Relationship between arthroscopic findings of synovitis and levels of tumour necrosis factor-alpha and matrix metalloproteinases in synovial lavage fluid from patients with unilateral high mandibular condyle fractures.

Arthrocentesis has an effect of washing out inflammatory products that accumulate in the joint compartment of a dysfunctional temporomandibular joint (TMJ). The procedure removes inflammatory cytokines, which are pain-causing substances, for early reduction of TMJ pain and quick recovery of jaw function, thus increasing the possibility of a successful rehabilitation. The aim of this study was to investigate the relationship between arthroscopy synovitis grade in patients with unilateral high condylar fractures and concentrations of the pro-inflammatory cytokines tumour necrosis factor (TNF)-alpha as well as of matrix metalloproteinases (MMPs) in washed-out synovial fluid (SF) samples obtained from those patients. A total of 26 patients with unilateral high condylar fractures who underwent arthrocentesis for a therapeutic purpose were examined. SF samples were collected before performing arthroscopy to determine synovitis grade. The detection rates and concentrations of TNF-alpha and MMPs were determined, and their association with synovitis grade was analysed. TNF-alpha was detected in 23 and MMP-3 in 22 of the TMJs. There was a correlation between synovitis grade and concentration of TNF-alpha in the fracture group. Furthermore, the concentrations of TNF-alpha and MMP-3 were significantly higher as compared to the control group, comprised of TMJs on the non-fracture side of the same patients, while a correlation was also noted between TNF-alpha concentration and synovitis grade in the fracture group. The present findings may provide a biological/biochemical rationale for arthrocentesis as a reasonable treatment modality for high condylar fractures.

Experimental comparison between tractional and compressive stress on temporomandibular joint.

OBJECTIVE: We experimentally compared the effects of compressive and tractional mechanical stress on the temporomandibular joint (TMJ) of rabbits to assess the etiology of progressive condylar resorption. MATERIALS AND METHODS: We performed a cortical osteotomy using custom-made devices that were lengthened by 0.25 mm every 12 h for 1 week after surgery. During this time, the rabbit TMJ was under compressive or tractional mechanical stress. The samples in each group were examined using micro-computed tomography and histological staining. RESULTS: Scores for the area of bone resorption were higher in the compressive group than in the tractional group. Moreover, scores for the depth of bone resorption were higher in the compressive group than those in the tractional group. We observed a significantly higher prevalence of resorption in the subcondylar bone in the compressive group than in the tractional group. There were substantially more cells that were positive for tartrate-resistant acid phosphatase in the compressive group than in the control and tractional groups. CONCLUSIONS: The outcomes here suggest that excessive mechanical stress, particularly compressive mechanical stress, may significantly affect morphological bone change findings in the TMJ.

Facilitation of bone resorption activities in synovial lavage fluid patients with mandibular condyle fractures.

The aim of this study was to investigate the bone resorption effect of the mediators delivered in joint cavity of patients with mandibular condyle fractures by detecting osteoclast markers using cellular biochemistry methods, and by analysing bone resorption activities via inducing osteoclast differentiation of the infiltrated cells from arthrocentesis. Sixteen joints in 10 patients with mandibular condyle fractures were evaluated. The control group consisted of synovial fluid (SF) samples from seven joints of four volunteers who had no clinical signs or symptoms involving the temporomandibular joint (TMJ) or disc displacement. We collected SF cells from all patients during therapeutic arthrocentesis. The infiltrating cells from TMJ SF were cultured, differentiated into tartrate-resistant acid phosphatase (TRAP)-positive osteoclast-like cells and examined bone resorption activities. We also investigated factors related to osteoclast induction of SF, using ELISA procedures. Osteoclast-like cells were induced from the SF cells obtained from all patients with condylar fractures. These multinucleated giant cells were positive for TRAP and actin, and had the ability to absorb dentin slices. The levels of macrophage colony-stimulating factor (M-CSF), prostaglandin E2 (PGE2), soluble form of receptor activator of nuclear factor kappa-B ligand (sRANKL) and osteoprotegerin (OPG), in SF samples from the patients, were significantly higher than in the controls. These findings indicate that bone resorption activities in SF from patients with mandibular condyle fractures were upregulated and may participate in the pathogenesis and wound healing.

Clinical comparison between arthrocentesis and conventional conservative treatment with maxillomandibular fixation for unilateral high condylar fractures.

