Was it Good for You? Gender Differences in Motives and Emotional Outcomes Following Casual Sex.

Casual sex, also referred to as a hookup, has been associated with a range of negative emotional outcomes for women, including regret, anxiety, depression and social stigma. However, it has been argued that it is the nature of the sexual motivation, not gender that influences the emotional outcome. This study was designed to ascertain what motivates people to have casual sex, what emotional outcomes follow casual sex and whether there are gender differences among these variables. Seven hundred and one participants (47% men and 52.8% women) completed a 44-item online survey. Gender differences were found for both sexual motivations and emotional outcomes of casual sex, with women generally having more negative emotional outcomes than men. Additionally, a principal components analysis uncovered four reliable principal motivations underlying engagement in casual sex, and three principal emotional outcomes of casual sex. Predictors of negative emotional outcomes included being motivated to regulate negative emotions and to achieve positive emotions. No predictors (apart from being a man) were found for a positive emotional outcome. While the stigma surrounding female sexual agency is diminishing, results generally support the presence of a sexual double-standard which encourages male promiscuity but dissuades female sexual autonomy.

Race-specific molecular alterations correlate with differential outcomes for black and white endometrioid endometrial cancer patients.

BACKGROUND: The objective of this study was to identify molecular alterations associated with disease outcomes for white and black patients with endometrioid endometrial cancer (EEC). METHODS: EEC samples from black (n = 17) and white patients (n = 13) were analyzed by proteomics (liquid chromatography-tandem mass spectrometry) and transcriptomics (RNA-seq). Coordinate alterations were validated with RNA-seq data from black (n = 49) and white patients (n = 216). Concordantly altered candidates were further tested for associations with race-specific progression-free survival (PFS) in black (n = 64) or white patients (n = 267) via univariate and multivariate Cox regression modeling and log-rank testing. RESULTS: Discovery analyses revealed significantly altered candidate proteins and transcripts between black and white patients, suggesting modulation of tumor cell viability in black patients and cell death signaling in black and white patients. Eighty-nine candidates were validated as altered between these patient cohorts, and a subset significantly correlated with differential PFS. White-specific PFS candidates included serpin family A member 4 (SERPINA4; hazard ratio [HR], 0.89; Wald P value = .02), integrin subunit α3 (ITGA3; HR, 0.76; P = .03), and Bet1 Golgi vesicular membrane trafficking protein like (BET1L; HR, 0.48; P = .04). Black-specific PFS candidates included family with sequence similarity 228 member B (FAM228B; HR, 0.13; P = .001) and HEAT repeat containing 6 (HEATR6; HR, 4.94; P = .047). Several candidates were also associated with overall survival (SERPINA4 and ITGA3) as well as PFS independent of disease stage, grade and myometrial invasion (SERPINA4, BET1L and FAM228B). CONCLUSIONS: This study has identified and validated molecular alterations in tumors from black and white EEC patients, including candidates significantly associated with altered disease outcomes within these patient cohorts. Cancer 2017;123:4004-12. © 2017 American Cancer Society.

Indocyanine green videoangiography-assisted prediction of flap necrosis in the rat epigastric flap using the flow® 800 tool.

BACKGROUND: The decision to re-operate on a potentially ischemic free flap remains challenging. Indocyanine green videoangiography (ICG) with the FLOW® 800 tool is a method which allows an immediate qualitative conclusion about the patency of an anastomosis. Is it also able to predict the outcome of potentially compromised vascular free flaps? MATERIALS AND METHODS: An epigastric flap was raised and repositioned in 79 rats. Intraoperative fluorescence angiography was performed using ICG videoangiography and the FLOW® 800 tool was applied. Six regions of interest were positioned systematically over the flap, changes of the ICG fluorescence were color coded with respect to time and 474 measurements were performed. The flap was clinically monitored for one week and the resulting necrotic areas were correlated with the ICG/FLOW® 800 results. RESULTS: Mean intensity of clinically vital areas was 83.39 ± 50.96 arbitrary units (AU) and 37.33 ± 15.14 AU in necrotic areas. The receiver operating characteristic curve and Youden-Index analysis revealed that the optimal cutoff for the maximal intensity of ICG after FLOW® 800 analysis was ≤ 61.733 for the prediction of flap necrosis and > 61.733 for the prediction of flap survival (P < 0.0001; 95% CI = 0.85-0.91; Youden-Index: 0.67). The maximal intensity of ICG angiography had a specificity of 96.1% and sensitivity of 71.4%. The positive predictive value was 97.46% and the corresponding negative predictive value was 61.34%. CONCLUSION: This demonstrates the potential additional value of ICG videoangiography including FLOW® 800 analyses in the postoperative monitoring of transplanted flaps. © 2016 Wiley Periodicals, Inc. Microsurgery 37:235-242, 2017.

