The aim of our study was to optimise an existing staining procedure: haematoxylin-eosin saffron (HES). The method follows the classical haematoxylin and eosin protocol with the addition of a staining step using natural saffron to better identify the collagen fibres.

The aim of our study was to optimise an existing staining procedure: haematoxylin-eosin saffron (HES). The method follows the classical haematoxylin and eosin protocol with the addition of a staining step using natural saffron to better identify the collagen fibres. The saffron solution was obtained by dissolving ground saffron stigmas in absolute alcohol. In order to test the HES method for its staining ability on four main types of collagen (I, II, III, and IV), specific tissues (skin, tooth, cartilage, aorta, spleen, and penis) were chosen. The procedure showed a sharp differentiation between muscle, stained red or pink, and connective tissue, stained bright yellow or orange. HES allows the diagnosis of reticulin fibrosis undetected in HE and in previous saffron staining procedures. HES represents an advantageous alternative to HE staining giving highly reproducible results with high diagnostic value.

The effects of mitomycin-C on intraocular pressure (IOP), facility of outflow (C), and Tenon's capsule fibrosis were studied over 60 days in 10 clinically normal dogs. A 1-piece, silicone glaucoma implant was surgically implanted into both eyes; the filtration site of one eye was treated with a single, 5-minute intraoperative application of mitomycin (0.5 mg/ml), and the fellow eye was treated in a similar manner with balanced salt solution. There were no significant differences in preoperative IOP or C-values between treatment groups. Mean IOP in eyes of both groups initially decreased from the preoperative value, but returned to the baseline value by day 21. Mean facility of aqueous outflow (C-value) increased in all eyes during the first 14 days (mitomycin-C-value = 2.26 +/- 0.72; control C-value = 2.38 +/- 0.81), then reached a plateau that was significantly higher than the baseline value in mitomycin (P = 0.039) and control (P = 0.041) eyes. Histologic evaluation revealed all implants surrounded by a connective tissue capsule composed of regular dense collagen and fibroblasts that was significantly (P = 0.003) thinner in the mitomycin-treated (scleral side = 167 +/- 62 micrometer; conjunctival side = 122 +/- 41 micrometer) than the control (scleral side = 261 +/- 92 micrometer; conjunctival side = 180 +/- 48 micrometer) group. There were, however, no significant differences in IOP or C-values between groups at any postoperative time interval. Results of this study indicate that intraoperative treatment with mitomycin suppresses, but does not prevent fibrosis around silicone filtering implants.

The relation between average duodenal mast cell count, duodenal mucosal mast cell numbers, duodenal connective tissue mast cell numbers, circulating basophil numbers, heterophil-to-lymphocyte ratio, and lesion score were studied to gain an understanding of the events that may lead to intestinal lesion formation associated with hemorrhagic enteritis virus (HEV) infection. Changes in vascular permeability in the duodenum in birds inoculated with HEV were examined, using colloidal carbon and ferritin as vascular markers. Turkeys inoculated with HEV had significantly (P < 0.05) higher duodenal mast cell counts than did noninfected controls. Birds inoculated with HEV had significantly (P < 0.05) more mucosal mast cells than did phosphate-buffered saline solution-inoculated birds. Connective tissue mast cell and basophil numbers were unaffected by viral inoculation. Thermal stress did not have significant effect on lesion severity, but did increase number of birds that developed the characteristic intestinal lesions. The heterophil-to-lymphocyte ratio was significantly (P < 0.05) higher in HEV-inoculated birds, compared with phosphate-buffered saline solution-inoculated controls. Increase in vascular permeability was only detected in HEV-inoculated birds with intestinal lesions. Results indicate that mast cells, and the vasoactive mediators contained within mast cells, may be important in the early manifestation of HEV infection. They also provide a possible mechanism through which biochemical and physiologic changes characteristic of HEV infection can occur.

Tendon in horse is the most important type of connective tissue which connects muscle to bone, constituting a vital component of the musculoskeletal system, by enabling movement. Tendons suffer from a wide range of disorders, which includes different types of mechanical injuries and degenerative diseases. The sample population was a deep digital flexure tendon (DDFT) at mid-metacarpal region of thirty adult horses examined using scanning electron microscopy (SEM). Sex distribution was not taken into account. Changes in its structural organisation due to aging following tendon degeneration was unknown. The complex structures of tendon and its distinctive characteristics have been well demonstrated by SEM techniques. The tendon structure under SEM showed a dense, regular connective tissue arrangement thatreflects the mechanical requirement of this tissue. It is defined by thick regular bundles of longitudinal collagen fibres arrangedin a zig-zag conformation. All tendons examined from these horses had no history of deep digital flexure tendinitis and were allmacroscopically normal. The tendon showed a hierarchical structure, with collagen molecules forming fibrils (50 nm to 500 nm), which in turn comprise a fascicle (50 μm to 300 μm), and fascicles were aligned along the longitudinal axis of the tendon.