This study aimed to compare the effects of arthrocentesis and conventional closed reduction for unilateral mandibular condyle fractures. A total of 30 patients with unilateral condylar fractures were evaluated. Patients with a high condylar fracture and magnetic resonance evidence of joint effusion (JE) were divided into two groups: those treated with intra-articular irrigation and betamethasone injection (group I) and those given conservative treatment and rigid maxillomandibular fixation (MMF) (group II). All patients were assessed for mandibular range of motion (ROM), protrusive movements, lateral excursion movements on the fractured and non-fractured sides, pain in the temporomandibular joint and malocclusion, both before and after treatment. There were no significant differences in regard to protrusion, lateral excursion movement and incidence of malocclusion at 12 months after treatment between the groups (P > 0.05). In group I, ROM and joint pain showed good improvement from the early stages of treatment, and those patients had better outcomes as compared to group II for those parameters at 1 and 3 months after injury. The present findings indicate that arthrocentesis may be more effective and provide faster healing than conventional closed reduction.

Pharmacokinetic evaluation of intraperitoneal doxorubicin in rats.

The intraperitoneal (ip) administration of doxorubicin (DOX) is considered to be an important approach for the treatment of peritoneal tumors, because the prognosis of peritoneal cancer is generally poor due to its refractoriness to conventional chemotherapy. In the present study, we examined the disposition behavior of DOX after ip administration in rats to evaluate the adequacy of the ip administration of DOX on the basis of pharmacokinetic aspects. By comparing the area under the serum concentration-time curve (AUC) after ip and intravenous (iv) dosing of 5 mg/kg DOX, the bioavailability of intraperitoneally administered DOX was estimated as 43.8%. This finding suggests that the majority of DOX remained in the abdominal cavity without being incorporated into the systemic circulation. The mean residence time (MRT) of DOX after its ip administration was about 80% longer than that after its iv administration, which indicated the slow absorption process associated with ip application. No significant difference was observed in the elimination rates of systemically absorbed DOX. These results indicate that the ip administration of DOX likely provided an adequate opportunity for it to interact with peritoneal tumors by maintaining sufficient DOX levels while reducing its systemic exposure

Weekly teriparatide injections successfully treated advanced bisphosphonate-related osteonecrosis of the jaws.

This study investigated whether weekly teriparatide (TPTD) injections are as effective as daily teriparatide injections for the treatment of stage 3 bisphosphonate-related osteonecrosis of the jaws (BRONJ) and compared serum markers of bone turnover between the two treatment regimens. Daily TPTD treatment has recently been reported to be effective for BRONJ, but there are no reports describing the effectiveness of weekly TPTD injections. We report two patients with stage 3 BRONJ. One patient was successfully treated with weekly TPTD injections and the other with daily TPTD injections. Changes in the levels of serum N-telopeptide of type I collagen (s-NTX) and serum N-terminal propeptide of type I collagen (P1NP) were studied. Two patients with stage 3 BRONJ that was refractory to conservative treatment were treated with TPTD. Their medical records were reviewed and the patients were interviewed. There was complete mucosal coverage of the intraoral defects after 3 months of TPTD treatment in both patients. Progressive bone regeneration in an area of mandibular fracture was identified after 4 months of treatment. The s-NTX level increased slightly in both patients. This is the first report of successful treatment of stage 3 BRONJ with weekly TPTD injections. Either daily or weekly TPTD injections may effectively treat stage 3 BRONJ and should be considered before or perhaps even in lieu of undertaking major resection and reconstruction.

Relationship between temporomandibular joint pain and magnetic resonance imaging findings in patients with temporomandibular joint disorders.

The purpose of this study was to evaluate abnormal magnetic resonance imaging (MRI) findings related to temporomandibular joint (TMJ) pain. This study included 245 joints of 152 patients with temporomandibular disorders with anterior disc displacement; of these, 129 joints had joint pain whereas 116 joints had no joint pain. MRI was used to evaluate the reduction of anterior disc displacement, joint effusion, mandible condylar morphology, bone marrow oedema of the mandibular condyle, and signal intensity of the posterior disc attachment (PDA) on fat-suppressed T2-weighted images. The odds ratio (OR) for each MRI variable for the pain group versus the no pain group was computed using logistic regression analysis. Univariate logistic regression analysis showed significant correlations between TMJ pain and all MRI findings. Multivariate logistic regression analysis showed significant correlations with joint effusion (P=0.03, OR 2.21), bone marrow oedema (P<0.001, OR 11.75), and signal intensity of the PDA (P<0.001, OR 6.21). These results suggest that bone marrow oedema, high signal intensity of the PDA on fat-suppressed T2-weighted images, and joint effusion, in descending order of influence, are factors related to TMJ pain.

