Development of a stable semi-continuous lipid production system of an oleaginous Chlamydomonas sp. mutant using multi-omics profiling.

BACKGROUND: Microalgal lipid production has attracted global attention in next-generation biofuel research. Nitrogen starvation, which drastically suppresses cell growth, is a common and strong trigger for lipid accumulation in microalgae. We previously developed a mutant Chlamydomonas sp. KAC1801, which can accumulate lipids irrespective of the presence or absence of nitrates. This study aimed to develop a feasible strategy for stable and continuous lipid production through semi-continuous culture of KAC1801. RESULTS: KAC1801 continuously accumulated > 20% lipid throughout the subculture (five generations) when inoculated with a dry cell weight of 0.8-0.9 g L-1 and cultured in a medium containing 18.7 mM nitrate, whereas the parent strain KOR1 accumulated only 9% lipid. Under these conditions, KAC1801 continuously produced biomass and consumed nitrates. Lipid productivity of 116.9 mg L-1 day-1 was achieved by semi-continuous cultivation of KAC1801, which was 2.3-fold higher than that of KOR1 (50.5 mg L-1 day-1). Metabolome and transcriptome analyses revealed a depression in photosynthesis and activation of nitrogen assimilation in KAC1801, which are the typical phenotypes of microalgae under nitrogen starvation. CONCLUSIONS: By optimizing nitrate supply and cell density, a one-step cultivation system for Chlamydomonas sp. KAC1801 under nitrate-replete conditions was successfully developed. KAC1801 achieved a lipid productivity comparable to previously reported levels under nitrogen-limiting conditions. In the culture system of this study, metabolome and transcriptome analyses revealed a nitrogen starvation-like response in KAC1801.

Enhancing carbohydrate repartitioning into lipid and carotenoid by disruption of microalgae starch debranching enzyme.

Light/dark cycling is an inherent condition of outdoor microalgae cultivation, but is often unfavorable for lipid accumulation. This study aims to identify promising targets for metabolic engineering of improved lipid accumulation under outdoor conditions. Consequently, the lipid-rich mutant Chlamydomonas sp. KOR1 was developed through light/dark-conditioned screening. During dark periods with depressed CO2 fixation, KOR1 shows rapid carbohydrate degradation together with increased lipid and carotenoid contents. KOR1 was subsequently characterized with extensive mutation of the ISA1 gene encoding a starch debranching enzyme (DBE). Dynamic time-course profiling and metabolomics reveal dramatic changes in KOR1 metabolism throughout light/dark cycles. During light periods, increased flux from CO2 through glycolytic intermediates is directly observed to accompany enhanced formation of small starch-like particles, which are then efficiently repartitioned in the next dark cycle. This study demonstrates that disruption of DBE can improve biofuel production under light/dark conditions, through accelerated carbohydrate repartitioning into lipid and carotenoid.

Hybridizing semiconductor nanocrystals with metal-organic frameworks for visible and near-infrared photon upconversion.

Hybrid materials consisting of semiconductor nanocrystals and metal-organic frameworks (MOFs) were prepared for the first time to achieve photon upconversion based on triplet-triplet annihilation (TTA-UC) in the solid-state, which allowed TTA-UC with large anti-Stokes shifts in the visible and near-infrared regions.

Efficacy and safety of oral propranolol for infantile hemangioma in Japan.

