Improvement in lip appearance (lateral and frontal aspects) following mandibular setback surgery.

INTRODUCTION: The post-operative facial profile is critical for patients who undergo orthognathic surgery. The present study investigated the improvement in lip appearance (lateral and frontal aspects) following mandibular setback surgery. MATERIAL AND METHODS: Thirty-one patients with mandibular prognathism underwent mandibular setback surgery. Lateral and posteroanterior cephalograms were obtained before surgery (T0) and more than 1 year after surgery (T1). The landmarks (soft and hard tissues) and linear distances were compared by statistical analysis. RESULTS: The lateral cheilion (Ch), point B (B), and pogonion (Pog) were significantly setbackin the horizontal plane: 5.59, 11.49, and 12.35 mm, respectively. In the vertical plane, B and Pog did not move significantly. The Ch moved significantly downward by 3.23 mm on average. The setback ratios of soft tissue/hard tissue, soft tissue of B/B, and soft tissue of Pog/Pog were 0.96. The Ch/Pog ratio was 0.45. The width of the frontal Ch was significantly reduced by 3.17 mm. CONCLUSIONS: The relationship between the corresponding soft and hard tissues of the chin was approximately 1. The relationship between the lip corner and chin bone was nearly 50%. The width of the lip corner was also significantly reduced.

Appearance on face reading (cheek line) after orthognathic surgery.

The cheek line (face reading) is an aesthetic element of the facial profile. The purpose of our study was to investigate the changes in the cheek line after mandibular setback surgery. Forty patients (20 female and 20 male, mean (SD) age 22 (5) years) were diagnosed with mandibular prognathism and treated by intraoral vertical ramus osteotomy alone. Cephalograms were obtained before operation (T1), at least a year postoperatively (T2), and final surgical changes over a year (T2-T1). The cheek line and landmarks (soft and hard tissues) were compared using the paired t test. The hypothesis was that the cheek line did not change significantly after mandibular setback. At the time of the final follow-up (T2-T1), the mean (SD) horizontal setback of pogonion (Pog) was 12.3 (3.5) mm for women and 11.7 (4.3) mm for men. The ratios of soft:hard tissue, labrale inferius:incisor inferius, labiomental sulcus:point B, soft tissue Pog:Pog, and cheek point:Pog in women were 0.96, 0.98, 0.98, and 0.08, and in men 0.91, 1.01, 0.94, and 0.13, respectively. The nasolabial and cervicomental angles in women were significantly increased by 11.1° and 11.4°, respectively, and in men the nasolabial angle was significantly increased by 11.1° and the mentolabial angle reduced by 9.9°. The cheek line (T2-T1) was moved significantly forwards. The hypothesis was therefore rejected. In conclusion, the cheek line was advanced significantly after isolated mandibular setback.

Zinc Finger Nucleases: Tailor-made for Gene Therapy.

Genome editing with the use of zinc finger nucleases has been successfully applied to variety of a eukaryotic cells. Furthermore, the proof of concept for this approach has been extended to diverse animal models from Drosophila to mice. Engineered zinc finger nucleases are able to target specifically and manipulate disease-causing genes through site-specific double strand DNA breaks followed by non-homologous end joining or homologous recombination mechanisms. Consequently, this technology has considerable flexibility that can result in either a gain or loss of function of the targeted gene. In addition to this flexibility, gene therapy by zinc finger nucleases may enable persistent long term gene modification without continuous transfection- a potential advantage over RNA interference or direct gene inhibitors. With systemic viral delivery systems, this gene-editing approach corrected the mutant factor IX in models of mouse hemophilia. Moreover, phase I clinical trials have been initiated with zinc finger nucleases in patients with glioblastoma and HIV. Thus, this emerging field has significant promise as a therapeutic strategy for human genetic diseases, infectious diseases and oncology. In this article, we will review recent advances and potential risks in zinc finger nuclease gene therapy.

Selective modification of HK peptides enhances siRNA silencing of tumor targets in vivo.