Form and Size Matter: Increased Risk of Thrombosis in Microvessels with Surgically Created Endothelial Lesions.

Background Atherosclerosis is a known risk factor for flap loss in microsurgery. Several microsurgical techniques, like plaque removal, have been proposed for atherosclerotic vessels, but these techniques often induce intimal injuries. The aim of this study was to investigate the impact of various endothelial defects on the risk of thrombosis in a rat acute intimal injury model. Methods Endothelial defects of various forms and sizes were created in the abdominal aorta of 30 male Wistar rats following a strict protocol. Defect sizes were measured and classified as round, horizontal, or vertical based on their configuration. An hour after reestablishing the blood flow, the abdominal aorta was harvested and the operation site was assessed for signs of thrombosis clinically and using light microscopy. Univariate and multiple linear regression analysis were performed to identify possible influencing factors on thrombosis. Results The mean defect size was 2.65 ± 1.19 mm2. Intimal lesions were classified as round in 36.7%, horizontal in 33.3%, and vertical in 30% of specimens. Thrombus formation was detected in 46.7% clinically and in 50% histologically. Univariate regression analysis revealed that defect size (p = 0.048) and vertical form (p = 0.017) were significantly associated with thrombus formation. Multiple regression analysis corroborated vertical defects as a risk factor for thrombosis (p = 0.03). Conclusion Endothelial injuries are associated with a high risk of thrombosis with highest risks associated with vertical defects. Arteries should be carefully examined for intimal defects before microvascular anastomosis, especially in the atherosclerotic patient.

Bisphosphonate and Medication-Related Osteonecrosis of the Jaw: A Review.

For patients with malignant disease taking bisphosphonates and denosumab, the incidence of medication-related osteonecrosis of the jaw (MRONJ) is up to 15% in contrast to 0.01% in patients with osteoporosis. Clinical presentation of MRONJ extends from asymptomatic exposure of bone in 94% of patients to severe cases of mandibular fractures in a minority of 4.5%. The strongest risk factors for MRONJ are invasive dental procedures and dental infections. Advances in imaging provide more preoperation information compared with panoramic radiograph. Prevention strategies are the elimination of potential risk factors leading to invasive dental procedures and maintenance of good oral hygiene prior to the administration of antiresorptive agents. Management of MRONJ depends on the underlying disease, extent of the necrosis, and the presence of contributing therapy. Conservative therapies include topical anti-infective rinses and systemic antibiotic therapy. The most important part of surgical therapy is to remove the exposed and necrotic bone. Several options for defect closure are possible from local tissue flaps to microvascular free flap procedures. The development of MRONJ in conjunction with dental implants is a severe side effect and should be avoided if potentially harmful medication has already been administered.

Mutations in the X-linked intellectual disability gene, zDHHC9, alter autopalmitoylation activity by distinct mechanisms.

Early onset intellectual disabilities result in significant societal and economic costs and affect 1-3% of the population. The underlying genetic determinants are beginning to emerge and are interpreted in the context of years of work characterizing postsynaptic receptor and signaling functions of learning and memory. DNA sequence analysis of intellectual disability patients has revealed greater than 80 loci on the X-chromosome that are potentially linked to disease. One of the loci is zDHHC9, a gene encoding a Ras protein acyltransferase. Protein palmitoylation is a reversible modification that controls the subcellular localization and distribution of membrane receptors, scaffolds, and signaling proteins required for neuronal plasticity. Palmitoylation occurs in two steps. In the first step, autopalmitoylation, an enzyme-palmitoyl intermediate is formed. During the second step, the palmitoyl moiety is transferred to a protein substrate, or if no substrate is available, hydrolysis of the thioester linkage produces the enzyme and free palmitate. In this study, we demonstrate that two naturally occurring variants of zDHHC9, encoding R148W and P150S, affect the autopalmitoylation step of the reaction by lowering the steady state amount of the palmitoyl-zDHHC9 intermediate.

