An Evaluation of Muscle Repair Techniques: Implications in Musculoskeletal Healing and Corollaries in Oral-Facial Clefting.

We performed an animal study to identify the techniques associated with the best muscle healing outcomes in cleft lip/palate surgery. The right triceps of thirty adult male Sprague-Dawley rats were cut and repaired by three different suture techniques: simple (n = 10), overlapping (n = 10), and splitting sutures (n = 10). Muscle tissues were isolated from 5 rats per group 1 and 8 weeks postoperation. The inflammatory response and muscle fiber healing were evaluated by hematoxylin and eosin (H&E) staining, Western blotting, immunohistochemistry for TNF-α and IL-1ß, and immunofluorescence for laminin and MyoD. Grip strength (N/100 g) and spatial gait symmetry were evaluated before surgery and 1, 2, 4 and 8 weeks postoperation. Eight weeks postoperation, grip force per weight was significantly higher in the simple suture (median, 3.49; IQR, 3.28-3.66) and overlapping groups (median, 3.3; IQR, 3.17-3.47) than the splitting group (median, 2.91; IQR, 2.76-3.05). There was no significant difference in range of motion between groups. The simple group exhibited significant remission of inflammation by H&E staining and lower expression of TNF-α and IL-1ß than the other groups by Western blotting and immunohistochemistry. Immunofluorescence revealed stronger expression of MyoD and weaker expression of laminin in the splitting group than in the other groups at week 8, indicating prolonged inflammation and healing followed by poor muscle fiber remodeling. Simple and overlapping sutures demonstrated similar functional healing, although greater inflammation and failure to maintain a thicker muscle belly were observed in the overlapping suture group compared with the simple suture group. Therefore, reconstruction of the philtral column with overlapping sutures alone may result in limited long-term fullness, and additional procedures may be needed.

Anthropometric Changes in the Upper Lip Between the Standing Position in the Clinic and Supine Position on the Operative Table in Secondary Cleft Lip Nasal Deformities.

BACKGROUND: We evaluated changes in lip height and length by position in cleft lip nasal deformity patients who underwent treatment with Millard technique. METHODS: A retrospective review of the medical records and an anthropometric analysis of photographs were performed in 209 cases of cleft lip nasal deformity. In the photogrammetric analysis, the lip height and length were calculated with respect to the intercanthal width. To evaluate the change in the difference between the cleft- and noncleft-side measurements between the standing and supine positions, the difference between the cleft- and noncleft-side measurements (Δnc-c) was calculated. We identified which patients and surgical factors affected the difference between the cleft- and noncleft-side measurements between the standing and supine positions (Δst-sp). RESULTS: From a total of 120 cases, cases with missing medical charts and clinical photographs were excluded, and 89 cases were included in this study. The discrepancy between the cleft-side lip height and noncleft-side lip height was significantly different between the standing and supine positions (P = 0.007). According to the univariate regression analysis, Δst-sp (sbal-cphi) had a significant positive correlation with body mass index (P = 0.044). In the final multiple linear regression analysis, body mass index had a significant positive correlation with Δst-sp (sbal-cphi, P = 0.044). CONCLUSION: In our results, the difference in vertical lip height is easy to underestimate in the supine position during surgery. Therefore, surgeons should verify lip height with the patient in the sitting or standing position during the operation to avoid undercorrecting lips with short heights.

Correlation between the inframammary fold and sixth rib: Application to breast reconstruction.

The inframammary fold (IMF) is an essential aesthetic element that influences the outcome of breast surgery. In this study, we introduced a new method for finding the IMF based on retrospective analysis of chest computed tomography (CT) scans of women's breasts. Sagittal sections of 80 female chest CTs were analyzed. Sagittal sections of 80 female chest CTs were analyzed by a single plastic surgeon. The distances between the fifth, sixth, and seventh ribs and the IMF were measured using Photoshop, and the location of the IMF was statistically analyzed according to age and height using paired t tests and Pearson's correlation test. As determined by our new method, the average distance between the IMF and the sixth rib was 5.33 mm, which was less than that between the IMF and any other rib. The IMF was closest to the sixth rib in all age groups (P < 0.001), and all IMFs were located between 39.5 mm superior and 24 mm inferior to the sixth rib. The sixth rib, one of the most visible reference structures in women's surface anatomy, is the "sentinel rib" for the IMF and the most predictive indicator of the location of the IMF. Knowing the original anatomic location of the IMF will enable better outcomes during breast surgery. Clin. Anat. 33:165-172, 2020. © 2019 Wiley Periodicals, Inc.