Our research has focused on systemic delivery of small interference RNA (siRNA) by branched peptides composed of histidine and lysine. After studying several histidine-lysine (HK) peptides, one four-branched peptide, H3K(+H)4b, with a predominant repeating pattern of -HHHK-, was found to be an effective carrier of siRNA. Although the unmodified H3K(+H)4b carrier of siRNA targeting an oncogene was previously shown to have promise in a tumor-bearing mouse model, we sought to develop a more effective HK carrier of siRNA in this study. Our primary goal was to determine whether different ligand (cyclic RGD)-pegylation patterns on the H3K(+H)4b peptide affect siRNA delivery in vitro and in vivo. We compared the unmodified H3K(+H)4b with two modified H3K(+H)4b peptides for their ability to deliver siRNA in a tumor-bearing mouse model; one modified HK peptide, (RGD-PEG)(4)-H3K(+H)4b, had four cyclic RGD-polyethylene glycol (cRGD-PEG) conjugates per molecule, whereas the other peptide, (RGD-PEG)-H3K(+H)4b, had one cRGD-PEG per molecule. Although the modified HK peptides by themselves did not form stable nanoplexes with siRNA, combination of a highly charged unmodified HK peptide, H2K4b, with either of the modified HK peptides did form stable siRNA nanoparticles. For in vitro experiments with MDA-MB-435 cells that expressed luciferase (Luc), the H3K(+H)4b siRNA nanoplexes targeting Luc decreased its activity by 90% compared with negligible downregulation by the modified H3K(+H)4b nanoplexes (P<0.01). In contrast, the two modified H3K(+H)4b siRNA nanoplexes administered intravenously were more effective than the H3K(+H)4b nanoplexes in silencing Luc in a tumor xenograft model. The Luc activity in tumor lysates of mice administered H3K(+H)4b, (RGD-PEG)-H3K(+H)4b and (RGD-PEG)(4)-H3K(+H)4b nanoplexes decreased by 18, 35 and 75%, respectively. Thus, the siRNA nanoplex incorporating the highly modified peptide, (RGD-PEG)(4)-H3K(+H)4b, was the most effective at silencing its target in vivo (P<0.01). These studies demonstrate that selectively modified HK polymers are promising candidates for targeting oncogenes with siRNA.

Protective effects of water extract of clam on normal and CCl4-induced damage in primary cultured rat hepatocytes.

The objective of this study was to investigate the effects of various concentrations and incubation times of water extract of clam (WEC) on glutathione, its antioxidant and the detoxification defense systems in normal and CCl4-induced oxidative damaged primary rat hepatocytes. This study showed that when the hepatocytes were treated with WEC (0.14 ~ 1.68 mg/ml), the intracellular glutathione (GSH) levels, GSH/GSSG ratio, and the activities of GSH-related enzymes (GPx, GRd, and GST) were higher than those in the control at 24 or 48 hour treatments. However, the lactate dehydrogenase (LDH) leakage and microscopic observations did not differ from those of the control. Yet, when the hepatocytes were pretreated with various concentrations of WEC for 24 hours and then exposed to 5 mM carbon tetrachloride (CCl4) for 1 hour, at concentrations of WEC between 0.42 ~ 1.68 mg/ml, the viabilities, intracellular GSH level, and activities of GST and GPx were significantly increased compared to those of the CCl4-treated control group (p < 0.05). In conclusion, WEC could improve the viability and the capabilities of detoxification and antioxidation in hepatocytes by increasing the GSH level and the activities of GSH-related enzymes.

Monte Carlo simulation of bacterial disinfection: nonlinear and time-explicit intensity of transition.

Bacteria being disinfected in fluid media are discrete entities and mesoscopic in size; moreover, they are incessantly as well as irregularly in motion and in collision among themselves or with the surrounding solid surfaces. As such, it is highly likely that some of the attributes of the bacterial population, for example, their number concentration, will fluctuate randomly. This is especially the case at the tail-end of disinfection when the population of bacteria is sparse. It might be effectual, therefore, to explore the resultant random fluctuations via a stochastic paradigm. Proposed herein is a Markovian stochastic model for the rate of bacterial disinfection, whose intensity of transition takes into account the contact time of the bacteria with the disinfecting agent to eliminate any given percentage of the bacteria in terms of a nonlinear function of time. The model's master equation has been simulated by resorting to the Monte Carlo method to circumvent the undue complexities in solving it analytically or numerically via conventional numerical techniques. For illustration, the mean, the variance (standard deviation), and the coefficient of variation of the number concentration of bacteria during disinfection have been estimated through Monte Carlo simulation. The results of simulation compare favorably with the available experimental data as well as with those computed from the corresponding deterministic model.

