In vitro measurement of rabbit corneal epithelial thickness using ultrahigh resolution optical coherence tomography

The objective of this study was to reproducibly measure corneal epithelial thickness centrally and at the limbus in the rabbit cornea using ultrahigh resolution optical coherence tomography (OCT). Twelve freshly enucleated New Zealand white rabbit eyes were kept in a moist chamber at 4 degrees C. An ultrahigh resolution OCT system with a spatial resolution of 1.3 microm was used to image the cornea and its component layers. The central and peripheral (limbal) regions of all the samples were scanned within 6 h of harvest in order to minimize the post-mortem degradation of the corneal epithelium. The thickness of the corneal epithelium was determined by measuring the pixel equivalents of the obtained image. Unpaired Student's t-test was used to evaluate differences. The epithelial thickness centrally was found to be 45.8 +/- 2.2 microm, and 37.6 +/- 1.4 microm at the limbus (P < 0.001). Rabbit corneal epithelium is thicker centrally than at the limbus when measured by ultrahigh resolution OCT. This technique will aid in delineating the pathophysiology of diseases of the anterior cornea.     

Relations between the goat MHC (GLA) and antibody response to diphtheria toxoid, human serum albumin and tetanus toxoid, using a twin model

SUMMARY: Norwegian dairy goats were tested for possible associations between the goat lymphocyte antigen system (GLA) and antibody response to the antigens diphtheria toxoid, human-serum albumin and tetanus toxoid. The serum titres to these antigens following immunization, and their GLA-specificities, were determined in 96 twin pairs. The antibody response and GLA specificity were compared between twins: twins sharing one or more GLA specificities showed a more similar primary-antibody response to diphtheria toxoid than twins with no common specificity. The main point in comparing twins is that the MHC specificity then serves as a marker for the complete haplotype, including MHC class-II genes. However, a gene substitution model that only tests the effect of the observed class-I alleles revealed that animals with GLA specificity Eu2 had a significantly higher anti-diphtheria response than Eu2-negative animals. ZUSAMMENFASSUNG: Untersuchungen über Beziehungen zwischen Ziegen MHC (GLA) und Antik?rperreaktion auf Diptherietoxin, humanes Serum Albumin und Tetanustoxin mit Zwillingsmodell Norwegische Milchziegen wurden in Hinblick auf m?gliche Assoziationen zwischen Ziegen Lymphozytenantigensystem (GLA) und Antik?rperreaktion auf Antigene Diptherie Toxin, humanes Serum Albumin una Tetanustoxin untersucht. Die Serumtiter nach Immunisation und die GLA-Spezifit?t wurden an 96 Zwillingspaaren erhoben und zwischen Zwillingen verglichen; Zwillinge mit ein oder mehr GLA Spezifit?ten zeigten ?hnlichere Prim?rantik?rperreaktion auf Diptherie Toxin als solche ohne gemeinsame Spezifit?t. Der wesentliche Vorteil der Zwillinge ist, da? MHC-Spezifit?t als Marker für den gesamten Haplotyp dienen kann, einschlie?lich MHC Klasse-II gene. Allerdings, ein Gen-Substitutionsmodell, das nur die Wirkungen der beobachteten Klasse-I Allele prüft, zeigte, da? Tiere mit GLA-Spezifit?t EU2 signifikant h?here Antidiptheriespiegel als EU2 negative Tiere hatten.     

Antigen-independent priming: a transitional response of bovine gammadelta T-cells to infection

