In-process monitoring of a tissue-engineered oral mucosa fabricated on a micropatterned collagen scaffold: use of optical coherence tomography for quality control.

Background: We previously reported a novel technique for fabricating dermo-epidermal junction (DEJ)-like micropatterned collagen scaffolds to manufacture an ex vivo produced oral mucosa equivalent (EVPOME) for clinical translation; however, more biomimetic micropatterns are required to promote oral keratinocyte-based tissue engineering/regenerative medicine. In addition, in-process monitoring for quality control of tissue-engineered products is key to successful clinical outcomes. However, evaluating three-dimensional tissue-engineered constructs such as EVPOME is challenging. This study aimed to update our technique to fabricate a more biomimetic DEJ structure of oral mucosa and to investigate the efficacy of optical coherence tomography (OCT) in combination with deep learning for non-invasive EVPOME monitoring. Methods: A picosecond laser-textured microstructure mimicking DEJ on stainless steel was used as a negative mould to fabricate the micropatterned collagen scaffold. During EVPOME manufacturing, OCT was applied twice to monitor the EVPOME and evaluate its epithelial thickness. Findings: Our moulding system resulted in successful micropattern replication on the curved collagen scaffold. OCT imaging visualised the epithelial layer and the underlying micropatterned scaffold in EVPOME, enabling to non-invasively detect specific defects not found before the histological examination. Additionally, a gradual increase in epithelial thickness was observed over time. Conclusion: These findings demonstrate the feasibility of using a stainless-steel negative mould to create a more biomimetic micropattern on collagen scaffolds and the potential of OCT imaging for quality control in oral keratinocyte-based tissue engineering/regenerative medicine.

Safer Vitrification of Mouse and Human Embryos Using the Novel Cryoroom Vitrification System for Assisted Reproductive Technology.

BACKGROUND: Vitrification is widely used for assisted reproductive technology (ART). Most vitrification devices require the skillful placement of embryos into the carrier and aspiration of excessive vitrification solution. OBJECTIVE: To evaluate the efficacy and safety of the Cryoroom as a vitrification device. MATERIALS AND METHODS: Mouse and human embryos were vitrified with Cryoroom or Cryotop, and the developmental potency was assessed in vitro. Mouse monozygotic twin blastocysts were vitrified with Cryoroom or Cryotop for microarray analysis. RESULTS AND DISCUSSION: In mouse and human embryos, there were no differences between the survival and developmental progress in each device. In silico, the Cryoroom device showed no changes, particularly in DNA methylation after vitrification compared with the Cryotop. These results showed that the form and function of the device may affect the gene expression levels in vitrified embryos. CONCLUSION: The Cryoroom represents a safe and potentially revolutionary vitrification device for ART.

Learning Promotes Subfield-Specific Synaptic Diversity in Hippocampal CA1 Neurons.

The hippocampus is functionally heterogeneous between the dorsal and ventral subfields with left-right asymmetry. To determine the possible location of contextual memory, we performed an inhibitory avoidance task to analyze synaptic plasticity using slice patch-clamp technique. The training bilaterally increased the AMPA/NMDA ratio at dorsal CA3-CA1 synapses, whereas the training did not affect the ratio at ventral CA3-CA1 synapses regardless of the hemisphere. Moreover, sequential recording of miniature excitatory postsynaptic currents and miniature inhibitory postsynaptic currents from the same CA1 neuron clearly showed learning-induced synaptic plasticity. In dorsal CA1 neurons, the training dramatically strengthened both excitatory and inhibitory postsynaptic responses in both hemispheres, whereas the training did not promote the plasticity in either hemisphere in ventral CA1 neurons. Nonstationary fluctuation analysis further revealed that the training bilaterally increased the number of AMPA or GABAA receptor channels at dorsal CA1 synapses, but not at ventral CA1 synapses, suggesting functional heterogeneity of learning-induced receptor mobility. Finally, the performance clearly impaired by the bilateral microinjection of plasticity blockers in dorsal, but not ventral CA1 subfields, suggesting a crucial role for contextual learning. The quantification of synaptic diversity in specified CA1 subfields may help us to diagnose and evaluate cognitive disorders at the information level.

XPS and NEXAFS studies of VUV/O3-treated aromatic polyurea and its application to microchip electrophoresis.