Peptide-based Antifungal Therapies against Emerging Infections.

Acquired drug resistance to mycotic infections is rapidly emerging as a major medical problem. Opportunistic fungal infections create therapeutic challenges, particularly in high risk immunocompromised patients with AIDS, cancer, and those undergoing transplantation. Higher mortality and/or morbidity rates due to invasive mycosis have been increasing over the last 20 years, and in light of growing resistance to commonly used antibiotics, novel antifungal drugs and approaches are required. Currently there is considerable interest in antifungal peptides that are ubiquitous in plant and animal kingdoms. These small cationic peptides may have specific targets or may be multifunctional in their mechanism of action. On the basis of recent advances in protein engineering and solid phase syntheses, the utility and potential of selected peptides as efficient antifungal drugs with acceptable toxicity profiles are being realized. This review will discuss recent advances in peptide therapy for opportunistic fungal infections.

Epidemiological investigation of a case of nosocomial Legionnaires' disease in Taiwan: implications for routine environmental surveillance.

An epidemiological investigation with Legionella and molecular subtyping was conducted to determine the source of a case of nosocomial Legionnaires' disease (LD) who was hospitalized in three hospitals within a month. Legionella pneumophila serogroup 3, an uncommon serogroup for infection, was isolated from the patient's sputum. Environmental surveillance revealed Legionella colonization in all three hospitals; the patient isolate matched the isolate from the first hospital by molecular typing. Culturing the hospital water supply for Legionella is a pro-active strategy for detection of nosocomial LD even in hospitals experiencing no previous cases.

Studies on the antioxidative activities of Hsian-tsao (Mesona procumbens Hemsl) leaf gum.

This study aimed at evaluating the antioxidative activity of crude hsian-tsao leaf gum extracted by sodium bicarbonate solutions and precipitated by 70% ethanol. The antioxidative activities, including the radical-scavenging effects, Fe(2+)-chelating ability, and reducing power as well as the inhibition of FeSO(4)-H(2)O(2)-induced malondialdehyde formation in rat tissue homogenate were studied in vitro. It was found that the antioxidative effect provided by hsian-tsao leaf gum was strongly concentration dependent. In general, the antioxidative activity increased with increasing gum concentration, to a certain extent, and then leveled off with further increase in gum concentration. A concentrtaion-dependent kinetics for the rate of change in antioxidative activity was proposed. The antioxidative activity constant (k) and the half-inhibition concentration (IC(50)) for each antioxidative reaction studied were calculated. From a comparison of the IC(50) values for different antioxidative reactions, it seemed that hsian-tsao leaf gum was more effective in scavenging superoxide radicals than chelating Fe(2+) or scavenging alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radicals. As compared to the commercial antioxidants, hsian-tsao leaf gum showed less scavenging effect on the DPPH radical and reducing power but better superoxide radical-scavenging effect and Fe(2+)-chelating ability than alpha-tocopherol and BHT.

Effect of nucleoside analogue therapy on duck hepatitis B viral replication in hepatocytes and bile duct epithelial cells in vivo.

