Effects of neolignans from the stem bark of Magnolia obovata on plant pathogenic fungi.

AIMS: To characterize antifungal principles from the methanol extract of Magnolia obovata and to evaluate their antifungal activities against various plant pathogenic fungi. METHODS AND RESULTS: Four neolignans were isolated from stem bark of M. obovata as antifungal principles and identified as magnolol, honokiol, 4-methoxyhonokiol and obovatol. In mycelial growth inhibition assay, both magnolol and honokiol displayed more potent antifungal activity than 4-methoxyhonokiol and obovatol. Both magnolol and honokiol showed similar in vivo antifungal spectrum against seven plant diseases tested; both compounds effectively suppressed the development of rice blast, tomato late blight, wheat leaf rust and red pepper anthracnose. 4-Methoxyhonokiol and obovatol were highly active to only rice blast and wheat leaf rust respectively. CONCLUSIONS: The extract of M. obovata and four neolignans had potent in vivo antifungal activities against plant pathogenic fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Neolignans from Magnolia spp. can be used and suggested as a novel antifungal lead compound for the development of new fungicide and directly as a natural fungicide for the control of plant diseases such as rice blast and wheat leaf rust.

Concomitant reconstruction of the medial collateral and posterior oblique ligaments for medial instability of the knee.

We present the operative technique and clinical results of concomitant reconstruction of the medial collateral ligament (MCL) and the posterior oblique ligament for medial instability of the knee using autogenous semitendinosus tendon with preservation of the tibial attachment. The semitendinosus tendon graft between the screw on the medial epicondyle and the tibial attachment of the graft was overlapped by the MCL, while the graft between the screw and the insertion of the direct head of the semimembranosus tendon was overlapped by the central arm of the posterior oblique ligament. Assessment was by stress radiograph and the Lysholm knee scoring scale. After a mean follow-up of 52.6 months (25 to 92), the medial joint opening of the knee was within 2 mm in 22 of 24 patients. The mean Lysholm score was 91.9 (80 to 100). Concomitant reconstruction of the MCL and posterior oblique ligament using autogenous semitendinosus tendon provides a good solution to medial instability.

Cellular characterization of multidrug resistance P-glycoprotein, alpha fetoprotein, and neovascular endothelium-associated antigens in canine hepatocellular carcinoma and cirrhotic liver.

P-glycoprotein (P-gp), which is encoded by the multidrug resistance gene (MDR-1); alpha fetoprotein (AFP); and vascular endothelium-associated antigens are well-known markers for human and canine hepatic diseases. We obtained liver tissues from 5 dogs with hepatocellular carcinoma (HCC) and 12 dogs with cirrhosis, and we performed histopathologic and immunohistochemical evaluations using anti-P-gp, anti-AFP, anti-CD31, and anti-CD34 antibodies. P-gp was expressed at higher levels in HCC than in cirrhotic livers ( P < .01), and was most commonly localized in biliary canaliculi and small ductuli. AFP was localized mainly in the cytoplasm in HCC ( P < .01) and in a few cases of cirrhosis. In both HCC and cirrhosis, the AFP-positive cells were morphologically similar to normal hepatocytes and showed an even cytoplasmic distribution of AFP. The endothelial markers CD31 and CD34 were used to investigate vascular distribution. CD31 was expressed strongly in the portal area and parenchyma in HCC, but it was rarely observed in the parenchyma in cirrhosis. CD34 expression could not be detected in both HCC and cirrhosis. This study constitutes the first comprehensive study of P-gp, AFP, and endothelial markers in canine HCC and cirrhosis. The importance of these markers in HCC and cirrhosis in dogs was demonstrated and provides a more accurate basis for a definitive diagnosis of HCC and cirrhosis in dogs.

Semitendinosus tenodesis for medial instability of the knee.

A new technique of semitendinosus tenodesis has been developed to treat medial instability of the knee. The semitendinosus tendon was sectioned at the musculotendinous junction. The accessory insertion of the tendon was dissected to be overlapping and parallel to the anterior band of the medial collateral ligament. An isometric point on the femur was located around the medial epicondyle. A bony trough was created just above the isometric point and a screw with washer was placed. The semitendinosus tendon was looped around the screw and the screw was tightened to hold the tendon in place. The free end of the tendon was pulled obliquely backward and passed through the insertion of the direct head of the semimembranosus tendon to reconstruct the posterior oblique ligament. The semitendinosus tendon reinforced both the medial collateral ligament and the posterior oblique ligament.

Arthroscopically assisted treatment of avulsion fractures of the posterior cruciate ligament from the tibia.

