Overlapping BXSB congenic intervals, in combination with microarray gene expression, reveal novel lupus candidate genes.

The BXSB mouse strain is an important model of glomerulonephritis observed in systemic lupus erythematosus (SLE). Linkage studies have successfully identified disease-susceptibility intervals; however, extracting the identity of the susceptibility gene(s) in such regions is the crucial next step. Congenic mouse strains present a defined genetic resource that is highly amenable to microarray analysis. We have performed microarray analysis using a series of chromosome 1 BXSB congenic mice with partially overlapping disease-susceptibility intervals. Simultaneous comparison of the four congenic lines allowed the identification of expression differences associated with both the initiation and progression of disease. Thus, we have identified a number of novel SLE disease gene candidates and have confirmed the identity of Ifi202 as a disease candidate in the BXSB strain. Sequencing of the promoter regions of Gas5 has revealed polymorphisms in the BXSB strain, which may account for the differential expression profile. Furthermore, the combination of the microarray results with the different phenotypes of these mice has allowed the identification of a number of expression differences that do not necessarily map to the congenic interval, but may be implicated in disease pathways.

Effect of ARA9 on dioxin receptor mediated transcription.

The dioxin (Aryl hydrocarbon) receptor (DR) is a unique bHLH transcription factor which is activated by binding of planar aromatic hydrocarbons typified by dioxin (TCDD). The active receptor is key to metabolism of aryl hydrocarbon xenobiotics by being a potent inducer of CYP1A1 gene activity. Chlorinated dioxins are inert to metabolism and initiate multifarious toxicities, including potent tumour promotion. These ill-effects are mediated by the activated DR and we are studying the mechanisms by which the ligand binding domain of the DR controls activity of the protein. The DR ligand binding domain resides within a PAS (Per/Arnt/Sim homology) region which is contiguous with the bHLH. The latent bHLH/PAS dioxin receptor (DR) is found in the cytoplasm of most mammalian cell types in a complex with heat shock protein 90, a novel immunophilin like protein termed ARA9/XAP2/AIP, and the co-chaperone p23. Here we use antisense ARA9 constructs to reveal that in the absence of ARA9, the DR is unable to form a transcriptionally active complex. Co-expression of antisense ARA9 with a form of the DR which is constitutively targeted to the nucleus leads to dramatically decreased levels of the nuclear DR protein, implying that ARA9 may function beyond its currently proposed role in cytoplasmic retention of the latent DR.

Flow pattern dynamics in convecting liquid helium.

We present experimental data which correlate thermal measurements and flow visualization in convecting liquid 4He. For a small range R(C)R1, generating thermal oscillations as in earlier reports. At higher R values the time dependence becomes aperiodic with the surprising appearance of spiral-defect chaos at an aspect ratio smaller than has previously been reported.

Multiple roles of ligand in transforming the dioxin receptor to an active basic helix-loop-helix/PAS transcription factor complex with the nuclear protein Arnt.

The dioxin receptor is a ligand-activated transcription factor belonging to an emerging class of basic helix-loop-helix/PAS proteins which show interaction with the molecular chaperone hsp90 in their latent states and require heterodimerization with a general cofactor, Arnt, to form active DNA binding complexes. Upon binding of polycyclic aromatic hydrocarbons typified by dioxin, the dioxin receptor translocates from the cytoplasm to the nucleus to allow interaction with Arnt. Here we have bypassed the nuclear translocation step by creating a cell line which expresses a constitutively nuclear dioxin receptor, which we find remains in a latent form, demonstrating that ligand has functional roles beyond initiating nuclear import of the receptor. Treatment of the nuclear receptor with dioxin induces dimerization with Arnt to form an active transcription factor complex, while in stark contrast, treatment with the hsp90 ligand geldanamycin results in rapid degradation of the receptor. Inhibition of degradation by a proteasome inhibitor allowed geldanamycin to transform the nuclear dioxin receptor to a heterodimer with Arnt (DR-Arnt). Our results indicate that unchaperoned dioxin receptor is extremely labile and is consistent with a concerted nuclear mechanism for receptor activation whereby hsp90 is released from the ligand-bound dioxin receptor concomitant with Arnt dimerization. Strikingly, artificial transformation of the receptor by geldanamycin provided a DR-Arnt complex capable of binding DNA but incapable of stimulating transcription. Limited proteolysis of DR-Arnt heterodimers indicated different conformations for dioxin versus geldanamycin-transformed receptors. Our studies of intracellular dioxin receptor transformation indicate that ligand plays multiple mechanistic roles during receptor activation, being important for nuclear translocation, transformation to an Arnt heterodimer, and maintenance of a structural integrity key for transcriptional activation.

Experimental cutaneous free flap transfers in the horse.

Equine limb wounds often heal slowly by epithelialization, and large scars are a frequent end result. In some ways, they resemble the wound associated with human tibial injuries. The literature indicates that previous investigators have failed to transfer free skin flaps successfully in the horse. In this paper, we review our experimental work with the deep circumflex iliac flap in the horse. Dissections of 20 cadavers confirmed the anatomical consistency of the flap. Four flaps survived well when elevated as island flaps, but five orthotopic and nine heterotopic free flap transfers all failed. The cause of failure is still unknown, but our experiments suggested that the horse must be highly susceptible to ischemic reperfusion injury.

Identification of a free skin flap from the region vascularized by the deep circumflex iliac artery of horses.

Areas of skin vascularized by large axial vessels potentially suitable for microvascular anastomosis were investigated in 10 horse cadavers. Eleven such areas were dissected, and the skin over the flank region vascularized by the deep circumflex iliac artery was most suitable. The anatomy of this area was further defined, using angiography and latex injection studies on 10 cadavers.

Transfer of deep circumflex iliac flaps to the tarsus by microvascular anastomosis in the horse.

Eighteen deep circumflex iliac flaps were elevated in healthy adult horses. Four flaps survived well when elevated as islands, but five orthotopic flaps and nine heterotopic flaps transferred to the tarsus and face failed. Technical reasons could explain the failure of the orthotopic flaps, but not the heterotopic flaps. Failure of the heterotopic flaps was apparently caused by the no-reflow phenomenon.

Culicoides hypersensitivity in the horse: 15 cases in southwestern british columbia.

The investigation of a chronic, seasonal dermatitis of horses in southwestern British Columbia is described. Typically the history indicated an insidious onset, followed by a gradual progression in the severity of the signs each year. Lesions appeared during the warmer months of the year and tended to regress during the winter. The clinical signs consisted of areas of pruritus and excoriation, affecting predominantly the ventral midline, mane and tailhead. In all cases corticosteroid therapy relieved the pruritus and allowed the lesions to heal.The salient pathological findings were hyperkeratosis, spongiosis and a dermal infiltration of eosinophils together with mononuclear cells. These changes are typical of an allergic dermatitis, which has been recognized in many parts of the world as a hypersensitivity reaction to the bites of Culicoides spp. In this instance, the epidemiological findings relating to the geographic area, the local insect population and the distribution of lesions implicated Culicoides obsoletus as the etiological agent.

Cutaneous onchocerciasis in the horse: five cases in southwestern british columbia.

Five horses were presented because of a dermatitis of the forehead. Unlike previous reports, ventral midline dermatitis was not the major problem, and was present in only two of five cases. All five horses responded to levamisole therapy at a daily dosage of 5.5 g for one week. Owners were cautioned that repeat therapy may be necessary.