Immunochemical and biochemical characterization of purified canine interferon-gamma. Production of a monoclonal antibody, affinity purification, and its effect on mixed lymphocyte culture and mixed lymphocyte kidney culture reactions.

We have advanced the hypothesis that the primary autolymphoproliferative response of dog T cells in mixed lymphocyte kidney cultures (MLKC) results from their recognition of tissue-specific (kidney-associated) antigen(s) presented in conjunction with class II MHC antigens. Lymphocyte culture-derived supernatants had been found previously to upregulate class II antigen expression on kidney cells and enhance T cell activation. In the present study we have isolated and characterized dog IFN-gamma, a class II-inducing substance that is secreted in the culture supernatant of activated T lymphocytes. Dog IFN-gamma was induced with A-23187 and PMA and purified stepwise using controlled-pore glass, Mono Q anion exchange chromatography, and Superose 6-gel filtration on FPLC. The purification resulted in two molecules of 42 Kd and 31 Kd molecular weights. An IgG1 monoclonal antibody was engendered to these molecules. With this mAb reagent, in immunochemical experiments, we have developed a sensitive ELISA and a method for purifying dog IFN-gamma by affinity chromatography. Species specificity studies indicated that purified dog IFN-gamma reacted with a polyclonal rabbit antihuman IFN-gamma, but not with a mAb to human IFN-gamma. However, the antidog IFN-gamma mAb that was generated also reacted with recombinant human IFN-gamma. In in vitro biological studies, the purified IFN-gamma (two mol. wt. species) upregulated the expression of canine class II MHC molecules on dog tubular epithelial cells and the dog kidney epithelial cell line (MDCK). The antidog IFN-gamma mAb blocked T cell proliferative response to kidney cell and, by inference, the interaction between endogenously released IFN-gamma in vitro with its cell surface receptor, thus inhibiting the induced upregulation of class II. Interestingly, although antidog IFN-gamma markedly blocked the MLKC (10 micrograms mAb/well), there was no effect on the allogeneic MLC. This observation indicates that the cytokine IFN-gamma may be a uniquely key substance amplifying the immune response of T cells to tissue-associated antigens on surrogate antigen-presenting cells that require induced upregulation of class II MHC antigen expression (MLKC), in contrast to reactions in which these antigens are already constitutively expressed on the antigen-presenting cells (mixed lymphocyte culture).     

Clinical cases in AIDS. 1.

As the human immunodeficiency virus (HIV) epidemic continues there are increasing numbers of patients with HIV-related disease. Doctors studying for the MRCP exam will need to be familiar with the common manifestations of HIV infection and acquired immunodeficiency syndrome.     

Depression symptoms in movement disorders: comparing Parkinson's disease, dystonia, and essential tremor.

Depression is common in Parkinson's disease (PD) and affects 30 to 50% of all patients. In contrast to the wealth of research on depression in PD, little is known about the occurrence of depression in other movement disorders. The primary objective of the current study was to determine whether the high prevalence of depression symptoms seen in PD is also found in other movement disorders, by directly comparing rates of specific depression symptoms and depression severity across PD, dystonia, and essential tremor (ET). Three hundred and fifty-four patients with PD, 83 patients with dystonia, and 53 patients with ET completed the Beck Depression Inventory (BDI). We found no significant between-groups differences for depression severity, frequency, or endorsement of specific depression symptoms. Forty-eight percent of PD patients, 37.3% of dystonia patients, and 34% of ET patients were found to be at least mildly depressed (BDI score of 10 or higher). The most commonly endorsed symptoms were fatigability, difficulty with work, anhedonia, and sleep disturbance. Clinicians should be aware that depression is a frequent problem in dystonia and ET, in addition to PD, and inquire about depression symptoms in these patients so that they can be appropriately treated.     

