Angiogenic conditioning of peripheral blood mononuclear cells promotes fracture healing.

OBJECTIVES: The objective of this study was to investigate the therapeutic effect of peripheral blood mononuclear cells (PBMNCs) treated with quality and quantity control culture (QQ-culture) to expand and fortify angiogenic cells on the acceleration of fracture healing. METHODS: Human PBMNCs were cultured for seven days with the QQ-culture method using a serum-free medium containing five specific cytokines and growth factors. The QQ-cultured PBMNCs (QQMNCs) obtained were counted and characterised by flow cytometry and real-time polymerase chain reaction (RT-PCR). Angiogenic and osteo-inductive potentials were evaluated using tube formation assays and co-culture with mesenchymal stem cells with osteo-inductive medium in vitro. In order to evaluate the therapeutic potential of QQMNCs, cells were transplanted into an immunodeficient rat femur nonunion model. The rats were randomised into three groups: control; PBMNCs; and QQMNCs. The fracture healing was evaluated radiographically and histologically. RESULTS: The total number of PBMNCs was decreased after QQ-culture, however, the number of CD34+ and CD206+ cells were found to have increased as assessed by flow cytometry analysis. In addition, gene expression of angiogenic factors was upregulated in QQMNCs. In the animal model, the rate of bone union was higher in the QQMNC group than in the other groups. Radiographic scores and bone volume were significantly associated with the enhancement of angiogenesis in the QQMNC group. CONCLUSION: We have demonstrated that QQMNCs have superior potential to accelerate fracture healing compared with PBMNCs. The QQMNCs could be a promising option for fracture nonunion.Cite this article: K. Mifuji, M. Ishikawa, N. Kamei, R. Tanaka, K. Arita, H. Mizuno, T. Asahara, N. Adachi, M. Ochi. Angiogenic conditioning of peripheral blood mononuclear cells promotes fracture healing. Bone Joint Res 2017;6: 489-498. DOI: 10.1302/2046-3758.68.BJR-2016-0338.R1.

Semiclassical catastrophe theory of simple bifurcations.

The Fedoriuk-Maslov catastrophe theory of caustics and turning points is extended to solve the bifurcation problems by the improved stationary phase method (ISPM). The trace formulas for the radial power-law (RPL) potentials are presented by the ISPM based on the second- and third-order expansion of the classical action near the stationary point. A considerable enhancement of contributions of the two orbits (pair consisting of the parent and newborn orbits) at their bifurcation is shown. The ISPM trace formula is proposed for a simple bifurcation scenario of Hamiltonian systems with continuous symmetries, where the contributions of the bifurcating parent orbits vanish upon approaching the bifurcation point due to the reduction of the end-point manifold. This occurs since the contribution of the parent orbits is included in the term corresponding to the family of the newborn daughter orbits. Taking this feature into account, the ISPM level densities calculated for the RPL potential model are shown to be in good agreement with the quantum results at the bifurcations and asymptotically far from the bifurcation points.

Effects of diabetes on tooth movement and root resorption after orthodontic force application in rats.

OBJECTIVE: To investigate the effects of diabetes on orthodontic tooth movement and orthodontically induced root resorption in rats. SETTING AND SAMPLE POPULATION: Twenty-three 10-week-old male Sprague-Dawley rats divided into control (n = 7), diabetes (n = 9), and diabetes + insulin (n = 7) groups. MATERIALS AND METHODS: Diabetes was induced by administering a single intraperitoneal injection of streptozotocin. Rats with a blood glucose level exceeding 250 mg/dl were assigned to the diabetes group. Insulin was administered daily to the diabetes + insulin group. A nickel-titanium closed-coil spring of 10 g was applied for 2 weeks to the maxillary left first molar in all rats to induce mesial tooth movement. Tooth movement was measured using microcomputed tomography images. To determine the quantity of root resorption, the mesial surfaces of the mesial and distal roots of the first molar were analyzed using both scanning electron microscopy and scanning laser microscopy. RESULTS: After 2 weeks, the amount of tooth movement in the diabetic rats was lower than that in the control rats. Root resorption was also significantly lower in the diabetic rats. These responses of the rats caused by diabetes were mostly diminished by insulin administration. CONCLUSIONS: Diabetes significantly reduced orthodontic tooth movement and orthodontically induced root resorption in rats. The regulation of blood glucose level through insulin administration largely reduced these abnormal responses to orthodontic force application.