Acral myxoinflammatory fibroblastic sarcoma of the foot.

We report a rare case of acral myxoinflammatory fibroblastic sarcoma (AMFS) in a 68-year-old woman. Tumor excision of a mass between the 1st and 2nd toe of the left foot was performed after a diagnosis of ganglion in February 2003. Examination of the surgical specimen confirmed AMFS. No recurrence or metastasis occurred during the follow-up period of 4 years. Clinical characteristics such as recurrence rate, metastasis rate and period of metastasis are unclear for AMFS. Long-term clinical follow-up is thus required.

Genetic analysis of high-metastatic clone of RCT sarcoma in mice, and its growth regression in vivo in response to angiogenesis inhibitor TNP-470.

Genetic analysis of a high-metastatic clone of RCT sarcoma (HM-RCT) was the aim of the study. HM-RCT was developed by the lung passage as well as limiting dilution method from the original RCT sarcoma, in which a tumor was spontaneously developed in a C3H/He mouse. HM-RCT expressed enhanced POU domain (class 2, associating factor 1), adenylate cyclase 7, procollagen type III (alpha), A kinase anchor protein 4 and Ehm (expressed on high-metastatic cells) and 11 expressed sequence tags (ESTs). compared with the original clone of RCT. Eighteen specific genes and 14 ESTs were underexpressed in HM-RCT. We investigated the effects of angiogenesis inhibitor TNP-470 on tumor growth and metastasis of this HM-RCT in vivo. In an experimental group, mice received TNP-470 (30 mg/kg) intraperitoneally every other day. After 5 weeks, the growth of the TNP-470-treated tumor was significantly suppressed in vivo, but did not affect the metastasis. The proportion of positive PCNA-stained cells and cellular telomerase activity was significantly low in response to TNP-470.

Experimental infection of Dirofilaria immitis in raccoon dogs.

Canine heartworm (Dirofilaria immitis) is a nematode that naturally parasitizes in the pulmonary arteries and the right ventricle of domestic dogs (Canis familiaris) as final hosts. Japanese raccoon dogs (Nyctereutes procyonoides viverrinus) also are known to be susceptible to infection by the parasite. However, prevalence of this infection among free-ranging raccoon dogs is low and so is the worm burden. To examine the susceptibility of the raccoon dog to D. immitis infection, 3 raccoon dogs and 2 beagles were inoculated 4 times with 25 third-stage larvae (L3s) of D. immitis at 3-wk intervals. Worms were recovered from 2 raccoon dogs and both domestic dogs. The average percentage of recovery (2.3%) of the raccoon dogs was almost 10 times lower (24.5%) than that of the domestic dogs, but there was no significant difference in the body length of worms recovered from 2 types of hosts. To examine microfilaremia, 2 raccoon dogs were infected with 100 L3s. Microfilaremia was observed for 180 days postinoculation (PI) but disappeared at about 300 days PI. The raccoon dog was mildly susceptible to infection with D. immitis, but surviving worms developed and matured normally.

Detection of the MYD88 mutation by the combination of the allele-specific PCR and quenching probe methods.

INTRODUCTION: The MYD88 missense mutation c.794T>C, p.Leu265Pro, is found in patients with Waldenstörm's macroglobulinemia and lymphoma. Direct sequencing, allele-specific PCR (AS-PCR), PCR-restriction fragment length polymorphism (PCR-RFLP), and high-resolution melting analysis (HRM) are currently used to detect the mutation; however, they are either time-consuming or have low detection sensitivity. Here, we developed a novel highly sensitive and rapid detection method based on the quenching probe (QP) technique and AS-PCR. METHOD: A lymphoma cell line heterozygous for the MYD88 mutation, two wild-type cell lines, and two samples from Waldenstörm's macroglobulinemia patients were analyzed by AS-PCR, PCR-RFLP, HRM, and QP, and their detection sensitivity was examined using the mixtures of the mutant and wild-type DNA. RESULTS: For mutation-carrying heterozygous samples, the QP method produced W-shaped melting profiles presenting curves derived from the wild-type and mutant alleles. The QP analysis was performed in 2 h and demonstrated the detection limit of 5%, which was similar to that of the other methods. However, the combination of AS-PCR and QP (AS-QP) improved the sensitivity to 0.62% of the mutant allele. CONCLUSION: The AS-QP analysis is rapid and minimally improves detection sensitivity compared to the AS-PCR.

