Multiple Epiphyseal Dysplasia With Knee Joint Locking Symptoms Caused by Intra-articular Loose Bodies.

Multiple epiphyseal dysplasia (MED) is a congenital disease causing epiphyseal dysplasia in long bones. Herein, we report a case of a middle-aged man with bilateral knee joint locking symptoms who was diagnosed with multiple epiphyseal dysplasia caused by Matrilin-3 (MATN3) pathogenic variants and was successfully treated with arthroscopic loose body removal. A 48-year-old man has had bilateral knee pain since his twenties and underwent loose body removal of both knees in his thirties. He visited our hospital for worsening locking symptoms in both knees. Twenty years ago, his son had been diagnosed with suspected multiple epiphyseal dysplasia. Genetic and imaging testing confirmed his diagnosis of multiple epiphyseal dysplasia due to Matrilin-3 pathogenic variants. Arthroscopic loose body removal was performed, and the locking symptoms disappeared after surgery. Arthroscopic loose body removal was effective for the locking symptoms in a mild adult case of multiple epiphyseal dysplasias caused by Matrilin-3 pathogenic variants.

Co-continuous structural effect of size-controlled macro-porous glass membrane on extracellular vesicle collection for the analysis of miRNA.

Recent studies have shown that extracellular vesicles (EVs) can be utilized as appropriate and highly specific biomarkers in liquid biopsy for the diagnosis and prognosis of serious illness. However, there are few methods that can collect and isolate miRNA in EVs simply, quickly and efficiently using general equipment such as a normal centrifuge. In this paper, we developed an advanced glass membrane column (AGC) device incorporating a size-controlled macro-porous glass (MPG) membrane with a co-continuous structure to overcome the limitations of conventional EV collection and miRNA extraction from the EVs. The size of macro-pores in the MPG membrane could be accurately controlled by changing the heating temperature and time on the basis of spinodal decomposition of B2O3, Na2O, and SiO2 in phase separation. The AGC device with an MPG membrane could collect the EVs simply and quickly (< 10 min) from cell culture supernatant, serum and urine. This AGC device could extract miRNA from the EVs captured in the MPG membrane with high efficiency when combined with a miRNA extraction solution. We suggest that the AGC device with an MPG membrane can be useful for the diagnosis and prognosis of serious illness using of EVs in various kinds of body fluids.

[Is Concomitant Therapy with Acetaminophen and Low-dose Aspirin a Risk Factor for CKD Progression? A 6-Year Cohort Study].

Concomitant therapy with acetaminophen (APAP) and low-dose aspirin is often used in clinical settings; however, it is unclear whether this combination is involved in the progression of chronic kidney disease (CKD). We hypothesized that concomitant therapy with APAP and low-dose aspirin may cause CKD progression. We carried out a retrospective 6-year cohort study that included all patients who received low-dose aspirin from January 2011 to December 2016 at Kaetsu Hospital. Primary outcome was defined as CKD progression at the end of the study compared with baseline. Among the 441 patients treated during the study period, we identified 89 cases of CKD progression. Multivariate regression analysis showed that exposure to APAP>50 g [odds ratio (OR), 2.68, 95% confidence interval (CI), 1.08-6.70], age increase by 1 year (OR, 1.05, 95% CI, 1.02-1.08), and diabetes mellitus (OR, 2.40, 95% CI, 1.41-4.08) had positive associations with CKD progression. Our findings suggested that concomitant therapy with APAP and low-dose aspirin increased the risk of CKD progression. Therefore, we recommend more thorough monitoring of serum creatinine when patients are on such concomitant therapy. Moreover, it is important to advise users of low-dose aspirin to avoid unnecessary use of APAP, in order to reduce the risk of CKD progression.

DoBISCUIT: a database of secondary metabolite biosynthetic gene clusters.

This article introduces DoBISCUIT (Database of BIoSynthesis clusters CUrated and InTegrated, http://www.bio.nite.go.jp/pks/), a literature-based, manually curated database of gene clusters for secondary metabolite biosynthesis. Bacterial secondary metabolites often show pharmacologically important activities and can serve as lead compounds and/or candidates for drug development. Biosynthesis of each secondary metabolite is catalyzed by a number of enzymes, usually encoded by a gene cluster. Although many scientific papers describe such gene clusters, the gene information is not always described in a comprehensive manner and the related information is rarely integrated. DoBISCUIT integrates the latest literature information and provides standardized gene/module/domain descriptions related to the gene clusters.

Outcome of transfibular ankle arthrodesis with Ilizarov apparatus.

