Magnetization transfer and spin lock MR imaging of patellar cartilage degeneration at 0.1 T.

PURPOSE: To investigate magnetization transfer (MT) parameters and rotating frame relaxation time T1 rho in patellar cartilage at different levels of degeneration. MATERIAL AND METHODS: Thirty cadaveric patellae were examined at 0.1 T using the time-dependent saturation-transfer MT technique and the spin lock (SL) technique. In an SL experiment, nuclear spins are locked with a radiofrequency (RF) field, and the locked nuclear magnetization relaxes along the magnetic component of the locking RF field. The specimens were divided into three groups according to the level of cartilage degeneration. MT parameters and T1 rho were measured. RESULTS: The MT effect was greater in degenerated cartilage than in normal cartilage. T1 rho was longer in advanced cartilage degeneration than in intermediate cartilage degeneration. CONCLUSION: The results suggest that more studies are needed to fully establish the value of SL imaging in cartilage degeneration.

Group II phospholipase A2 in nasal fluid, mucosa and paranasal sinuses.

The aim of this study was to determine the type of phospholipase A2 (PLA2) in nasal fluid and to demonstrate its cellular origin. The concentration of group II PLA2 was high (591.5 micrograms/l) in nasal fluid compared with serum level (10.8 micrograms/l) and the fluid of paranasal sinuses (10.6 micrograms/l). Methacholine stimulated nasal fluid contained only small amounts (19.1 micrograms/l) of group II PLA2 when the flow of tear fluid through the nasolacrimal duct was obstructed. Occasional glands secreting group II PLA2 were found in nasal and paranasal mucosa by immunohistochemistry. Lysozyme was found in the majority of mucosal glands. It was concluded that nasal and paranasal mucosal glands contain group II PLA2. In nasal fluid, however, PLA2 is mainly derived from tear fluid.

Lacrimal bone thickness at the lacrimal sac fossa.

BACKGROUND AND OBJECTIVE: Because laser dacryocystorhinostomy techniques have become more popular during the past few years, interest has grown concerning the anatomic structures that need to be penetrated in these procedures. The authors therefore studied the thickness and the histologic type of the lacrimal bone at the lacrimal sac fossa. PATIENTS AND METHODS: The thickness of 69 lacrimal bones at the lacrimal sac fossa from 48 patients was measured. RESULTS: The mean thickness was 106 microns. In 67% of the patients the mean thickness of individual lacrimal bone was less than 100 microns and in 4% it was more than 300 microns. The thinnest measured cross section of the lacrimal bone sample was 11 microns and the thickest was 722 microns. The lacrimal bone was composed of a thin plate of lamellar bone. CONCLUSION: In most cases the lacrimal bone at the lacrimal sac fossa is so thin that it can be easily penetrated with most surgical instruments.

Synthesis of group II phospholipase A2 and lysozyme in lacrimal glands.

PURPOSE: The aim of this study was to demonstrate the synthesis and cellular distribution of group II phospholipase A2 and lysozyme in the main and accessory lacrimal glands. METHODS: The authors studied samples of normal main lacrimal glands of seven autopsied subjects and accessory lacrimal glands of eight patients who underwent ptosis surgery. The specimens were immunostained with a rabbit antiserum against group II phospholipase A2 and a monoclonal antibody against lysozyme. Expression of group II phospholipase A2 gene was shown using Northern hybridization and in situ hybridization. RESULTS: Lysozyme was present in the secretory granules of most acini, whereas group II phospholipase A2 was seen in a minority of acinar cells, primarily in the central parts of lobules in the main and accessory lacrimal glands. Synthesis of group II phospholipase A2 in the glandular cells was confirmed by Northern hybridization and by in situ hybridization. CONCLUSIONS: There are two specialized cell types in the main and accessory lacrimal glands, one synthesizing group II phospholipase A2 and the other synthesizing lysozyme. These enzymes are important nonspecific antibacterial factors in tears.

Bone formation in rabbit cancellous bone defects filled with bioactive glass granules.