Probing novel elements for protein splicing in the yeast Vma1 protozyme: a study of replacement mutagenesis and intragenic suppression.

Protein splicing is a compelling chemical reaction in which two proteins are produced posttranslationally from a single precursor polypeptide by excision of the internal protein segment and ligation of the flanking regions. This unique autocatalytic reaction was first discovered in the yeast Vma1p protozyme where the 50-kD site-specific endonuclease (VDE) is excised from the 120-kD precursor containing the N- and G-terminal regions of the catalytic subunit of the vacuolar H(+)-ATPase. In this work, we randomized the conserved valine triplet residues three amino acids upstream of the C-terminal splicing junction in the Vma1 protozyme and found that these site-specific random mutations interfere with normal protein splicing to different extents. Intragenic suppressor analysis has revealed that this particular hydrophobic triplet preceding the C-terminal splicing junction genetically interacts with three hydrophobic residues preceding the N-terminal splicing junction. This is the first evidence showing that the N-terminal portion of the V-ATPase subunit is involved in protein splicing. Our genetic evidence is consistent with a structural model that correctly aligns two parallel beta-strands ascribed to the triplets. This model delineates spatial interactions between the two conserved regions both residing upstream of the splicing junctions.

Protective immunity against Brugia malayi infective larvae in mice. I. Parameters of active and passive immunity.

Protective immunity against infective larvae of Brugia malayi was studied in different strains of mice using various sources of antigens. The following strains of mice were susceptible to infective larvae development for 2 weeks after primary ip challenge: BALB/c, C3H/HeJ, C3H/NeN, C3H/HeJms, C57BL/6Jms, and DDD. In comparison to gerbils, BALB/c mice developed stronger resistance to infective larvae after immunization with irradiation attenuated larvae or with killed microfilariae (mf). However, killed mf failed to enhance resistance in C3H/HeJ mice, although C3H/HeN mice were strongly protected and C3H/HeJms mice were protected to a lesser degree by this antigen. Extracts of mf with phosphate buffered saline and sodium dodecyl sulfate both induced high levels of resistance in BALB/c mice. Transfer of resistance from BALB/c mice immunized with attenuated infective larvae to naive mice was accomplished at a high level at protection with nylon wool nonadherent spleen cells (T cells) but not with adherent cells treated with anti-Thy 1.2 serum and complement. In contrast, sera from immunized mice were much less protective.

Dirofilaria immitis: experimental infection of rabbits with immature fifth-stage worms.

To establish an animal model for human pulmonary dirofilariasis, we experimentally infected nine rabbits with immature fifth-stage worms of Dirofilaria immitis. The rabbits were infected by subcutaneous transplantation with various numbers of immature worms collected from 110- and 120-day-old infections of dogs. Four of seven rabbits infected with up to four larvae possessed encapsulated worms in the lungs at 196 or 308 days post-transplantation. Two rabbits transplanted with eight worms died of pulmonary hemorrhagic infarction 18 and 28 days post-transplantation. Marked histopathologic changes were observed in the lungs at the site of degenerating worms, which were encapsulated by a fibrous wall. Severe to mild infiltrations with eosinophils, heterophils, lymphocytes, plasma cells, and histiocytes were found in granulomas and their surrounding areas. The findings in these rabbits resemble those reported for human cases of pulmonary dirofilariasis.

Involvement of macrophage scavenger receptors in protection against murine malaria.

Macrophage scavenger receptor A (MSR-A) deficient mice MSR-A(-/-) were infected by the intraperitoneal injection of the Plasmodium berghei NK65 strain in the erythrocytic stage. The MSR-A(-/-) mice died significantly earlier than the control mice (P=0.060). In the surviving mice, two peaks of parasitemia were observed: the first 5-7 days and the second at 2-3 weeks after infection. Death of all MSR-A(-/-) mice occurred at either peak of parasitemia, suggesting that MSR-A protects mice from severe infection. This model may be useful for the study of molecular mechanisms of macrophage functions in malaria infection.