BACKGROUND: The attachment of the posterior cruciate ligament to the posterior intercondylar fossa of the tibia is in a location that is difficult to access for arthroscopic surgical procedures. This report presents a variety of arthroscopically assisted reduction and fixation methods for managing avulsion fractures of the posterior cruciate ligament from the tibia. METHODS: Thirteen patients (fourteen knees) who had an avulsion fracture of the posterior cruciate ligament were treated with an arthroscopic procedure. Eleven patients underwent the operation in the acute phase (four to ten days after the injury), and two patients had delayed surgery (at nineteen and twenty months after the injury) because of nonunion. The choice of fixation method was based on the size of the avulsed fragment. Six knees that had a small bone fragment (<10 mm) with comminution were fixed with use of multiple sutures. Two knees that had a small bone fragment without comminution were fixed with 23-gauge wires. Two knees that had a medium-sized fragment (10 to 20 mm) were fixed with Kirschner wires. Four knees that had a large single fragment of bone (>20 mm) that involved the condyles were fixed with one or two cannulated screws. RESULTS: All patients had osseous union as determined on radiographs. Three injured knees in two patients showed limitation of motion after the operation. These patients had been immobilized for two or three months after the surgery because of concomitant fractures. The eleven patients who had undergone the operation in the acute phase, including two in whom postoperative arthrofibrosis had developed, showed no or trace posterior instability following the procedure. However, the two patients in whom the surgery had been delayed had residual grade-I posterior instability. The postoperative side-to-side differences, when measured with use of the KT-2000 arthrometer and posterior stress radiographs, showed better results in the patients in whom the surgery had been performed in the acute phase than in the patients in whom the operation had been delayed. CONCLUSION: Arthroscopic procedures can be used to treat tibial avulsion fractures of the posterior cruciate ligament.

Medial and lateral discoid meniscus in the same knee.

To our knowledge, only 1 case of medial and lateral discoid meniscus in the same knee has been reported. We report on 2 cases of medial and lateral discoid meniscus in the same knee.

Arthroscopic treatment for localized pigmented villonodular synovitis of the knee.

This study investigated 11 patients with localized pigmented villonodular synovitis of the knee that was diagnosed and treated by arthroscopic technique. There were six male and five female patients between the ages of 15 and 59 years (mean, 34.6 years). Seven patients reported extension limitation without joint line tenderness. Four of the 11 patients had a history of trauma before the onset of knee symptoms. All patients were treated by arthroscopic resection with partial synovectomy. The most common involved site was the anteromedial synovium near the anterior horn of the medial meniscus (five patients). The remaining cases were located in the anterior fat pad (two patients), suprapatellar pouch, posteromedial compartment, medial gutter, and the anterior horn of the lateral meniscus. Nine patients had one mass, and the remaining patients each had two or three masses. There was no evidence of recurrence at followup for an average of 29.9 months (range, 24-48 months). Arthroscopy is effective in the diagnosis of localized pigmented villonodular synovitis with minimal morbidity, and complete arthroscopic excision can be considered the definitive treatment for localized pigmented villonodular synovitis.

Tibial tuberosity avulsion fracture combined with meniscal tear.

Avulsion fractures of the tibial tuberosity are uncommon injuries. They usually occur during athletic activities in adolescents. The classification of these injuries has been divided into three types. Only two cases of avulsion fractures of tibial tuberosity have previously been reported with associated damages to menisci. We report a type III fracture of the tibial tuberosity associated with tear of the medial meniscus.

Operative hip arthroscopy.

Twenty patients underwent operative arthroscopic procedures of the hip joint. All procedures were performed with the patient in the supine position on a standard fracture table using fluoroscopy through three arthroscopic portals (anterolateral, anterior paratrochanteric, and posterior paratrochanteric). The initial indications were therapeutic in 16 patients: loose bodies in four, synovial chondromatosis in three, rheumatoid arthritis in five, ankylosing spondylitis in one, septic arthritis in one, avascular necrosis of femoral head in one, and primary osteoarthritis in one. In four patients who had unexplained hip pain, the initial indications were diagnostic: minimal synovial change was seen in two patients, a synovial chondromatosis was present in another, and a tear of the acetabular labrum and hypertrophy of ligament teres were present in a fourth patient. In one patient who had primary osteoarthritis, the insertion of the arthroscopic instrument into the hip joint failed because of profuse osteophytes along the acetabular rim. Twelve of the 19 patients showed significant improvement of the symptoms after the arthroscopic procedure, but seven patients had no benefit from the procedure. One patient had a postoperative reflex sympathetic dystrophy.

Arthroscopic reduction and fixation of bony avulsion of the posterior cruciate ligament of the tibia.

Bony avulsion fractures of the posterior cruciate ligament of the tibia have commonly been treated by open reduction and internal fixation using the posterior approach. However, this approach, using the prone position, makes it difficult to investigate and treat other combined injuries of the knee joint. We report a case of posterior cruciate ligament avulsion of the tibia that was arthroscopically reduced and firmly fixed with two cannulated screws. The posterior sag was absent after the operation and the result was excellent. By arthroscopy, we got rigid fixation of the avulsed fragment for early rehabilitation, and detection of a concomitant injury was also possible.

Tenosynovial giant-cell tumor in the knee joint.

We report the case of a tenosynovial giant-cell tumor causing limitation of extension of the knee. The tumor was treated successfully by arthroscopic removal.

Protein SP40,40-like Immunoreactivity in the Rat Brain: Progressive Increase With Age.

