Bilaterally existing sternoclavicularis anticus muscles

A 92-year-old Japanese woman was found to have the sternoclavicularis anticus muscles existing bilaterally during the course of educational dissection at Nagoya City University Medical School. It was confirmed that these muscles received the nerve supply from small branches of the lateral pectoral nerves from the brachial plexus. Herein is documented the precise gross anatomical findings with some morphometric measurements. Moreover, the morphology of this muscle is discussed in relation to previously described variations of the pectoral sheet of muscles.     

Regulatory T cells for tolerance

Regulatory T cells (Tregs) are critical mediators of immune homeostasis and hold significant promise in the quest for transplantation tolerance. Progress has now reached a critical threshold as techniques for production of clinical therapies are optimised and Phase I/II clinical trials are in full swing. Initial safety and efficacy data are being reported, with trials assessing a number of different strategies for the introduction of Treg therapy. It is now more crucial than ever to elucidate further the function and behaviour of Tregs in vivo and ensure safe delivery. This review will discuss the current state of the art and future directions in Treg therapy.     

Histological study of the developing pterygoid process of the fetal mouse sphenoid

The pterygoid process undergoes ossification of both the cartilage and membrane. However, few studies have attempted to explore the sequential development of the pterygoid process. Using histological examination, we performed morphological observations of the pterygoid process and surrounding tissue. ICR mice at embryonic days 13.5–18.0 and postnatal day 0 were used for morphological observations of the pterygoid process. By embryonic day 14.5, a mesenchymal cell condensation forming the anlage of the future medial pterygoid process differentiated into osteoid-like tissue and cartilage. At embryonic days 15.5–16.5, cartilage cells were clearly evident in the medial pterygoid process. In the medial pterygoid process, a bone collar was evident and calcified bone tissue surrounded the cartilage. At this point, a mesenchymal cell condensation formed the anlage of the pterygoid hamulus. At embryonic days 17.0–18.0, the cartilages were located along the lower and posterior border of the medial pterygoid process. A metachromatically stained matrix first became detectable around cells located in the pterygoid hamulus. On the other hand, at embryonic day 13.5, a metachromatically stained matrix was already evident in the space between the flattened cells in the lateral pterygoid process. At embryonic day 17.0, a hypertrophic cell zone had clearly formed in the diaphysis. On the basis of our present investigation, the lateral pterygoid process can be classified as primary cartilage, whereas the medial pterygoid process can be classified as secondary cartilage. Furthermore, it was found that the pterygoid hamulus is formed latest in the medial pterygoid process.     

Cells transplanted onto the surface of the glial scar reveal hidden potential for functional neural regeneration

