Adipose Derived Stromal Vascular Fraction and Mesenchymal Stem Cells Improve Angiogenesis in a Rat Hindlimb Ischaemia Model.

OBJECTIVE: This study aimed to investigate the effect of human adipose tissue derived stromal vascular fraction (AD-SVF) and mesenchymal stem cells (AD-MSCs) on blood flow recovery and neovascularisation in a rat hindlimb ischaemia model. METHODS: SVF was isolated using an automated centrifugal system, and AD-MSCs were obtained from adherent cultures of SVF cells. Rats were divided into four groups of six rats each: non-ischaemia (Group 1); saline treated ischaemia (Group 2); SVF treated ischaemia (Group 3); and AD-MSC treated ischaemia (Group 4). Unilateral hindlimb ischaemia was induced in Sprague-Dawley rats via femoral artery ligation. Saline, SVF, or AD-MSCs were injected intramuscularly into the adductor muscle intra-operatively. Cell viability was calculated as the percentage of live cells relative to total cell number. Blood flow improvement, muscle fibre injury, and angiogenic properties were validated using thermal imaging and histological assessment. RESULTS: The viabilities of SVF and AD-MSCs were 83.3% and 96.7%, respectively. Group 1 exhibited no significant temperature difference between hindlimbs, indicating a lack of blood flow changes. The temperature gradient gradually decreased in SVF and AD-MSC treated rats compared with saline treated rats. In addition, only normal muscle fibres with peripherally located nuclei were observed in Group 1. Groups 3 and 4 exhibited significantly fewer centrally located nuclei, indicating less muscle damage compared with Group 2. Regarding angiogenic properties, CD31 staining of endothelial cells showed similar patterns among all groups, whereas expression of vascular endothelial growth factor, as a crucial angiogenesis factor, was enhanced in the SVF and AD-MSC treated groups. CONCLUSION: SVF and AD-MSCs improved blood flow and neovascularisation in a rat hindlimb ischaemia model, suggesting their potential ability to promote angiogenesis. Further extensive research is warranted to explore their potential applications in the treatment of severe lower extremity arterial disease.

Performance Assessment of the Post-Tensioned Anchorage Zone Using High-Strength Concrete Considering Confinement Effect.

Post-tensioned anchorage zones need enough strength to resist large forces from jacking forces from prestress and need spiral reinforcement to give confinement effect. High-strength concrete (HSC) has high-strength and brings the advantage of reducing material using and simplifying reinforcing. We tested strain stabilization, load-displacement, and strain of lateral reinforcements. Specimens that used one and two lateral reinforcements without spiral reinforcement did not satisfy the strain stabilization. Load capacity also did not satisfy the condition of 1.1 times the nominal tensile strength of PS strands presented in ETAG 013. On the other hand, specimens that used three and four lateral reinforcements without spiral reinforcement satisfied the strain stabilization but did not satisfy 1.1 times the nominal tensile strength of PS strands. However, the secondary confinement effect could be confirmed from strain stabilization. In addition, the affection of HSC characteristics could be confirmed from a reinforcing level comparing other studies. The main confinement effect could be confirmed from the reinforcement strain results; there was a considerable difference between with and without spiral reinforcement at least 393 MPa. Comprehensively, main and secondary confinement effects are essential in post-tensioned anchorage zones. In addition, the performance of the anchorage zone could be increased by using HSC that the combination of high-strength and confinement effect.

Genetic Inheritance of Late-Onset, Down-Sloping Hearing Loss and Its Implications for Auditory Rehabilitation.

OBJECTIVES: Late-onset, down-sloping sensorineural hearing loss has many genetic and nongenetic etiologies, but the proportion of this commonly encountered type of hearing loss attributable to genetic causes is not well known. In this study, the authors performed genetic analysis using next-generation sequencing techniques in patients showing late-onset, down-sloping sensorineural hearing loss with preserved low-frequency hearing, and investigated the clinical implications of the variants identified. DESIGN: From a cohort of patients with hearing loss at a tertiary referral hospital, 18 unrelated probands with down-sloping sensorineural hearing loss of late onset were included in this study. Down-sloping hearing loss was defined as a mean low-frequency threshold at 250 Hz and 500 Hz less than or equal to 40 dB HL and a mean high-frequency threshold at 1, 2, and 4 kHz greater than 40 dB HL. The authors performed whole-exome sequencing and segregation analysis to identify the genetic causes and evaluated the outcomes of auditory rehabilitation in the patients. RESULTS: There were nine simplex and nine multiplex families included, in which the causative variants were found in six of 18 probands, demonstrating a detection rate of 33.3%. Various types of variants, including five novel and three known variants, were detected in the MYH14, MYH9, USH2A, COL11A2, and TMPRSS3 genes. The outcome of cochlear and middle ear implants in patients identified with pathogenic variants was satisfactory. There was no statistically significant difference between pathogenic variant-positive and pathogenic variant-negative groups in terms of onset age, family history of hearing loss, pure-tone threshold, or speech discrimination scores. CONCLUSIONS: The proportion of patients with late-onset, down-sloping hearing loss identified with potentially causative variants was unexpectedly high. Identification of the causative variants will offer insights on hearing loss progression and prognosis regarding various modes of auditory rehabilitation, as well as possible concomitant syndromic features.

