Testicular leiomyoma in a ram.
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Biomedical subjects
Publications and source records attributed to R A Foster.
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One of the initial events required for regeneration of periodontal tissues lost due to disease is the establishment of connective tissue attachment to root surfaces. Thus, considerable research efforts have focused on developing reliable procedures to gain new connective tissue attachment. Our studies focus on evaluating agents for their ability to promote cell attachment and spreading using an in vitro assay. For these studies human gingival fibroblasts (GF) and human periodontal ligament (PDL) cells, after exposure to fibronectin; 44 kilodalton bone phosphoprotein (44K BPP-osteopontin) or guanidine EDTA extracts of bone, cementum, or dentin, were compared as to degree of cell attachment and spreading. Fibronectin equally enhanced attachment and spreading PDL cells and GF. In contrast, 44K BPP, as well as guanidine EDTA extracts of bone and cementum, preferentially promoted attachment of GF when compared with attachment of PDL cells. For both PDL cells and GF the attached cells exhibited spreading. The guanidine EDTA extract of dentin did not promote attachment of either cell type. These results suggest that PDL cells and GF have different attachment properties which need to be considered for investigations directed at developing regenerative periodontal treatments.
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Thirteen rams with serological evidence of Brucella ovis exposure (CFT of 1:8 or greater), but with no or only mild epididymitis, were selected from a ram flock. Serum, semen, preputial washings and fluids from the accessory sex glands (ASGF) and testis and epididymis (TEF) were examined and immunoglobulin (Ig) concentrations estimated. Genital tissues were examined histologically and the percentages of class specific immunoglobulin containing cells (ICC) determined. Eleven of these rams had histological evidence of active inflammation consistent with B. ovis infection; the organisam was cultured from the semen of 7. IgA concentration was high in semen (mean +/- standard deviation of 5.03 +/- 1.78 mg/ml) and ASGF (9.18 +/- 7.28 mg/ml). These levels were much higher than those recorded in noninfected rams. IgA concentration was low in serum (0.78 +/- 0.55 mg/ml) and TEF (0.59 +/- 0.78 mg/ml). The concentrations of IgG1, IgG2 and IgM were low in all genital fluids sampled and not significantly different from those recorded in noninfected rams. This indicated that infection with B. ovis results in a pronounced IgA response in secretions, mostly from the accessory sex glands. Examinations of ICC, however, revealed that the plasma cell infiltrates of the epididymis, vas deferens, ampulla and seminal vesicle were predominantly IgG-containing (92.4, 97.2, 79.4 and 91.9% respectively). Fewer IgM-containing cells were scattered throughout these tissues, constituting 3.9, 6.3, 0.3 and 6.5% of all ICC, respectively. IgA-containing cells were most frequently seen in the ampulla (9.6% of ICC) where they were located directly beneath the epithelium, suggesting the ampulla as the most prominant location for the local production of IgA.(ABSTRACT TRUNCATED AT 250 WORDS)
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A total of 40 mature or aged rams with spontaneous varicocele detected by scrotal palpation were subjected to detailed necropsy examination. Varicocele was bilateral in 22 rams or was located on the left or right sides only, in 8 and 10 rams, respectively. This distribution contrasted with varicocele in man in which the left side is involved in 70 to 100% of cases. Mean sizes of varicoceles in mm (length x diameter) were 117 x 46 and 104 x 45 for the left and right sides, respectively, and they were located high in the pampiniform plexus, approximately 100 mm from the dorsal pole of the corresponding testis. All varicoceles were thrombosed. Changes associated with large varicoceles included testicular mineralisation and occluding thrombosis of testicular vessels. Total testis weight-bodyweight ratio in rams with varicocele (5.8 to 6.4 x 10(-3] was significantly less (P less than 0.05) than in normal (control) rams from the same flocks (7.9 x 10(-3] suggesting that some degree of testicular atrophy resulted from presence of a varicocele.