Macrophage migration inhibition test of peritoneal exudate cells of Litomosoides carinii-infected cotton rats by direct method in agar plate.

The macrophage migration inhibition test (MIT) was successfully made in agar plates using peritoneal exudate cells (PEC) of cotton rats, Sigmodon hispidus, during infection with Litomosoides carinii. It was found that 1% Bacto agar plate containing heat-inactivated serum was most suitable for the migration of PEC of the cotton rat induced by liquid paraffin injection. Ten percent normal cotton rat serum mixed in agar plate was best for PEC migration as compared with the various concentrations of horse, fetal and newborn calf sera. Migration areas of PEC from normal cotton rats increased with the number of cells per well and 2 X 10(6) cells/well was optimal. With concentrations of greater than or equal to 20 micrograms/ml of adult L. carinii antigen, significant migration inhibition of PEC from infected animals was observed from 18 hours after incubation at 37 degrees C with 5% CO2 gas phase, as compared with the migration of PEC from non-infected animals. Comparative analyses of the different aspects of infection showed that there was no correlation between the degree of migration inhibition and the duration of infection or the number of live worms recovered from the pleural cavity. Cell-mediated immunity against L. carinii as indicated by MIT still existed in 18 months after infection, although adult worms had died and were absorbed in the pleural cavity of the cotton rat.

Quantitative analysis of microfilarial periodicity of Dirofilaria immitis in cats.

Microfilarial periodicity of Dirofilaria immitis in the venous blood of infected cats was analyzed by a trigonometric model. Cats were infected by subcutaneous transplantation with 120-day-old juvenile D. immitis. Microfilariae in the blood were first observed 98 days after transplantation. Blood was collected at 4h intervals for a 24h period, and examinations were repeated five times in two cats. The calculated periodicity index was 75.1 and 50.3 in these two cats. The estimated hour of peak microfilarial density ranged from 1.00 to 2.84h. Thus, the periodicity of microfilariae of D. immitis in the blood of cats was characterized as nocturnally sub-periodic.

Histopathology of the lungs of rabbits experimentally infected with Dirofilaria immitis.

The lungs of rabbits experimentally infected with Dirofilaria immitis were examined histopathologically, to compare the changes with those seen in human pulmonary dirofilariasis. Eight rabbits were infected subcutaneously with two to eight immature worms to induce pulmonary dirofilariasis. Obstructive changes, similar to those reported in human pulmonary dirofilariasis, were observed in the blood vessels surrounding the worms. A form of arteritis, similar to occlusive arteritis, and periarteritis were also observed in the lungs of the rabbits. These results suggest that experimentally induced dirofilariasis in the rabbit is a useful animal model for human pulmonary dirofilariasis.

Preservation of feline anti-Toxoplasma gondii antibody activity using blood absorbed on filter paper stored under different conditions.

The present study deals with the successful long-term preservation of feline anti-Toxoplasma gondii antibody activity on filter paper strips stored with silica gel. When filter paper strips were kept at 25 degrees C with silica gel, antibody activity in blood samples was preserved for at least six months.

Prevalence of Dirofilaria immitis infection in cats in Saitama, Japan.

To clarify Dirofilaria immitis infection among cats in Saitama Prefecture, Japan, 1,840 cats were examined postmortem for adult worms and microfilariae in the blood from 1989 to 1995. As a reference control, 500 dogs from the same area were examined in the same way and period. D. immitis worms were found in 15 cats, one of which had microfilariae in the blood. Prevalence rate of D. immitis infection was 0.8% (15/1,840) in cats and 46.8% (234/500) in dogs examined, whereas it was 4.1% and 64.6% in cats and dogs, respectively, aged 2 years and over. Worm burden per positive cat was 1.5 +/- 0.7 (mean +/- SD), the maximum number of worm was 3 in 2 cats, and 10 cats had a single worm each. All the worm-positive cats were tested for antibodies to feline immunodeficiency virus (FIV) and antigens of feline leukemia virus (FeLV) in sera. Positive rates of coinfection with D. immitis were 26.7% and 13.3% for FIV and FeLV, respectively.