Cell transplantation therapy has long been investigated as a therapeutic intervention for neurodegenerative disorders, including spinal cord injury, Parkinson’s disease, and amyotrophic lateral sclerosis. Indeed, patients have high hopes for a cell-based therapy. However, there are numerous practical challenges for clinical translation. One major problem is that only very low numbers of donor cells survive and achieve functional integration into the host. Glial scar tissue in chronic neurodegenerative disorders strongly inhibits regeneration, and this inhibition must be overcome to accomplish successful cell transplantation. Intraneural cell transplantation is considered to be the best way to deliver cells to the host. We questioned this view with experiments in vivo on a rat glial scar model of the auditory system. Our results show that intraneural transplantation to the auditory nerve, preceded by chondroitinase ABC (ChABC)-treatment, is ineffective. There is no functional recovery, and almost all transplanted cells die within a few weeks. However, when donor cells are placed on the surface of a ChABC-treated gliotic auditory nerve, they autonomously migrate into it and recapitulate glia- and neuron-guided cell migration modes to repair the auditory pathway and recover auditory function. Surface transplantation may thus pave the way for improved functional integration of donor cells into host tissue, providing a less invasive approach to rescue clinically important neural tracts.Cells transplanted into the nervous system could cure various forms of neurodegenerative disease and neural injury (1, 2), but most donor cells die without functional integration (3), and glial scar tissue is strongly inhibitory to axon regeneration (4). Glial scar tissue forms following conditions such as ischemia or mechanical trauma when reactive astrocytes increase proliferation, become hypertrophic, and up-regulate glial fibrillary acidic protein (GFAP) (5, 6). In this study, however, we demonstrate that if donor cells are delivered appropriately, they can interact successfully with the glial scar and restore lost neuronal function. We developed an in vivo model of the chronic gliotic environment in clinical patients by compressing the rat auditory nerve without breaching the fluid spaces that contain the sensory structures inside the cochlea (7–10). In this model, spiral ganglion cells (the auditory neurons) degenerate selectively, but hair cells are preserved both morphologically and functionally (7, 8, 10, 11). We then waited 5 wk before cell transplantation to allow the formation of a glial scar with the progressive degeneration of auditory neurons. We characterized the glial scar with several experimental measures. This characterization is an important element because experimental models for cell-based therapies for neurodegenerative diseases should include the chronic gliotic environment to simulate the appropriate clinical condition in patients.To compress the auditory nerve, the CNS portion was atraumatically exposed in the cerebellopontine angle cistern through right suboccipital craniectomy, a nerve hook was placed into the internal auditory meatus, and the nerve was injured by a single compression (Fig. 1 A and B and Fig. S1A). We used a murine auditory neuroblast cell line for transplantation (US/VOT-N33) (12). This cell line was derived from the ventral otocyst (inner ear anlage) of a mouse embryo at embryonic day ED10.5 and was selected because it expresses key markers for auditory sensory neurons and differentiates with the appropriate bipolar morphology in the ear both in vitro and in vivo (11, 12).Open in a separate windowFig. 1.Glial scar formation after auditory nerve compression. (A) The CNS portion (CNS-P) of the normal auditory nerve protrudes into the auditory nerve trunk and the transitional zone (TZ) at the boundary with the PNS is within the internal auditory canal (IAC). Mechanical compression (red arrow) applied to the CNS-P induced a glial scar. The red circle corresponds to that in Fig. S1A. BS, brainstem; CN, cochlear nucleus; Fs, fundus of the internal auditory canal; HC, hair cell; HP, habenula perforata; IAM, internal auditory meatus; PNS-P, PNS portion of the auditory nerve; SGC, spiral ganglion cells. (B) Auditory nerve (AuN) in sham-operated and compressed (Comp) ears. The compression site is marked with a double arrow. Distal extension of the GFAP domain is indicated by arrowheads. Spiral ganglion cells, labeled with Tuj1, were lost following compression (multiple single arrows). Asterisks indicate glial scar processes projecting into small bony canals beyond the fundus of the IAC. (C) Fluorescence image of glial scar formed 5 wk after auditory nerve injury, ChABC treated (Upper). The boxed area is enlarged in lower panels where confocal microscopy disclosed fine glial scar processes (arrowheads) past Rosenthal’s canal (R). The margin of the limbus was nonspecifically stained (arrows). bsl, mdl, ap: basal, middle, apical cochlear turn, respectively. (D) Expression of GFAP. PPA, positive pixel area (n = 4). (E) Western blot for GFAP. (F) Relative expression of GFAP mRNA after 1, 4, and 25 wk. (G) Expression of 1G2 (Neurocan) (n = 4). (H) Relative expression of Neurocan mRNA. (I) Relative expression of Nestin mRNA. (J) Nestin expression at compression site. Most of the glial scar processes were Nestin positive (arrowheads) and some were GFAP positive (arrows). The boxed area in the left panel is enlarged in the right. The asterisk indicates tissue loss due to compression injury. (K) The ChABC-digested auditory nerve was 2B6-positive (n = 4). (Scale bars, 200 μm in B and C, Upper; 100 μm in J; and 20 μm in C, Lower, D, G, and K.) All values represent mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.Open in a separate windowFig. S1.Techniques to induce glial scar in rat auditory system to transplant donor cells to gliotic auditory nerve and the primary antibodies used in this study. (A) Operative view of auditory nerve compression in the cerebellopontine angle (CPA). Red circle corresponds to that in Fig. 1A. Fundus, fundus of the internal auditory canal; IAM, internal auditory meatus; PB, petrous bone. (B) Donor cells were transplanted to gliotic auditory nerve by two different techniques: the intraneural method and the surface transplantation method. The tips of the syringes are indicated by circles. (C) Primary antibodies used to identify relevant endogenous and exogenous components of the auditory system (SI Materials and Methods).In the first experiments, we delivered cells into the auditory nerve via a thin fused silica tube (Fig. S1B), treating the nerve topically with chondroitinase ABC (ChABC) (13) soaked in a gelatin sponge at the time of nerve injury and then again immediately after cell transplantation. Three months later, we assessed the structural and functional recovery of the auditory nerve.During these experiments, we noticed that donor cells that spilled onto the surface of the nerve were apparently able to enter the ChABC-treated tissue and to survive. Thus, we reasoned that cell delivery onto the surface of the glial scar might be successful and less invasive. In this context, we designed an experiment to compare two methods of cell delivery. In the first, we applied the conventional method of intraneural infusion, and in the second, we delivered cells to the surface of the nerve without mechanical damage to its structure. We then compared the results in terms of cell morphology and functional recovery of the auditory system.     

Development of less invasive neuromuscular electrical stimulation model for motor therapy in rodents

Background

Combination therapy is essential for functional repairs of the spinal cord. Rehabilitative therapy can be considered as the key for reorganizing the nervous system after spinal cord regeneration therapy. Functional electrical stimulation has been used as a neuroprosthesis in quadriplegia and can be used for providing rehabilitative therapy to tap the capability for central nervous system reorganization after spinal cord regeneration therapy.

Objective

To develop a less invasive muscular electrical stimulation model capable of being combined with spinal cord regeneration therapy especially for motor therapy in the acute stage after spinal cord injury.

