliferation in mice immunized with IRBP. These findings suggest that suppressive ODN affects both afferent and efferent limbs of the immunopathogenic process and may be of use in the treatment of autoimmune ocular inflammation.

Induction of suppressors of cytokine signaling (SOCS) in the retina during experimental autoimmune uveitis (EAU): potential neuroprotective role of SOCS proteins.

Suppressors of cytokine signaling (SOCS) are implicated in immunopathogenic mechanisms of autoimmune diseases. We show here that SOCS expression in retina is temporarily correlated with progression of experimental autoimmune uveitis (EAU), an organ-specific autoimmune disease that serves as model of human uveitis. Peak of EAU correlates with highest SOCS genes expression while disease resolution coincides with their down-regulation. Surprisingly, SOCS5 is constitutively expressed in retina. SOCS5 expression increases significantly during EAU and remains elevated even after disease resolution. Our data suggest that cytokine-inducible SOCS members may be involved in negative regulation of inflammatory cytokines activities during EAU, while constitutively expressed SOCS5 may have neuroprotective functions.

High-dose rabeprazole-amoxicillin versus rabeprazole-amoxicillin-metronidazole as second-line treatment after failure of the Japanese standard regimen for Helicobacter pylori infection.

BACKGROUND: There is currently no optimal second-line treatment after failure of Helicobacter pylori triple therapy. AIM: To determine effective salvage therapy after failure of lansoprazole-amoxicillin-clarithromycin. METHODS: After failure of lansoprazole-amoxicillin-clarithromycin 123 out-patients were randomized to receive either 2-week rabeprazole (20 mg b.d.) + amoxicillin (1000 mg b.d.) (RA group) or 1-week rabeprazole (10 mg b.d.) + amoxicillin (750 mg twice b.d.) + metronidazole (250 mg b.d.) (RAM group). Eradication was assessed by the 13C-urea breath test. We also evaluated cytochrome p450 (CYP) 2C19 genotype status, determined by polymerase chain reaction - restriction fragment length polymorphism, and susceptibility to clarithromycin and metronidazole. RESULTS: On an intention-to-treat basis, H. pylori infection cure was achieved in 37 of 63 (59%) patients in the RA group and in 49 of 60 (82%) patients in the RAM group. Per protocol-based eradication rates in the RA and RAM groups were 66% (37/56) and 88% (49/56), respectively. In both analytic sets there were significant differences between the treatment groups (P < 0.01 in each). Mild adverse events were observed in eight and five patients from the RA and RAM groups, respectively. Genetic predisposition of CYP2C19 and antibiotic resistance did not influence the treatment outcome either regimen. CONCLUSIONS: The rabeprazole + amoxicillin + metronidazole therapy yielded satisfactory results. In contrast, the cure rate in high-dose rabeprazole + amoxicillin was below an acceptable level.

Application of denaturing gradient gel electrophoresis (DGGE) to the analysis of microbial communities of subgingival plaque.

OBJECTIVES: Denaturing gradient gel electrophoresis (DGGE) was applied to the microbiologic examination of subgingival plaque. MATERIALS AND METHODS: The PCR primers were designed from conserved nucleotide sequences on 16S ribosomal RNA gene (16SrDNA) with GC rich clamp at the 5'-end. Polymerase chain reaction (PCR) was performed using the primers and genomic DNAs of typical periodontal bacteria. The generated 16SrDNA fragments were separated by denaturing gel. RESULTS: Although the sizes of the amplified DNA fragments were almost the same among the species, 16SrDNAs of the periodontal bacteria were distinguished according to their specific sequences. The microflora of clinical plaque samples were profiled by the PCR-DGGE method, and the dominant 16SrDNA bands were cloned and sequenced. Simultaneously, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia were detected by an ordinary PCR method. In the deep periodontal pockets, the bacterial community structures were complicated and P. gingivalis was the most dominant species, whereas the DGGE profiles were simple and Streptococcus or Neisseria species were dominant in the shallow pockets. The species-specific PCR method revealed the presence of A. actinomycetemcomitans, P. gingivalis and P. intermedia in the clinical samples. However, corresponding bands were not always observed in the DGGE profiles, indicating a lower sensitivity of the DGGE method. CONCLUSION: Although the DGGE method may have a lower sensitivity than the ordinary PCR methods, it could visualize the bacterial qualitative compositions and reveal the major species of the plaque. The DGGE analysis and following sequencing may have the potential to be a promising bacterial examination procedure in periodontal diseases.