BACKGROUND: Both transfibular ankle arthrodesis with fibular onlay grafting and ankle arthrodesis using the Ilizarov apparatus have resulted in acceptable fusion rates. This study analyzed outcomes in the authors' practice for all patients who underwent transfibular ankle arthrodesis with fibular onlay grafting using the Ilizarov apparatus. MATERIALS AND METHODS: A retrospective review was conducted of all patients who experienced severe ankle pain associated with osteoarthritis, rheumatoid arthritis, talar osteonecrosis, or septic arthritis and underwent transfibular ankle arthrodesis with an Ilizarov apparatus between 1995 and 2010. Postoperative clinical outcomes were evaluated for 42 consecutive patients (43 feet) who received primary or revision ankle arthrodesis using the Ilizarov technique at two centers. Mean duration of follow-up was 4.1 (range, 1.5 to 13) years. Outcomes were assessed using the ankle-hindfoot scale of the American Orthopaedic Foot and Ankle Society. RESULTS: All ankles fused successfully. Mean clinical outcome improved significantly from 36.2 (range, 8 to 64) preoperatively to 77.4 (range, 0 to 86; p < .05) postoperatively. CONCLUSION: Transfibular ankle arthrodesis using the Ilizarov external fixation system and fibular onlay strut grafting can achieve a very high rate of bony union and lead to general improvements in clinical outcome. Forefoot stabilization with a forefoot ring achieved rigid stabilization resulting in a shorter duration of external fixation.

Deciphering the genome of polyphosphate accumulating actinobacterium Microlunatus phosphovorus.

Polyphosphate accumulating organisms (PAOs) belong mostly to Proteobacteria and Actinobacteria and are quite divergent. Under aerobic conditions, they accumulate intracellular polyphosphate (polyP), while they typically synthesize polyhydroxyalkanoates (PHAs) under anaerobic conditions. Many ecological, physiological, and genomic analyses have been performed with proteobacterial PAOs, but few with actinobacterial PAOs. In this study, the whole genome sequence of an actinobacterial PAO, Microlunatus phosphovorus NM-1(T) (NBRC 101784(T)), was determined. The number of genes for polyP metabolism was greater in M. phosphovorus than in other actinobacteria; it possesses genes for four polyP kinases (ppks), two polyP-dependent glucokinases (ppgks), and three phosphate transporters (pits). In contrast, it harbours only a single ppx gene for exopolyphosphatase, although two copies of ppx are generally present in other actinobacteria. Furthermore, M. phosphovorus lacks the phaABC genes for PHA synthesis and the actP gene encoding an acetate/H(+) symporter, both of which play crucial roles in anaerobic PHA accumulation in proteobacterial PAOs. Thus, while the general features of M. phosphovorus regarding aerobic polyP accumulation are similar to those of proteobacterial PAOs, its anaerobic polyP use and PHA synthesis appear to be different.

Complete genome sequence of phototrophic betaproteobacterium Rubrivivax gelatinosus IL144.

Rubrivivax gelatinosus is a facultative photoheterotrophic betaproteobacterium living in freshwater ponds, sewage ditches, activated sludge, and food processing wastewater. There have not been many studies on photosynthetic betaproteobacteria. Here we announce the complete genome sequence of the best-studied phototrophic betaproteobacterium, R. gelatinosus IL-144 (NBRC 100245).

Complete genome sequence of Sphingobium sp. strain SYK-6, a degrader of lignin-derived biaryls and monoaryls.

Sphingobium sp. strain SYK-6 is able to grow on an extensive variety of lignin-derived biaryls and monoaryls, and the catabolic genes for these compounds are useful for the production of industrially valuable metabolites from lignin. Here we report the complete nucleotide sequence of the SYK-6 genome which consists of the 4,199,332-bp-long chromosome and the 148,801-bp-long plasmid.

Whole-genome analysis of Salmonella enterica serovar Typhimurium T000240 reveals the acquisition of a genomic island involved in multidrug resistance via IS1 derivatives on the chromosome.

Salmonella enterica serovar Typhimurium is frequently associated with life-threatening systemic infections, and the recent global emergence of multidrug resistance in S. enterica isolates from agricultural and clinical settings has raised concerns. In this study, we determined the whole-genome sequence of fluoroquinolone-resistant S. enterica serovar Typhimurium T000240 strain (DT12) isolated from human gastroenteritis in 2000. Comparative genome analysis revealed that T000240 displays high sequence similarity to strain LT2, which was originally isolated in 1940, indicating that progeny of LT2 might be reemerging. T000240 possesses a unique 82-kb genomic island, designated as GI-DT12, which is composed of multidrug resistance determinants, including a Tn2670-like composite transposon (class 1 integron [intI1, bla(oxa-30), aadA1, qacEΔ1, and sul1], mercury resistance proteins, and chloramphenicol acetyltransferase), a Tn10-like tetracycline resistance protein (tetA), the aerobactin iron-acquisition siderophore system (lutA and lucABC), and an iron transporter (sitABCD). Since GI-DT12 is flanked by IS1 derivatives, IS1-mediated recombination likely played a role in the acquisition of this genomic island through horizontal gene transfer. The aminoglycoside-(3)-N-acetyltransferase (aac(3)) gene and a class 1 integron harboring the dfrA1 gene cassette responsible for gentamicin and trimethoprim resistance, respectively, were identified on plasmid pSTMDT12_L and appeared to have been acquired through homologous recombination with IS26. This study represents the first characterization of the unique genomic island GI-DT12 that appears to be associated with possible IS1-mediated recombination in S. enterica serovar Typhimurium. It is expected that future whole-genome studies will aid in the characterization of the horizontal gene transfer events for the emerging S. enterica serovar Typhimurium strains.