Septal Integration Graft, a Modified Direct Type of Septal Extension Graft for the Treatment of Unilateral Cleft Lip Nose Deformity: An Anthropometric Analysis.

BACKGROUND: The purpose of this study was to evaluate the cosmetic outcomes and stability of septal integration grafts (SIGs), a modified direct type of septal extension graft, in patients with cleft lip nose deformity. METHODS: Seventy-three patients underwent SIGs between January 2002 and July 2017. The exclusion criteria were patients who lacked clinical photographs preoperatively or postoperatively. We reviewed data regarding the patient demographics, combined procedures, and complications as well as preoperative and postoperative clinical photography. Eleven measurement items were evaluated from basal and right lateral views with photogrammetry and 3-dimensional anthropometry using standardized clinical photographic techniques. RESULTS: Sixty-one patients were analyzed in this study. Twelve patients were excluded owing to a lack of postoperative photographs. The mean ± SD follow-up period was 26.70 ± 29.43 months (range, 6-134 months). Compared with the preoperative value, the nasal tip projection index, height of nose index, and nostril axis inclination significantly improved. The columellar length (Rt-Lt) index significantly decreased, which implied improved columellar deviation. Complications included hematoma (n = 1), wound dehiscence (n = 1), tearing of septal mucosa (n = 3), and silicone implant deviation (n = 2). CONCLUSIONS: We used SIGs to correct nasal deformity in patients with cleft lip nose deformity. The SIG can minimize the amount of the graft used for septal extension and can effectively correct nasal tip and columella deformities.

Fragmentation of kidney epithelial cell primary cilia occurs by cisplatin and these cilia fragments are excreted into the urine.

The primary cilium, which protrudes from the cell surface, is associated with the pathogenesis of various diseases, including acute kidney injury (AKI). Primary cilium length dynamically changes during the progression of diseases. However, its relevance in disease and the underlying mechanism are largely unknown. In this study, we investigated the role of primary cilia in AKI induced by cisplatin, an effective anticancer drug, and the underlying mechanisms. In addition, we evaluated the usefulness of length alteration and deciliation of primary cilia into the urine for the diagnosis of AKI. Cisplatin induced shortening, elongation, and normalization of the primary cilia in kidney epithelial cells over time. During shortening, primary cilia fragments and ciliary proteins were excreted into the urine. During deciliation, cell proliferation and the expression of cyclin-dependent kinase inhibitor and proliferating cell nuclear antigen were not significantly changed. Shortening and deciliation of primary cilia were observed before significant increases in plasma creatinine and blood urea nitrogen concentration occurred. Pretreatment with Mito-Tempo, a mitochondria-targeted antioxidant, prevented cisplatin-induced primary cilium shortening and inhibited the increases in superoxide formation, lipid peroxidation, blood urea nitrogen, and tissue damage. In contrast, isocitrate dehydrogenase 2 (Idh2) gene deletion, which results in defect of the NADPH-associated mitochondrial antioxidant system, exacerbated cisplatin-induced changes in mice. Taken together, our findings demonstrate that cisplatin induces deciliation into the urine and antioxidant treatment prevents this deciliation, renal dysfunction, and tissue damage after cisplatin injection. These results suggest that cisplatin-induced AKI is associated with primary cilia and urine primary cilia proteins might be a non-invasive biomarker of kidney injury.

Personal technique for definite repair of complete unilateral cleft lip: modified Millard technique.