The value of perioperative antibiotics on the success of oral free flap reconstructions.

The subject of the presented study was to monitor and compare problems and outcomes of reconstructive surgery with microvascular free flaps in the head and neck region between groups of patients treated with perioperative antibiotics and a group of patients without antibiotics. Patients requiring oral reconstructive surgery following cancer resections with microvascular free flaps were prospectively evaluated (2007-2012). Antibiotic therapy was started 30 min before the operation and administered for 10 days. Three hundred and fifty patients were included (208 male, 59.4%; 142 female, 40.6%; mean age 59.8 ± 13.2 years). 330 patients received perioperative antibiotics. Twenty patients (5.7%) who received no antibiotics for specific reasons served as the control group. Wound infections developed in 33 of 122 patients (27%) who received benzylpenicillin, 17 of 88 patients (19.3%) who received amoxicillin combined with sulbactam and 25 of 120 patients (20.8%) who received cefuroxime. Ten patients (50%) who did not receive antibiotics developed wound infections. Receiving no antibiotics or penicillin showed no benefit (P = 0.11). Those receiving cefuroxime showed significantly lower incidence of wound infections (P = 0.034; risk decreased by the factor 2.88). The use of amoxicillin combined with sulbactam showed the lowest rate of wound infections (P = 0.018; risk decreased by the factor 3.46). The use of amoxicillin combined with sulbactam appears to be the most appropriate prophylactic antibiotic followed by cefuroxime in oral microsurgical free flap reconstructions. These data may serve as a guide until a controlled multicenter prospective trial is performed comparing newer antibiotics against current standards.

Ketamine-Xylazine Anesthesia in Rats: Intraperitoneal versus Intravenous Administration Using a Microsurgical Femoral Vein Access.

BACKGROUND: Ketamine-xylazine is a frequently used combination for anesthesia in microsurgically operated rats and can be administered by intraperitoneal (IP) or intravenous (IV) injection. Both methods are associated with relatively high misadministration rates. In the present study, we want to introduce a femoral vein access that requires microsurgical cannulation but enables a 100% administration rate. METHODS: In this study, the maximal time of anesthesia was analyzed, time to response, latency time, and the total need for anesthetic agents in IP (n = 200) and IV (n = 40) anesthesia in Wistar rats for the purpose of microvascular operations. IV injections were achieved with an inserted microcatheter that was applied via a microsurgical femoral vein access. RESULTS: The time needed for the placement of the microcatheter was 5.76 ± 0.89 minutes. Maximal time of anesthesia (IP vs. IV) was 7.02 ± 1.92 versus 21.78 ± 5.77 hours (p < 0.0001), time to response was 137.5 ± 3.91 versus 18 ± 2.18 seconds (p < 0.0001), latency time 35.53 ± 3.21 versus 27.8 ± 2.88 minutes (p < 0.0001), and total volume of anesthetic 1.42 ± 0.39 versus 2.78 ± 0.73 mL (p < 0.0001), respectively. CONCLUSION: IV administration using the microsurgical femoral vein access is a feasible method with a quicker response rate and a 100% administration rate. Furthermore, it enables longer anesthesia, for example, complex microsurgical or other experimental procedures in the rat.

Incidence and outcome for patients with occult lymph node involvement in T1 and T2 oral squamous cell carcinoma: a prospective study.

