Efficacy and Safety of Biosimilar Filgrastim for Cord Blood Transplantation: A Single-Institution Retrospective Analysis.

BACKGROUND: Filgrastim has been widely used for hematopoietic recovery after hematopoietic stem cell transplantation. Recently, biosimilar filgrastim (BF) has been approved for the same indications as for the originator filgrastim (OF). However, evidence of the efficacy and safety of BF for unrelated cord blood transplant (CBT) is unreported. Therefore, we evaluated the efficacy and safety of BF and OF (historical control) for CBT. METHODS: Twenty-two consecutive patients with hematologic malignant tumors were assessed. Patients received BF (n = 12) or OF (n = 10) from day 1 after CBT for hematopoietic recovery. The time to hematopoietic recovery, total filgrastim dose, duration of filgrastim administration, total transfusion units, incidences of engraftment, documented infection, febrile neutropenia, acute and chronic graft-vs-host disease, incidence and severity of adverse events, hospitalization duration, and 100-day and 1-year overall survival were evaluated. RESULTS: The median total dose of BF and OF used for hematopoietic recovery were 9.68 and 10.80 mg, respectively. There were no significant between-group differences in time to hematopoietic recovery, total filgrastim dose, duration of filgrastim administration, total transfusion units, incidences of engraftment, documented infection, febrile neutropenia, acute and chronic graft-vs-host disease, incidence and severity of adverse events, hospitalization duration, and overall survival. Multivariate analysis demonstrated that filgrastim type was not a significant factor for neutrophil recovery. Median total filgrastim costs per patient were 446,405 and 910,320 yen for BF and OF, respectively. CONCLUSIONS: BF is as safe and effective as OF for hematopoietic recovery after CBT. BF is a useful option for CBT owing to its economic benefits.

Cyclooxygenase-2 activity is important in craniofacial fracture repair.

The aim of this study was to examine the effect of cyclooxygenase (COX)-2 on bone repair after craniofacial fracture in mice. A 4-mm fracture was created in the parietal bone of 8-week-old male COX-2 wild-type (COX-2(+/+)) and knockout (COX-2(-/-)) mice. Ribonucleic acid was extracted from the fractured bone and analysed. For morphological and histological analysis, the mice were killed 8 and 12 weeks after treatment, and sections were prepared. Three-dimensional computed tomography was performed, and the sections were stained with hematoxylin-eosin for histological examination. Expression of COX-2 messenger ribonucleic acid was induced in COX-2(+/+) mice, but not in COX-2(-/-) mice. Ossification at the fracture site was almost complete 12 weeks after fracture in COX-2(+/+) mice. In COX-2(-/-) mice, incomplete union had occurred at the fracture site. In both types of mice, the fracture site contained no cartilaginous tissue, and the callus formed from the periosteal side. These results suggest that COX-2 plays an important role in craniofacial fracture repair and that COX-2-selective non-steroidal anti-inflammatory drugs might interfere with fracture repair of the membranous viscerocranium in the clinical setting.

Unilateral expansion of a narrow mandibular dental arch combined with bimaxillary osteotomies in a patient with hypoglossia.

A 23-year-old female with hypoglossia, who had a narrow mandibular dental arch, was treated using the gradual expansion technique. Three lower incisors were missing and the right molar occlusion showed a scissor bite. Her speech was acceptable. Gradual unilateral expansion of the mandibular alveolar bone was performed. Orthodontic tooth alignment was performed prior to surgical treatment. A tooth-borne expander was devised using a hyrax-type screw to move the inclined right alveolar bone into an upright position. Alveolar bone osteotomies were performed under general anesthesia and the expander was placed in the mandibular dental arch. After a 5-day latency period, the screw was activated for 21 days. After expansion, the width of the mandibular dental arch increased by 10mm at the first molar region and the right molars were moved to an upright position. After a consolidation period of 7 days, simultaneous two-jaw surgery that combined Le Fort I osteotomy and intraoral vertical ramus osteotomies was performed to obtain a stable occlusion. After post-surgical orthodontic and prosthodontic treatment, her occlusion improved without deterioration of her speech. The results indicate that this technique is useful for unilateral expansion of distorted mandibular alveolar process.

Influence of arthroscopically observed fibrous adhesions before and after joint irrigation on clinical outcome in patients with chronic closed lock of the temporomandibular joint.