BACKGROUND: Millard's rotation-advancement repair, which is used by many surgeons, can make a natural philtral column, but most surgeons use a modification of the rotation-advancement flap. The purpose of this article is to introduce a modification utilized by the authors and to provide detailed surgical procedure. METHODS: We retrospectively reviewed 82 patients' medical records and presented surgical technique and outcomes. The main features of the authors' strategy are emphasizing horizontal length of the lip, orbicularis oris muscle duplication for improving the definition of the philtral column, overcorrection of domal portion than the non-cleft side in order to compensate for the recurrence during growth. Two judges rated two times the appearance of the patients' nose and lip using Asher-McDade aesthetic index. Intra- and interobserver reliabilities were determined using Cohen's kappa statistics. RESULTS: All patients recovered eventually after surgery; however, two patients have a minor complications (wound infection in one patient, wound disruption due to trauma in the other patient). The improvement of the aesthetic results can be achieved with this modified Millard technique. Total mean scores of the Asher-McDade index was 2.08, fair to good appearance. The intraobserver reliabilities were substantial to almost perfect agreement and the interobserver reliabilities were moderate to almost perfect agreement. CONCLUSION: We modified Millard method for repair of complete unilateral cleft lip. The surgical outcomes were favorable in long-term follow-up. We hope our technique will serve as a guide for those new to the procedure.

Evaluation of the Effectiveness of an Alar Transfixion Suture for the Correction of a Vestibular Web and Alar-Facial Groove: A Photogrammetric Analysis.

BACKGROUND: Alar transfixion sutures are commonly used for vestibular web correction. The purpose of this study was to evaluate the long-term results of the use of alar transfixion sutures in patients with a unilateral cleft lip nasal deformity using photogrammetric analysis. METHODS: The study included 42 patients who were divided into child and adult groups. A total of 4 measurement items were evaluated from a basal view by photogrammetry using standardized clinical photographic techniques preoperatively, immediately postoperatively, 3 months postoperatively, and 6 months postoperatively. RESULTS: When the preoperative and last postoperative values were compared, no significant changes in any measurement items were noted in the adult group. In the child group, the proportional index (the ratio of the cleft side to the noncleft side) of the alar slope line inclination was significantly increased, but other measurement items showed no significant change. When the measurement items were compared between time points, no significant changes in any measurement items were noted in the adult group. In the child group, the proportional indexes of the alar length, the width between the subnasale and the alare, and the webbing degree were significantly decreased immediately postoperatively compared with the preoperative values. However, these significant changes were diminished at 3 months postoperatively. The proportional index of the alar slope line inclination was significantly increased at 3 months postoperatively compared with the preoperative value, but the significant change was diminished at 6 months postoperatively. CONCLUSIONS: The alar transfixion suture procedure is not effective for correcting a vestibular web and alar-facial groove.

Prdx1 (peroxiredoxin 1) deficiency reduces cholesterol efflux via impaired macrophage lipophagic flux.

Oxidative stress activates macroautophagy/autophagy and contributes to atherogenesis via lipophagic flux, a form of lipid removal by autophagy. However, it is not known exactly how endogenous antioxidant enzymes are involved in lipophagic flux. Here, we demonstrate that the antioxidant PRDX1 (peroxiredoxin 1) has a crucial role in the maintenance of lipophagic flux in macrophages. PRDX1 is more highly expressed than other antioxidant enzymes in monocytes and macrophages. We determined that Prdx1 deficiency induced excessive oxidative stress and impaired maintenance of autophagic flux in macrophages. Prdx1-deficient macrophages had higher intracellular cholesterol mass and lower cholesterol efflux compared with wild type. This perturbation in cholesterol homeostasis was due to impaired lipophagic cholesterol hydrolysis caused by excessive oxidative stress, resulting in the inhibition of free cholesterol formation and the reduction of NR1H3 (nuclear receptor subfamily 1, group H, member 3) activity. Notably, impairment of both lipophagic flux and cholesterol efflux was restored by the 2-Cys PRDX-mimics ebselen and gliotoxin. Consistent with this observation, apoe -/- mice transplanted with bone marrow from prdx1-/-apoe-/- mice had increased plaque formation compared with apoe-/- BM-transplanted recipients. This study reveals that PRDX1 is crucial to regulating lipophagic flux and maintaining macrophage cholesterol homeostasis against oxidative stress. We suggest that PRDX1-dependent control of oxidative stress may provide a strategy for treating atherosclerosis and autophagy-related human diseases.