BACKGROUND: There have been few reports on the efficacy and safety of oral propranolol at 3 mg/kg/day for infantile hemangioma (IH) in Japanese patients. METHODS: A multicenter, open-label phase III study was conducted to evaluate the efficacy and safety of oral propranolol solution in Japanese infants aged 35-150 days with proliferating IH. Thirty-two patients were enrolled in the study, received propranolol solution for 24 weeks at 3 mg/kg/day, and completed the study. RESULTS: The success rate (complete or nearly complete resolution) at week 24 (primary endpoint) was 78% (95%CI: 60-91%). The improvement rate since the previous visit was 100% (32/32) after week 5. Overall, the IH surface area, maximum diameter, and color intensity all decreased over time. Consistency in assessment between the centralized and the investigator on-site assessments was observed in 26 patients. Of the 32 patients, 11 needed further treatment other than the study drug. The incidence of adverse events (AE) and drug-related AE was 97% and 31%, respectively. AE that occurred in ≥two patients were either typical of propranolol use (such as blood pressure decrease) or common events in infants. AE that resulted in dose reduction were observed in two patients, but no serious AE or AE that led to study drug discontinuation were observed. CONCLUSION: Oral propranolol solution at 3 mg/kg/day is effective and safe in Japanese IH patients.

A novel active intraoral appliance for presurgical orthopaedic treatment in patients with complete bilateral cleft lip and palate.

BACKGROUND: Management of the protruding/deviated premaxilla in patients with complete bilateral cleft lip and palate is a challenging problem for surgeons and orthodontists. Various passive and active methods have been developed for the presurgical orthopaedic treatment. However, most of these treatments are complicated and laborious for the patient's parents and clinicians. Here, we describe our original active intraoral appliance comprising two components, that is, the premaxillary and palatine process plates, connected with two elastic chains, and we assess its therapeutic efficacy. PATIENTS AND METHODS: We retrospectively evaluated 15 patients treated using this appliance during 2006-2012, followed up for an average of 60.3 months (range, 18-97 months). We analysed the cleft widths and maxillary size, obtained pretreatment, post-treatment and pre-palatoplasty. RESULTS: Cleft widths and premaxillary protrusion were significantly decreased post treatment; however, the transverse dimensions were not significantly altered. In all cases, the protruding/deviated premaxilla was set into a suitable position within 1 month, and we could perform one-stage cheiloplasty using the modified Mulliken method with low tension. CONCLUSION: Our appliance is technically simple to use, less invasive to the skin and bone and cost-effective, with reliable and predictable outcomes. In the follow-up period, we observed no detrimental growth of the maxilla or dentition. Therefore, we consider our appliance to be useful for application in presurgical orthopaedic treatments of complete bilateral cleft lip and palate.

Simultaneous correction of deviated columella and wide nostril floor using the Y-V advancement in unilateral cleft lip nasal deformities.

Here, we describe an operative method for the simultaneous correction of deviated columella and wide nostril floor in unilateral cleft lip nasal deformities. The V-shaped flap of the affected nostril floor was inserted into the side wall of the columella using the Y-V advancement. By suturing the anterocaudal septum, which is often deviated to the unaffected side, with the subcutaneous tissue of the affected alar base, it was possible to simultaneously reposition the columellar base in the midline as well as narrow the wide nostril floor. This method was technically simple, with low invasiveness for the septal cartilage. Furthermore, as noted in one of the two cases described, it would enable the reconstruction of the natural shape of the nostril sill.

A novel pre-surgical maxillary orthodontic device using ß-titanium wire for wide unilateral cleft lip and palate patients: preliminary study of its efficacy and impact for the maxillary formation.

BACKGROUND AND AIM: For patients with a wide, complete, unilateral cleft lip and palate, pre-surgical maxillary orthodontic treatments have been used to reduce the alveolar gap before cheiloplasty. However, most of these treatments are complicated and laborious for patients and for medical professionals. Thus, we developed an original pre-surgical orthodontic device made with two separate acrylic resin plates connected with a spring-shaped ß-titanium wire (ß-TW). When the device was applied on the palate, each segment of the maxilla was automatically aligned for our target formation with the elastic force of ß-titanium alloy. This study aimed to evaluate the efficacy of the new device and the size of the maxilla in comparison with the conventional Hotz procedures. PATIENTS AND METHODS: A total of 47 patients with a wide unilateral cleft lip and palate were retrospectively evaluated; 33 patients were treated with our new device (ß-TW plate group) and 14 were treated with a Hotz plate (HP group). We evaluated the alveolar gap reduction and the size of the maxilla between the two groups, obtaining intraoral maxillary impressions at birth, at 3 months and 1 year. RESULTS: The width of the alveolar gap in the ß-TW plate group was significantly reduced compared with that in the HP group 1 month after the treatment (p < 0.001). The alveolar gap reduction continued until the age of 1 year (p = 0.02). By contrast, no significant difference in the maxillary size was observed between the two groups at any examination period. CONCLUSION: Our treatment protocol using the ß-TW plate was not only easy and simple to apply but it was also cost-effective, with highly predictable outcomes. Moreover, it provided the ideal alveolar cleft reduction without detrimental collapse of the alveolar segments. Therefore, we consider our ß-TW plate device to be useful for application in pre-surgical orthodontic treatments.