Prevalence of Respiratory Viral Infections in Korean Adult Asthmatics With Acute Exacerbations: Comparison With Those With Stable State.

PURPOSE: Viral infections are involved in ~50% of exacerbations among Caucasian adult asthmatics. However, there have been few reports on the causative virus of exacerbations in Korean adult asthmatics. Thus, we compared frequencies and types of viruses between lower respiratory tract illnesses (LRTIs) with exacerbations (exacerbated LRTIs) and those without exacerbations (stable LRTIs) to evaluate contribution of respiratory viruses to exacerbations. METHODS: Viral RNA was extracted from sputum using the Viral Gene-spin™ Kit. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect adenovirus (ADV), metapneumovirus (MPV), parainfluenza virus (PIV) 1/2/3, influenza virus (IFV) A, IFV B, respiratory syncytial virus (RSV) A/B, and rhinovirus (RV) A. RESULTS: Among the 259 patients, 210 underwent a single sputum examination, and the remaining 49 underwent 2 to 4 sputum examinations. Virus was detected in 68 of the 259 exacerbated episodes and in 11 of the 64 stable episodes. Among the exacerbated episodes, RV was the most frequently detected virus, followed by influenza A, parainfluenza, RSV A/B, and ADV. Among the 11 stable episodes, RV was most frequently detected. Detection rates of these viruses did not differ between the 2 groups (P>0.05). Thirty-five patients underwent the virus examination at 2 episodes of exacerbation, while 14 patients underwent at each time of exacerbated and stable episodes. Virus detection rate at the second examination was significantly higher in cases with 2 exacerbation episodes than in those with initial exacerbation and sequential stable episodes (P=0.003). A seasonal pattern was noted in the detection rates of RV (September to December), IFV (January to April), PIV (May to September), and RSV A/B (September to April). CONCLUSIONS: Respiratory viruses were identified in approximately 20% of LRTI irrespective of the presence of asthma exacerbation. RV and IFV A/B were most frequently detected. A group of patients experienced frequent viral infections followed by asthma exacerbations.

Simple realization of efficient barrier performance of a single layer silicon nitride film via plasma chemistry.

Due to the problem of degradation by moisture or oxygen, there is growing interest in efficient gas diffusion barriers for organic optoelectronic devices. Additionally, for the continuous and long-term operation of a device, dedicated flexible thin film encapsulation is required, which is the foremost challenge. Many efforts are being undertaken in the plasma assisted deposition process control for the optimization of film properties. Control of the plasma density along with the energy of the principal plasma species is critical to inducing alteration of the plasma reactivity, chemistry, and film properties. Here, we have used the radio frequency (RF) plasma enhanced chemical vapor deposition (PECVD) technique to deposit amorphous silicon nitride (SiNx) barrier films onto a plastic substrate at different pressures. A large part of our efforts is devoted to a detailed study of the process parameters controlling the plasma treatment. Numerous plasma diagnostic techniques combined with various characterization tools are purposefully used to characterize and investigate the plasma environment and the associated film properties. This contribution also reports a study of the correlations between the plasma chemistry and the chemical, mechanical, barrier, and optical properties of the deposited films. The data reveal that the film possesses a very low stress for the condition where the net energy imparted on the substrate is at a minimum. Simultaneously, a relatively high ion flux and high energy of the ions impinging on the film growth surfaces are crucial for controlling the film stress and the resulting barrier properties.

Single-bundle anterior cruciate ligament reconstruction in active young men using bone-tendon achilles allograft versus free tendon achilles allograft.