The aim of this study was to investigate the changes in the state of arthroscopically observed fibrous adhesions (FA) after visually guided irrigation (VGIR) and the influence of FA on clinical outcome in patients with chronic closed lock of the temporomandibular joint (TMJ). Forty-eight TMJs of 48 patients with unilateral chronic closed lock were enrolled in this study. All 48 joints underwent VGIR twice. After the first VGIR (immediately before the second VGIR), clinical outcome was assessed as regards maximal interincisal opening (MIO) and self-evaluated TMJ pain (VAS). Thirty patients were symptom-free (good outcome group) and the remaining 18 patients had symptoms (poor outcome group). In each group, the changes of the MIO, VAS and severity of FA (FA score) after the first VGIR were studied. The influence of FA score in the first and second VGIR on clinical outcome was analyzed by logistic regression analysis. There was no joint with disappearance or reduction of FA after the first VGIR. In both groups, MIO and VAS were significantly improved after the first VGIR even though the state of FA became significantly worse. The multivariate logistic regression analysis showed that the risk of poor outcome for FA scores in the first and second VGIR were 0.89-times (95% CI: 0.33-2.40, P=0.82) and 1.76-times (95% CI: 0.54-5.73, P=0.35), respectively. The dose-response relationships between FA scores in the first or second VGIR were not significant. In conclusion, our results indicate that the presence of FA or a postoperative worsening of FA (including postoperative new FA formation) seems not to affect the clinical outcome as regards MIO and VAS in patients with chronic closed lock of the TMJ.

Immunohistochemical localization of cyclooxygenase-1 and -2 in synovial tissues from patients with internal derangement or osteoarthritis of the temporomandibular joint.

This study examined the immunohistochemical expression and localization of cyclooxygenase-1 and -2 (COX-1 and COX-2) in synovial tissues from patients with internal derangement (ID) or osteoarthritis (OA) of the temporomandibular joint (TMJ). Synovial tissues from patients with condylar fractures of the mandible were studied as control. Synovial tissues from 13 TMJs of 10 patients with ID or OA and from 5 TMJs of 4 patients with fractures were examined for COX-1 and COX-2 expression by immunohistochemical staining using two monoclonal antibodies. In addition, whether the COX-2 expression grade correlated with the synovitis score and clinical findings was assessed. COX-2 was expressed in the synovial lining, infiltrating mononuclear cells, fibroblast-like cells, and blood vessels, including CD31-positive endothelial cells, in the synovium of patients with ID or OA. Expression levels of COX-1 in synovial lining cells and endothelial cells were similar in the specimens obtained from the patients with ID or OA and those obtained from the controls. The expression of COX-2 positively correlated with arthroscopic findings of synovitis (p = 0.55, P = 0.023) and with joint pain (p = 0.56, P = 0.021). These results suggest that up-regulation of COX-2 in synovium may play a part in the pathogenesis of synovitis in patients with ID or OA of the TMJ.

Suppression of the morphine-induced rewarding effect and G-protein activation in the lower midbrain following nerve injury in the mouse: involvement of G-protein-coupled receptor kinase 2.

The present study was designed to investigate whether a state of neuropathic pain induced by sciatic nerve ligation could alter the rewarding effect, antinociception, and G-protein activation induced by a prototype of mu-opioid receptor agonist morphine in the mouse. The sciatic nerve ligation caused a long-lasting and profound thermal hyperalgesia. Under this neuropathic pain-like state, an i.c.v. morphine-induced place preference was observed in sham-operated mice but not in sciatic nerve-ligated mice. However, no differences in the antinociceptive effect of i.c.v.-administered morphine were noted between the groups. The increases in the binding of guanosine-5'-o-(3-[(35)S]thio)triphosphate induced by morphine in lower midbrain membranes including the ventral tegmental area, which contributes to the expression of the rewarding effect of opioid, were significantly attenuated in sciatic nerve-ligated mice. On the other hand, there were no differences in the stimulation of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding to pons/medulla membranes, which plays an important role in the antinociception of mu-opioid receptor agonists, between the groups. In addition, no changes in levels of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding by either the selective delta- or kappa-opioid receptor agonists were noted in membrane of the lower midbrain and limbic forebrain membranes obtained from sciatic nerve-ligated mice. Reverse transcription-polymerase chain reaction analysis showed that sciatic nerve ligation did not alter the mRNA product of mu-opioid receptors in the lower midbrain, indicating that a decrease in some mu-opioid receptor functions may result from the uncoupling of mu-opioid receptors from G-proteins. We found a significant increase in protein levels of G-protein-coupled receptor kinase 2, which causes receptor phosphorylation in membranes of the lower midbrain but not in the pons/medulla, obtained from mice with nerve injury, whereas there were no changes in the protein level of phosphorylated-protein kinase C in the lower midbrain. These results suggest that the uncoupling of mu-opioid receptors from G-proteins by G-protein-coupled receptor kinase 2 in the lower midbrain may, at least in part, contribute to the suppression of the rewarding effect of morphine under neuropathic pain.