We examined new bone formation after filling cancellous bone defects with bioactive glass (BG) in granular form. Cylindrical defects in the trochanter area of 18 rabbit femora were filled with BG granules (diameter 600-830 microns) and compared with similar defects filled with morcellized autogenous bone. New bone formation and surface reaction of BG particles were evaluated by light microscopy, histomorphometry, and scanning electron microscopy. The chemical profile at the bone- material interface was studied by energy dispersive x-ray analysis (EDXA). In the BG group, 41, 32, and 38 percent of the defects were filled with new bone after 3, 6, and 12 weeks, respectively. The corresponding figures for the autogenous bone group were 36, 29, and 34 percent. The thickness of the reaction layer on the glass surface increased from 82 to 163 microns during the observation periods. An intimate contact without intervening soft tissue between new bone lamellae and BG granules was a constant finding. EDXA showed a chemical continuum between the granules and the new bone. No adverse reactions related to BG were observed. BG is a promising material for filling cancellous bone defects.

Acid cysteine proteinase inhibitor in cutaneous lymphocytic infiltrates.

Acid cysteine proteinase inhibitor (ACPI, cystatin A) is normally present in squamous epithelium and dendritic cells of lymphoid follicles. Its expression is altered both in proliferative and malignant squamous epithelium and in neoplastic lymphoid follicles. The expression of ACPI in the lymphoid infiltrates of cutaneous psuedolymphomas and B-cell lymphomas was studied. Eighteen pseudolymphomas from 15 patients were divided into three groups according to the proportion of B and T lymphocytes. The B-cell-type lesions with well-developed follicles and germinal centers showed a pronounced ACPI expression in dendritic cells. Varying amounts of ACPI-positive cells were present in the mixed B- and T-cell-type and also in the T-cell-type lesions. The labeled cell population was distinct from the factor XIIIa-positive dermal dendrocytes, S-100-positive histiocytes, and HAM 56-positive histiocytes. Malignant lymphomas contained a few haphazardly arranged ACPI-positive cells with short dendrites and granular cytoplasm. It was concluded that follicular dendritic cells can be reliably labeled with ACPI antiserum in cutaneous pseudolymphomas. The structure and distribution of ACPI-containing cells in malignant cutaneous B-cell lymphomas is altered when compared with pseudolymphomas.

Isolation of bone from muscles prevents the development of experimental callus-like heterotopic bone. A study of the interaction of bone and muscle in new bone formation.

Heterotopic bone (HB) has previously been provoked in the compartment of the profundus part of the vastus intermedius muscle (VIP) by manipulation of the rabbit knee immobilized in extension. The authors isolated the femoral bone of adult rabbits with a permeable or an impermeable polymer membrane to investigate the interactions of bone, periosteum, and muscle in callus-like HB formation, using the HB model. Heterotopic bone developed beside but not over the membrane if isolation was carried out before the immobilization-manipulation period, regardless of the type of membrane used. In cases where the femoral bone was isolated from the muscle by insertion of the tubular membrane one to two weeks after the immobilization-manipulation had begun, there was HB formation over and beside the membrane. No HB developed under the membrane in any of the cases of femoral bone isolation. Thus direct and extensive contact between bone and muscle appears to be essential at the onset of formation of experimental HB. An intertissue exchange of soluble substances derived from bone or muscle, however, does not seem to play an essential role here. After the onset of HB formation, its further development will not be prevented by subsequent isolation of muscle and bone. The study elucidates the interaction between bone and adjacent muscles and the results may be valuable in further investigations on HB and fracture healing.

Secretion of group 2 phospholipase A2 by lacrimal glands.