PURPOSE: The purpose was to prospectively compare the outcomes of anterior cruciate ligament (ACL) reconstruction using bone-tendon Achilles allograft with those of free tendon Achilles allograft in 2 groups of active young men. METHODS: We enrolled 72 subjects in the study. Of these, 33 subjects who had ACL reconstruction with Achilles allograft with calcaneal bone block on its end (group I) and 34 who had ACL reconstruction with free tendon Achilles allograft (group II) were followed up for a minimum of 2 years. The Lachman test, pivot-shift test, and 1-leg hop test were evaluated at the last follow-up. The International Knee Documentation Committee classification, Lysholm score, and side-to-side difference were assessed preoperatively and at the last follow-up. The Tegner activity scale was evaluated before injury and at the last follow-up. RESULTS: The mean follow-up period was 30.9 months (range, 26 to 38 months). At the last follow-up, there were no statistically significant differences between the 2 groups for the Lachman test, pivot-shift test, 1-leg hop test, International Knee Documentation Committee classification, Lysholm score, and side-to-side difference (P > .05). Three subjects in group I and three in group II showed an activity level 2 steps below the preinjury activity level. The rest of the subjects returned to the preinjury activity level or showed a level 1 step below the preinjury activity level. CONCLUSIONS: There were no significant differences in clinical outcomes of ACL reconstructions using Achilles allograft between the bone-tendon group and the free tendon group in active young men. Free tendon Achilles allograft may be considered an option for ACL reconstruction. LEVEL OF EVIDENCE: Level II, therapeutic study, randomized controlled trial.

Outcomes of operative treatment of unstable ankle fractures: a comparison of metallic and biodegradable implants.

BACKGROUND: Biodegradable implants for internal fixation of ankle fractures may overcome some disadvantages of metallic implants, such as imaging interference and the potential need for additional surgery to remove the implants. The purpose of this study was to evaluate the outcomes after fixation of ankle fractures with biodegradable implants compared with metallic implants. METHODS: In this prospectively randomized study, 109 subjects with an ankle fracture underwent surgery with metallic (Group I) or biodegradable implants (Group II). Radiographic results were assessed by the criteria of the Klossner classification system and time to bone union. Clinical results were assessed with use of the American Orthopaedic Foot & Ankle Society (AOFAS) ankle-hindfoot scale, Short Musculoskeletal Function Assessment (SMFA) dysfunction index, and the SMFA bother index at three, six, and twelve months after surgery. RESULTS: One hundred and two subjects completed the study. At a mean of 19.7 months, there were no differences in reduction quality between the groups. The mean operative time was 30.2 minutes in Group I and 56.4 minutes in Group II (p < 0.001). The mean time to bone union was 15.8 weeks in Group I and 17.6 weeks in Group II (p = 0.002). The mean AOFAS score was 87.5 points in Group I and 84.3 points in Group II at twelve months after surgery (p = 0.004). The mean SMFA dysfunction index was 8.7 points in Group I and 10.5 points in Group II at twelve months after surgery (p = 0.060). The mean SMFA bother index averaged 3.3 points in Group I and 4.6 points in Group II at twelve months after surgery (p = 0.052). No difference existed between the groups with regard to clinical outcomes for the subjects with an isolated lateral malleolar fracture. CONCLUSIONS: The outcomes after fixation of bimalleolar ankle fractures with biodegradable implants were inferior to those after fixation with metallic implants in terms of the score on the AOFAS scale and time to bone union. However, the difference in the final AOFAS score between the groups may not be clinically important. The outcomes associated with the use of biodegradable implants for the fixation of isolated lateral malleolar fractures were comparable with those for metallic implants.

Achilles tendon allograft with its bony attachment to repair rupture and extensive degeneration of the heel cord.

An Achilles tendon allograft with its bony calcaneal attachment was successfully used to repair a spontaneous rupture of a severely degenerative and irreparable Achilles tendon. After 18 months of follow-up, the patient is satisfied and the Achilles tendon power has returned to a normal level.

Hybrid tibial fixation for anterior cruciate ligament reconstruction with Achilles tendon allograft.

