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Enterohepatic circulation of 12α-hydroxylated (12αOH) bile acid (BA) is enhanced depending on the energy intake in high-fat diet-fed rats. Such BA metabolism can be reproduced using a diet supplemented with cholic acid (CA), which also induces simple steatosis, without inflammation and fibrosis, accompanied by some other symptoms that are frequently observed in the condition of non-alcoholic fatty liver in rats. We investigated whether supplementation of the diet with raffinose (Raf) improves hepatic lipid accumulation induced by the CA-fed condition in rats. After acclimation to the AIN-93-based control diet, male Wistar rats were fed diets supplemented with a combination of Raf (30 g/kg diet) and/or CA (0·5 g/kg diet) for 4 weeks. Dietary Raf normalised hepatic TAG levels (two-way ANOVA P < 0·001 for CA, P = 0·02 for Raf and P = 0·004 for interaction) in the CA-supplemented diet-fed rats. Dietary Raf supplementation reduced hepatic 12αOH BA concentration (two-way ANOVA P < 0·001 for CA, P = 0·003 for Raf and P = 0·03 for interaction). The concentration of 12αOH BA was reduced in the aortic and portal plasma. Raf supplementation increased acetic acid concentration in the caecal contents (two-way ANOVA P = 0·001 as a main effect). Multiple regression analysis revealed that concentrations of aortic 12αOH BA and caecal acetic acid could serve as predictors of hepatic TAG concentration (R 2 = 0·55, P < 0·001). However, Raf did not decrease the secondary 12αOH BA concentration in the caecal contents as well as the transaminase activity in the CA diet-fed rats. These results imply that dietary Raf normalises hepatic lipid accumulation via suppression of enterohepatic 12αOH BA circulation.

Bifidobacterium is a natural inhabitant of the human gastrointestinal (GI) tract. We studied the role of the extracellular sialidase (SiaBb2, 835 amino acids [aa]) from Bifidobacterium bifidum ATCC 15696 in mucosal surface adhesion and carbohydrate catabolism. Human milk oligosaccharides (HMOs) or porcine mucin oligosaccharides as the sole carbon source enhanced B. bifidum growth. This was impaired in a B. bifidum ATCC 15696 strain harboring a mutation in the siabb2 gene. Mutant cells in early to late exponential growth phase also showed decreased adhesion to human epithelial cells and porcine mucin relative to the wild-type strain. These results indicate that SiaBb2 removes sialic acid from HMOs and mucin for metabolic purposes and may promote bifidobacterial adhesion to the mucosal surface. To further characterize SiaBb2-mediated bacterial adhesion, we examined the binding of His-tagged recombinant SiaBb2 peptide to colonic mucins and found that His-SiaBb2 as well as a conserved sialidase domain peptide (aa 187 to 553, His-Sia) bound to porcine mucin and murine colonic sections. A glycoarray assay revealed that His-Sia bound to the α2,6-linked but not to the α2,3-linked sialic acid on sialyloligosaccharide and blood type A antigen [GalNAcα1-3(Fucα1-2)Galβ] at the nonreducing termini of sugar chains. These results suggest that the sialidase domain of SiaBb2 is responsible for this interaction and that the protein recognizes two distinct carbohydrate structures. Thus, SiaBb2 may be involved in Bifidobacterium -mucosal surface interactions as well as in the assimilation of a variety of sialylated carbohydrates. IMPORTANCE Adhesion to the host mucosal surface and carbohydrate assimilation are important for bifidobacterium colonization and survival in the host gastrointestinal tract. In this study, we investigated the mechanistic basis for B. bifidum extracellular sialidase (SiaBb2)-mediated adhesion. SiaBb2 cleaved sialyl-human milk oligosaccharides and mucin glycans to produce oligosaccharides that supported B. bifidum growth. Moreover, SiaBb2 enhanced B. bifidum adhesion to mucosal surfaces via specific interactions with the α2,6 linkage of sialyloligosaccharide and blood type A antigen on mucin carbohydrates. These findings provide insight into the bifunctional role of SiaBb2 and the adhesion properties of B. bifidum strains. Adhesion to the host mucosal surface and carbohydrate assimilation are important for bifidobacterium colonization and survival in the host gastrointestinal tract. In this study, we investigated the mechanistic basis for B. bifidum extracellular sialidase (SiaBb2)-mediated adhesion. SiaBb2 cleaved sialyl-human milk oligosaccharides and mucin glycans to produce oligosaccharides that supported B. bifidum growth. Moreover, SiaBb2 enhanced B. bifidum adhesion to mucosal surfaces via specific interactions with the α2,6 linkage of sialyloligosaccharide and blood type A antigen on mucin carbohydrates. These findings provide insight into the bifunctional role of SiaBb2 and the adhesion properties of B. bifidum strains.