The relationship of muscle and bone structure to limb weakness was examined in 60 Duroc pigs from 3 lines divergently selected for thoracic limb weakness. The lines were designated high, control, or low, with the low line having inferior thoracic limb structure. At approximately 100 kg, 10 pigs of each line and gender were scored for thoracic limb structure and movement. Right and left thoracic limbs were collected at slaughter. A computerized morphometric image analysis system was used to determine cross-sectional areas of muscles, bones, and soft tissues at levels through the brachium, antebrachium, metacarpus, and digits. The statistical model that was used to analyze the data included the effects of line, sire, gender, and side (left vs right), with weight as a covariate. Total bone area was similar for all 3 lines of pigs at all cross-sectional levels, but significant differences in muscle and other soft tissue areas were observed, including significantly greater extensor area for the antebrachium (P less than 0.001) in low-line pigs than in control- and high-line pigs, smaller total area (P less than 0.05) of the metacarpus in low-line pigs than in control and high-line pigs, and less total area of the medial digit (P less than 0.01) in low-line pigs than in control- or high-line pigs. Total area of bone and soft tissue for each cross-sectional region was significantly greater (P less than 0.05) in boars than in gilts. Side differences also were observed in total cross-sectional areas of bone and soft tissue of the antebrachium, metacarpus, and digits. Pigs selected for inferior thoracic limb structure had less total soft tissue cross-sectional areas in distal limb regions than did control- or high-line pigs. Limb weakness may be related to altered distribution of soft tissue supporting structures of the thoracic limb.

The connective tissue structures commonly referred to as the periorbita, orbital septum, muscular fasciae, and vagina bulbi or collectively, as the orbital fasciae were dissected then illustrated and described. Two sheets (layers) of the periorbita (endorbita) were found in our dogs. The periorbita should be renamed endorbita because of its anatomic relations. The periorbita did not always fuse with the periosteum of frontal and sphenoid bones. Rather, the periorbita and the periosteum were often distinct and separate; only medioventrally did several fibrous bands unite the superficial sheet of the endorbita with the periosteum. Two layers of the endorbita fused with the periosteum of the margin of the bony orbit and with the orbital ligament. The muscular fasciae were divided into 3 layers. The superficial layer extended caudally from the orbital septum, was thick, and was pierced by arteries, veins, and nerves. The middle layer was attached to the sclerocorneal junction and, at the temporal canthus of the eye, was divided into superficial and deep sheets. The deep portion was attached to the lateral angle of the third eyelid, similar to a strong ligament. The deep layer of the muscular fasciae extended caudally from the sclerocorneal junction in intimate contact with recti and oblique muscles of the eyeball. The deep portion of the deep muscular fascia covered the deep surface of all recti muscles and separated them from the retractor bulbi muscle. Intermuscular septa were observed between middle and deep muscular fascia layers. The body of the third eyelid was located between superficial and middle muscular fascia layers and was fixed ventrally to the lateral angle of the eye by the deep sheet of the middle muscular fascia.

A number of studies have measured collagen fibers and collagen&#xD;deposition in wound healing process with advances imaging techniques. However, these are performed by complicated methods and need specific tools. In search of the easier ways in routine histopathological laboratory, collagen measurement and staining pattern of wound healing process were observed in wounded skin of Sprague Dawley’s rat by using two different stains which are standard haematoxylin and eosin (H&E) and modified Masson’s&#xD;trichrome staining (MT). The comparison between these staining in wounded tissues was made to evaluate the advantages and disadvantages of both staining in wound healing study for 21 days postwounding. Tissues which stained with MT staining was then evaluated its collagen re-organization and density by using polarized light microscope with the aid of image analyzer software. Results showed that tissues stained with standard H&E could not be used to measure and differentiate the collagen deposition which is contradictory to MT staining. Wounded tissue stained with MT staining has showed a clear view of collagen fibers deposition&#xD;and re-organisation compared to H&E staining. This finding could validate the using of modified MT staining which leads to accurate histopathological analysis and observation in wound healing study.