PURPOSE: To compare clinical outcomes of tibial fixation between a biodegradable interference screw only and a biodegradable interference screw supplemented by a post-tie using a washer screw in single-bundle anterior cruciate ligament (ACL) reconstruction with 2-strand free tendon Achilles allograft in active young men. METHODS: A prospective study was performed in 80 subjects who underwent single-bundle ACL reconstruction. A 2-strand free tendon Achilles allograft fixed with an EndoButton (Smith & Nephew, Andover, MA) in the femoral tunnel and with a Bio-Interference screw (Arthrex, Naples, FL) in the tibial tunnel was used in group I. Supplementary fixation was performed with a post-tie using a washer screw in group II. The Lachman test, pivot-shift test, International Knee Documentation Committee classification, Lysholm score, range of knee motion, and side-to-side difference were evaluated preoperatively and at the last follow-up. The Tegner activity scale was evaluated before injury and at the last follow-up. The 1-leg hop test was assessed at the last follow-up. RESULTS: Of the patients, 36 in group I and 35 in group II could be followed up for at least 2 years. At the last follow-up, 7 patients in group I and 1 in group II showed a 2+ or 3+ on the Lachman test (P = .027). The mean side-to-side difference was 2.9 ± 2.3 mm in group I and 1.4 ± 1.6 mm in group II at the last follow-up (P = .002). The other clinical outcomes were not significantly different between the 2 groups. CONCLUSIONS: Supplementary tibial fixation with a post-tie in ACL reconstruction using 2-strand free tendon Achilles allograft was more effective than a biodegradable interference screw only in restoration of the anterior stability of the tibia. LEVEL OF EVIDENCE: Level II, randomized controlled trial.

Synovialization on second-look arthroscopy after anterior cruciate ligament reconstruction using Achilles allograft in active young men.

PURPOSE: This study is to assess the relationship between clinical outcomes and the extent of synovialization through second-look arthroscopy for anterior cruciate ligament (ACL) reconstruction using Achilles allograft in active young men. METHODS: Ninety-five subjects were diagnosed with ACL rupture and underwent reconstruction with Achilles allograft. Median age was 22 years old (range, 17-40). Mean period from injury to operation was 15.1 ± 18.2 weeks. Lysholm score, IKDC classification, range of motion (ROM) of knee, and side-to-side difference (SSD) in anterior instability were evaluated preoperatively and at the last follow-up. Tegner activity scale was evaluated before injury and at the final follow-up. The extent of synovialization of the graft was evaluated under second-look arthroscopy at least 1 year after surgery. Formation of the synovial membrane was divided into four groups-group 1 for 25% or less, group 2 for 25-50%, group 3 for 50-75%, and group 4 for more than 75%. Outcomes were compared between each group. RESULTS: Lysholm score and IKDC classification were improved after surgery (P < 0.05). Most subjects had full ROM at the final follow-up except three subjects that showed flexion deficit of 5 degrees or less. Mean SSD in anterior instability was 9.0 ± 2.1 mm preoperatively and 1.6 ± 2.0 mm at the final follow-up (P < 0.001). Median Tegner activity scale was 7 before injury and 7 at the final follow-up (P < 0.001). Twelve subjects were in group 1, 10 in group 2, 14 in group 3, and 59 in group 4. Clinical outcomes depending on the extent of synovialization of the grafts were different between each group (P < 0.05). The average period from injury to reconstruction in each group was significantly different (P < 0.001). Correlation coefficient between the period from injury to reconstruction and the extent of synovialization was -0.411 (P < 0.001). CONCLUSION: The extent of the synovialization is positively correlated with clinical outcomes and is negatively correlated with the period from injury to reconstruction.

Estimation of inhomogeneous zeta potential using velocity measurements of EOF.

We have developed a method of estimating the zeta potential distribution along the microchannel wall using velocity measurements of the EOF. The relevant inverse problem is solved through the minimization of a performance function utilizing the conjugate gradient method. Employing a set of simulated velocity measurements, which is constructed by adding random noise to the computed exact velocity, the present method is found to estimate the distribution of the zeta potential along the channel wall with reasonable accuracy.

Naltrexone influences protein kinase Ce and integrin alpha7 activity in SH-SY5Y neuroblastoma cells.

Alcohol influences the neuroadaptation of brain cells where receptors and enzymes like protein kinase C (PKC) exist. Naltrexone acts on opioid receptors. However, other mechanisms of action remain unknown. We prepared SH-SY5Y neuroblastoma cells, and fed them with 150 mM ethanol for 72 h followed by treatment with naltrexone for 24 h. We performed microarray analysis and reverse transcriptase-polymerase chain reaction. Our results showed that PKCepsilon increased 1.90 times and showed an overall decreasing pattern as time increased. Phosphorylated ERK also increased 2.0 times according to the change of PKCepsilon. Integrin alpha7 increased 2.32 times and showed an increasing pattern as time increased. In conclusion, naltrexone influences PKCepsilon neuronal signaling system and endothelial adhesion molecule integrin alpha7 in addition to the well-known opioid system.