Selective tubular activation of hypoxia-inducible factor-2α has dual effects on renal fibrosis.

Hypoxia-inducible factor (HIF) is a key transcriptional factor in the response to hypoxia. Although the effect of HIF activation in chronic kidney disease (CKD) has been widely evaluated, the results have been inconsistent until now. This study aimed to investigate the effects of HIF-2α activation on renal fibrosis according to the activation timing in inducible tubule-specific transgenic mice with non-diabetic CKD. HIF-2α activation in renal tubular cells upregulated mRNA and protein expressions of fibronectin and type 1 collagen associated with the activation of p38 mitogen-activated protein kinase. In CKD mice, activation of HIF-2α at the beginning of CKD significantly aggravated renal fibrosis, whereas it did not lead to renal dysfunction. However, activation at a late-stage of CKD abrogated both renal dysfunction and fibrosis, which was associated with restoration of renal vasculature and amelioration of hypoxia through increased renal tubular expression of VEGF and its isoforms. As with tubular cells with HIF-2α activation, those under hypoxia also upregulated VEGF, fibronectin, and type 1 collagen expressions associated with HIF-1α activation. In conclusion, late-stage renal tubular HIF-2α activation has protective effects on renal fibrosis and the resultant renal dysfunction, thus it could represent a therapeutic target in late stage of CKD.

Modified Direct-Type Septal Extension Grafts: Their Stability and Usefulness in Asian Rhinoplasty.

BACKGROUND: In Asian rhinoplasty, many autogenous cartilage grafts are required for correction of the nasal tip and columella, but the amount has limitations. A modified direct-type septal extension graft, in continuity with the entire caudal border of the septal cartilage with an edge-to-edge coaptation, can effectively and concomitantly correct the nasal tip and columella deformities using a limited amount of septal cartilage graft. The purpose of this study was to evaluate long-term cosmetic outcomes and stability from the modified direct extension grafts. METHODS: Fifty-seven patients with a follow-up of more than 1 year were enrolled in the study. A total of 11 measurement items were evaluated from basal and right lateral views by photogrammetry using standardized clinical photographic techniques. RESULTS: The overall mean follow-up period was 20.4 months. When comparing the preoperative and postoperative values, the nasal tip projection, nasal bridge length, nasal tip angle, height of nose, and the columellar labial angle increased significantly; additionally, the soft nose width index, width between ac-ac index, nostril axis inclination, columellar length (Rt-Lt), and the alar length (Rt-Lt) decreased significantly. No resorption, buckling, or displacement of the graft was observed during the follow-up period. CONCLUSIONS: The modified direct extension graft demonstrated a marked aesthetic improvement in the nasal tip and columella, and it provided long-term stability. Therefore, the modified direct extension graft is useful for correction of the nasal tip and columella in Asian rhinoplasty.

Long-term results of secondary alveolar bone grafting using a technique to harvest pure calvarial cancellous bone: Evaluation based on plain radiography and computed tomography.

BACKGROUND: We introduce a method to extract the maximum amount of cancellous bone from the calvarium and analyze the outcomes using plain radiography and computed tomography. METHODS: We retrospectively analyzed the medical records of patients who underwent secondary alveolar bone grafts using a maximal harvesting technique for calvarial cancellous bone. Bone survival was determined based on Enemark's grading system using plain radiographs. Alveolar thickness and height were analyzed by computed tomography and compared with radiographic measurements. RESULTS: Our study enrolled 27 patients, 22 unilateral and 5 bilateral, for a total of 32 alveolar clefts. The mean age at surgery was 9.04 ± 0.41 years (range: 6-12 years). Donor sites were successfully recovered without complications, such as hematomas, seromas, infections, or tears in the dura mater. The mean follow-up duration for obtaining plain radiographs was 6.90 ± 0.61 years (range: 2.0-18.50 years). The mean follow-up duration for computed tomography was 6.51 ± 0.99 years (range: 2.0-14.83 years). A canine tooth erupted in 30 cases. Success (Enemark's grades I or II) was achieved in 26 cases. Alveolar height was strongly correlated with the Enemark scale results (γ = 0.878, p < 0.001), but alveolar thickness was not correlated (γ = 0.4575, p = 0.08). CONCLUSION: Our cancellous bone harvesting technique resulted in successful graft survival without serious complications, including dural or brain injuries. Although the ilium is widely accepted as the optimal donor site for secondary alveolar bone grafting, the calvarium could be an alternative donor site that results in equivalent outcomes.