Methods

The tibialis anterior and gastrocnemius motor points were identified in intact anesthetized adult female Fischer rats, and stimulation needle electrodes were percutaneously inserted into these points. Threshold currents for visual twitches were obtained upon stimulation using pulses of 75 or 8 kHz for 200 ms. Biphasic pulse widths of 20, 40, 80, 100, 300, and 500 µs per phase were used to determine strength–duration curves. Using these parameters and previously obtained locomotor electromyogram data, stimulations were performed on bilateral joint muscle pairs to produce reciprocal flexion/extension movements of the ankle for 15 minutes while three-dimensional joint kinematics were assessed.

Results

Rhythmic muscular electrical stimulation with needle electrodes was successfully done, but decreased range of motion (ROM) over time. High-frequency and high-amplitude stimulation was also shown to be effective in alleviating decreases in ROM due to muscle fatigue.

Conclusions

This model will be useful for investigating the ability of rhythmic muscular electrical stimulation therapy to promote motor recovery, in addition to the efficacy of combining treatments with spinal cord regeneration therapy after spinal cord injuries.     

A multicenter early phase II study of high-dose chemotherapy with autologous peripheral blood stem cell transplantation for treatment of intermediate-grade non-Hodgkin's lymphoma. Japan Blood Cell Transplantation Study Group

We conducted a multicenter early phase II study to evaluate the feasibility and therapeutic efficacy of high-dose chemotherapy supported by autologous peripheral blood stem cell transplantation (auto-PBSCT) for the treatment of intermediate grade non-Hodgkin's lymphoma (IG-NHL). High-dose etoposide or cyclophosphamide followed by G-CSF was used for PBSC mobilization, and a sufficient number of CD34+ cells (> 1 x 10(6)/kg) were collected. Out of 81 enrolled patients, 50 received high-dose chemotherapy with auto-PBSCT; Hematologic recovery after transplantation was rapid. The incidence of grade III/IV toxicity (Bearman) was about 6%; treatment-related mortality was 6% (3/50). The Disease-free survival rate for the patients in complete remission who received high-dose chemotherapy with auto-PBSCT was better than that for the patients who were treated with conventional chemotherapy (57% vs 35%). These preliminary results indicated that high-dose chemotherapy with auto-PBSCT is feasible and effective. A prospective randomized phase III clinical trial will be required to assess the efficacy of high-dose chemotherapy with auto-PBSCT as a post-remission therapy for IG-NHL.     

An early phase II study of autologous peripheral blood stem cell transplantation for acute myelogenous leukemia in first remission. Japan Blood Cell Transplantation Study Group

We performed a multicenter, an early phase II clinical trial to evaluate the feasibility, safety and efficacy of myeloablative therapy supported by autologous peripheral blood stem cell transplantation (auto-PBSCT) for the treatment of acute myelogenous leukemia (AML) in first remission. A total of 105 patients were enrolled in the study, and 56 patients in first complete remission received auto-PBSCT. The median age was 44 years. Of the 56 patients, 34 (60.7%) had M2 or M3 AML by the French-American-British Classification system. The median concentration of infused CD34+ cells was 2.3 x 10(6)/kg by recipient body weight. Median days to reach an absolute neutrophil count > 500/microliter and a platelet count > 20000/microliter were 14 and 16, respectively. The median disease-free survival rate was estimated to be 62.0% at a median follow-up time of 534 days. Although the study enrolled a small number of patients and the follow-up period was relatively short, the preliminary results were encouraging and indicated that myeloablative chemotherapy with auto-PBSCT is feasible and can be performed safely as a post-remission therapy for AML. A prospective randomized clinical trial of auto-PBSCT versus standard chemotherapy alone will be necessary to assess the efficacy of high-dose therapy facilitated by auto-PBSCT as a post-remission therapy for AML.     

Long-term pathophysiological studies on loop ileostomy and ileoanal anastomosis

Mucosal proctectomy with ileoanal anastomosis (IAA) had been performed on 37 patients with adenomatosis coli and 16 with ulcerative colitis between 1978 and 1987. These patients were followed up for a mean of 7.5 years. In 38 cases (73%), this procedure was completed by closure of loop ileostomy. The mean number of bowel movements per 24 hours was 6.0. Seven patients had occasional episodes of minor nocturnal soiling. The postoperative maximum resting anal pressure was the most important parameter reflecting clinical results and it rose from 72 to 92 cm of water during 5.3 year follow-up period after IAA. The pathophysiological studies on loop ileostomy and IAA were performed in these patients. Postoperative small bowel transit time evaluated by radioopaque markers was shortened. Daily output of water and sodium, and Na/K ratio in the ileal excreta increased and total counts of anaerobes in feces decreased. On the other hand, daily volume, Na/K ratio and PH of urine fell significantly. These phenomena were remarkable in patients who received loop ileostomy with about 60 cm defunctioning terminal ileum. These results indicate that it is necessary to maintain intestinal continuity in the ileal pouch-anal procedures.