Genetic polymorphisms of alcohol and aldehyde dehydrogenase, dopamine and serotonin transporters in familial and non-familial alcoholism.

One hundred and eleven male patients with alcohol dependence and 123 nonalcoholic healthy men were tested for the genetic polymorphisms of alcohol dehydrogenase 2 (ADH2), aldehyde dehydrogenase 2 (ALDH2), serotonin transporter (5-HTT) and dopamine transporter (DAT1). There were significant differences in genotype frequencies of ADH2 C992G and A13543G SNPs between alcoholic patients with family history of alcohol dependence (familial) and alcoholic patients without family history (non-familial). Genotype and allele frequencies of ALDH2 G1951A SNP in familial or non-familial alcoholic patients differ from normal controls. Neither 5-HTTLPR L/S nor DAT1 G2319A SNP genotypes nor alleles discriminated alcoholic patients from normal controls. These findings suggest that the genetic characteristics of alcohol metabolism in non-familial alcoholics fall between non-alcoholism and familial alcoholics.

Scanning of genetic effects of alcohol metabolism gene (ADH1B and ADH1C) polymorphisms on the risk of alcoholism.

Alcoholism is a multifactorial and polygenic disorder involving complex gene-to-gene and gene-to-environment interactions. Alcohol metabolism is one of the biological determinants that could significantly be influenced by genetic polymorphisms in alcohol-metabolism genes. These genetic polymorphisms are believed to influence drinking behavior and development of alcoholism. Direct DNA sequencing of whole ADH1B and ADH1C genes revealed 36 sequence variants, including six nonsynonymous and 14 novel polymorphisms. Seventeen polymorphisms among them were selected for genotyping in a larger study (n = 352) based on linkage disequilibria (LDs) among SNPs, locations, and frequencies. Hardy-Weinberg equilibrium (HWE) analyses of polymorphisms revealed severe deviations only in alcoholics, which strongly suggest that a selection bias (or pressure) may be involved. The analyses of genotype distribution in alcoholics (n = 106) and normal controls (n = 246) showed dramatic associations with the risk of alcoholism. Fourteen polymorphisms in ADH1C and ADH1B showed a series of different strengths of association and magnitudes of risk. Based on referent and subgroup analysis, it was strongly suggested that the genetic effects come from the ADH1B*47Arg/*47Arg genotype, and that the positive signals from other sites are just tracking the genetic effect of ADH1B His47Arg. In this article we present summaries of previous studies and of the present study, to give an overview of the worldwide effects of ADH1B His47Arg on the risk of alcoholism. The information derived from this study could be valuable for understanding the genetic factors involved in the risk of alcoholism and facilitate further investigation in other ethnic groups.

Fluoxetine-induced up-regulation of 14-3-3zeta and tryptophan hydroxylase levels in RBL-2H3 cells.

The primary mechanisms of antidepressants are based on the monoamine depletion hypothesis. However, we do not yet know the full cascade of mechanisms responsible for the therapeutic effect of antidepressants. To identify the genes involved in the therapeutic mechanism of the selective serotonin reuptake inhibitor, fluoxetine, we used a cDNA microarray analysis with RBL-2H3 cells. We observed the transcriptional changes of several tens of genes containing the 14-3-3zeta gene in the fluoxetine-treated RBL-2H3 cells. Real-time RT-PCR and Western blotting confirmed changes in the expression of the gene and protein. The increase of 14-3-3zeta mRNA was observed at 72 h in the fluoxetine-treated RBL-2H3 cells. The increase of 14-3-3zeta protein was observed at 48 and 72 h. In this study, the expressions of the 14-3-3zeta gene and the protein were up-regulated at 72 h. In addition, the increase of TPH mRNA was observed at 12, 24 and 72 h in the fluoxetine-treated RBL-2H3 cells. We conclude that fluoxetine induces increases of 14-3-3zeta mRNA, 14-3-3zeta protein and TPH mRNA at 72 h in the RBL-2H3 cells. This suggests that the 14-3-3zeta and TPH genes may play a role in the molecular mechanism of fluoxetine. To date, no cases of 14-3-3zeta alterations by antidepressants and specifically by fluoxetine have been reported.