Functional folate receptor beta-expressing macrophages in osteoarthritis synovium and their M1/M2 expression profiles.

OBJECTIVE: The distribution of folate receptor (FR)-ß+ macrophages and their M1/M2 expression profiles were examined in osteoarthritis (OA) synovial tissues, and compared to those in rheumatoid arthritis (RA) synovial tissues and CD163+ macrophages in both OA and RA synovial tissues. METHOD: The phenotypes and fluorescein isothiocyanate (FITC)-folate uptake of FR-ß+ synovial macrophages were analysed by flow cytometry. The distribution of FR-ß+ macrophages in OA and RA synovial tissues was examined by immunofluorescent microscopy. Tumour necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), interleukin (IL)-10, and transforming growth factor (TGF)-ß expression in FR-ß+ macrophages was detected by double-immunostaining in both OA and RA synovial tissues. RESULTS: FR-ß+ macrophages were predominantly present in the synovial lining layer in OA patients. The proportion of CD163-FR-ß+ cells in synovial mononuclear cells (MNCs) was increased in OA compared to RA synovial tissues. FR-ß(high) macrophages from OA synovial tissues represented the majority of folic acid-binding cells. Although FR-ß+ or CD163+ macrophages in the synovial tissues of OA and RA patients expressed a mixed pattern of M1 and M2 macrophage markers, there were more M2 markers expressing synovial macrophages in OA than in RA patients. CONCLUSIONS: The distribution and M1/M2 expression profiles of FR-ß+ synovial macrophages were different between OA and RA synovial tissues. Thus, the findings underscore that the M1/M2 paradigm using surface markers FR-ß and CD163 is an oversimplification of macrophage subsets. Functional FR-ß present on OA synovial macrophages provides a potential tool for the diagnosis and treatment of OA.

Stability of MALAT-1, a nuclear long non-coding RNA in mammalian cells, varies in various cancer cells.

Recent large-scale transcriptome analyses have revealed a large number of transcripts with low protein-coding potential, known as non-coding RNAs (ncRNAs). Many studies revealed that several long ncRNAs are involved in the regulation of genome organization and gene expression, or in the structural components of functional domains in the nucleus. As regulation of mRNA decay in the cytoplasm is crucial for controlling the abundance of cellular transcripts and the levels of protein expression, so regulation of long non-coding RNA decay in the nucleus is considered to be important for biological function. Although enzymatic pathways involved in cytoplasmic mRNA decay have been studied extensively, far less is known about those in nuclear long ncRNA decay. Here, we have investigated decay of metastasis associated lung adenocarcinoma transcript 1 (MALAT-1), which is a long (~ 8 kb) ncRNA that is misregulated in many human cancers and was shown to be retained specifically in the nucleus in nuclear speckles, as a model of nuclear long ncRNA in mammalian cells. We have found that the half-life of MALAT-1 ranges from ~ 9 h to > 12 h in various cancer cells. Moreover, Xrn2, PM/Scl-75, PARN, and Mtr4, known nuclear RNases or RNA helicases, did not affect MALAT-1 degradation or single knockdown of these components did not change the MALAT-1 decay rate.

Reduced expression of Sytl 1 and Ccdc21 and impaired induction of Mt I by oxidative stress in SII-K1 knockout mice.

SII-K1 is a member of the transcription elongation factor S-II family. In the mouse, SII-K1 is expressed exclusively in the liver, kidney, heart, and skeletal muscle. Here, we report that deletion of the SII-K1 gene in mice resulted in the downregulation of the synaptotagmin-like 1 (Sytl 1) gene in liver and of the coiled-coil domain-containing 21 (Ccdc21) gene in liver and kidney. Moreover, the induction of the metallothionein I (Mt I) gene in SII-K1-deficient mice liver was impaired in diethyl maleate-induced oxidative stress conditions. Our results suggest that SII-K1 regulates these genes in vivo.

Traumatic spinal-cord herniation associated with pseudomeningocele after lower-thoracic nerve-root avulsion.