PURPOSE: Tears are known to have antimicrobial properties. The authors investigated the presence of the antibacterial enzyme phospholipase A2 (PLA2) in tears and lacrimal glands. METHODS: The catalytic activity of PLA2 and the amount of pancreatic group 1 PLA2 and nonpancreatic group 2 PLA2 were measured in homogenates of eight human lacrimal glands from autopsied subjects and in tears from four healthy volunteers. The localization of PLA2s in lacrimal gland sections was studied by immunohistochemistry. Skeletal muscle was used as a control. RESULTS: The catalytic activity of PLA2 was significantly higher in lacrimal glands than in skeletal muscle. Immunochemical analysis showed significantly higher amounts of group 2 PLA2 in lacrimal gland than in skeletal muscle homogenates. Group 1 PLA2 was present in trace amounts only. The concentration of group 2 PLA2 in tears was high (1451.3 micrograms/l) compared to that in the serum of healthy individuals (3.7 micrograms/l). By immunohistochemistry, a granular reaction of group 2 PLA2 was localized in the glandular cells of lacrimal glands. The apical cytoplasm of many duct cells also was labeled. CONCLUSIONS: The lacrimal glands secreted nonpancreatic group 2 PLA2, which most likely acts as an antiinfectious factor in tears.

Bioactive glass versus hydroxylapatite in reconstruction of osteochondral defects in the rabbit.

We studied osseointegration of a bioactive glass (BG) and hydroxylapatite (HA) in rabbit femur epiphyseal and metaphyseal regions. 17 BG and 24 HA cones implanted in defects through arthrotomy were analyzed. The holes for implants were drilled through distal femur joint surfaces. The cartilage wound repaired generally by fibrous tissue. Histomorphometry showed that 61, 78, and 79 percent of BG surface was covered by bone at 3, 6, and 12 weeks, respectively. The corresponding figures for HA were 47, 67, and 78 percent. Chemical bonding between bone and implants of both types was confirmed by scanning electron microscopy (SEM) and energy-dispersive x-ray analysis (EDXA). Formation of a calcium phosphate-rich layer on the surface BG implant was demonstrated by EDXA. Our results indicate that the osseointegration rate of bioactive glass does not differ from that of hydroxylapatite.

Pancreatic phospholipase A2 in cerulein-induced acute pancreatitis in the rat.

We investigated the concentration of immunoreactive pancreatic phospholipase A2 (pan-PLA2) and the catalytic activity of phospholipase A2 (CA-PLA2) in plasma and pancreases of rats with cerulein-induced acute pancreatitis. Edematous pancreatitis with ascites and fat necroses in the abdominal cavity developed after 8 h infusion of cerulein (5 micrograms/kg/h). Large vacuoles were found in acinar cells and there were small areas of acinar cell necrosis. No pathological changes were seen in saline-infused control animals. Pancreatic PLA2 was localized by immunohistochemistry in pancreatic acinar cells in both groups of animals and in the proximal tubular cells of the kidney in cerulein-infused animals. The lungs and kidneys appeared normal by light microscopy in all animals. The pan-PLA2 values increased markedly, whereas the CA-PLA2 values did not change during the cerulein-infusion. The CA-PLA2 values in the homogenates of pancreatic tissue of cerulein-infused animals did not differ significantly from those of saline-infused controls. The results indicate that the CA-PLA2 in plasma is independent from the concentration of pan-PLA2 in cerulein-induced acute pancreatitis in rat.

Morphology of osteogenesis in bioactive glass interface.

Bone formation around bioactive glass implants (S56.5P4) in the trabeculous subchondral bone in the distal femur of rabbits was studied by histology and scanning electron microscopy. Three types of tissue: bone, connective and hematopoietic tissue developed around the implants resulting in lamellar new bone covering 76% of the surface of the implants at twelve weeks. Bone formation around implants began as woven bone changing mainly to lamellar, osteon like new bone in contact with the S56.5P4 surface. Endochondral ossification was absent. In the area of bone containing hematopoietic tissue, new bone grew often as a thin layer along the implant surface. However, bone seemed to form adjacent to the implant surface through osteo-conduction. Only 21% of the implant surface was covered by loose connective tissue. Some proteoglycan containing thin fluid filled spaces were seen ten days after implantation. In few areas with apparent breakdown of the implant surface decreased amount or no bone formation was observed. Von Kossa method stained the reaction layer as two parallel dark brown lines, toluidine blue as two blue stripes, whereas van Gieson did not stain the reaction layer at all. In conclusion, the present histological results indicate bone bonding, which is a physico-chemical process observed between S56.5P4 implant and host bone.

Cholinergic hypothesis of alcoholic pancreatitis.