An anatomical study in the oriental nose of the location of the vibrissae-bearing area in relation to the lower lateral cartilage.

BACKGROUND: There is no well-known indicator that can assist with a precise intranasal incision during open rhinoplasty on the caudal border of the lower lateral cartilage. However, the vibrissae-bearing area is clinically known as a good landmark for cartilage. The aim of this study was to investigate the features of the vibrissae-bearing area in relation to the lower lateral cartilage. METHODS: Twenty-four heminoses of fixed Japanese cadavers were dissected to clarify the anatomical location of the vibrissae-bearing area in relation to the lower lateral cartilage. RESULTS: The medial part of the vibrissae-bearing area was precisely located on the medial crus of the lower lateral cartilage. Via a transitional state at the dome, the lateral part was located cephalic to the lateral crus in a manner in which the vibrissae-bearing area was adjacent to the lateral crus (adjacent type) in 22 cases, whilst the vibrissae-bearing area overlapped the lateral crus to some extent (overlap type) in two cases. CONCLUSIONS: The anatomical location of the vibrissae-bearing area in relation to the lower lateral cartilage is almost uniform, suggesting its utility as an open rhinoplasty incision landmark.

"Bibloc advancement" with a combination of internal and external distracters.

Frontofacial monobloc advancement is one of the most rewarding techniques for correcting aesthetic and functional problems of patients with severe craniofacial synostosis, which can advance the upper and middle third of the face simultaneously. Application of gradual distraction technique has been reported to reduce notorious risks after a frontofacial monobloc advancement. The so-called "bibloc advancement" is a derivative of the frontofacial monobloc advancement. "Facial bloc" is horizontally divided into 2 different components: fronto-orbital component and maxillozygomatic component. From a different angle, it can be described as a combination of fronto-orbital advancement and Le Fort III advancement. Two pairs of distracters (1 internal for the supraorbital area and 1 external for the maxillozygomatic area with a pair of cross-facial pinning) were applied after the so-called bibloc osteotomy. Advancement of the upper and middle third of the face was done individually. This technique can be a good option for treating infants with severe syndromic craniofacial synostosis.

One-stage vermilion switch flap procedure for the correction of thin lips in patients with bilateral cleft lips.

In this article, we describe the operative method for correction of thin lips in patients with bilateral cleft lips. Two transverse triangular flaps were designed in the lower vermilion and elevated along with the labial arteries and the surrounding tissues. These flaps were inserted into the recipient site on the upper vermilion through the subcutaneous tunnels at the oral commissure through a one-stage process; the donor site was closed directly. Two patients with bilateral cleft lip underwent this operation for thin lips, and satisfying results with less morbidity were obtained. The upper lip retained its volume postoperatively, and the donor-site scar was inconspicuous. Our method enabled both the augmentation of the upper vermilion and the reduction of the lower vermilion in a one-stage procedure.

[Multidisciplinary treatment for severe syndromic craniosynostosis].