BACKGROUND: Traumatic spinal-cord herniation after nerve root avulsion is rare. We report on the first patient with spinal-cord herniation associated with pseudomeningocele in the lower conus medullaris region after nerve avulsion. CASE: This 72-year-old man presented with progressive pain in the left leg and motor weakness after two traumatic accidents. Constructive interference in steady-state (CISS) imaging showed the attachment of the spinal cord to the wall of a herniated pseudomeningocele and associated syringomyelia at the level of T12. At the time of surgery, a herniated pseudomeningocele was observed. The lateral portion of the spinal cord that had herniated into the pseudomeningocele was detached from its wall; this was followed by repair of the dural defect. A redundant nerve root was observed inside the pseudomeningocele, suggesting nerve root avulsion as the primary lesion. To facilitate cerebrospinal fluid drainage from the syringomyelia, we next performed dorsal root entry zone (DREZ)tomy to the pseudomeningocele. Postoperatively, he manifested significant clinical improvement. CONCLUSIONS: This is the first report of spinal cord herniation after nerve root avulsion in the conus medullaris region. CISS imaging is highly useful for the demonstration of spinal cord herniation, syringomyelia and pseudomeningocele. To restore neurological function in patients with progressive symptoms, we recommend surgical treatment.

Defining the roles of inflammatory and anabolic cytokines in cartilage metabolism.

In osteoarthritis (OA), adult articular chondrocytes undergo phenotypic modulation in response to alterations in the environment owing to mechanical injury and inflammation. These processes not only stimulate the production of enzymes that degrade the cartilage matrix but also inhibit repair. With the use of in vitro and in vivo models, new genes, not known previously to act in cartilage, have been identified and their roles in chondrocyte differentiation during development and in dysregulated chondrocyte function in OA have been examined. These new genes include growth arrest and DNA damage (GADD)45beta and the epithelial-specific ETS (ESE)-1 transcription factor, induced by bone morphogenetic protein (BMP)-2 and inflammatory cytokines, respectively. Both genes are induced by NF-kappaB, suppress COL2A1 and upregulate matrix meatalloproteinase-13 (MMP-13) expression. These genes have also been examined in mouse models of OA, in which discoidin domain receptor 2 is associated with MMP-13-mediated remodelling, in order to understand their roles in physiological cartilage homoeostasis and joint disease.

Delayed segmental motor paralysis following laminoplasty: two case reports.

STUDY DESIGN: Two patients who experienced the onset of segmental motor paralysis several years after laminoplasty are presented. OBJECTIVES: To discuss the mechanism of development of delayed segmental motor paralysis following laminoplasty. SETTING: A department of orthopaedic surgery in Japan. METHODS: One patient experienced motor weakness in his deltoid and biceps muscles on the left side 5 years after laminoplasty. The other patient noticed motor weakness in his deltoid and biceps on the right side 7 years after laminoplasty. CT myelography revealed posterior spur formation and hypertrophic facet joints on the hinged side at the C4-C5 level in both patients. RESULTS: Posterior foraminotomy was performed at the C4-C5 level on the hinged side in both patients. Postoperatively, motor weakness in the deltoid and biceps muscles was improved in both patients. CONCLUSIONS: Although spondylotic changes, including spur formation and disc herniation, have occasionally developed in operated segments after laminoplasty, few patients have required additional surgery for treatment of relapse of neurological deficits. It has been believed that spinal cord is rarely compressed by the spondylotic changes since it shifts posteriorly in the enlarged spinal canal. However, laminoplasty disturbs the facet joints since the medial portion of dorsal cortex and cancellous bone in facet joints is drilled out to make a trough. Facet joints disturbed in this fashion undergo degeneration over time after surgery. Nerve roots may occasionally be compressed by degenerated facet joints and spurs that have developed at the entrance of root canal, resulting in segmental motor paralysis several years after laminoplasty. Careful long-term observation is necessary after this procedure.

Increased expression of humanin peptide in diffuse-type pigmented villonodular synovitis: implication of its mitochondrial abnormality.