BACKGROUND: The evidence base to inform the decision making process in patients with early stage oral cancer and a clinical and radiological N0 neck remains insufficient to answer the question when it is safe to "watch and wait" and when to proceed with a selective neck dissection. METHODS: A total of 327 consecutive cases of histopathologically staged T1-2, N0-1 and M0, but clinically N0, squamous cell carcinoma of the tongue were prospectively analysed. Univariate and multivariate analyses were used for statistical analysis and are represented as Kaplan-Meier analyses or Cox proportional hazard regression analysis. RESULTS: In 61 patients (18.65%) lymph node involvement was found in the histopathological processing. The mean survival of all patients was 73.3 ± 48.6 months. The 2-year and 5-year overall survival rates of all patients were 87.5% and 68.4%, respectively. The 2-year and 5-year survival rates for stage N0 were 89.1% and 70.7% compared to 83.3% and 62.9% in N1 situations. The 2-year and 5-year survival rates for stage T1 were 87.9% and 73.6% compared to 87.2% and 65.3% in stage T2, respectively. The time to recurrence in stage N0 was 35.1 ± 30.5 months compared to 25.63 ± 24.6 months in cases with N1 disease. Stage T1 was associated with a time to recurrence of 38.1 ± 33.9 months compared with 27.2 ± 22.7 months in patients classified T2.Variables found to be strongly associated with survival in the univariate analysis included older age, higher tumour and N stage, and grading. Age, tumour stage (p = 0.011, 95% CI, 1.09 to 2.0), nodal stage (p = 0.038, 95% CI, 1.02 to 2.07), and recurrence were independently and significantly associated with survival in the multivariate analysis. CONCLUSIONS: This confirms a high overall disease free survival for patients with T1 and N0 treated with single modality surgery and in common with the literature confirms the poor impact on prognosis of the N positive neck.

A fluorescence-based assay to monitor autopalmitoylation of zDHHC proteins applicable to high-throughput screening.

Palmitoylation, the posttranslational thioester-linked modification of a 16-carbon saturated fatty acid onto the cysteine residue of a protein, has garnered considerable attention due to its implication in a multitude of disease states. The signature DHHC motif (Asp-His-His-Cys) identifies a family of protein acyltransferases (PATs) that catalyze the S-palmitoylation of target proteins via a two-step mechanism. In the first step, autopalmitoylation, palmitate is transferred from palmitoyl-CoA to the PAT, creating a palmitoyl:PAT intermediate and releasing reduced CoA. The palmitoyl moiety is then transferred to a protein substrate in the second step of the reaction. We have developed an in vitro, single-well, fluorescence-based enzyme assay that monitors the first step of the PAT reaction by coupling the production of reduced CoA to the reduction of NAD(+) using the α-ketoglutarate dehydrogenase complex. This assay is suitable for determining PAT kinetic parameters, elucidating lipid donor specificity and measuring PAT inhibition by 2-bromopalmitate. Finally, it can be used for high-throughput screening (HTS) campaigns for modulators of protein palmitoylation.

Opened end-to-side technique for end-to-side anastomosis and analyses by an elastic true-to-scale silicone rubber model.

The end-to-side anastomosis is frequently used in microvascular free flap transfer, but detailed rheological analyses are not available. The purpose of this study was to introduce a new modified end-to-side (Opened End-to-Side, OES-) technique and compare the resulting flow pattern to a conventional technique. The new technique was based on a bi-triangulated preparation of the branching-vessel end, resulting in a "fish-mouthed" opening. We performed two different types of end-to-side anastomoses in forty pig coronary arteries and produced one elastic, true-to-scale silicone rubber model of each anastomosis. Then we installed the transparent models in a circulatory experimental setup that simulated the physiological human blood flow. Flow velocity was measured with the one-component Laser-Doppler-Anemometer system, recording flow axial and perpendicular to the model at four defined cross-sections for seven heart cycles in each model. Maximal and minimal axial velocities ranged in the conventional model between 0.269 and -0.122 m/s and in the experimental model between 0.313 and -0.153 m/s. A less disturbed flow velocity distribution was seen in the experimental model distal to the anastomosis. The OES-technique showed superior flow profiles distal to the anastomosis with minor tendencies of flow separation and represents a new alternative for end-to-side anastomosis.

Identification of perioperative risk factor by laser-doppler spectroscopy after free flap perfusion in the head and neck: a prospective clinical study.