We describe the treatment of patients having syndromic craniosynostosis with severe craniofacial abnormality. One patient had Cruzon's syndrome, one had Beare-Stevenson cutis gyrata syndrome, and four had Pfeiffers syndrome. Anterior cranial deformity in all patients was treated using fronto-orbital advancement (FOA) by gradual distraction. Initially, the frontal and supraorbital bones were removed and remodeled. Next, the frontal bones were fixed loosely to the supraorbital bones with absorbable threads. Then, the supraorbital and temporal bones were connected using distraction devices on both sides. The temporal bones were thereafter reinforced with titanium plates. Distraction was started one week postoperation, and the mean amount of elongation was 28.9 mm. Distraction devices were removed one to five months after the operation. One case required FOA by the traditional method ten months after the initial operation. Local infection was observed in three cases, but there were no majors complications. Posterior cranial remodelings were performed in five cases, with one requiring a second operation. We chose the appropriate procedure according to the degree of cranial deformity and operative findings. We performed decompression of the foramen magnum in five cases and laminectomy of the atlas in two cases. Ventriculoperitoneal shunt for hydrocephalus and tracheotomy for airway obstruction were performed in all cases. Cranial remodeling for treating severe craniofacial abnormality requires careful inspection of abnormalities, proper timing, close attention to the procedure and adequate perioperative care. Multidisciplinary therapy is essential for treating severe syndromic craniosynostosis due to systematic osseocartilaginous aplasia.

Lateral orbital expansion and gradual fronto-orbital advancement: an option to treat severe syndromic craniosynostosis.

In some patients with severe syndromic craniosynostosis, bony orbits are so small and shallow that the eyeballs dislocate. Dry cornitis and conjunctivitis can be seen often. When conventional fronto-orbital advancement is attempted in these cases, side walls of the orbit cannot go forward, because the width of bony orbit is smaller than the eyeball. To expand bony orbits and cranial volume, supralateral rim of the orbit was expanded laterally at the time of operation and gradually advanced foward postoperatively. With a coronal skin incision approach, frontal bone was taken off. Supralateral orbital rim bone was detached and cut at the centers of the orbits. Lateral expansion, 5 to 10 mm, was made and fixed with polylactate plates. A pair of distraction devices was fixed between the orbital rim and the temporal bone. Frontal bone was let floating on the dura mater and tied loosely with the orbital rim. Advancement of 1 to 1.5 mm/d was carried out, and the devices were taken off after 1-month consolidation period. Five patients with Pfeiffer syndrome, 1 with Crouzon, and 1 with Beare-Stevenson cutis gyrata syndrome were treated with this method. Procedure, outcomes, and complications are discussed.

Stabilization of a mobile premaxilla by cementing a bite splint with 1-stage bilateral alveolar bone grafting.

This study describes a method for stabilizing maxillary segments by performing bilateral alveolar bone grafting with an acrylic splint with a flat undersurface to reduce the occlusal pressure to the premaxilla. After premaxilla repositioning and bilateral alveolar bone grafting, the splint was attached to the maxillary teeth with glass ionomer cement. In 6 bilateral cleft cases, premaxilla repositioning and bilateral alveolar bone grafting were performed in 1-stage manner, and successful results were obtained. This easy and cost-effective procedure enabled rigid fixation of the insecure premaxilla with no complications during the treatment period.

Early maxillary alignment of unilateral cleft lip and palate using beta-titanium wire.

The aim of this article is to describe early maxillary alignment in patients with unilateral cleft lip and palate using a new orthopedic appliance incorporating beta-titanium wire. The new orthopedic appliance was made with acrylic resin and a spring-shaped beta-titanium wire, and the appliance was fixed onto the hard palate with microscrews under general anesthesia for approximately 1 month. At the end of treatment, both the major and minor segments rotated and moved into a suitable maxillary arch form by the special property of shape memory alloy, resulting in alignment of the cleft palate gap and satisfactory reconstruction of the maxillary arch.

Simultaneous concentration of platelets and marrow cells: a simple and useful technique to obtain source cells and growth factors for regenerative medicine.