Semiclassical trace formula for the two-dimensional radial power-law potentials.

The trace formula for the density of single-particle levels in the two-dimensional radial power-law potentials, which nicely approximate up to a constant shift the radial dependence of the Woods-Saxon potential and its quantum spectra in a bound region, was derived by the improved stationary phase method. The specific analytical results are obtained for the powers α=4 and 6. The enhancement of periodic-orbit contribution to the level density near the bifurcations are found to be significant for the description of the fine shell structure. The semiclassical trace formulas for the shell corrections to the level density and the energy of many-fermion systems reproduce the quantum results with good accuracy through all the bifurcation (symmetry breaking) catastrophe points, where the standard stationary-phase method breaks down. Various limits (including the harmonic oscillator and the spherical billiard) are obtained from the same analytical trace formula.

Entry point to the Sylvian fissure for the pterional transsylvian approach.

BACKGROUND: Although the anatomy of the Sylvian fissure is understood, there is little information on where to start its dissection in the pterional transsylvian (PT-TS) approach. At small craniotomy using the PT-TS approach, we set the entry point to the Sylvian fissure at 15 mm behind the anterior edge of the craniotomy along the Sylvian fissure and designated this site "point 15." Here we compared the utility of "point 15" with the Sylvian point (point on the Sylvian fissure giving rise to the horizontal and anterior ascending rami) that had been recommended earlier as the entry site for opening the Sylvian fissure. MATERIALS AND METHODS: This study includes 16 patients with 7 ruptured and 9 unruptured anterior circulation aneurysms. We evaluated the usefulness of "point 15" in the PT-TS approach for aneurysmal neck clipping with respect to the adequacy of anatomical exposure and low invasiveness. RESULTS: In 12 patients "point 15" provided for excellent anatomical exposure of the Sylvian fissure; complete neck clipping was possible with minimal brain retraction and damage. In two patients with ruptured aneurysms and thick subarachnoid hemorrhage and in two patients with unruptured aneurysms, the dissection had to be enlarged 3 to 4 mm distally without reaching the Sylvian point. In the latter two patients the Sylvian veins were tethered to frontal and temporal lobes. CONCLUSIONS: The "point 15" was an easily set entry point to the Sylvian fissure. It provided for sufficient anatomical exposure at surgery for anterior circulation aneurysms; additional posterior dissection was required in rare cases. We found that "point 15" was useful in small craniotomies using the PT-TS approach.

Low distribution of synaptic vesicle protein 2A and synaptotagimin-1 in the cerebral cortex and hippocampus of spontaneously epileptic rats exhibiting both tonic convulsion and absence seizure.

The spontaneously epileptic rat (SER) is a double mutant (zi/zi, tm/tm) which begins to exhibit tonic convulsions and absence seizures after 6 weeks of age, and repetitive tonic seizures over time induce sclerosis-like changes in SER hippocampus with high brain-derived neurotrophic factor (BDNF) expression. Levetiracetam, which binds to synaptic vesicle protein 2A (SV2A), inhibited both tonic convulsions and absence seizures in SERs. We studied SER brains histologically and immunohistochemically after verification by electroencephalography (EEG), as SERs exhibit seizure-related alterations in the cerebral cortex and hippocampus. SERs did not show interictal abnormal spikes and slow waves typical of focal epilepsy or symptomatic generalized epilepsy. The difference in neuronal density of the cerebral cortex was insignificant between SER and Wistar rats, and apoptotic neurons did not appear in SERs. BDNF distributions portrayed higher values in the entorhinal and piriform cortices which would relate with hippocampal sclerosis-like changes. Similar synaptophysin expression in the cerebral cortex and hippocampus was found in both animals. Low and diffuse SV2A distribution portrayed in the cerebral cortex and hippocampus of SERs was significantly less than that of all cerebral lobes and inner molecular layer (IML) of the dentate gyrus (DG) of Wistar rats. The extent of low SV2A expression/distribution in SERs was particularly remarkable in the frontal (51% of control) and entorhinal cortices (47%). Lower synaptotagmin-1 expression (vs Wistar rats) was located in the frontal (31%), piriform (13%) and entorhinal (39%) cortices, and IML of the DG (38%) in SER. Focal low distribution of synaptotagmin-1 accompanying low SV2A expression may contribute to epileptogenesis and seizure propagation in SER.