Molecular evolutionary analysis of the influenza A(H1N1)pdm, May-September, 2009: temporal and spatial spreading profile of the viruses in Japan.

BACKGROUND: In March 2009, pandemic influenza A(H1N1) (A(H1N1)pdm) emerged in Mexico and the United States. In Japan, since the first outbreak of A(H1N1)pdm in Osaka and Hyogo Prefectures occurred in the middle of May 2009, the virus had spread over 16 of 47 prefectures as of June 4, 2009. METHODS/PRINCIPAL FINDINGS: We analyzed all-segment concatenated genome sequences of 75 isolates of A(H1N1)pdm viruses in Japan, and compared them with 163 full-genome sequences in the world. Two analyzing methods, distance-based and Bayesian coalescent MCMC inferences were adopted to elucidate an evolutionary relationship of the viruses in the world and Japan. Regardless of the method, the viruses in the world were classified into four distinct clusters with a few exceptions. Cluster 1 was originated earlier than cluster 2, while cluster 2 was more widely spread around the world. The other two clusters (clusters 1.2 and 1.3) were suggested to be distinct reassortants with different types of segment assortments. The viruses in Japan seemed to be a multiple origin, which were derived from approximately 28 transported cases. Twelve cases were associated with monophyletic groups consisting of Japanese viruses, which were referred to as micro-clade. While most of the micro-clades belonged to the cluster 2, the clade of the first cases of infection in Japan originated from cluster 1.2. Micro-clades of Osaka/Kobe and the Fukuoka cases, both of which were school-wide outbreaks, were eradicated. Time of most recent common ancestor (tMRCA) for each micro-clade demonstrated that some distinct viruses were transmitted in Japan between late May and early June, 2009, and appeared to spread nation-wide throughout summer. CONCLUSIONS: Our results suggest that many viruses were transmitted from abroad in late May 2009 irrespective of preventive actions against the pandemic influenza, and that the influenza A(H1N1)pdm had become a pandemic stage in June 2009 in Japan.

The effect of intraosseous graft length on tendon-bone healing in anterior cruciate ligament reconstruction using flexor tendon.

The current study was performed to understand the relationship between graft length placed within the bone tunnel and intraosseous graft healing in anterior cruciate ligament (ACL) reconstruction. Twenty-four adult beagle dogs were divided into two groups of 12 animals each. In each animal, ACL reconstruction using a 4-mm diameter autogenous flexor tendon graft was done in the left knee. In groups I and II, the graft having a length of 15 and 5 mm, respectively, was placed within the tibial tunnel. The proximal end of the graft was placed through the over-the-top route in all animals. In each group, five animals were sacrificed immediately after surgery, and the remaining seven were sacrificed at 6 weeks postoperatively. Biomechanical and histologic evaluations were performed. In pull out testing, the ultimate failure load and the linear stiffness of the graft-tibia complex harvested at 6 weeks were significantly greater than those harvested at the time-zero period. There were no significant differences in those parameters between groups I and II at 6 weeks. In each group, the perpendicular collagen fibers connecting the tendon to the bone tunnel wall were observed only in the narrow area located close to the intra-articular tunnel outlet. In conclusion, excessively long placement of the flexor graft within the bone tunnel does not result in an additional increase of anchoring strength and stiffness of the graft in ACL reconstruction.

The effect of transforming growth factor-beta1 on intraosseous healing of flexor tendon autograft replacement of anterior cruciate ligament in dogs.