Platelet-rich plasma (PRP) has attracted attention as a safe and cost-effective source of growth factors that stimulate cells to regenerate tissue. Bone marrow aspirate was processed with the same protocol to obtain PRP from peripheral blood. This concentrate contained condensed nucleated bone marrow cells, which are useful for regenerative medicine, as well as condensed platelets. In PRP derived from bone marrow aspirate, the density of platelets and levels of growth factors (platelet-derived growth factor and transforming growth factor-beta) were the same as in PRP derived from peripheral blood. Condensation of nucleated cells, especially small-sized cells, was confirmed. With a simple and cost-effective technique, source cells and growth factors can be obtained at the same time. This simultaneous concentration of platelets and bone marrow cells has great potential as a source of materials for regenerative medicine.

Soft palate mucosal adhesion as a preparation for Furlow's double-opposing Z-palatoplasty.

BACKGROUND: Furlow's double-opposing Z-palatoplasty enables both soft palate lengthening and palatal muscle reorientation, and outstanding speech results have been reported. However, Z-plasty often results in horizontal stress and requires relaxing incisions in some cases. This article presents the authors' novel two-stage Furlow double-opposing Z-palatoplasty used in 13 cleft lip-cleft palate patients. METHODS: During cheiloplasty, partial soft palate adhesion was performed. Only mucosal tissue was approximated to prevent damage to the palatal muscle. The residual palatal cleft decreased gradually, and palatoplasty was scheduled at the age of 1 year. Adhesive soft palate tissue was incised and Furlow's double-opposing Z-palatoplasty was performed in the usual manner. RESULTS: Eleven patients underwent Furlow's palatoplasty without relaxing incisions, and in two patients relaxing incisions were required along the inside of the alveolus. Relaxing incisions around the maxillary tuberosities were not required in any of the 13 cases. Analysis of the palatal cast models was carried out on the adhesion group (n = 13) and the control group (n = 9). In both groups, the mean distance between the tuberosities increased, and cleft widths at the alveolus and the middle of the hard palate were decreased. Furthermore, cleft width at the junction of the hard and soft palates was decreased significantly in the adhesion group compared with the control group. CONCLUSIONS: As a preparation for Furlow's palatoplasty, soft palate adhesion induced a decrease of cleft width without interfering with the transverse growth of the posterior maxilla and enabled Z-plasty of an adequate size without relaxing incisions.

Gradual distraction fronto-orbital advancement with 'floating forehead' for patients with syndromic craniosynostosis.

Eleven patients with syndromic craniosynostosis were treated with gradual distraction fronto-orbital advancement using "floating forehead." The frontal and supraorbital area was cut and remolded. Bony orbits were widened in three patients. Frontal bone was let floating on the dura and fixed loosely with absorbable threads to remolded supraorbital bone. A pair of distracters with hinge plates (A.V.D. system, Bear Medic Corp, Tokyo, Japan) was fixed between the temporal area and supraorbital bone. Distraction was begun 5 to 7 days after the surgery, and 1.8 to 3.2 cm advancement was obtained. Distracters were taken off after 3 to 7 weeks of consolidation periods. Although no major complication was encountered, some minor complications related to the devices were experienced.

Efficacy of platelet-rich plasma in alveolar bone grafting.

PURPOSE: In this study, we performed alveolar bone grafting with autologous iliac cancellous bone incorporation with platelet-rich plasma (PRP) and evaluated its efficacy in osteoregeneration. MATERIALS AND METHODS: Seven alveolar cleft patients with adult dentition (average age, 16.1 years) underwent iliac bone grafting with PRP. Quantitative evaluation of regenerated bone was made with 3-dimensional computed tomography scans and compared with controls. RESULTS: The average of the volume ratio of regenerated bone to alveolar cleft in cases with PRP was higher than in controls (P <.05). There were no complications from the blood draw or PRP. CONCLUSION: PRP was a safe and cost-effective source for growth factors and was easy to extract. It could enhance the osteogenesis of alveolar bone grafting in cleft lip and palate patients and may useful for subsequent orthodontic therapy.