In this study, the authors performed X-ray photoelectron spectroscopy (XPS) and near-edge X-ray absorption fine structure (NEXAFS) studies of vacuum ultraviolet (VUV)/O3-treated aromatic polyurea films to investigate their treatment effects. XPS and NEXAFS spectra indicate that the benzene ring was cleaved after treatment and that carboxyl, hydroxyl, ketone and aldehyde groups were formed at the cleaved sites. The VUV/O3-treated polyurea film was applied to a polymethylmethacrylate (PMMA) microchip for microchip electrophoresis (MCE) of bovine serum albumin (BSA). Fast electro-osmotic mobility of 4.6×10(-4) cm²/V/s as well as reduction of the BSA adhesion was achieved. This functional surface is useful for high-speed MCE analysis.

Immunohistochemistry of symptomatic hypertrophy of the posterior longitudinal ligament with special reference to ligamentous ossification.

STUDY DESIGN: Immnunohistochemical staining of the thickened posterior longitudinal ligament of the cervical spine. OBJECTIVES: To clarify the histological characteristics of hypertrophy of the posterior longitudinal ligament (HPLL) of the cervical spine and the relationship between HPLL and ossification of the posterior longitudinal ligament (OPLL). SETTING: Aichi Medical University, Aichi, Japan. METHODS: Eight specimens of HPLL and two of OPLL were obtained during anterior decompressive surgery on the cervical spine from patients with myelopathy. Hematoxylin and eosin staining, alcian blue staining and immunohistochemical staining with antibodies against bone morphogenetic protein (BMP), transforming growth factor (TGF)-beta, proliferating cell nuclear antigen (PCNA), alkaline phosphatase (ALP) and osteopontin (OPN) were carried out on the specimens. RESULTS: HPLL showed hyalinoid degeneration, the proliferation of chondrocytes and fibroblast-like spindle cells, infiltration of vessels and small ossification. In four cases, chondroid tissue was prominent with chondrocytes, which were expressed by ALP and OPN. The cells in HPLL were weakly or moderately stained by BMP, TGF-beta and PCNA. Their expression was similar to that of OPLL. Immunohistochemical staining was negative for all cells in the control cases. CONCLUSIONS: Histological and biochemical evidence supports the hypothesis that HPLL transforms into OPLL. The positive expression of BMP and TGF-beta in HPLL cells of myelopathic patients, and their similarity to OPLL, suggest that these cells have the potential to differentiate into osteogenic cells. Of note, neither BMP nor TGF-beta was demonstrated in the PLL of control subjects. Furthermore, the expression of chondrocytes by ALP and OPN in cartilage-prominent HPLL suggests that the cartilage can be replaced by new bone.

Symptomatic ossification of the anterior longitudinal ligament with stenosis of the cervical spine: a report of seven cases.

Seven men with a mean age of 63.9 years (59 to 67) developed dysphagia because of oesophageal compression with ossification of the anterior longitudinal ligament (OALL) and radiculomyelopathy due to associated stenosis of the cervical spine. The diagnosis of OALL was made by plain lateral radiography and classified into three types; segmental, continuous and mixed. Five patients had associated OALL in the thoracic and lumbar spine without ossification of the ligamentum flavum. All underwent removal of the OALL and six had simultaneous decompression by removal of ossification of the posterior longitudinal ligament or a bony spur. All had improvement of their dysphagia. Because symptomatic OALL may be associated with spinal stenosis, precise neurological examination is critical. A simultaneous microsurgical operation for patients with OALL and spinal stenosis gives good results without serious complications.

Postmortem study of the spinal cord showing snake-eyes appearance due to damage by ossification of the posterior longitudinal ligament and kyphotic deformity.

UNLABELLED: CLINICAL DESIGN: A case report. OBJECTIVES: To elucidate the clinical role of snake-eyes appearance in this case, correlation between radiological, clinical and postmortem study was performed. SETTING: Aichi, Japan. CASE REPORT: A 73-year-old man developed weakness and pain in the upper limbs due to kyphotic deformity secondary to laminectomy for cervical ossification of the posterior longitudinal ligament. Axial magnetic resonance imaging revealed snake-eyes appearance from C4 to C6. He died of acute myocardial infarction 3 months after anterior decompressive surgery. RESULTS: A postmortem examination of the cervical spinal cord showed small cystic six necrotic areas at the junction of the central gray matter and the ventrolateral posterior column, one in the right and one in the left, in association with neuronal loss in the anterior horn. CONCLUSIONS: Bilateral small intramedullary high-signal areas known as 'snake-eyes appearance' located around the central gray matter and the ventrolateral posterior column, are associated with neuronal loss in the compressed anterior horn that played an important role in worsening weakness of the upper limbs.