S-nitrosoglutathione-containing hydrogel accelerates rat cutaneous wound repair

BACKGROUND: Nitric oxide (NO) plays a key role in wound repair and S-nitrosothiols like S-nitrosoglutathione (GSNO) are well known NO donors. METHODS: Animals were separated in two groups and submitted to excisional wounds on the dorsal surface at the first day. GSNO (100 microm)-containing hydrogels were topically applied on the wound bed in the GSNO group, daily, during the first 4 days. Control group was topically treated with hydrogel without GSNO for the same period. Wound contraction and re-epithelialization were measured. Animals were sacrificed 21 days after wounding. Samples of lesion and normal tissue were formalin-fixed, paraffin embedded for histological analysis. RESULTS: Wound contraction, measured 14 and 21 days after wounding, was greater in the GSNO group than in the control group (P<0.05 for both). The re-epithelialized wound area, measured 14 days after wounding, was higher in the GSNO group than in the control group (P<0.05). A higher amount of inflammatory cells was observed in superficial and deep areas of the granulation tissue of the control group compared to the GSNO group. Twenty-one days after wounding, thin red-yellow collagen fibers arranged perpendicularly to the surface were found in the granulation tissue of the control group, whereas in the GSNO-treated group collagen fibers were thicker and arranged parallel to the surface. Increased number of mast cells was observed in the GSNO group compared with that in the control group. Vascularization and myofibroblast distribution were similar in both groups. CONCLUSION: Topical application of GSNO-containing hydrogel during the early phases of rat cutaneous wound repair accelerates wound closure and re-epithelialization and affects granulation tissue organization.     

Cataract surgery takes longer in functionally monocular patients.

BACKGROUND AND OBJECTIVES: To determine differences in the amount of work involved in caring for functionally monocular patients who undergo cataract surgery compared with binocularly sighted patients. PATIENTS AND METHODS: This was a retrospective study that included 100 consecutive functionally monocular patients and 100 binocularly sighted control patients matched by age (+/-5 years) and date of surgery ( +/-1 year). Office records were reviewed to determine several measures of office and operating room work effort. RESULTS: There was no difference between groups in preoperative telephone calls (P = .136), postoperative telephone calls (P = .580), preoperative office visits (P = .875), postoperative office visits (P = .601), or the number of times surgery was scheduled (P = 1.00). Monocular patients required more time for surgery (37.4 minutes) than binocular patients (32.4 minutes) (P= .010). CONCLUSIONS: It takes longer to perform cataract surgery on functionally monocular patients than on binocularly sighted patients.     

The proliferation and differentiation of the bone-lining cell in estrogen-induced osteogenesis

The purpose of this study was to determine the capacity of the bone-lining cell, which covers most nonremodeling bone surfaces in the adult skeleton, to proliferate and contribute to the rapid endosteal osteogenic response following estrogen treatment in male Japanese quail. In control, untreated birds bone-lining cells cover most of the femoral endosteal surface. Bone-lining cells are thinly extended along bone surfaces, have flat nuclei, and account for about 77% of the cells adjacent to the bone surface. The lineal density of bone surface cells in the controls was about 21 cells/mm of bone surface. By 16-20 h after estrogen administration cells with larger, round nuclei were seen on the bone surface and some of these cells contained a 3H-thymidine (3H-TdR) label. Within the first 24 h after estrogen administration the lineal bone surface cell density had increased to about 38 cells/mm. At 20-30 h after estrogen administration, most of the cells adjacent to the bone surface were round. The peak 3H-TdR labeling of cells immediately adjacent to the bone surface occurred about 36 h after estrogen administration. By 48 h the bone surface was covered with osteogenic cells and developing medullary bone. These results suggest that the bone-lining cell in the adult appears to retain some proliferative capability and osteogenic potential. However, because a rapid increase in cell density began prior to the rapid increases in 3H-TdR labeling, as well as the appearance of mitotic figures on the bone surface, another source of cells may have also contributed to the osteogenic response induced by estrogen.