Interleukin-23 is critical for full-blown expression of a non-autoimmune destructive arthritis and regulates interleukin-17A and RORγt in γδ T cells

Introduction  

Interleukin (IL)-23 is essential for the development of various experimental autoimmune models. However, the role of IL-23 in non-autoimmune experimental arthritis remains unclear. Here, we examined the role of IL-23 in the non-autoimmune antigen-induced arthritis (AIA) model. In addition, the regulatory potential of IL-23 in IL-17A and retinoic acid-related orphan receptor gamma t (RORγt) expression in CD4⁺ and TCRγδ⁺ T cells was evaluated systemically as well as at the site of inflammation.     

HistoChoice® as an alternative to formalin fixation of undecalcified bone specimens

We compared histochemical and immunohistochemical staining as well as fluorochrome labeling in murine bone specimens that were fixed with 10% neutral buffered formalin to those fixed with HistoChoice®. We showed that sections from undecalcified tibiae fixed for 4 h in HistoChoice® resulted in enhanced toluidine blue and Von Kossa histochemical staining compared to formalin fixation. HistoChoice® produced comparable or improved staining for alkaline phosphatase. Acid phosphatase localization was better in formalin fixed specimens, but osteoclasts were visuralized more easily in HistoChoice® fixed specimens. As expected, immunohistochemical labeling was antibody dependent; some antibodies labeled better in HistoChoice® fixed specimens while others were better in formalin fixed specimens. Toluidine blue, Von Kossa, and alkaline phosphatase staining of sections fixed for 12 h produced sections that were similar to 4 h fixed sections. Fixation for 12 h preserved acid phosphatase activity better. Increasing fixation to 12 h affected immunolocalization differentially. Bone sialoprotein labeling in HistoChoice® fixed specimens was comparable to formalin fixed samples. On the other hand, after 12 h formalin fixation, osteocalcin labeling was comparable to HistoChoice®. For most histochemical applications, fixing murine bone specimens for 4 h with HistoChoice® yielded superior staining compared to formalin fixation. If immunohistochemical localization is desired, however, individual antibodies must be tested to determine which fixation process retains antigenicity better. In addition, there was no detectable difference in the intensity of fluorochrome labeling using either fixative. Finally, fixation duration did not alter the intensity of labeling.     

Proliferative activity in the cerebellar external granular layer evaluated by bromodeoxyuridine labeling

We have optimised an indirect immunoperoxidase technique demonstrating bromodeoxyuridine (BrdU) incorporation into dividing cells for cerebellar tissue sections of four-day-old rats injected with this marker. This permits confident identification of granule-cell precursors engaged in DNA synthesis in the external granular layer of the developing cerebellum. Preservation of BrdU immunoreactivity is attained using methanol/acetic acid fixation and different pretreatments before immunostaining, while unlabeled nuclei can be recognized clearly after Feulgen or hematoxylin counterstaining. We established conditions to ensure satisfactory BrdU uptake without affecting cell-cycle progression during the postlabeling time period. The dose of BrdU employed provides saturation S-phase labeling from at least 1 h after BrdU delivery. Various kinetic parameters and phase durations have been determined in experiments involving a single injection or cumulative labeling sequences, and the cycle time was calculated based on two models of generative behavior: steady-state and exponential growth. The working hypothesis of steadystate kinetics can be adopted successfully if the existence of neuroblasts with different proliferation rates is taken into account.     

Electrophysiological identification of the stimulatory and interactive components of a complex odorant

We are studying the olfactory system of the spiny lobster, Panulirusargus, to understand how chemical mixtures are coded. By monitoringthe activity of high-order interneurons which carry olfactoryinformation out of the brain, we have identified the componentsof a natural food of lobsters which contribute to the activityof the mixture by being either stimulatory by themselves orinteractive (suppressive or synergistic) with other componentsin the mixture. The results demonstrate that virtually all ofthe activity of this complex odorant resides in 15 stimulatoryand suppressive components, and that mixture suppression isa prevalent feature of chemosensory processing in the olfactorypathway of the spiny lobster.     

Differential distribution of factors involved in pre-mRNA processing in the yeast cell nucleus.

The yeast cell nucleus has previously been shown to be divided into two regions by a variety of microscopic approaches. We used antibodies specific for the 2,2,7-trimethylguanosine cap structure of small nuclear ribonucleic acids (snRNAs) and for a protein component of small nuclear ribonucleoprotein particles to identify the distribution of small nuclear ribonucleoprotein particles within the yeast cell nucleus. These studies were performed with the fission yeast Schizosaccharomyces pombe and the budding yeast Saccharomyces cerevisiae. By using immunofluorescence microscopy and immunoelectron microscopy, most of the abundant snRNAs were localized to the portion of the nucleus which has heretofore been referred to as the nucleolus. This distribution of snRNAs is different from that found in mammalian cells and suggests that the nucleolar portion of the yeast nucleus contains functional domains in addition to those associated with RNA polymerase I activity.     

Responses of olfactory receptor neurons in the spiny lobster to binary mixtures are predictable using a noncompetitive model that incorporates excitatory and inhibitory transduction pathways

Coding of binary mixtures by a population of olfactory receptor neurons in the spiny lobster (Panulirus argus) was examined. Extracellular single-unit responses of 50 neurons to seven compounds and their binary mixtures were recorded. The ability of a noncompetitive model with correction for binding inhibition to predict responses to mixtures based on responses to their components was compared with the predictive abilities of other models. This model assumes that different compounds activate different transduction processes in the same neuron leading to excitation or inhibition, and it includes a term quantifying the degree to which binding of an odorant to its receptor sites is inhibited by other compounds. The model accurately predicted the absolute response magnitude of the population of neurons for 13 of 15 mixtures assessed, which is superior to the predictive power of any of the other models. The model also accurately predicted the across neuron patterns generated by the binary mixtures, as evaluated by multidimensional scaling analysis. The results suggest that there is no emergence of unique qualities for binary mixtures relative to components of these mixtures.Abbreviations AMP or A adenosine-5-monophosphate - ANP across neuron pattern - ARM absolute response magnitude - ASW artificial sea water - Bet or B betaine - Cys or C L-cysteine - Glu or G L-glutamate - MDS multidimensional scaling - MID mixture interaction distance - NC model noncompetitive model - NCBI model noncompetitive model with correction for binding inhibition - C model competitive model - CBI model competitive model with correction for binding inhibition - MEC more effective component - NH ₄ or N ammonium chloride - ORN olfactory receptor neuron - Suc or S DL-succinate - Tau or T taurine