Application of linear solvation energy relationships to polymeric pseudostationary phases in micellar electrokinetic chromatography.

Polymerized sodium 11-acrylamidoundecanoate (poly(Na 11-AAU)) was used as a pseudostationary phase (PSP) for micellar electrokinetic chromatography to separate uncharged compounds. The polymer PSP showed signifcantly different solute migration behaviors from conventional micelles including sodium dodecyl sulfate and poly (sodium 10-undecylenate), giving high separation efficiencies (>200000 theoretical plates/m). Linear solvation energy relationships were used to evaluate and characterize the chemical interactions that influence the retention behavior in the poly (Na 11-AAU) micellar system. It was found that the solute volume and solute hydrogen bond basicity mainly influenced the retention. The characteristic feature of the poly (Na 11-AAU) micellar system is that the micelle has a significantly higher capacity for dipole-dipole and dipole-induced dipole interactions as well as a slightly higher capacity for electron pair interactions than the aqueous phase. Due to its unique selectivity, the poly(Na 11-AAU) micellar system would become an attractive new option for selectivity optimization on methods development.

Macromolecular surfactant as a pseudo-stationary phase in micellar electrokinetic capillary chromatography.

We examined polymers of sodium 11-acrylamidoundecanoate [poly(Na 11-AAU)] with a very high molecular mass (>10(6)) for their potential use as a pseudo-stationary phase in micellar electrokinetic capillary chromatography (MEKC). Size-exclusion chromatography and capillary electrophoresis studies reveal that the polymers are highly charged, and have a densely packed chain structure. For aromatic compounds, the polymeric surfactant showed significantly different selectivity than sodium dodecyl sulfate (SDS). It was suggested that one molecule of poly(Na 11-AAU) forms one micelle. The structural stability of this pseudo-stationary phase permitted its use with relatively high percentages of organic modifiers in the buffer medium, allowing the separation of highly hydrophobic compounds which are difficult to analyze by conventional MEKC with SDS.

Negative effects of chronic inflammatory periodontal disease on diabetes mellitus.

Periodontal disease is the result of a complex interplay of bacterial infection and host responses, and is often modified by various systemic diseases such as diabetes mellitus. Such diseases are capable of affecting the periodontium and/or the treatment of periodontal disease. However, recent research has changed our concept of how periodontal disease should be treated. Here we present several concerns directed towards the periodontal therapy of patients with diabetes mellitus based on our studies. When treating periodontitis patients who have diabetes mellitus it is important to consider the type of diabetes. Patients with non-insulin dependent diabetes mellitus can be further classified according to the degree of insulin resistance, since recent epidemiological studies have suggested that successful anti-microbial therapy might result in improved insulin resistance in highly insulin resistant patients. Because the major contributing factor for insulin resistance is currently considered to be the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha), and because periodontal surgery may cause transient bacteremia which may up-regulate the serum TNF-alpha level, which in turn suppresses insulin action, patients should be strictly treated non-surgically and their serum TNF-alpha levels should be periodically monitored. On the other hand, diabetic patients positive for serum anti-glutamate decarboxylase auto-antibody should be examined for the source of this antibody, since 1) gingival and periodontal ligament fibroblasts were found to express glutamate decarboxylase, and 2) some otherwise healthy periodontitis patients develop anti-glutamate decarboxylase antibody. Thus, chronic periodontitis may influence the level of this antibody which is widely used as a predictive marker for slowly progressive insulin dependent diabetes mellitus. Not only is periodontal disease thereby affected by systemic diseases, but carefully managed periodontal therapy may also have a positive effect on the general health of patients with systemic diseases.

Dynamic contrast-enhanced MRI of Implanted VX2 tumors in rabbit muscle: comparison of Gd-DTPA and NMS60.

We studied the dynamics of injected contrast enhancement in implanted VX2 tumors in rabbit thigh muscle. We compared two contrast agents Gd-DTPA and NMS60, a novel gadolinium containing trimer of molecular weight 2.1 kd. T1-weighted spin echo images were acquired preinjection and at 5-60 min after i.v. injection of 0.1 mmol/kg of agent. Dynamic T1-weighted SPGR images (1.9 s/image) were acquired during the bolus injection. Male NZW rabbits (n = 13) were implanted with approximately 2 x 10(6) VX2 tumor cells and grew tumors of 28+/-27 mL over 12 to 21 days. NMS60 showed significantly greater peak enhancement in muscle, tumor rim, and core compared to DTPA in both T1-weighted and SPGR images. NMS60 also showed delayed peak enhancement in the dynamic scans (compared to Gd-DTPA) and significantly reduced leakage rate constant into the extravascular space for tumor rim (K21 = 5.1 min(-1) vs. 11.5 min(-1) based on a 2 compartment kinetic model). The intermediate weight contrast agent NMS60 offers greater tumor enhancement than Gd-DTPA and may offer improved regional differentiation on the basis of vascular permeability in tumors.