Microvascular decompression for trigeminal neuralgia due to compression by the vertebral artery: report of 3 cases.

BACKGROUND: Trigeminal neuralgia elicited by the vertebral artery is unusual. According to a large trigeminal neuralgia series, only 4 of 1,404 (0.3%) consecutive patients with typical trigeminal neuralgia presented with vertebral artery compression. In such cases the vertebrobasilar system tends to be atherosclerotic, ectatic, and tortuous, requiring, in addition to an ordinary microvascular decompression method, technical modifications of this procedure. We report on 3 patients with trigeminal neuralgia due to compression by a tortuous vertebral artery. PATIENTS: All 3 patients underwent microvascular decompression via a small lateral suboccipital craniotomy. Operative exposure demonstrated that the root of the trigeminal nerve was compressed directly and stretched by a loop of the vertebral artery. The compression was successfully released by dislocation of the loop using Teflon (polytetrafluoroethene) slings. Immediately after the operation all 3 patients became pain-free. CONCLUSION: Among the surgical procedures used in microvascular decompression surgery, dislocation of the offending vessel with Teflon slings is a useful surgical technique to treat trigeminal neuralgia due to a tortuous vertebral artery.

Paraumbilical peritoneal incision using the little finger in shunt operations for hydrocephalus: technical note.

INTRODUCTION: The shunt operation remains the standard procedure for the treatment of hydrocephalus. We describe a simple minilaparotomy method that involves perforation of the peritoneum with the surgeon's little finger. TECHNIQUE: After placing a small paraumbilical incision at the skin and fascia, the little finger is introduced through the incision to perforate the pre-peritoneal fat and peritoneum. The finger should be inserted at a 30-45° angle to the horizontal plane to avoid injuring the underlying viscera and major blood vessels and to put sufficient shear force on the peritoneum. A catheter is inserted into the abdominal cavity after visual confirmation of proper perforation. CONCLUSION: As the paraumbilical wound is not noticeable postoperatively due to the presence of the natural umbilical skin fold, this method yields a cosmetically appealing result.

New insight into mechanisms of pruritus from molecular studies on familial primary localized cutaneous amyloidosis.

Macular and lichen amyloidosis are common variants of primary localized cutaneous amyloidosis (PLCA) in which clinical features of pruritus and skin scratching are associated with histological findings of deposits of amyloid staining on keratinous debris in the papillary dermis. Most cases are sporadic, but an autosomal dominant family history may be present in up to 10% of cases, consistent with a genetic predisposition in some individuals. Familial PLCA has been mapped to a locus on 5p13.1-q11.2 and in 2008 pathogenic heterozygous missense mutations were identified in the OSMR gene, which encodes oncostatin M receptor beta (OSMRbeta), an interleukin (IL)-6 family cytokine receptor. OSMRbeta is expressed in various cell types, including keratinocytes, cutaneous nerves and nociceptive neurones in dorsal root ganglia; its ligands are oncostatin M and IL-31. All pathogenic mutations are clustered in the fibronectin-III repeat domains of the extracellular part of OSMRbeta, sites that are critical for receptor dimerization (with either gp130 or IL-31RA), and lead to defective signalling through Janus kinase-signal transducers and activators of transcription, extracellular signal-regulated protein kinase 1/2 and phosphoinositide 3 kinase/Akt pathways. Elucidating the molecular pathology of familial PLCA provides new insight into mechanisms of pruritus in human skin, findings that may have relevance to developing novel treatments for skin itching. This review provides a clinicopathological and molecular update on familial PLCA.