BACKGROUND: Recent studies have implicated bile duct epithelial cells (BDEC) as a reservoir of hepatitis B virus (HBV) infection that may be particularly important in the development of post-liver transplant recurrence of hepatitis B. The aim of this study was to compare the effects of antiviral therapy on duck HBV (DHBV) expression in hepatocytes and BDEC and to determine if this was affected by biliary hyperplasia. METHODS: Ducklings congenitally infected with DHBV received penciclovir (10 mg/kg per day) treatment from 9 days of age. In order to mimic the biliary hyperplasia that often accompanies severe post-liver transplant HBV recurrence, half the animals underwent bile duct ligation. Duck HBV-DNA in serum was measured at day 1, and serum and liver DHBV-DNA were determined when the animals were killed on day 17. Intrahepatic expression of viral preS1 antigen and DHBV-DNA was measured by immunohistochemistry and in situ hybridization, respectively. RESULTS: Viraemia became undetectable in the penciclovir-treated animals at day 17, following 8 days of therapy. Examination of liver tissue revealed that all hepatocytes and the majority of BDEC contained DHBV preS1 antigen and DHBV-DNA. Penciclovir greatly reduced the intrahepatic viral burden, but there was no antiviral effect on viral markers within BDEC. Despite the increased number of BDEC after bile duct ligation, the same proportion of BDEC was seen to be infected, and this was unaffected by antiviral therapy. CONCLUSIONS: In the duck model with and without biliary hyperplasia, penciclovir controls DHBV replication and reduces viral burden in hepatocytes, but not in BDEC. The BDEC appear to be an important reservoir of virus that is relatively unaffected by antiviral treatment, and may play an important role in disease persistence and relapse following cessation of therapy.

Effect of age and sex on plasma total homocysteine in Taiwanese subjects.

Plasma total homocysteine (tHcy) is now established as a clinical risk factor for coronary artery disease, as well as for other arterial and venous occlusive diseases. Therefore, we measured the plasma tHcy concentrations in 385 healthy Chinese subjects in Taiwan and in 40 patients with occluded coronary artery disease or maintenance hemodialysis. The plasma tHcy levels in Taiwanese male and female volunteers were found to increase gradually with age (age group: 20-29, 30-39, 40-49, 50-59, and >60; mean +/- SD 8.22 +/- 2.00, 8.51 +/- 2.67, 8.87 +/- 2.22, 11.41 +/- 2.50 and 13.28 +/- 2.31 microM for male volunteers and 6.49 +/- 1.75, 7.15 +/- 1.20, 7.40 +/- 1.30, 9.57 +/- 3.01 and 10.95 +/- 2.11 microM for female volunteers). At the same age, male volunteers were shown to have higher tHcy levels than female volunteers. In addition, the mean concentrations of plasma tHcy in occluded coronary artery disease (13.62 +/- 5.43 microM) or in maintenance hemodialysis (21.28 +/- 4.32 microM) were statistically higher than in age-matched normal subjects (11.02 +/- 2.85 microM). This study emphasizes the significance of age and sex-associated difference in the plasma tHcy levels, and underlines the importance of the range for plasma homocysteine in normal Taiwanese subjects.

MRP1 not MDR1 gene expression is the predominant mechanism of acquired multidrug resistance in two prostate carcinoma cell lines.

Multidrug resistant prostate cancer cell lines DU 0.03 and PC 0.03 were established from the parental prostate cancer cell lines DU145 and PC-3 respectively by stepwise selection in doxorubicin (DOX) from 0.001 to 0.03 &mgr;g/ml. As cells adapted to each concentration of DOX. the drug concentration was increased by 0.001 &mgr;g/ml. The chemosensitivity of each line was determined by growth inhibition assay. The DU 0.03 and PC 0.03 lines exhibit a 5-10-fold and 1.3-2.8-fold increase in resistance to anthracyclines, vinblastine (VLB) and mitozantrone (Mito), respectively. Verapamil (5 &mgr;M) partially reversed the resistance to the anthracycline and completely reversed the resistance to VLB and Mito. Drug kinetic studies measured by intracellular accumulation of (3)H-daunorubicin demonstrated a 3 fold decrease in the level of intracellular (3)H-daunorubicin in the PC 0.03 and DU 0.03 resistant lines compared with their respective parental line. This effect was partially reversed by 5 &mgr;M verapamil. The expression of MDR1 and MRP genes was analysed by Northern blotting and RT-PCR. P-glycoprotein (Pgp) and MRP protein were tested by immunocytochemistry staining using the monoclonal antibodies J-SB1. C219 and MRK16 (Pgp) and MRPm6 and MRPr1 (MRP). Neither Northern blot analysis nor the more sensitive RT-PCR demonstrated detectable MDR1 transcripts in any of the prostate cancer cell lines and the three Pgp monoclonal antibodies failed to reveal expression of Pgp. A 2-4-fold increase in MRP1 mRNA levels in the drug resistant DU 0.03 and PC 0.03 lines were demonstrated by both Northern blotting and RT-PCR consistent with the findings observed after staining by the two specific monoclonal antibodies, MRPm6 and MRPr1. Southern blot analysis demonstrated a 2-fold increase in the MRP1 gene copy number in the PC 0.03 line but not in the DU 0.03 line, suggesting that the overexpression of the MRP gene was regulated at the level of transcription in the latter line. We conclude that MRP1 not MDR1 overexpression. contributes to acquired drug resistance in these two prostate cancer cell lines. Prostate Cancer and Prostatic Diseases (2000) 3, 66-75