Dentoalveolar reconstruction of a missing premaxilla using bone graft and endosteal implants.

We report here on a patient with bilateral cleft lip and palate (BCLP) and a missing premaxilla, who underwent dentoalveolar reconstruction of the cleft and premaxillary alveolus using endosteal implants after bone grafting. The patient, whose maxillary incisors and premaxilla were missing, had corticocancellous bone grafting from the iliac crest, followed by excellent bone formation at the anterior alveolus. After the placement of the endosteal implants and the completion of the pre-surgical orthodontic alignment, orthognathic surgery was performed for the restoration of a Class III open bite. After post-operative orthodontic preparation, the final fixed prostheses were completed. This treatment procedure offers an option of dentoalveolar reconstruction for BCLP patients with an excised premaxilla.

Partial inferior turbinectomy during secondary alveolar bone grafting.

This study examined the characteristics and outcome of patients undergoing partial inferior turbinectomy during secondary alveolar bone grafting. Thirty-three of 55 patients with cleft lip and palate or cleft lip and alveolus who underwent secondary alveolar bone grafting concurrently received partial inferior turbinectomy to ensure that the height of the nasal floor was similar on the cleft side and non-affected side. At the time of surgery, patients who underwent turbinectomy were significantly older than those who did not undergo the procedure. The proportion of patients who underwent turbinectomy was significantly higher among patients with cleft lip and palate than among those with cleft lip and alveolus. These differences apparently reflected the developmental stage of the inferior turbinate and the relative severity of alveolar and palatal defects. In most patients who underwent partial inferior turbinectomy, postoperative X-ray films revealed excellent bone formation at the graft site. Our findings suggest that partial inferior turbinectomy during secondary alveolar bone grafting is a very useful procedure that facilitates dissection to the height of the nasal floor, reconstruction of the mucosal nasal floor, and formation of a sufficient bone bridge. It also promotes alveolar cleft closure, especially in patients with wide bone defects.

Glia-synapse interaction through Ca2+-permeable AMPA receptors in Bergmann glia.

Glial cells express a variety of neurotransmitter receptors. Notably, Bergmann glial cells in the cerebellum have Ca2+-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) assembled without the GluR2 subunit. To elucidate the role of these Ca2+-permeable AMPARs, we converted them into Ca2+-impermeable receptors by adenoviral-mediated delivery of the GluR2 gene. This conversion retracted the glial processes ensheathing synapses on Purkinje cell dendritic spines and retarded the removal of synaptically released glutamate. Furthermore, it caused multiple innervation of Purkinje cells by the climbing fibers. Thus, the glial Ca2+-permeable AMPARs are indispensable for proper structural and functional relations between Bergmann glia and glutamatergic synapses.

Ca(2+)-sensor region of IP(3) receptor controls intracellular Ca(2+) signaling.

Many important cell functions are controlled by Ca(2+) release from intracellular stores via the inositol 1,4,5-trisphosphate receptor (IP(3)R), which requires both IP(3) and Ca(2+) for its activity. Due to the Ca(2+) requirement, the IP(3)R and the cytoplasmic Ca(2+) concentration form a positive feedback loop, which has been assumed to confer regenerativity on the IP(3)-induced Ca(2+) release and to play an important role in the generation of spatiotemporal patterns of Ca(2+) signals such as Ca(2+) waves and oscillations. Here we show that glutamate 2100 of rat type 1 IP(3)R (IP(3)R1) is a key residue for the Ca(2+) requirement. Substitution of this residue by aspartate (E2100D) results in a 10-fold decrease in the Ca(2+) sensitivity without other effects on the properties of the IP(3)R1. Agonist-induced Ca(2+) responses are greatly diminished in cells expressing the E2100D mutant IP(3)R1, particularly the rate of rise of initial Ca(2+) spike is markedly reduced and the subsequent Ca(2+) oscillations are abolished. These results demonstrate that the Ca(2+) sensitivity of the IP(3)R is functionally indispensable for the determination of Ca(2+) signaling patterns.

Retrovirus insertion and transcriptional activation of the multidrug-resistance gene in leukemias treated by a chemotherapeutic agent in vivo.