The levels of the immunoglobulins IgA, IgG1, IgG2, and IgM were measured in serum and fluid from various locations in the reproductive tract of normal rams. These fluids included semen, preputial washings, and fluid from the accessory sex glands (ASG), vasa deferens, rete testes, and tissue fluid from the seminal vesicles, bulbourethral glands, epididymal tails and efferent ducts. In addition, the prevalence of specific Ig-containing cells (ICC) was measured in sections of formalin fixed tissues stained by an indirect peroxidase-antiperoxidase labelling technique. Mean IgA levels in semen (1.23 mg/ml) and ASG fluid (0.46 mg/ml), were higher than in serum (0.19 mg/ml) and were at levels higher than IgG1 or IgG2 levels in semen, ASG fluid, and preputial washings, thus confirming the existence of a local immune system primarily in the ASG of ram genitalia. Relatively low concentrations of IgA and IgG in other genital fluids and IgG levels in these fluids were consistent with diffusion from serum. The relatively high prevalence of IgA-containing cells in bulbourethral (56% of all ICC) and prostate (49%) glands confirmed these tissues as major sites of local Ig production. ICC were also found in large numbers beneath pelvic urethral and preputial epithelia, but these were predominantly IgG-containing (88 and 72% respectively).
Both periodontal ligament and gingival tissue are thought to harbor cells with the ability to stimulate periodontal regeneration, i.e., formation of new bone, cementum, and connective tissue attachment. To understand further the role of these cells in the regenerative process, we compared human periodontal ligament cells and gingival fibroblasts, both derived from the same patient, same passage, in vitro. Protein and collagen production was significantly greater in periodontal ligament cells when compared with that of gingival fibroblasts. In addition, periodontal ligament cells had higher alkaline phosphatase levels when compared with those of gingival fibroblasts.
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While the exact mechanisms regulating bone homeostasis are unknown, it is generally accepted that factors with the capacity to regulate cell attachment and spreading play a role in osteogenesis. A 44 kDa bone phosphoprotein (44K BPP), isolated from rat bone and synthesized by osteoblasts, was evaluated for its role in attachment and spreading of fibroblasts. In uncoated plates, enhanced cell attachment and spreading were observed when fibroblasts were exposed to the 44K BPP. The attachment properties of the bone phosphoprotein are different from those of fibronectin, in that the 44K BPP did not promote cell attachment in type I collagen wells, as was seen with fibronectin. Also, 44K BPP continued to enhance cell attachment up to 24 h, whereas cell attachment declined in time with cells exposed to fibronectin. Cycloheximide did not alter 44K BPP promotion of cell attachment, indicating that de novo protein synthesis was not required. These studies suggest that the 44K BPP is important in the regulation of cell attachment and spreading at sites of mineralization.
Non-confluent cell cultures were exposed to both guanidine and guanidine-EDTA extracts of cementum, dentine and alveolar bone, at concentrations from 2 to 50 micrograms/ml for 48 h. The cells were radioactively labelled during the last 24 h. Total protein production was measured via incorporation of radioactive proline; collagen production was estimated by digestion of the radioactive protein mixture with bacterial collagenase. All guanidine-EDTA extracts elicited statistically-significant increases in total protein production when compared to controls. At 50 micrograms/ml of extract, the increase in protein production was 340, 143 and 338 per cent for bone, cementum and dentine, respectively. Similar results were obtained for collagen production. Guanidine-EDTA extracts also stimulated an increase in the production of specific proteins, as ascertained by gel electrophoresis. In contrast, the guanidine extracts had no effect on either protein or collagen production. Thus the functions of gingival fibroblasts can be altered by proteins from associated mineralized tissues. Identification of such proteins and their biological functions would enhance knowledge of the mechanisms that regulate connective-tissue regeneration.
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A primary objective in the treatment of periodontal diseases is attachment of fibrous tissue to root surfaces. An in vitro method was used to evaluate agents for their potential in enhancing this attachment process. Guanidine EDTA protein extracts of alveolar bone, cementum, and dentin in culture media were added to petri dishes and incubated at 37 degrees C for 1 hour. Human gingival fibroblasts were added to the dishes and incubated for an additional 90 minutes. Following incubation, cells attaching to the dish were quantified electronically, using a Coulter Counter. Extracts of alveolar bone and cementum enhanced cell attachment while dentin extract had no effect. These results indicate that both cementum and bone contain attachment proteins which could prove beneficial for attachment of fibrous tissue to root surfaces.
The percentage of synthesis dedicated to collagen is elevated in low-density cultures of human gingival fibroblasts, as is per-cell total protein synthetic activity and glycosaminoglycan accumulation. These observations can be explained, in part, by a decrease in membrane transport of precursor substance in high-density cultures. Synthetic activity by human fibroblasts can be reliably assayed in vitro using as few as 500 cells sparsely seeded. Such low-cell number assay is essential for study of single-cell clones, where replicative life span is limited.
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