C7 fracture treated with a pedicle screw system under navigation guidance.

A 51-year-old woman with traumatic fracture-subluxation of C6-C7 vertebrae was treated by pedicle screw fixation. Among several methods for surgical treatment of the cervical spine, this technique provides a more rigid anchor to prevent collapsing or instability of the spinal column, particularly at the cervicothoracic junction. However, the risk of injuring the adjacent neurovascular structures cannot be completely eliminated. The characteristics of the cervicothoracic junction, surgical approaches and pedicle screw fixation techniques are emphasised.

Load independence of temperature-dependent Ca2+ recirculation fraction in canine heart.

Intramyocardial Ca(2+) recirculation fraction (RF) critically determines the economy of excitation-contraction coupling. RF is obtainable from the exponential decay of the postextrasystolic potentiation of left ventricular (LV) contractility. We have shown that RF remains unchanged despite increasing LV volume (LVV) at normothermia, but decreases with increasing temperature at a constant LVV. However, it remains unknown whether the temperature-dependent RF was not due to the simultaneously changed peak LV pressure (LVP) at a constant LVV. We hypothesized that this temperature-dependent RF would be independent of the simultaneous change in LVP. We used nine excised, cross-circulated canine hearts and allowed their LVs to contract isovolumically. During stable regular beats at 500 msec intervals, we inserted an extrasystolic beat at 360 msec interval followed by the postextrasystolic beats (PESs) at 500 msec intervals. We equalized the temperature-dependent peak LVPs of the regular beats at 36 degrees C and 38 degrees C to the peak LVP level of the stable regular beat at 33 degrees C by adjusting LVV. We fitted the same equation: nEmax = a.exp[-(i - 1)/tau(e)] + b.exp[-(i - 1)/tau(s)]cos[pi(i - 1)] + 1, used before to the normalized Emax (maximum elastance) values of PESi (i = 1-6) relative to the regular beat Emax. RF given by exp(-1/tau(e)) decreased by 19% to 38 degrees C from 33 degrees C. The temperature coefficient (Q(10)) of 1/RF was significantly greater than 1.3. The present results indicated a similar temperature dependence of RF and its Q(10) to those we observed previously without equalizing peak LVP. Thus, the temperature-dependent RF is independent of ventricular loading conditions.

Cervical amyotrophy caused by hypertrophy of the posterior longitudinal ligament.

STUDY DESIGN: A case report. OBJECTIVES: To report a case of cervical amyotrophy caused by hypertrophy of the posterior longitudinal ligament (HPLL). SETTING: Department of Neurological Surgery, Aichi Medical University, Aichi, Japan. METHODS: The patient had severe muscular atrophy in the deltoid and triceps with slight localized hypesthesia in the C5 area and severely unstable gait due to diminished vibration sense in the knees and ankles. Magnetic resonance imaging (MRI) showed expanded cord compression from C4 to C6 with intramedullary high-signal intensity due to HPLL. Transverse image MRI was useful to identify the HPLL. RESULTS: Resection of HPLL was achieved by an anterior approach. Histological findings of the surgical specimens showed thickening of the ligamentous tissue with proliferation of chondrocytes. CONCLUSIONS: HPLL should be included as a causative pathology of cervical spondylotic amyotrophy. Careful neurological examination including sensory examination of the lower limbs should be performed to avoid confusion with motor neuron disease.

Lunatomalacia in association with congenital synostosis between the capitate and the hamate.

This report describes a case of lunatomalacia associated with congenital synostosis between the capitate and the hamate, an association not previously reported. The case was examined in detail, and compared to the findings in the literature including observation regarding prognosis of lunatomalacia.

Combined anterior and posterior instrumentation in severe fracture-dislocation of the lower cervical spine with help of navigation: a case report.

The authors report a case of severe fracture-dislocation of C6-C7 with dural and spinal cord damage causing quadriplegia. The patient was treated surgically with circumferential stabilisation. Intense spinal instrumentation with pedicle screw fixation in addition to anterior decompression and plate fixation was essential for restoring the original vertebral column. This technique of internal fixation provided a more rigid anchor, however the risk to the neurovascular structures could not be completely eliminated. Therefore, the combined anterior and posterior stabilisation assisted with a navigation system was a safer and reasonable surgical treatment for this patient with severe cervical injuries.

[Usefulness of epidural administration of ketamine for relief of postherpetic neuralgia].