nm23 protein expression and p53 immunoreactivity in cutaneous fibrohistiocytic tumors.

Recent data indicate that reduced expression of the 17-kD protein encoded by the nm23 gene may be important in the pathogenesis of several types of human tumors. Immunohistochemistry was performed using a murine monoclonal antibody, NCL-nm23 (Novocastra, 1:150 dilution) to investigate nm23 protein immunoreactivity in a group of locally aggressive cutaneous fibrohistiocytic tumors; dermatofibrosarcoma protuberans (DFSP) (n = 14) and atypical fibroxanthoma (AFX) (n = 7). Cases of dermatofibroma (DF) (n = 17) formed the benign control group. Comparison with p53 protein immunoreactivity in the same cases studied previously was made. Strong immunohistological expression of the nm23 protein was seen in most of the cases of DF (n = 15; 88%) in the form of strong cytoplasmic immunolabelling without nuclear staining. However, strong nm23 immunoreactivity was observed in only a minority of the cases of DFSP (n = 5; 36%) and AFX (n = 2; 29%). Statistically significant differences in nm23 immunoreactivity were found between DFSP and DF (p = 0.008, chi 2 test with continuity correction) and between AFX and DF (p = 0.015; chi 2 test with continuity correction). No significant difference was seen between DFSP and AFX (p = 0.87, chi 2 test with continuity correction). There was inverse correlation between nm23 and p53 immunoreactivity (r = 0.331; r2 = 0.109; p = 0.046; simple regression analysis). In summary, nm23 protein immunoreactivity is reduced in DFSP and AFX but not in dermatofibroma suggesting that reduced expression of the protein may be important in influencing the behavior of fibrohistiocytic tumors, although this is not well characterised. nm23 protein expression is also found to be inversely related to p53 immunohistological expression in these tumors.

Parathyroid hormone-related protein mRNA and protein expression in multiple myeloma: a case report.

Multiple myeloma frequently leads to complications, such as osteolytic lesions, hypercalcemia, and pathological fractures. Increased bone resorption in myeloma is due to osteoclast activation. The nature of the osteoclast activator(s) remains unclear. We describe a case of multiple myeloma with marked hypercalcemia and skeletal complications that progressed rapidly despite chemotherapy. The patient had marked hypercalcemia at diagnosis (4.5 mmol/l), and elevated parathyroid hormone-related protein (PTHrP) levels were found in plasma. Analysis of the bone marrow trephine biopsy showed PTHrP gene transcription and protein in myeloma cells. These results provide strong evidence for the production of significant amounts of PTHrP by human myeloma cells. PTHrP has been measured as elevated in the plasma of patients with myeloma and might be an important contributor to the skeletal complications in this disease.

A histological study of Denonvilliers' fascia and its relationship to the neurovascular bundle.

OBJECTIVE: To evaluate the anatomical relationship between the nerves and fasciae at the site of dissection in a radical retropubic prostatectomy. MATERIALS AND METHODS: Eight adult male (autopsy) en bloc specimens of bladder and rectum were examined histologically after staining with either haematoxylin and eosin or S-100 protein (as a specific nerve stain). RESULTS: All specimens showed Denovilliers' fascia to have no clearly defined layers and no definable lateral edge. No distinct neurovascular bundle was seen but nerves were scattered throughout the fasciae, including medially towards the midline. CONCLUSION: In radical retropubic prostatectomy, a piece of Denonvilliers' fascia is taken with the specimen, thus removing these nerves. The loss of these nerve fibres may explain the significant rate of erectile dysfunction after 'nerve-sparing' surgery.

p53 protein immunoreactivity in fibrohistiocytic tumors of the skin.