PURPOSE: To clarify the effect of transforming growth factor (TGF)-beta1 on intraosseous healing of the flexor tendon autograft in anterior cruciate ligament (ACL) replacement. TYPE OF STUDY: Randomized trial in animals. METHODS: Twenty-one adult beagle dogs were divided into 3 groups of 7 animals each. The ACL was replaced with a flexor tendon autograft in the left knee. The distal end of each graft was placed in the tibial bone tunnel. In group I, no treatment was applied in the bone tunnel. In group II, 0.1 mL of fibrin sealant was applied in the tendon-bone gap. In group III, 2 ng of TGF-beta1 mixed with 0.1 mL fibrin sealant was applied in the gap. Each animal was euthanized 3 weeks after surgery. RESULTS: In pullout testing, the ultimate load of the graft-tibia complex of group III (188.2 +/- 51.4 N) was significantly greater than that of groups I (86.7 +/- 36.5 N) and II (99.0 +/- 18.7 N) (P = .005). There were no significant differences between groups I and II. Histologically, the perpendicular collagen fibers connecting the tendon to the bone were narrowly seen in groups I and II, whereas the fibers were richly generated in group III. CONCLUSIONS: In ACL replacement using flexor tendon autograft, exogenous administration of TGF-beta1 significantly increased the bonding strength of the graft to the tunnel wall at 3 weeks, although the long-term effects are unknown. CLINICAL RELEVANCE: Intraosseous healing of the flexor tendon autograft in ACL reconstruction may be enhanced by a use of some growth factors involving TGF-beta1 in the future.

The effect of growth factors on biomechanical properties of the bone-patellar tendon-bone graft after anterior cruciate ligament reconstruction: a canine model study.

BACKGROUND: No studies have dealt with the effect of growth factors on the free tendon autograft in anterior cruciate ligament reconstruction. HYPOTHESIS: Application of exogenous transforming growth factor-beta and epidermal growth factor may affect the structural properties and histology of the bone-patellar tendon-bone autograft after anterior cruciate ligament reconstruction. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty dogs underwent anterior cruciate ligament reconstruction with the autogenous bone-patellar tendon-bone graft in bilateral knees. In 10 animals, 12 ng transforming growth factor-beta and 300 ng epidermal growth factor mixed with fibrin sealant of 0.6 mL were applied to the left knee. In the remaining 10 dogs, fibrin sealant alone was applied to the left knee. No additional treatments were applied to the right knee. RESULTS: The growth factor application increased the stiffness and maximum failure load of the femur-graft-tibia complex at 12 weeks (P =.016 and P =.012, respectively); the sham treatment did not significantly affect them. Histologically, most of the cells in the grafts treated with growth factors had spindle-shaped nuclei; cells in the other grafts had round-shaped nuclei. CONCLUSIONS: Application of transforming growth factor-beta and epidermal growth factor improves the structural properties of the autograft after anterior cruciate ligament reconstruction in the canine model. CLINICAL RELEVANCE: Application of growth factors is a possible strategy to prevent graft deterioration in anterior cruciate ligament reconstruction.

Juvenile Blount's disease: bilateral case with asynchronous onset.

We encountered a rare case of bilateral juvenile Blount's disease diagnosed as unilateral at the onset. The patient was a 9-year-old girl who visited our outpatient clinic with pain and deformity in the left knee. Examination of the left knee showed severe varus deformity. Correction of the varus deformity and tibial shortness was carried out at the same time by the callotasis method using an Orthofix external fixator. During follow-up the varus deformity in her right knee gradually progressed. At 18 months after the initial surgery, the right varus deformity and tibial shortness were corrected at the same time by the callotasis method using an Orthofix external fixator; proximal epiphysiodesis was also performed. The coronal-plane alignment of both knees remains good. In cases with bilateral involvement of Blount's disease, the onset of the deformity is usually nearly synchronous. In some cases, however, it begins unilaterally, as with this patient, so it is essential to check both sides by means of tomography or magnetic resonance imaging and not just radiography. Moreover, we must be careful to plan the treatment in such a case so as to anticipate possible leg-length discrepancy problems.

The effect of graft-tunnel diameter disparity on intraosseous healing of the flexor tendon graft in anterior cruciate ligament reconstruction.

BACKGROUND: Graft-to-tunnel healing is a significant factor in anterior cruciate ligament reconstruction, but there have been few studies on the effect of graft-tunnel diameter disparity on intraosseous healing of the flexor tendon graft. HYPOTHESIS: Graft-tunnel diameter disparity of 2 mm has no effect on the pull-out strength of the graft from the bone tunnel. STUDY DESIGN: Controlled laboratory study. METHODS: Forty-two beagle dogs were divided into three groups. In each animal, reconstruction was performed in the left knee by using a 4-mm diameter autogenous flexor tendon for groups 1 and 2 and by using a 4-mm wide bone-patellar tendon-bone graft in group 3. A 4-mm diameter tunnel was drilled in the tibia of groups 1 and 3 and a 6-mm diameter tunnel, in group 2. In each group, seven animals were sacrificed at 3 and 6 weeks. RESULTS: The perpendicular fibers connecting the graft to the bone were generated in groups 1 and 2, and the number appeared to be higher in group 2, where the space was greater. There was no significant difference in the ultimate failure load between groups 1 and 2 at each period. CONCLUSION: Graft-tunnel diameter disparity of up to 2 mm may not adversely affect intraosseous healing of the flexor tendon graft. CLINICAL RELEVANCE: Surgeons need not be overly concerned about minor graft-tunnel diameter disparities.