Analysis of global gene expression in immune cells has provided unique insights into immune system function and response to infection. Recently, we applied microarray and serial analysis of gene expression (SAGE) techniques to the study of gammadelta T-cell function in humans and cattle. The intent of this review is to summarize the knowledge gained since our original comprehensive studies of bovine gammadelta T-cell subsets. More recently, we have characterized the effects of mucosal infection or treatment with microbial products or mitogens on gene expression patterns in sorted gammadelta and alphabeta T-cells. These studies provided new insights into the function of bovine gammadelta T-cells and led to a model in which response to pathogen-associated molecular patterns (PAMPs) induces 'priming' of gammadelta T-cells, resulting in more robust responses to downstream cytokine and/or antigen signals. PAMP primed gammadelta T-cells are defined by up-regulation of a select number of cytokines, including MIP1alpha and MIP1beta, and by antigens such as surface IL2 receptor alpha (IL-2Ralpha) and CD69, in the absence of a prototypic marker for an activated gammadelta T-cell, IFN-gamma. Furthermore, PAMP primed gammadelta T-cells are more capable of proliferation in response to IL-2 or IL-15 in the absence of antigen. PAMPs such as endotoxin, peptidoglycan and beta-glucan are effective gammadelta T-cell priming agents, but the most potent antigen-independent priming agonists defined to date are condensed oligomeric tannins produced by some plants.     

Engraftment of canine peripheral blood lymphocytes into nonobese diabetic-severe combined immune deficient IL-2R common gamma chain null mice

To study the canine immune system we generated a mouse model engrafted with canine lymphocytes using NOD SCID IL2R common gamma chain ?/? (NSG) mice as recipients (Ca-PBL-SCID). Engraftment of canine peripheral blood lymphocytes (PBLs) was determined post-injection with 10⁷ peripheral blood mononuclear cells (PBMCs) into irradiated NSG mice using flow cytometry and fluorescently labeled antibodies specific to canine helper T cells (CD45⁺ CD4⁺), cytotoxic lymphocytes (CD45⁺ CD8⁺), regulatory T cells (CD45⁺ CD4⁺ Foxp3⁺), and B cells (CD45⁺ Ig⁺ CD21_lo). Canine CD45⁺ lymphocytes were detectable as early as day 1 in the peritoneal cavity, and beginning at 9 days in the blood, bone marrow, and spleen. CD4⁺ T cells, of which Foxp-3⁺ CD25_hi cells constituted a minor percentage, were the predominant lymphocyte population at 9 days post engraftment contrasting with increasing proportions of CD8⁺ CTL's and Ig⁺ B cells beginning at 16 days. Canine immunoglobulin was initially detected in the serum of Ca-PBL-SCID mice at 9 days post-engraftment and peaked in concentration at day 36. From day 28 to 52 post-engraftment 30% of the Ca-PBL-SCID mice became markedly anemic and thrombocytopenic, yet gross and histopathologic examination of bone marrow, kidneys, spleen, liver, and intestine revealed no obvious lesions. Blood smear evaluation revealed agglutination of mature red blood cells, reticulocytes and a regenerative anemia. These findings demonstrate that NSG mice are capable of engraftment of canine PBLs yet develop graft versus host disease similar to Hu-PBL-SCID mice.     

Islet cell autoantibodies in African patients with Type 1 and Type 2 diabetes in Dar es Salaam Tanzania: a cross sectional study

Background

The aim of the present study was to assess the occurrence of glutamic acid decarboxylase autoantibodies (GADA) and insulinoma antigen 2 autoantibodies (IA2A) among patients of African origin in Dar es Salaam, Tanzania and to compare the occurrence of autoimmune mediated Type 1 diabetes with findings previously reported from the same place and from other African diabetic populations.

Methods

Two hundred and forty five patients from the diabetic clinic at Muhimbili Hospital were recruited for a cross sectional study. Patients were clinically classified into groups with Type 1 (T1D) and Type 2 diabetes (T2D); there were 94 patients with T1D and 151 with T2D. Autoantibodies for GAD and IA2 were measured with an assay based on radioligand binding. Fasting and random blood glucose, HbA1c, and C-peptide levels were also determined.

Results

Of the patients with T1D, 28 (29.8%) were GADA positive and 20 (21.3%) were IA2A positive. The overall occurrence of any autoantibody was 42.6%. The GAD and IA2 autoantibodies were detected more frequently among patients with T1D than among patients with T2D (P < 0.001). A higher autoantibody prevalence was observed with combined GADA and IA2A measurements compared to individual autoantibody measurements; 40 (42.6%) patients with T1D versus 11 (7.3%) with T2D had at least one positive autoantibody titer. There was no correlation between duration of disease and detection of autoantibodies in patients with T1D. There was a strong association with family history of diabetes among the autoantibody positive versus autoantibody negative patients with T1D (p < 0.01).