Four patients with postherpetic neuralgia had their pain alleviated by epidural administration of ketamine. No oral non-steroidal anti-inflammatory drugs and anti-depressant drugs were effective in all cases. Lidocaine or bupivacaine was administered epidurally to all four patients. When these patients stated that they did not feel pain reduced, they received epidural infusion of ketamine at doses from 5 mg to 20 mg with lidocaine or bupivacaine and their postherpetic neuralgia was controlled. Therefore with these cases, we suspect that epidural administration of ketamine, an antagonist for N-methyl-D-aspartic acid receptor, could be an effective and useful alternative treatment in a patient with refractory postherpetic neuralgia.

Primary amyloidoma of the thoracic spine presenting with acute paraplegia.

BACKGROUND: Primary solitary amyloidoma of the spine is a rare disease characterized by localized deposition of amyloid. To the best of our knowledge, there have been only 14 cases previously reported in the literature. Patients with focal spinal amyloidoma usually have relatively long symptomatic periods preoperatively, ranging from 3 weeks to 6 years (mean: 12 months). Only two reported patients had acute paraplegia. We add a third case of a thoracic spine amyloidoma presenting with acute paraplegia. CASE DESCRIPTION: A 65-year-old man presented with a three-day history of progressive paraplegia and urinary retention. He was found to have severe cord compression at T2 on magnetic resonance imaging. He underwent emergent decompressive laminectomy with instrumentation for spinal stabilization. Histopathology revealed abundant amyloid deposits. A systemic work-up was negative for amyloidosis. The patient showed marked neurological improvement with residual mild spastic gait after 1 year. CONCLUSIONS: Primary spinal amyloidoma with acute paraplegia is rare. One-stage surgery combining prompt decompression and stabilization of the spinal column is mandatory in cases of spinal amyloidoma with acute myelopathy, because primary solitary amyloidoma carries a good prognosis.

[Stump pain relieved by continuous intravenous ketamine infusion therapy].

We experienced a case of stump pain relieved by continuous intravenous ketamine infusion therapy. A 59-year-old male had his left first through fourth toes amputated because a giant iron plate at work fell on his left foot fifteen years ago. Thereafter he had refractory spontaneous burning pain and night pain on his stump. On examination, we found his left foot skin hard, lustrous, and with sweating disturbance, allodynia and hyperpathia. As intravenous administrations of ketamine 10 mg and thiamylal 50 mg were positive as a drug challenge test, we performed intravenous ketamine infusion at 1 mg.kg-1.hr-1 for 1 hour and a half. After this treatment, his visual analogue scale (VAS) improved dramatically to 0 mm, and night pain, allodynia and hyperpathia disappeared for three days. Thereafter stump pain was relieved to the level of VAS 20 mm. Therefore we diagnosed his stump pain as central pain of neuropathic origin. We suspect that continuous intravenous infusion of ketamine, a noncompetitive blocker of N-methyl-D-aspartic acid receptor, might be an effective and useful alternative treatment in a patient with refractory stump pain.

Two-staged operation for thoracolumbar osteomyelitis following methicillin-resistant staphylococcus aureus infection of a craniectomy wound--case report.

A previously healthy 53-year-old woman developed pyogenic vertebral osteomyelitis (PVO) manifesting as progressive lumbago following wound infection of a decompressive craniectomy performed for brain contusion caused by a traffic accident. Magnetic resonance imaging disclosed vertebral osteomyelitis at T-12 and L-1 with paravertebral abscess. Anterior debridement and fusion using autografts were performed at the first operation. Methicillin-resistant Staphylococcus aureus (MRSA) was cultured from the abscess specimen. Antibiotic therapy resolved the infection. Pedicle screw fixation was performed at the second operation. The patient became free from back pain and no recurrence of infection was seen. The diagnosis of PVO is frequently observed or delayed because of the nonspecific symptomatic presentation in the early stage. Coexistent infection or trauma makes early diagnosis more difficult. Indications and timing of instrumentation for the spinal column infected with MRSA is difficult. Two-staged operation with anterior debridement and posterior instrumentation after eradication of the infection is a safe and effective procedure for MRSA vertebral osteomyelitis.

[Usefulness of epidural infusion of ketamine for relief of localized superficial pain].