Chronic alcohol intake interferes especially with the two main pathways regulating exocrine pancreatic secretion: the cholinergic and the pancreozymin pathway. Recently, a new theory of the pathogenesis of alcoholic pancreatitis was proposed emphasizing disordered agonist-receptor interaction at the level of pancreatic acinar cells. Accordingly, alcohol-induced alterations in the control of exocrine pancreatic secretion result in hyperstimulation of pancreatic acinar cells and their muscarinic receptors, mimicking the mechanism of acute pancreatitis caused by scorpion sting, intoxication with an anti-acetylcholinesterase-containing insecticide or supramaximal doses of secretagogues. The present review emphasizes the role of these alcohol-induced secretory alterations in the pathogenesis of alcoholic pancreatitis.

Acantholytic squamous cell carcinoma of the uterine cervix with amyloid deposition.

An 84-year-old woman suffered for 1 year from intermittent vaginal bleeding. Clinical examination revealed a large ulcerative cervical tumor that was histologically classified as well-differentiated squamous cell carcinoma. Acantholytic areas, apoptotic cell death, and pronounced amyloid deposition characterized the tumor. No evidence of papilloma virus infection was found in immunohistochemical examination or in nucleic acid in situ hybridization. Amyloid formed globular structures surrounded by neoplastic cells that reacted with cytokeratin antibodies. Although the amyloid itself was not labeled, electron microscopy showed filamentous degeneration of the squamous cells analogous to that described in different types of cutaneous keratin-derived amyloidoses. It was concluded that similar pathogenetic mechanisms are involved both in the cutaneous amyloidosis and in the amyloid deposition of squamous cell carcinoma.

Pancreatic phospholipase A2 in proximal tubules of rat kidney in experimental acute pancreatitis and after intravenous injection of the enzyme.

Kinetics and distribution of i.v. human pancreatic phospholipase A2 (h-PLA2) were determined in intact and nephrectomized rats, and tissue localization of rat pancreatic PLA2 (r-PLA2) was studied by immunohistochemistry in experimental acute pancreatitis. The concentration of h-PLA2 and the catalytic activity of phospholipase A2 in plasma decreased exponentially in intact and nephrectomized animals after the injection. The initial 15-min half-life was considerably longer in nephrectomized animals, and higher h-PLA2 concentrations and PLA2 catalytic activities were found in plasma. h-PLA2 was localized in endocytotic vesicles and apical cytoplasmic vacuoles in proximal tubule cells of the kidney. The intensity of the immunoreaction decreased considerably between 15 and 50 min in these cells. No signs of tubular damage were seen by light microscopy. Neither immunoreactive h-PLA2 nor PLA2 catalytic activity was found in urine. r-PLA2 was observed in proximal tubule cells 15 min after an injection of sodium taurocholate (necrotizing pancreatitis group) or saline (edematous pancreatitis group) into the pancreatic duct. Signs of tubular damage were present in necrotizing pancreatitis, but tubular morphology was normal in the animals with edematous pancreatitis. We conclude that the proximal tubule cells of the kidney participate in the metabolism of circulating pancreatic PLA2, and considerably higher PLA2 levels persist in plasma in nephrectomized animals. Endogenous pancreatic PLA2 is detected in kidneys in acute pancreatitis.

Standards of morphological evaluation and histological grading in experimental acute pancreatitis.

Acute pancreatitis is characterized morphologically by edema, hemorrhages, parenchymal necrosis and fat necrosis. The inflammation is accompanied by infiltration of polymorphonuclear leukocytes. According to the absence or presence of necrosis the disease can be divided into interstitial (or edematous) pancreatitis and hemorrhagic-necrotizing pancreatitis. The severity of disease can be graded in the histological sections either by giving scores to the different types of morphological alterations or by determining the proportion of necrotic tissue of the total lobular parenchyma. The former method is based on subjective assessment of histological slides and is suitable for the evaluation of both edematous and necrotizing pancreatitis. Histometric measurement of necrotic parenchyma can be used only in the necrotizing forms of experimental pancreatitis, e.g. in those induced by intraductal injection of bile, bile salts or digestive enzymes, and in the dietary ethionine-induced pancreatitis. Grading of the tissue damage is essential when the effects of different therapies are evaluated.