Fritless packed columns for capillary electrochromatography: separation of uncharged compounds on hydrophobic hydrogels.

A novel column is described that does not require frits to keep packing material within a capillary. A continuous bed is prepared in situ in aqueous solution by radical copolymerization of N-isopropylacrylamide and 2-acrylamido-2-methylpropanesulfonic acid (the resultant gel is denoted poly(AMPS-co-IPAAm). N,N'-Methylenebisacrylamide is used for cross-linking. On the application of an electrical field, electroosmotic flow (EOF) is developed in the bed along the capillary, where fluid propulsion would be otherwise difficult to achieve. The resultant EOF transports neutral compounds through the column without forcing the gel out of the capillary. Examination of the fluid motion in the continuous bed using a video microscope system and an image processor shows a relatively flat flow profile of EOF. The bed functions as the stationary phase for reversed-phase capillary electrochromatography (CEC). This new approach is an alternative to packed capillary columns which have been used previously in CEC. A high efficiency is obtained for a steroid which is separated on a 4.0% total monomer concentration (T), 10.0% degree of cross-linking (C), and 10.0% mole fraction of AMPS in the total monomer (S), poly(AMPS-co-IPAAm) column. A mixture of polyaromatic hydrocarbons is separated on a 6.9% T, 5.8% C, and 5.5% S poly(AMPS-co-IPAAm) column. The capacity factor of benzo[a]pyrene increases from 0.63 to 1.91 as the acetonitrile content in a Tris-boric acid buffer is decreased from 45 to 30% (v/v). The run-to-run RSD of analyte migration time is less than 0.73%, and the day-to-day RSD is acceptable. Potential benefits of this approach are also mentioned.

Noradrenergic and neuropeptide Y-immunoreactive nerves in the pancreatic islets of spontaneously hypertensive rats.

Innervation by noradrenergic (NA) and neuropeptide Y-immunoreactive (NPY-IR) nerves to the pancreatic islets and the arteries supplying the islets was studied by immunocytochemistry in 6- and 20-week-old spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). Since both the NA and NPY-IR nerves follow the intra- and interlobular small arteries and innervate the islets, we examined mainly the innervation of intra- and interlobular small arteries by these nerves. The NA nerve density of the intralobular small arteries supplying the islets was significantly more extensive in 6-week-old SHR than in age-matched WKY (7.11 +/- 1.78 vs. 4.04 +/- 0.71 x 10(-2) micron/micron2, P < 0.001), and the NPY-IR nerve density of the intralobular small arteries was significantly more extensive in SHR than in WKY at 6 weeks of age (7.68 +/- 2.28 vs. 5.69 +/- 0.91 x 10(-2) micron/micron2, P < 0.05). The interlobular small arteries branching off the intralobular small arteries in SHR were also more innervated by NA (P < 0.05) and NPY-IR nerves (P < 0.001) than by those in WKY at 6 weeks of age. At 20 weeks of age, the NA and NPY-IR nerve densities of the intra- and interlobular small arteries were higher in SHR than in WKY, and showed an increase with age in both strains. In addition, the islet innervation by these nerves in cross-section appeared denser in SHR than in WKY.(ABSTRACT TRUNCATED AT 250 WORDS)

Phlegmonous enteritis in alcoholic fatty liver.

Phlegmonous enteritis is a rare inflammatory bowel disease. A 52-yr-old man with a history of alcoholic abuse was admitted to the hospital for an acute abdomen and died of septicemia and its complications. Autopsy revealed phlegmonous inflammation of the ileum and severe fatty liver. Numerous Gram-negative rod bacilli were demonstrated in the ileal mucosa. Shortened villi and decreased lysozyme activity of Paneth cells in the small intestine might be results of chronic alcohol ingestion. The relationship between phlegmonous enteritis and alcoholic abuse was strongly suggested in this case.