Novel mutation p.Gly59Arg in GJB6 encoding connexin 30 underlies palmoplantar keratoderma with pseudoainhum, knuckle pads and hearing loss.

BACKGROUND: Connexins, components of the gap junction, are expressed in several organs including the skin and the cochlea. Mutations in connexin genes including GJB2 (Cx26), GJB3 (Cx31), GJB4 (Cx30.3), GJB6 (Cx30) and GJA1 (Cx43) are responsible for various dermatological syndromes and/or inherited hearing loss, frequently showing overlapping phenotypes. OBJECTIVES: To clarify the spectrum of clinical phenotypes caused by connexin mutations. METHODS: We report a 32-year-old Japanese woman with mild palmoplantar keratoderma (PPK) with severe sensorineural hearing loss, knuckle pads and pseudoainhum of her toes. RESULTS: Direct sequencing revealed no mutation in GJB2, but a novel heterozygous missense mutation p.Gly59Arg in GJB6. Electron microscopy revealed no apparent morphological abnormality of gap junctions in the patient's lesional epidermis. CONCLUSIONS: The patient harboured the novel GJB6 missense mutation p.Gly59Arg in the first extracellular loop of Cx30. Mutations in glycine 59 of Cx26 are associated with PPK-deafness syndrome, and the similar phenotype here supports the observed heteromeric channel formation; the dominant nature of the mutation suggests an effect on gap junctions similar to that of the comparable mutation in Cx26.

Colocalization of kindlin-1, kindlin-2, and migfilin at keratinocyte focal adhesion and relevance to the pathophysiology of Kindler syndrome.

Kindler syndrome (KS) results from pathogenic loss-of-function mutations in the KIND1 gene, which encodes kindlin-1, a focal adhesion and actin cytoskeleton-related protein. How and why abnormalities in kindlin-1 disrupt keratinocyte cell biology in KS, however, is not yet known. In this study, we identified two previously unreported binding proteins of kindlin-1: kindlin-2 and migfilin. Co-immunoprecipitation and confocal microscopy studies show that these three proteins bind to each other and colocalize at focal adhesion in HaCaT cells and normal human keratinocytes. Moreover, loss-of-function mutations in KIND1 result in marked variability in kindlin-1 immunolabeling in KS skin, which is mirrored by similar changes in kindlin-2 and migfilin immunoreactivity. Kindlin-1, however, may function independently of kindlin-2 and migfilin, as loss of kindlin-1 expression in HaCaT keratinocytes by RNA interference and in KS keratinocytes does not affect KIND2 or FBLIM1 (migfilin) gene expression or kindlin-2 and migfilin protein localization. In addition to identifying protein-binding partners for kindlin-1, this study also highlights that KIND1 gene expression and kindlin-1 protein labeling are not always reduced in KS, findings that are relevant to the accurate laboratory diagnosis of this genodermatosis by skin immunohistochemistry.

Atypical teratoid/rhabdoid tumour in sella turcica in an adult.

Although atypical teratoid rhabdoid tumours preferentially arise in the posterior fossa of infants, we encountered a 56 year old woman with an atypical teratoid rhabdoid tumour located in the sella. She presented with right abducent and oculomotor nerve paresis. Magnetic resonance imaging demonstrated an intrasellar tumour impinging on the right cavernous sinus. Microscopically, the tumour was composed of cells with rhabdoid features; we observed atypia, eccentric nuclei, and intracytoplasmic inclusion bodies. The Ki-67 labeling index was around 30%. The tumour cells were positive for vimentin, epithelial membrane antigen, and neurofilament, but negative for INI1. Despite extended local brain and whole-spine irradiation she died of neural axis dissemination.