Purpose This study aimed to examine the associations between sonographic measurements of the abductor pollicis brevis (APB), grip and pinch strength, and distal motor latency (DML) in patients with carpal tunnel syndrome (CTS) before and after surgery. Methods We prospectively studied patients (46 hands) who underwent 1 year of postoperative follow-up after endoscopic carpal tunnel release. The patients underwent ultrasound (US) scans, grip and pinch strength assessment, a nerve conduction study, and patient-reported outcome measures (Carpal Tunnel Syndrome Instrument and Michigan Hand Outcomes Questionnaire) before and 1 year after surgery. The standardized response mean was calculated to compare the sensitivity of clinical changes in these measurements. Results US measurements (thickness of the APB and the cross-sectional area of the APB) and muscle strength (grip strength, key pinch, and tip pinch) were greater, and DML was reduced after surgery compared with those before surgery (all P  < 0.05). Patient-reported outcome measures also showed clinical improvement 1 year after surgery ( P  < 0.05). US measurements of the APB were significantly correlated with grip and pinch strength (all P  < 0.05), but not with DML, before surgery and 1 year after surgery. The standardized response mean showed a large responsiveness for US measurements of the APB and patient-reported outcome measures. Conclusion US evaluation of the APB after CTS can complement the evaluation of grip and pinch strength in the clinical setting. Postoperative recovery of the APB leads to improved motor dysfunction in CTS. Therefore, US measurement of the APB could be a useful tool for evaluating motor function.

The human gut houses bile acid 7α-dehydroxylating bacteria that produce secondary bile acids such as deoxycholic acid (DCA) from host-derived bile acids through enzymes encoded by the bai operon. While recent metagenomic studies suggest that these bacteria are highly diverse and abundant, very few DCA producers have been identified. Here, we investigated the physiology and determined the complete genome sequence of Eubacterium sp. c-25, a DCA producer that was isolated from human feces in the 1980s. Culture experiments showed a preference for neutral to slightly alkaline pH in both growth and DCA production. Genomic analyses revealed that c-25 is phylogenetically distinct from known DCA producers and possesses a multi-cluster arrangement of predicted bile-acid inducible (bai) genes that is considerably different from the typical bai operon structure. This arrangement is also found in other intestinal bacterial species, possibly indicative of unconfirmed 7α-dehydroxylation capabilities. Functionality of the predicted bai genes was supported by the induced expression of baiB, baiCD, and baiH in the presence of cholic acid substrate. Taken together, Eubacterium sp. c-25 is an atypical DCA producer with a novel bai gene cluster structure that may represent an unexplored genotype of DCA producers in the human gut.

Purpose This study sought to compare ultrasound-guided measurements of the abductor pollicis brevis (APB) using the water bath technique (WBT) and the direct contact method (DM) and investigate whether the DM can reproduce the measurements that would be obtained with a non-contact method, such as the WBT. Methods The APB muscles of 80 hands (40 healthy adults) were measured. The WBT was performed in a plastic container filled with water. The probe was placed adjacent to the skin surface without contact. In the DM, sonographic images were obtained with the probe and skin separated by sufficient transmission gel. The muscle thickness and cross-sectional area (CSA) were calculated with both methods. All subjects were examined three times by two examiners to estimate the inter- and intra-observer reliability. Bland–Altman analysis was performed to examine the agreement between the methods. Results No significant differences in the thickness or CSA of the APB were found. The interclass correlation coefficients for the WBT and DM showed almost perfect intra- and inter-observer reliability (range 0.87–0.94). There was no systematic bias between the techniques in the Bland–Altman analysis. Conclusion Similar to the WBT, the DM provides measurements of the APB thickness and CSA without causing morphometric changes.

Branched fatty acid esters of hydroxy fatty acids (FAHFAs) are novel endogenous lipids with important physiological functions in mammals. We previously identified a new type of FAHFAs, named short-chain fatty acid esterified hydroxy fatty acids (SFAHFAs), with acetyl or propyl esters of hydroxy fatty acids of carbon chains, C ≥ 20. However, sensitive determination of SFAHFAs is still a challenge, due to their high structural similarity and low abundance in biological samples. This study employs one-step chemical derivatization following total lipid extraction using 2-dimethylaminoethylamine (DMED) for enhanced detection of SFAHFAs. The labeled extracts were subjected to ultrahigh performance liquid chromatography coupled to linear ion trap quadrupole-Orbitrap mass spectrometry (UHPLC/LTQ-Orbitrap MS). Our results demonstrated that the detection sensitivities of SFAHFAs increased after DMED labeling, and is highly helpful in discovering six additional novel SFAHFAs in the cecum and colon contents of WKAH/HKmSlc rats fed with normal and high-fat diet (HFD). The identified DMED labeled SFAHFAs were characterized by their detailed MS/MS analysis, and their plausible fragmentation patterns were proposed. The concentrations of SFAHFAs were significantly reduced in the cecum of HFD group compared to the control. Hence, the proposed method could be a promising tool to apply for the enhanced detection of SFAHFAs in various biological matrices, which in turn facilitate the understanding of their sources, and physiological functions of these novel lipids.