MR imaging of patellar cartilage degeneration at 0.02 T. Study of 23 cadaveric patellae.

MR imaging with a 0.02 T resistive magnet was used to establish the correlation between the histologic grading of patellar cartilage degeneration and fat water separation images or T1- and T2-relaxation times. We examined 23 cadaveric patellae. There was a positive correlation between histologically graded cartilage degeneration and T1-relaxation time. Patellar cartilage was well differentiated from surrounding structures on chemical shift water proton images, and an evaluation of cartilage degeneration was possible. No correlation was found between cartilage damage and T2-relaxation time. Chemical shift imaging at 0.02 T is easy to perform and gives further information of cartilage disorders.

Ultrastructural signs of altered intracellular metabolism in acral persistent papular mucinosis.

An asymptomatic 24-year-old woman presented with multiple discrete papules on the extensor surfaces of the hands and wrists. Light microscopy revealed focal increase in the amount of dermal fibroblasts as well as deposition of hyaluronidase-labile mucoid substance. The collagen and elastin were decreased. The changes were consistent with acral persistent papular mucinosis (APPM). In electron microscopy, the intercellular glycosaminoglycans showed small ruthenium red-positive granules and thin filaments indicating normal morphology. The fibroblastic cells, however, were conspicuously altered. Endoplasmic reticulum was dilated, cytoplasm contained large amounts of osmiophilic, concentric lysosomal structures, and there was distinct fibrous lamina in the nuclear membrane. It was concluded that the primary event in APPM probably affects the intracellular metabolism of the dermal fibroblast. The accumulation of lysosomal structures may be a distinct feature of APPM differentiating it from the other reactive cutaneous mucinoses, or it may only reflect nonspecific degeneration in a long-standing lesion.

Macrophages in trauma-induced myositis ossificans.

The basic cellular mechanisms of different forms of myositis ossificans are poorly known. In the current experiment the nature of the early (24-168 h) inflammatory cell reaction preceding trauma-induced myositis ossificans was studied. New bone formation was induced in the vastus intermedius region of the rabbit quadriceps muscle by means of immobilization and daily passive mobilization. Before the start of treatment, a cell harvesting device (viscose cellulose sponge in a silastic tube) was implanted in the region of interest. The opposite intermedius muscle and a standardized surgical skin wound served as the control sites. The results showed a significantly prolonged invasion of macrophages into the ossifying intermedius muscle as compared with the control intermedius muscle. It is hypothesized that microinjury and subsequent muscle necrosis cause the invasion of macrophages, and these cells respond to the conditions of the traumatized muscle under passive mobilization by releasing osteogenic growth factors.

The role of phospholipase A2 in pancreatic acinar cell injury.

The integrity of rat pancreatic acinar cells under the influence of human phospholipase A2 (PLA2) was studied. Isolated pancreatic acini showed no increased discharge of aspartylaminotransferase (ASAT) when incubated either in solutions containing human pancreatic PLA2 or the bile salt sodium deoxycholate (DEC), the latter in concentrations that augment PLA2 activity but have no destructive detergent effect. When human pancreatic PLA2 was injected into the rat pancreatic duct, uneven distribution was observed at 15 min and 3 h in immunohistochemical sections. Edema and a mild inflammatory reaction were the main changes in the pancreas. The necrotic areas seen by light and electron microscopy were quite small and located mostly at the periphery of lobules corresponding the spread of the injected material. Necrosis was of the coagulation type and showed equal extent after the injection of PLA2 with or without DEC. Internalized human pancreatic PLA2 was present already 15 min after the injection in the cytoplasm of some intact acinar cells, indicating a functioning protective mechanism. It was concluded that pancreatic acinar cells are quite resistant to PLA2-catalyzed hydrolysis of membrane phospholipids in vitro, but additional trauma, e.g., pressure caused by intraductal injection, and tissue related factors, such as the mediators of the inflammatory reaction, make acinar cells susceptible to the effect of PLA2.