To understand the molecular basis for multidrug-resistant (MDR) cancer cells in vivo, this study analyzed molecular changes of the mdr1a gene region in leukemia cells in mice during continuous treatment with vincristine. An inverse insertion of murine leukemia retrovirus (MuLV) into the 5'-flanking region of the mdr1a gene was found. This insertion was concomitantly accompanied by up-regulation of the mdr1a gene and the loss of chemosensitivity. Deletion of long-terminal repeat (LTR) sequences dramatically decreased the mdr1a promoter-driven reporter activity. The MuLV LTR insertion appears to exert its enhancer activity on mdr1a transcription during the appearance of MDR leukemia cells. Two mechanisms were postulated to explain the mdr1a gene activation by retrovirus insertion during in vivo chemotreatment: de novo insertion of MuLV induced by vincristine treatment and selection of a small fraction of pre-existing cells carrying MuLV insertion during vincristine treatment. No rearranged sequence was detected by polymerase chain reaction in parental cells. This result argued for the first mechanism. The randomly altered distribution of MuLV during repetitive chemotreatment might also be consistent with this hypothesis. On the other hand, the retrovirus insertion was detected at the same site of the mdr1a promoter region in 2 independent experiments, which suggests the second mechanism. It should be noted that in vivo chemotreatment using vincristine could generate the mdr1a-overexpressing cells through retrovirus insertion and the enhancer effect of the LTR.

Evaluation of distal and proximal axonal degeneration in patients with carpal tunnel syndrome.

In patients with carpal tunnel syndrome, varying degrees of demyelination and axonal degeneration occur in the median nerve. Only a few studies have examined axonal degeneration produced at proximal to the lesion. In this study proximal axonal degeneration was evaluated and compared with other parameters. In 40 consecutive CTS patient hands, distal latency (DL), compound muscle action potential amplitude (CMAP) and motor conduction velocity (MCV) were analyzed by conventional motor nerve conduction studies. Intrafascicular compound nerve action potential amplitude (N-CNAP) at the elbow after wrist simulation and its nerve conduction velocity (NCV) between wrist and elbow were also analyzed. The negative correlation of DL with CMAP was statistically significant (r = 0.577, p < 0.001). CMAP was correlated with either MCV (r = 0.537, p < 0.001) or N-CMAP (r = 0.710, p < 0.001). A significant correlation of MCV with NCV (r = 0.517, p < 0.001) was also indicated. There were no any other significant correlation among the parameters. In CTS the degree of demyelination and axonal degeneration influence the prognosis for nerve recovery after decompressive surgery. DL is mainly influenced by demyelination that results in conduction block and slowing at the carpal tunnel. CMAP and N-CNAP indicate the degree of axonal degeneration at distal and proximal to the compression site. As in electrophysiologic evaluation of mononeuropathies, proximal axonal degeneration is best assessed by both stimulation and recording electrode locationing proximal to the lesion. Recording of intrafascicular nerve action potential was a little invasive method, but it provided important informations. The negative correlation between DL and CMAP implies that distal axonal degeneration can occur in proportion to the conduction disturbance. Moreover, N-CNAP had a higher correlation with CMAP. The greater the distal axonal degeneration, the more the proximal axonal degeneration. Conduction velocity represents the velocity of the fastest conduction fiber, not the degree of axonal degeneration.

Synthesis and structure--activity analysis of novel dihydropyridine derivatives to overcome multidrug resistance.

The structure activity relationships were studied on newly synthesized 1,4-dihydropyridine derivatives possessing a 1-pentyl group at the 4-position, and 3-pyridylpropylester was found to be one of the effective fragments for overcoming P-glycoprotein mediated multidrug-resistance (MDR) in cultured human cancer cells, in vitro. 3-Pyridylpropylester was also found to be one of the effective fragments for increasing the life span of P-glycoprotein overexpressing MDR P388 leukemia-bearing mice, in vivo. All compounds had weak calcium antagonistic activities, but there appeared no relationship between MDR reversing effect and calcium antagonistic activity.

Blue-light-dependent osmoregulation in protoplasts of Phaseolus vulgaris Pulvini.

Blue light was found to induce shrinkage of the protoplasts isolated from first-leaf lamina pulvini of 18-day-old Phaseolus vulgaris. The response was transient following pulse stimulation, while it was sustainable during continuous stimulation. No apparent difference was found between flexor and extensor protoplasts. Protoplasts of the petiolar segment located close to the pulvinus showed no detectable response. In the plants used, the pulvinus was fully matured and the petiole was ceasing its elongation growth. When younger, 12-day-old, plants were used, however, the petiolar protoplasts did respond to blue light. The pulse-induced response was similar to that in pulvinar protoplasts, although the response to continuous stimulation was transient and differed from that in pulvinar protoplasts. No shrinkage was induced in pulvinar protoplasts when the far-red-light-absorbing form of phytochrome was absent for a period before blue-light stimulation, indicating that the blue-light responsiveness is strictly controlled by phytochrome. Inhibitors of anion channels and H(+)-ATPase abolished the shrinking response, supporting the view that protoplasts shrink by extruding ions. The response of pulvinar protoplasts is probably involved in the blue-light-induced, turgor-based movement of pulvini. The blue-light responding system in pulvini is suggested to have evolved from that functioning in other growing organs.