Transposon mutagenesis systems are still under development in bifidobacteria, partly because intrinsic active insertion sequences are not well characterized in bifidobacteria. Here, we isolated an active insertion sequence, ISBlo11, from Bifidobacterium longum 105-A using a sacB-based counterselection system, which is generally used to screen for active insertion sequences from bacterial genomes. ISBlo11 is 1432 bp long and belongs to the IS3 family. It has a single ORF encoding a transposase and 25-bp inverted repeats at its termini. Full-length copies of ISBlo11 are specifically conserved among certain B. longum genomes and exist in different sites. Transposition analysis of an artificial ISBlo11 transposon using an Escherichia coli conjugation system revealed that ISBlo11 has adequate transposition activity, comparable to the reported activity of IS629, another IS3 family element initially isolated from Shigella sonnei. ISBlo11 also showed low transposition selectivity for non-conserved 3- or 4-bp target sequences. These characteristics of ISBlo11 seem suitable for the development of a new transposon mutagenesis system in bifidobacteria. A novel active insertion sequence was isolated from bifidobacteria and its transposition properties were characterized aimed to develop transposon mutagenesis system in bifidobacteria.

Bifidobacteria are one of the major components in human gut microbiota and well-known as beneficial microbes. However, clarification of commensal mechanisms of bifidobacteria in the intestines is still ongoing, especially in the presence of the gut microbiota. Here, we applied recombinase-based in vivo expression technology (R-IVET) using the bacteriophage P1 Cre/loxP system to Bifidobacterium longum subsp. longum 105-A (B. longum 105-A) to identify genes that are specifically expressed in the gastrointestinal tract of conventionally raised mice. Oral administration of the genomic DNA library of B. longum 105-A to conventionally raised mice resulted in the identification of 73 in vivo-induced genes. Four out of seven tested genes were verified in vivo-specific induction at least in the cecum by quantitative reverse transcription PCR. Although there is still room for improvement of the system, our findings can contribute to expanding our understanding of the commensal behavior of B. longum in the gut ecosystem.

The Sauvé-Kapandji procedure (SK), which combines distal radioulnar joint arthrodesis with creation of an ulnar pseudarthrosis, achieves good outcomes with few complications for patients with distal radioulnar joint instability or arthritis. The authors describe a case of atypical fracture of the radial diaphysis that occurred in a 79-year-old woman with rheumatoid arthritis who had undergone SK 18 years earlier. The patient had taken alendronate for osteoporosis for 13 years but had discontinued treatment 2 years before onset of symptoms. Retrospective review of serial radiographs revealed focal cortical thickening at the fracture site beginning 18 months before the onset of prodromal pain and becoming more evident over time. The patient underwent surgical repair of the fracture, which had occurred at the attachment of the pronator teres. Histopathological examination of bone excised from the fracture site and from the iliac graft used for fracture repair revealed markedly fewer trabecular osteoblasts than normal and no osteoclasts, indicating severe suppression of bone turnover. It is important to evaluate the radial diaphysis on radiographs taken after SK for early signs of atypical fracture, especially in patients with rheumatoid arthritis and a history of bisphosphonate use. Given the patient's history of bisphosphonate use and the similar radiographic appearance to atypical femoral fractures, the authors applied the term atypical radial fracture to this case. [Orthopedics. 20XX;XX(X):xx–xx.]

To clarify how the peripheral nerve adapts to elongation during gradual limb lengthening, electrophysiological and histomorphometric examinations were performed on the sciatic nerves in 18 rabbits. External fixators were used to lengthen the right femora by 30 mm (30%), at a daily rate of 0.5 mm (Group 1) or 2.0 mm (Group 2). Examinations were performed immediately after the limb lengthening procedure. Electrophysiologically, mild conduction slowing was observed in Group 1; a conduction block was evident in Group 2. Histologically, the mean diameter of myelinated fibers was unchanged in Group 1, but a significantly decreased diameter was observed in Group 2. Electron microscopy revealed that mild degenerative change of unmyelinated axons occurred sporadically in two cases in Group 2, but neither group showed evidence of thinning of myelin sheath of myelinated fibers. The mean internodal length (between nodes of Ranvier) of teased fibers was 1216±295 μ m in the control contralateral side, 1484±347 μ m in Group 1, and 1467±322 μ m in Group 2. Thus the internodes were lengthened by 22.1% (Group 1) and 20.7% (Group 2) in comparison with those of the controls. Straightening of the geometry of paranodal myelin sheath was significantly correlated with the rate of distraction. These results indicate that myelinated nerve fibers adapt to gradual elongation by lengthening each Schwann cell body, not by proliferation of Schwann cells.