Conclusion

The prevalence of GAD and IA2 autoantibodies among African patients with T1D in Dar es Salaam was the same as that reported previously for South Africa and Ethiopia. It was much higher than the prevalence of islet cell autoantibodies (ICA) reported from the same clinic about 15 years ago. For unknown reasons the prevalence of pancreatic related autoantibodies in this African population is lower than the prevalence found among Caucasian populations.

     

Occurrence of occult bacteriuria in healthy cats

Knowledge of the occurrence of bacteriuria in adult, healthy cats is scarce in the scientific literature. A study was designed to investigate the occurrence of bacteriuria in healthy cats without current or previous signs of lower urinary tract disease. The study included 108 cats, 53 males (49.5%) and 55 females (50.5%). The cats ranged in age between 7 months and 18 years, with a mean age of 4.4 years and a median age of 4.0 years. Urine was obtained by cystocentesis from all the cats, and was submitted for bacteriological analyses. Urine and urine sediment was cultured on separate blood agar plates for quantification and species identification by standard procedures. Detection of ≥10(3)colony forming units (cfu) per ml urine was defined as significant bacteriuria. Significant bacteriuria exceeding 10(5) cfu/ml was detected in one sample with a combination of Enterococcus species and Staphylococcus species. There was no bacterial growth in the urine samples from 107 cats (99.1%). Results from our study indicate that the prevalence of bacteriuria in clinically healthy, adult cats is low. Also, that contamination of samples is rare when urine is collected by cystocentesis.     

An hPer2 phosphorylation site mutation in familial advanced sleep phase syndrome

Familial advanced sleep phase syndrome (FASPS) is an autosomal dominant circadian rhythm variant; affected individuals are "morning larks" with a 4-hour advance of the sleep, temperature, and melatonin rhythms. Here we report localization of the FASPS gene near the telomere of chromosome 2q. A strong candidate gene (hPer2), a human homolog of the period gene in Drosophila, maps to the same locus. Affected individuals have a serine to glycine mutation within the casein kinase Iepsilon (CKIepsilon) binding region of hPER2, which causes hypophosphorylation by CKIepsilon in vitro. Thus, a variant in human sleep behavior can be attributed to a missense mutation in a clock component, hPER2, which alters the circadian period.     

Ceramic thin-film formation on functionalized interfaces through biomimetic processing

Processing routes have been developed for the production of thin ceramic films through precipitation from aqueous solutions. The techniques are based on crystal nucleation and growth onto functionalized interfaces. Surface functionalization routes have been developed by the mimicking of schemes used by organisms to produce complex ceramic composites such as teeth, bones, and shells. High-quality, dense polycrystalline films of oxides, hydroxides, and sulfides have now been prepared from "biomimetic" synthesis techniques. Ceramic films can be synthesized on plastics and other materials at temperatures below 100 degrees C. As a low-temperature process in which water rather than organic solvents is used, this synthesis is environmentally benign. Nanocrystalline ceramics can be produced, sometimes with preferred crystallite orientation. The direct deposition of high-resolution patterned films has also been demonstrated. The process is well suited to the production of organic-inorganic composites.     

BIOECONOMICS OF CAPTURE-BASED AQUACULTURE OF COD (GADUS MORHUA)

This study formulates and parameterizes a bioeconomic model of capture-based aquaculture (CBA) of cod (Gadus morhua). The model is solved for the optimal harvest pattern and calculates economic profit and net present value for a model farm. The biological sub-model incorporates knowledge from interviews with existing farmers, research trials and existing cod aquaculture literature. Economic components are obtained from interviews and sales statistics from exporters. A farm of the modeled scale is likely to influence market prices, hence sales prices were estimated assuming a supply response based on the price elasticity. Taking into account the opportunity cost of selling the fish directly, NPV is found to be marginally positive. Sensitivity analysis revealed that profitability is sensitive to changes in several parameters. Hence, further research is valuable and care should be taken when considering investments in cod CBA.