Long-term outcomes of simplified gingivoperiosteoplasty performed at the time of hard palatal closure: A review of 55 alveolar clefts.

Few studies have examined the long-term outcomes of gingivoperiosteoplasty at the age of permanent canine eruption. The success rates of gingivoperiosteoplasty vary significantly depending on the surgeon because of the difficulty of the surgical technique. Therefore, the authors utilized a simplified gingivoperiosteoplasty technique by inserting one or two large, triangular-shaped mucoperiosteal flaps on the opposite side in a tongue-in-groove or interdigitation fashion. The purpose of this study was to evaluate the long-term outcomes of this simplified gingivoperiosteoplasty technique at the age of permanent canine eruption. Forty nonsyndromic cleft lip and palate patients who had undergone simplified gingivoperiosteoplasty were retrospectively reviewed. Canine eruption and the bone formation rate of the interalveolar septum using the Bergland method and a modified Long et al. method were evaluated. A total of 55 alveolar cleft sites were treated. Simplified gingivoperiosteoplasty was performed at a mean age of 13.4 months, and the mean age at the time of follow-up was 14.3 years. The overall success rate of simplified gingivoperiosteoplasty was 50.90%. With regard to cleft types, clefts of the primary palate and unilateral alveolar clefts showed a significantly higher success rate than clefts of the primary and secondary palates and bilateral alveolar clefts, respectively. Our gingivoperiosteoplasty technique is a simple surgical procedure and is performed at 12 months of age, at which time the cleft alveolus has grown sufficiently, to ensure the success of the delicate surgical technique. Therefore, we could obtain favorable outcomes.

Asian Rhinoplasty: Correction of the Short Nose with a Septal Integration Graft.

Correction of the short nose is the most challenging part in a rhinoplasty. It is even more difficult in Asians compared with Caucasians due to the thicker skin soft tissue envelope and weaker cartilaginous frameworks. Additionally, most of the Asian patients need augmentation of the nasal dorsum, and projection of the nasal tip at the same time of short nose surgery, which makes short nose correction more complex. Septal extension grafts can effectively and concomitantly correct a short nose. However, the amount of the septal cartilage is not sufficient as a source for the graft often; therefore, the authors devised a septal integration graft in which a septal cartilage graft was fixed to the caudal septum (columellar L-strut) in an end-to-end position to save an amount of the septal cartilage for correction of the short nose. A septal integration graft application resulted in lengthening of the short nose. In addition, the retracted columella and the acute columellar labial angle were improved aesthetically. Therefore, the authors suggest a septal integration graft, a modification of the septal extension graft, for correction of the short nose in an Asian rhinoplasty.

Differentiated tonsil-derived mesenchymal stem cells embedded in Matrigel restore parathyroid cell functions in rats with parathyroidectomy.

Parathyroid cells release parathyroid hormone (PTH), which controls calcium homeostasis. Loss of parathyroid cells results in hypoparathyroidism and consequent low-turnover bone disease. Here, we investigated whether our recently-established human tonsil-derived mesenchymal stem cells (TMSC) restore in vivo parathyroid cell function in rats with parathyroidectomy (PTX). Compared with undifferentiated control TMSC, TMSC differentiated with activin A and soluble sonic hedgehog induced a significant release of PTH as early as day 7, with increased PTH release occurring in response to lower calcium levels and vice versa. Released PTH increased osteocalcin expression and alizarin red S staining in preosteoblastic cells, indicating its functional activity. PTX rats fed calcium-free diet only survived for ∼10 days. Subcutaneous injection with TMSC alone did not increase their survival rates, regardless of differentiation. However, survival rates increased for up to 28 days in response to TMSC embedded in Matrigel (TMSC-MA), showing 40% and 80% in control and differentiated TMSC-MA, respectively. When compared with continuous increases by control TMSC-MA, stable levels of secreted PTH and serum ionized calcium were found in PTX rats with differentiated TMSC-MA. This is the first report that differentiated TMSC resemble parathyroid cells and, if embedded in Matrigel, restore in vivo parathyroid function.