OBJECTIVES: To define the pathogenesis of pigmented villonodular synovitis (PVNS), by searching for highly expressed genes in primary synovial cells from patients with PVNS. METHODS: A combination of subtraction cloning and Southern colony hybridisation was used to detect highly expressed genes in PVNS in comparison with rheumatoid synovial cells. Northern hybridisation was performed to confirm the differential expression of the humanin gene in PVNS. Expression of the humanin peptide was analysed by western blotting and immunohistochemistry. Electron microscopic immunohistochemistry was performed to investigate the distribution of this peptide within the cell. RESULTS: 68 highly expressed genes were identified in PVNS. Humanin genes were strongly expressed in diffuse-type PVNS, but were barely detected in nodular-type PVNS, rheumatoid arthritis, or osteoarthritis. Humanin peptide was identified in synovium from diffuse-type PVNS, and most of the positive cells were distributed in the deep layer of the synovial tissue. Double staining with anti-humanin and anti-heat shock protein 60 showed that humanin was expressed mainly in mitochondria. Electron microscopy disclosed immunolocalisation of this peptide, predominantly around dense iron deposits within the siderosome. CONCLUSIONS: Increased expression of the humanin peptide in mitochondria and siderosomes is characteristic of synovial cells from diffuse-type PVNS. Humanin is an anti-apoptotic peptide which is encoded in the mitochondrial genome. Present findings suggest that mitochondrial dysfunction may be the principal factor in pathogenesis of diffuse-type PVNS and that humanin peptide may play a part in the neoplastic process in this form of PVNS.

Primary malignant peripheral nerve sheath tumor of the cauda equina in a child case report.

STUDY DESIGN: A case report of primary malignant peripheral nerve sheath tumor (MPNST) of the cauda equina in a child is presented, and the literature is reviewed. OBJECTIVE: To discuss the problems involved in the treatment of primary intradural MPNSTs. SETTING: A department of orthopaedic surgery in Japan. METHODS: A 4-year-old boy complained of low-back pain radiating to the left calf. MRI revealed an intradural tumor at L3-L5 level. Following laminectomy of L3, L4 and L5, the tumor was removed en bloc. Based on pathological and immunohistological findings, the tumor was diagnosed as an MPNST. RESULTS: Although adjuvant chemotherapy was administered local recurrence and cerebral and spinal metastases of the tumor were found 6 months after the operation. Following additional incomplete removal of the recurrent tumor, radiation therapy was administered. Although recurrent and metastatic tumors disappeared or diminished in size by radiation, tumors increased in size thereafter, despite additional adjuvant chemotherapy. At 21 months after the first operation, he died of pneumonia. CONCLUSIONS: Reported clinical outcomes for patients with primary intradural MPNST are very poor. Although no gold standard for the treatment of tumors has been established yet, surgical removal of tumors combined with postoperative high-dose radiation may be recommended.

Use of an otolith-deficient mutant in studies of fish behavior in microgravity.

The mutant strain (ha) of medaka (Oryzias latipes) lack utricular otoliths as fry, and some never form otoliths for life. The cross (F1 generation) between the strain having good eyesight and another strain having ordinary eyesight augmented visual acuity of the F1 generation. Crossing the good eyesight strain and ha mutant produced fish having good eyesight and less sensitivity to gravity in the F2 population. Their tolerance to microgravity was tested by parabolic flight using an airplane. The fish exhibited less looping and no differences in degree of looping between light and dark conditions, suggesting that loss of eyesight (in darkness) is not a direct cause for looping behavior in microgravity. The ha embryos could not form utricular otoliths. They did form saccular otoliths, but with a delay. Fry of the mutant fish lacking the utricular otoliths are highly dependent on light upon hatching and exhibit a perfect dorsal-light response (DLR). As they grow, they eventually shift from being light-dependent to being gravity-dependent. Continuous treatment of the fry with altered light direction suppressed this shift to gravity dependence. Being less dependent on gravity, these fish can serve as models in studying the differences expected for the vestibular system of fish reared in microgravity. When these fish were exposed to microgravity (parabolic flights) of an airplane, they spent far less time looping than fish reared in an ordinary light regimen.

Otolith formation in a mutant Medaka with a deficiency in gravity-sensing.

Mutant Medaka ha exhibit spontaneous mutation that is characterized by frequent inhibition or perturbation in the formation of utricular otoliths and/or semicircular canals. Three major features of otolith morphogenesis were observed in ha strain: 1) The initial appearance of otoliths was delayed, mispositioned, and malformed compared to normal embryos. 2) No utricular otoliths appeared on macula of any ha fry just after hatching. A symmetric state of otoliths was seen only when saccular otoliths were situated on macula in both inner ears. 3) In some fry, formation of utricular otoliths was observed in their later development. However, no new utricular otoliths appeared after fish were seventy or more days old after hatching. These observations show that otolith morphogenesis in ha is very different from that of wild-type. In this study, we classified adult ha into four different phenotypes using the existence or absence of utricular otoliths as our criteria. We concluded that dysfunction of utricular otoliths and semicircular canals cause a defect that affects the gravity-sensing abilities of medaka ha.