Abnormal expression of the 53 kDa nuclear phosphoprotein produced by the p53 gene is observed in many human cancers. p53 nuclear immunoreactivity is found commonly in tumor cells. Immunohistochemistry was performed using a monoclonal antibody, DO-7 (DAKO, Denmark; cat. no. M7001; 1:100 dilution), to investigate p53 protein immunoreactivity in a group of cutaneous fibrohistiocytic tumors that are known to be locally aggressive. The study group consisted of dermatofibrosarcoma protuberans (DFSP) (n = 14) and atypical fibroxanthoma (AFX) (n = 7). Cases of dermatofibroma (DF) (n = 16) formed the benign control group. Intense nuclear immunostaining for p53 protein was observed in 71% of DFSP and 86% of AFX. None of the dermatofibromas showed strong p53 nuclear immunostaining. Statistical analyses revealed significant differences in p53 immunoreactivity between DFSP and DF (P = 0.0001, chi 2 test) and between AFX and DF (P = 0.0001, chi 2 test). In conclusion, increased p53 protein immunoreactivity is found in DFSP and AFX but not in DF. These differences in p53 immunoreactivity suggest that increased expression of the protein may be important in the pathogenesis of the more aggressive group of fibrohistiocytic tumors.

Temporal expression of PTHrP during endochondral bone formation in mouse and intramembranous bone formation in an in vivo rabbit model.

Expression of parathyroid hormone-related protein (PTHrP) messenger RNA (mRNA) and protein was investigated throughout the developmental progression of endochondral bone formation in mouse and intramembranous bone formation in an in vivo model in rabbit, using in situ hybridization and immunohistochemistry. Endochondral bone formation was investigated in a developing embryo, newborn, and adult mouse. In fetal long bones through to newborn (day 7), PTHrP mRNA and protein were consistently expressed in chondrocytes within the proliferative, transitional, and hypertrophic zones. In addition, high levels of PTHrP were also detected in osteoblasts on the surface of trabecular bone surfaces. Similarly, at the adult stage (week 7), PTHrP mRNA and protein were consistently expressed in chondrocytes at epiphyseal ends of the subarticular cartilage, within cortical periosteum, as well as in osteoblasts located at the metaphyseal trabecular bone surfaces. Using an in vivo intramembranous bone formation model in rabbits, expression of PTHrP mRNA and protein was demonstrated in preosteoblasts prior to trabecular bone formation (1-week bone harvest). As bone formed (2-, 3-, and 4-week bone tissue harvests), PTHrP mRNA and protein were highly expressed in actively synthesizing osteoblasts and in those osteocytes embedded within the superficial layers of the bone matrix. Lining osteoblasts and osteocytes buried deeply in the bone matrix displayed weak or no signal for PTHrP. The pattern of spatial and temporal expression of PTHrP demonstrated in cartilage cells and osteoblasts in the two systems suggests an important role of PTHrP in both endochondral and intramembranous bone formation.

Evaluation of extracellular lipid peroxidation in brain cortex of anaesthetized rats by microdialysis perfusion and high-performance liquid chromatography with fluorimetric detection.

A method for in vivo evaluation of lipid peroxidation in the extracellular space of anaesthetized rat brain cortex was developed. This method involved the use of microdialysis perfusion and high-performance liquid chromatography. The microdialysates, eluted from implanted probes, were reacted with thiobarbituric acid (TBA) prior to analysis by an HPLC system equipped with a fluorescence detector (excitation and emission wavelengths were 515 and 550 nm, respectively). Lipid peroxidation in the extracellular space was evaluated as the concentration of malondialdehyde, a lipid peroxidation end product which reacts with TBA to form a fluorescent conjugate. Significantly increased production of malondialdehyde following hydrogen peroxide perfusion (0.03%, 0.3% at a flow-rate of 1 microl/min) was observed in the brain cortex of anaesthetized rats.