Curvilinear transformation of z-shaped upper lip scar by diamond-shaped excision in secondary cleft lip deformities: a photogrammetric evaluation.

Purpose : The visible Z-shaped upper lip scar that occurs after the Tennison and Randall triangular flap technique remains a cleft stigma. Herein, we present our curvilinear transformation technique for the Z-shaped upper lip scar by diamond-shaped excision and evaluate the results using photogrammetric analyses. Patients and Methods : From 1997 to 2006, 23 patients with secondary cleft lip deformity with the visible Z-shaped upper lip scar underwent correction with the technique. The scar was excised in the diamond shape above the muscle. After curvilinear closure, the elongated length of the upper lip was excised just below the nostril sill, as the measured Cupid's bow height discrepancy. The result was assessed by the authors' standardized photogrammetry technique. Results : There was a statistically significant decrease between the preoperative central limb of the Z-shaped scar and the width of the postoperative curvilinear upper lip scar. The pre- and postoperative Cupid's bow height differences were not statistically significant. Conclusions : The curvilinear transformation of the Z-shaped scar is an efficient procedure that provides (1) a significant decrease in the width of an upper lip scar to make it less conspicuous, (2) incorporation of the scar into the philtral column, (3) a biconcave natural philtral column shape in frontal view, and (4) formation of the natural concave philtral contour in profile view.

Novel reconstructive methods of the conchal central strut using an absorbable plate after total harvesting of the conchal cartilage.

PURPOSE: Conchal cartilage is widely used in the field of plastic surgery, but donor-site morbidity is inevitable when all of the conchal cartilage is harvested. To maintain ear shape, the authors introduce a new method using an absorbable plate to reconstruct the conchal central strut after total harvesting of the conchal cartilage. METHODS: In total, 18 subjects underwent surgery, and 14 subjects who were followed up for more than 12 months were included in this article. Before the total harvesting of the conchae, an absorbable plate was adjusted to the proper curvature and length. After harvesting, the central strut was reconstructed by the precrafted absorbable plate. When the harvested cartilage was too large or one implant was not sufficient to reconstruct the central strut, an additional implant was added to the cymba conchae.Outcomes were evaluated by photogrammetry and questionnaires. Seven items were selected to evaluate the shape of the ear and conchal cavity. The authors compared preoperative and postoperative photographs, looking at proportion indices of 7 items using Photoshop. Ten assessors who did not participate in the operation were also shown preoperative and postoperative photographs and answered questionnaires about the shapes of the ear, conchal cavity, and conchal central strut. RESULTS: Except for the effective conchal cavity height index, no statistically significant differences were observed between preoperative and postoperative ear shapes. Additionally, the questionnaire showed excellent assessments for all items. CONCLUSIONS: Reconstruction of the central strut using an absorbable plate after total harvest of the conchal cartilage was a simple and effective method to prevent ear collapse.

Changes of eyebrow muscle activity with aging: functional analysis revealed by electromyography.

BACKGROUND: This study evaluated the activity of the frontalis muscle, the corrugator supercilii muscle, and the orbicularis oculi muscle according to eyebrow movement and aging. METHODS: Two random cohorts of women aged 20 to 30 years (young group, n = 20; mean age, 24.8 years) and 50 to 70 years (old group, n = 20; mean age, 55.8 years) were recruited prospectively. Surface electromyography was used to evaluate motor unit action potential for each muscle in each of six eyebrow movements. Details on statistical analysis are described in the text. RESULTS: In both age groups, corrugator supercilii muscle activity was the highest for all six movements. Frontalis muscle activity was highest with maximal frowning but not with maximal eye opening. Orbicularis oculi muscle activity was significantly greater in the older age group than in the younger age group for most actions. When the motion proportion of each muscle was compared, the respective orbicularis oculi muscle and corrugator supercilii muscle indices were higher in the older age group than in the young group. CONCLUSIONS: Eyebrows are maintained by the dynamic balance of frontalis, corrugator supercilii, and orbicularis oculi muscles, and various combinations of motor recruitment of these muscles determine the eyebrow position and shape. For youthful eyebrows, attenuation of the depressor muscle may restore the muscle balance in treatments for eyebrow rejuvenation.