LY309887, antifolate via the folate receptor suppresses murine type II collagen-induced arthritis.

OBJECTIVE: To examine the effect of LY309887, an inhibitor of glycinamide ribonucleotide formyltransferase in de novo purine biosynthesis on murine type collagen-induced arthritis (CIA). METHODS: CIA was induced by immunization with bovine type II collagen in adjuvant. The expression of folate receptors was examined in dissected synovial tissues and bone marrow cells from arthritic and non-arthritic mice by the semi-quantitative reverse transcription-polymerase chain reaction. LY309887 was administered to CIA mice after the onset of arthritis. Mice were monitored for arthritis index for 21 days. Levels of IgG1 and IgG2a antibodies against bovine type II collagen were measured in sera from CIA mice with or without LY309887 treatment by the enzyme-linked immunosorbent assay. Histologic analyse were also performed in synovial tissues from arthritic joints with or without LY309887 treatment. RESULTS: Levels of mRNA of folate receptor beta (FR-beta) were elevated in arthritic joints from CIA mice, compared with those in nonarthritic joints. The expression of mRNA of FR-beta was dominant in bone marrow cells of CIA mice. The administration of LY309887 suppressed the disease progression of CIA mice as defined by the lower arthritis index, and decreased production of serum IgG1 and IgG2a anti-type II collagen antibody, and the damage to cartilage or bone. CONCLUSION: The administration of LY309887 was effective on CIA mice. It was suggested that LY309887 might be useful in the treatment of rheumatoid arthritis.

Aquatic animal research in space station and its issues--focus on support technology on nitrate toxicity.

We studied the effects of accumulated nitrate in water on the spawning, hatching and development of medaka using a simple nitrifying filter and a combined filter having both nitrifying and denitrifying capabilities. A nitrate concentration of 100 mg NO3(-)-N/L was clearly of lethal toxicity to fish when they were exposed to nitrate in both adult and the growing phases. A nitrate concentration of 75 mg NO3(-)-N/L reduced the fertilization rate, delayed the hatching time and reduced the hatching rate of the eggs laid by adults and decreased the growth rate of juveniles. In addition, nitrate accumulations as low as 50 mg NO3(-)-N/L remarkably retarded spawning and lowered the number of eggs laid by fish exposed in the juvenile phase. The effects on the reproduction system may be initiated by a low concentration, approximately 30 mg NO3(-)-N/L.

Swimming behavior of larval Medaka fish under microgravity.

Fish exhibit looping and rolling behaviors when subjected to short periods of microgravity during parabolic flight. Strain-differences in the behavioral response of adult Medaka fish (Oryzias latipes) were reported previously, however, there have been few studies of larval fish behavior under microgravity. In the present study, we investigated whether microgravity affects the swimming behavior of larvae at various ages (0 to 20 days after hatching), using different strains: HNI-II, HO5, ha strain, and variety of different strains (variety). The preliminary experiments were done in the ground laboratory: the development of eyesight was examined using optokinetic response for the different strains. The visual acuity of larvae improved drastically during 20 days after hatching. Strain differences of response were noted for the development of their visual acuity. In microgravity, the results were significantly different from those of adult Medaka. The larval fish appeared to maintain their orientation, except that a few of them exhibited looping and rolling behavior. Further, most larvae swam normally with their backs turning toward the light source (dorsal light response, DLR), and the rest of them stayed with their abdomen touching the surface of the container (ventral substrate response, VSR). For larval stages, strain-differences and age-differences in behavior were observed, but less pronounced than with adult fish under microgravity. Our observations suggest that adaptability of larval fish to the gravitational change and the mechanism of their postural control in microgravity are more variable than in adult fish.

[Aquatic animals and use of centrifuge in space experiments].

There are some experiments which do not need a centrifuge apparatus in space, however, for other space experiments the use of centrifuge is indispensable as the control. The characteristics of these two types of space experiments are explained. Generally, the more quantitatively the phenomenon is analyzed, the greater the need of centrifuge apparatus becomes. The centrifuge in space can realize various gravity values ranging from 0 to 1 G, and this will extend the present biology to a more universal biology.