The pattern of distribution of SP40,40-like immunoreactive structures has been studied in the rat brain using a well-characterized polyclonal antibody raised against the SP40,40 protein. Protein SP40,40 is the human counterpart of the rat sulphated glycoprotein 2, whose mRNA shows widespread expression in the developing and mature brain. In young adult rats few immunoreactive structures were observed. Some immunoreactive neurons were found in the cingulate cortex, the arcuate and perifornical hypothalamic nuclei, as well as glial labelling in the hypothalamus. A striking increase in the number of immunoreactive cells was observed as a function of age. In 20 - 22-month-old rats, numerous immunoreactive cells were observed in the cingulate cortex, several thalamic and hypothalamic nuclei, the red nucleus, olivary nuclei, superior colliculus, and many cranial nerve nuclei. Whereas the immunoreactivity was restricted to a diffuse labelling of the cell bodies and processes in young rats, other forms of labelling were observed in aged rats: punctate cytoplasmic labelling and intensely stained granules with no visible cell membrane. A further increase in the density of the immunoreactive material was observed in 30 - 31-month-old rats. Double labelling experiments demonstrated that the SP40,40 immunoreactivity was almost exclusively located in neurons and not in glial cells (with the exception noted above). The distribution of SP40,40 immunoreactivity in aged rats did not coincide with the distribution of the microtubule-associated tau protein, OX42 or lipofuscin.

Isolation of two forms of decay-accelerating factor (DAF) from human urine.

Decay-accelerating factor (DAF) was purified from human pooled urine by conventional techniques. The urine DAF was separated into two peaks, pool I and pool II, by gel chromatography. DAF-U1 was isolated from pool I by hydrophobic chromatography, and DAF-U2 from pool II by anti-DAF IgG column. The specific activities of DAF-U1 and DAF-U2 to decay membrane-phase C5 convertase were about 3% and 70% of membrane form DAF, respectively. However, both urine DAFs revealed a similar activity to each other and slightly higher activity than that of membrane form DAF in decay-accelerating fluid-phase C3 convertase of the alternative pathway.

Functional and structural domains of the sixth component (C6) of human complement.

The effects of serine protease inhibitors, diisopropyl fluorophosphate (DFP) and phenylmethanesulfonyl fluoride (PMSF), on hemolytic activity of C6 were reinvestigated. C6 was inactivated in a range of 1-10 mM by both of the inhibitors as previously reported. Limited proteolytic digestion was also studied to elucidate the functional and structural domains of C6. The major fragments produced by trypsin, plasmin, or lysyl endopeptidase could not be separated unless disulfide bonds were disrupted, but Staphylococcus aureus V8 protease yielded several fragments, each of which was not linked by disulfide bond. When C6 labeled with [3H]DFP was subjected to limited digestion with V8 protease, a fragment with a molecular weight of 38 kilodaltons (kDa) was mainly labeled and other fragments of 53 kDa and 26.4 kDa were also faintly labeled, while fragment 35 kDa wasn't labeled, indicating specific domains reactive with DFP. On the other hand, when C6 with or without DFP treatment was digested with V8 protease and those fragments were incubated with C5 and subjected to sucrose density ultracentrifugation, fragments 53, 38, 35 and 27.5 kDa interacted with C5 in both cases. These results suggest that C6 modified by DFP can interact with C5, and the amino-terminal sequences of fragment 38 and 35 kDa suggest the binding domain of C6 with C5 takes place within the two short consensus repeats.

Assignment of a human serum glycoprotein SP-40,40 gene (CLI) to chromosome 8.

SP-40,40 is a serum glycoprotein consisting of two different subunits (alpha and beta) assembled into a dimer by disulfide bonds. Northern blot hybridization, using total RNA from several cell lines, showed that SP-40,40 is expressed in glioblastoma and testicular tumor cells, as well as hepatoma cells. Spot blot hybridization of flow-sorted human chromosomes, using a SP-40,40 cDNA fragment as a probe, localized the gene for SP-40,40 to human chromosome 8. This gene has been given the designation CLI, for complement lysis inhibitor, by the Human Gene Nomenclature Committee.

Sandwich ELISA assay for quantitative measurement of SP-40,40 in seminal plasma and serum.

SP-40,40 was purified from human plasma by PEG fractionation, DEAE-Sephacel, Phenyl-Toyopearl 650M, Bio-Gel A-0.5m and hydroxylapatite chromatographies. Three monoclonal antibodies (IF12, IID9 and IVF4) to this protein were prepared: IF12 and IID9 were specific for the beta subunit and IVF4 for the alpha subunit. The concentrations of SP-40,40 in seminal plasmas and sera were determined using a sandwich ELISA method. The results showed that the average concentrations of SP-40,40 were 438 +/- 285 micrograms/ml in seminal plasmas and 111 +/- 50 micrograms/ml in sera of normal donors. SP-40,40 concentrations in seminal plasmas of Klinefelter and excretory azoospermia patients were similar to those of normal donors. However, those of oligozoospermia and idiopathic azoospermia patients were about half the normal value.