The effect of centrifugation condition on mature adipocytes and adipose stem cell viability.

Different researchers have recommended different lipoaspirate centrifugation speeds and times, probably due to the limits in fat cell viability assays. We assessed fat cell viability using a fluorescein diacetate and propidium iodide (FDA-PI) stain and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay after harvesting syringe liposuction and spun with different centrifugation speeds to determine the optimal conditions. Lipoaspirates, harvested from 13 donors, were transferred into a centrifuge tube and spun at 1000, 3000, and 4000 rpm for 3 minutes. Mature adipocytes and adipose stem cells were isolated and tested with a direct counting of FDA-PI-stained cells under fluorescence microscope and XTT assay. We incubated adipocytes and adipose stem cells for 1 day and 3 days, and we compared both of them with fresh samples to evaluate the influence of culturing condition on fat cell viability. Centrifugation speeds from 1000 rpm to 4000 rpm for 3 minutes showed no change in the percentage of adipocytes and adipose stem cell viability not only in the fresh samples but also in the cultured samples (1 day and 3 days). Centrifugation speeds under 4000 rpm do not change the percentage of fat cell viability. To differentiate viable cells from dying or dead mature adipocytes and oil accurately, combinations of viability tests are essential.

Antineoplastic effect of WIN 55,212-2, a cannabinoid agonist, in a murine xenograft model of gastric cancer.

BACKGROUND: We have previously reported the antineoplastic effects of a cannabinoid agonist in gastric cancer cells. Our aim was to evaluate this in a murine xenograft model. METHODS: Animal models were created after injecting AGS gastric cancer cells subcutaneously into the flank of male BALB/c-nude mice. A cannabinoid agonist, WIN 55,212-2 (7 mg/kg body weight) or vehicle was injected around the tumor subcutaneously every 24 h for 14 days. Tumors were explanted for analysis. RESULTS: Tumor volume decreased by 30% in the WIN 55,212-2-treated group compared to the group treated with vehicle (p < 0.05). Apoptotic cells were found more commonly in the WIN 55,212-2 treatment group than in the control on immunohistochemistry. Compared to the control, WIN 55,212-2 treatment significantly increased caspase-3 cleavage and decreased MMP-2, MMP-7 and MMP-9 protein levels significantly (all p < 0.05). VEGF-A protein level was not different between the 2 groups. CONCLUSION: WIN 55,212-2 has antineoplastic effect on the gastric cancers in in vivo model.

Expression of the rhesus glycoproteins, ammonia transporter family members, RHCG and RHBG in male reproductive organs.

The rhesus glycoproteins, Rh B glycoprotein (RHBG) and Rh C glycoprotein (RHCG), are recently identified ammonia transporters. Rhcg expression is necessary for normal male fertility, but its specific cellular expression is unknown, and Rhbg has not been reported to be expressed in the male reproductive tract. This study sought to determine the specific cellular expression of Rhcg, to determine whether Rhbg is expressed in the male reproductive tract, and, if so, to determine which cells express Rhbg using real-time RT-PCR, immunoblot analysis, and immunohistochemistry. Both Rhbg and Rhcg were expressed throughout the male reproductive tract. In the testis, high levels of Rhbg were expressed in Leydig cells, and Rhcg was expressed in spermatids during the later stages of their maturation (steps 13-16) in stages I-VIII of the seminiferous epithelium cycle. In the epididymis, basolateral Rhbg was present in narrow cells in the initial segment, in principal cells in the upper corpus, and in clear cells throughout the epididymis. Apical Rhcg immunolabel was present in principal cells in the caput and upper corpus epididymidis and in clear cells in the middle and lower corpus and cauda epididymidis. In the vas deferens, apical Rhcg immunolabel and basolateral Rhbg immunolabel were present in some principal cells and colocalized with H(+)-ATPase immunolabel. We conclude that both Rhbg and Rhcg are highly expressed in specific cells in the male reproductive tract where they can contribute to multiple components of male fertility.