Nonsurgically managed patients with degenerative spondylolisthesis: a 10- to 18-year follow-up study.

OBJECT: Controversy exists concerning the indications for surgery and choice of surgical procedure for patients with degenerative spondylolisthesis. The goals of this study were to determine the clinical course of nonsurgically managed patients with degenerative spondylolisthesis as well as the indications for surgery. METHODS: A total of 145 nonsurgically managed patients with degenerative spondylolisthesis were examined annually for a minimum of 10 years follow-up evaluation. Radiographic changes, changes in clinical symptoms, and functional prognosis were surveyed. Progressive spondylolisthesis was observed in 49 patients (34%). There was no correlation between changes in clinical symptoms and progression of spondylolisthesis. The intervertebral spaces of the slipped segments were decreased significantly in size during follow-up examination in patients in whom no progression was found. Low-back pain improved following a decrease in the total intervertebral space size. A total of 84 (76%) of 110 patients who had no neurological deficits at initial examination remained without neurological deficit after 10 years of follow up. Twenty-nine (83%) of the 35 patients who had neurological symptoms, such as intermittent claudication or vesicorectal disorder, at initial examination and refused surgery experienced neurological deterioration. The final prognosis for these patients was very poor. CONCLUSIONS: Low-back pain was improved by restabilization. Conservative treatment is useful for patients who have low-back pain with or without pain in the lower extremities. Surgical intervention is indicated for patients with neurological symptoms including intermittent claudication or vesicorectal disorder, provided that a good functional outcome can be achieved.

Results of a longer than 10-year follow-Up of patients with rheumatoid arthritis treated by occipitocervical fusion.

STUDY DESIGN: Evaluation of results a longer than 10-year follow-up of patients with upper cervical lesions due to rheumatoid arthritis who underwent occipitocervical fusion. OBJECTIVE: To determine the final outcome of patients with upper cervical lesions due to rheumatoid arthritis treated by occipitocervical fusion. SUMMARY OF BACKGROUND DATA: There are few studies reporting the final outcome of patients with rheumatoid arthritis treated by occipitocervical fusion and observed for longer than 10 years. METHODS: The subjects were 16 patients with rheumatoid arthritis with myelopathy who underwent occipitocervical fusion with a rectangular rod more than 10 years ago. All 16 patients had irreducible atlantoaxial dislocation, and 11 also had vertical dislocation of the axis. All patients had preoperative nuchal pain, and were classified into Class II (two patients), Class IIIA (nine patients), and class IIIB (five patients) according to Ranawat's preoperative neurologic classification. RESULTS: The atlas-dens interval remained the same as immediately after surgery. Vertical dislocation returned to the preoperative condition, despite successful surgical correction. Preoperative occipital pain disappeared or was reduced in all cases. Myelopathy improved in 12 of the 16 patients (75%) by more than one class in the Ranawat preoperative neurologic classification. Survival rate at 10 years after surgery was 38%; mean age at death was 70.7 years. The postoperative periods during which patients could walk by themselves ranged from 6 months to 13 years (mean, 7.5 years). CONCLUSIONS: Occipitocervical fusion for patients with rheumatoid arthritis is useful for decreasing nuchal pain, reducing myelopathy, and improving prognosis.

Mutant Medaka fish having a deficiency in otolith-vestibular system and their behavior in microgravity.

Vestibular and visual senses are two major factors fish use for controlling their posture under 1 G conditions. From various studies in the past (parabolic flights and some ground-laboratory experiments), I came to have a hypothesis that "heavily eye-dependent" fish are tolerant to microgravity (micro-G). Theoretically, for "heavily eye-dependent" fish, the following three types may exist. (a) Fish having good eyesight, and having an ordinary sensitivity to gravity, (b) Fish having an ordinary eyesight, and having less sensitivity to gravity, (c) Fish having good eyesight, and having less sensitivity to gravity. In Medaka fish (Oryzias latipes), these three types have successfully been obtained. In this paper, the behavior of such different types of Medaka fish under micro-G will be studied by use of parabolic flights of an airplane. The ground-laboratory experiments are also reported, which focus on the contribution of both vestibular and visual senses to the posture control for each of three types of Medaka fish.