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo.

We have developed a method for visualizing phosphorylation of proteins in living cells using a novel fluorescent indicator composed of two green fluorescent protein (GFP) variants joined by the kinase-inducible domain (KID) of the transcription factor cyclic adenosine monophosphate (cAMP)-responsive element binding protein (CREB). Phosphorylation of KID by the cAMP-dependent protein kinase A (PKA) decreased the fluorescence resonance energy transfer (FRET) among the flanking GFPs. By transfecting COS-7 cells with an expression vector encoding this indicator protein (termed ART for cAMP-responsive tracer), we were able to visualize activation dynamics of PKA in living cells.

Histological evaluation of autogenous iliac particulate cancellous bone and marrow grafted to alveolar clefts--a preliminary report of five young adult cases.

OBJECTIVE: We histologically evaluated iliac particulate cancellous bone and marrow (PCBM) grafted to alveolar clefts in five young adults with cleft lip and palate. PATIENTS AND METHODS: Five young adults with cleft lip and palate underwent secondary alveolar bone grafting. The mean age of the patients was 21.2 years (range = 17 to 27 years). Bone specimens were taken from the graft site 5 to 10 months after the surgery simultaneously with implant fixture placement (two cases), vestibuloplasty (two cases), and rebone grafting (one case). These five bone specimens were prepared and stained with hematoxylin and eosin and examined microscopically. RESULT: One specimen, taken 5 months after surgery, showed immature trabecular bone partially lined by osteoblasts. The remaining four specimens showed well-mineralized trabeculae and fatty change in the marrow space. CONCLUSION: In young adult cases, PCBM remodeled after grafting and became mature bone about 5 to 6 months after the surgery.

Antibody studies of factor VIII inhibitor in a case with Waldenström's macroglobulinemia.

We report a case of Waldenström's macroglobulinemia with prominent bleeding tendency; laboratory investigation revealed an elevated activated partial thromboplastin time. Further laboratory evaluation showed circulating factor VIII anticoagulant, deemed polyclonal IgG, with a titer of 700 Bethesda Units/ml. The factor VIII inactivation kinetics of the patient plasma were identical to those of a type II inhibitor, and the inhibitor was found to recognize the A2 domain of the factor VIII heavy chain. Apparently, paraprotein is not always the cause of reduced activity of coagulation factors in neoplastic dysproteinemias.

Dynamic Ca2+ signalling in rat arterial smooth muscle cells under the control of local renin-angiotensin system.

1. We visualized the changes in intracellular Ca2+ concentration ([Ca2+]i), using fluo-3 as an indicator, in individual smooth muscle cells within intact rat tail artery preparations. 2. On average in about 45 % of the vascular smooth muscle cells we found spontaneous Ca2+ waves and oscillations ( approximately 0.13 Hz), which we refer to here as Ca2+ ripples because the peak amplitude of [Ca2+]i was about one-seventh of that of Ca2+ oscillations evoked by noradrenaline. 3. We also found another pattern of spontaneous Ca2+ transients often in groups of two to three cells. They were rarely observed and are referred to as Ca2+ flashes because their peak amplitude was nearly twice as large as that in noradrenaline-evoked responses. 4. Sympathetic nerve activity was not considered responsible for the Ca2+ ripples, and they were abolished by inhibitors of either the Ca2+ pump in the sarcoplasmic reticulum (cyclopiazonic acid) or phospholipase C (U-73122). 5. Both angiotensin antagonists ([Sar1,Ile8]-angiotensin II and losartan) and an angiotensin converting enzyme inhibitor (captopril) inhibited the Ca2+ ripples. 6. The extracellular Ca2+-dependent tension borne by unstimulated arterial rings was reduced by the angiotensin antagonist by approximately 50 %. 7. These results indicate that the Ca2+ ripples are generated via inositol 1,4, 5-trisphosphate-induced Ca2+ release from the intracellular Ca2+ stores in response to locally produced angiotensin II, which contributes to the maintenance of vascular tone.