Three patients with localized superficial pain had their pain alleviated by single epidural infusion with low dose ketamine. The patients are as follows: a 62-year-old female with herpetic neuralgia on her right sixth thoracic nerve area; a 52-year-old male whose left shoulder, anterior chest and abdomen had been burned by acetylene gas; and a 49-year-old male whose bilateral hands suffering from frostbite by propane gas. Epidural tube insertion to administer a single dose of 10 mg ketamine with lidocaine or bupibacaine was performed in all the three patients. They were administered single epidural infusion of 10 mg ketamine with lidocaine or bupibacaine everyday and they continued to receive epidural block with lidocaine or bupivacaine including buprenorphine or morphine. Therefore, we suspect that single epidural infusion of ketamine, an antagonist for N-methyl-D-aspartic acid receptor, could be an effective and useful alternative treatment in patients with various refractory localized superficial pain of either acute or chronic nature.

[Treatment with stellate ganglion block, continuous epidural block and ulnar nerve block of a patient with postherpetic neuralgia who developed complex regional pain syndrome (CRPS)].

We present a case of a 46-year-old female patient with systemic lupus erythematosus who developed herpes zoster of the right eighth cervical nerve. Her whole right forearm, hand and the first through fifth fingers were coated with some gel and protected against pain. She had been suffering from continuous and spasmodic burning pain, hyperalgesia, allodynia, drop in skin temperature, sudmotor disturbance, edema, constructure of the joints, muscle atrophy and bone atrophy of her right upper extremity probably due to postherpetic neuralgia (PHN) associated with complex regional pain syndrome (CRPS). She received right stellate ganglion block (SGB), continuous cervical epidural block and right ulnar nerve block. Reduction of pain and edema as well as improvement in mobility of each joint of her right upper extremity was observed. We suspect that SGB, continuous cervical epidural block and ulnar nerve block are effective and useful alternative treatments in a patient with PHN associated with CRPS of the eighth cervical nerve.

[Convulsion following the combination of single preoperative oral administration of enoxacine and single postoperative intravenous administration of flurbiprofen axetil].

We experienced a case of convulsion following the combination of single oral administration of enoxacine before an emergency operation and single postoperative intravenous administration of flurbiprofen axetil. The patient was an 87-year-old female referred to our hospital for severe abdominal pain. She was diagnosed as having the strangulation ileus, then underwent the emergent operation of partial resection of the necrotic small intestine under general anesthesia. Unfortunately we did not know that she had temporarily received oral enoxacine 200 mg, a new quinolone, administered by the previous doctor on the day before the operation. After the operation, flurbiprofen axetil 50 mg, a nonsteroidal anti-inflammatory drug, was given intravenously in thirty seconds due to postoperative pain. One minute after administration of the drug, she immediately developed a convulsive fit, severe disturbance of consciousness and apnea. We then administered at once, a single dose of diazepam intravenously for convulsion treatment, kept her airway open and controlled her ventilation. Convulsion disappeared in a minute and her condition improved gradually. We suspect that convulsive seizure may have been induced by the drug interaction between single oral dose of enoxacine before the operation and single intravenous dose of flurbiprofen axetil after the operation. We also suspect that the serum concentration of enoxacine was kept high because of metabolic disturbance and renal dysfunction resulting from her old age and dehydration. This case suggests that medication before the emergency operation must be considered in anesthetic management because of the possible side effect such as convulsion induced by the drug interaction between neuquinolones and anti-inflammatory drugs.

Outcome analysis of anterior decompressive surgery and fusion for cervical ossification of the posterior longitudinal ligament: report of 107 cases and review of the literature.

OBJECT: This study was designed to determine the surgical technique and surgery-related outcome, fusion rate, and complication of anterior decompression and fusion (with various graft materials) performed in patients with ossification of the posterior longitudinal ligament (OPLL) of the cervical spine. METHODS: Between 1980 and 1998, 107 patients with radiologically proven OPLL underwent surgery via the anterior approach for direct removal of the ossified mass. Graft materials included iliac crest in 45 cases, vertebral body (VB) in 37 cases; and interbody fusion cages in 25 cases. In four patients with three-level VB grafts and one with a two-level VB graft, anterior plates were placed. Surgery-related outcome was excellent or good in 89% and fair in 11%. This clinical improvement correlated well with the severity of preoperative myelopathy. Only one patient with severe myelopathy due to extensive mixed-type OPLL developed a segmental weakness of the bilateral upper extremities. The overall fusion rate was 97%. Three patients with obvious spinal instability due to pseudarthrosis required reoperation. Of the graft materials used in this series, VB grafts were the most fragile. CONCLUSIONS: The anterior approach is an effective route for decompressing the cervical cord with OPLL. Slight asymptomatic kyphotic deformity may be encountered. Of the graft materials used in our series, VB graft was considered most fragile, and thus least optimal.