The aim of this study was to evaluate perioperative flap perfusion using noninvasive monitoring with a laser-Doppler flowmetry and spectrophotometry unit (O2C) and identify whether perioperative blood flow, velocity, hemoglobin level (Hb), and oxygen saturation (SO2 ) measured could be used as indicators of free flap success. Measurements of blood flow, velocity, Hb, and SO2 were performed in 196 microvascular flaps, which had been transferred into the oral cavity to reconstruct ablative defects after surgery for oral cancer. The values were calculated superficially on the skin surface and at a depth of 8 mm. The results showed that perioperative absolute values measured were not associated with an increased rate of microvascular revisions or free flap failure. Independent predictors of microvascular revisions at the first postoperative day were the development of a falling trend in superficial and deep blood flow, and velocity in comparison with baseline values of variables measured. On day 2, all superficial and deep values of Hb, flow, and velocity were independent prognostic factors (P < 0.01), demonstrated as a downward trend were associated with a need for revision. The superficial and deep values of SO2 (P = 0.59 and 0.43, respectively) were not associated with ultimate free flap failure. This is the first clinical study to demonstrate that during early free flap integration to the recipient site different parameters of perfusion and oxygenation play an important role at different points of time. Within the first two postoperative days, changes in these parameters can help influence the decision to revise microvascular anastomoses.

The Erf4 subunit of the yeast Ras palmitoyl acyltransferase is required for stability of the Acyl-Erf2 intermediate and palmitoyl transfer to a Ras2 substrate.

Protein S-palmitoylation is a posttranslational modification in which a palmitoyl group is added to a protein via a thioester linkage on cysteine. Palmitoylation is a reversible modification involved in protein membrane targeting, receptor trafficking and signaling, vesicular biogenesis and trafficking, protein aggregation, and protein degradation. An example of the dynamic nature of this modification is the palmitoylation-depalmitoylation cycle that regulates the subcellular trafficking of Ras family GTPases. The Ras protein acyltransferase (PAT) consists of a complex of Erf2-Erf4 and DHHC9-GCP16 in yeast and mammalian cells, respectively. Both subunits are required for PAT activity, but the function of the Erf4 and Gcp16 subunits has not been established. This study elucidates the function of Erf4 and shows that one role of Erf4 is to regulate Erf2 stability through an ubiquitin-mediated pathway. In addition, Erf4 is required for the stable formation of the palmitoyl-Erf2 intermediate, the first step of palmitoyl transfer to protein substrates. In the absence of Erf4, the rate of hydrolysis of the active site palmitoyl thioester intermediate is increased, resulting in reduced palmitoyl transfer to a Ras2 substrate. This is the first demonstration of regulation of a DHHC PAT enzyme by an associated protein.

Objective qualitative and quantitative assessment of blood flow with near-infrared angiography in microvascular anastomoses in the rat model.

Intraoperative near-infrared indocyanine-green (ICG) angiography enables the visualization of microvascular perfusion and may help in the early detection of complications. The purpose of the present study was to examine whether the effect of microvascular stenoses can be quantitatively assessed by analysis of ICG-angiography in a microvascular model. Graded stenoses and total vessel occlusion of the carotid, aorta, and femoral arteries were created in 25 Wistar rats. Stenoses were graded to reduce arterial flow by 25%, 50%, 75%, and 100% of baseline flow as measured by transit-time flowmeter analyzing the emission signal of the ICG detected and investigated by the mathematical software tool (FLOW 800). ICG angiography was performed to assess vessel perfusion and flow curves were analyzed and correlated with the stenosis rate. A total of 576 investigations were performed. The area under the curve (P < 0.001), first and second maximum (P < 0.001), and the maximum slope to the first maximum (P < 0.001) were found to be of high prognostic value in evaluating the different flow patterns. Differences were displayed in comparisons by the maximum intensity of the ICG-concentrations. The maximum slope to the second maximum was found to be predictive in selected vessel types, and specific changes of the flow curve were found to indicate compromised vascular flow. The FLOW 800 tool applied for ICG angiography has shown to be a quick and reliable method for assessing blood flow in vessels in this study. The dynamic assessment of the ICG signal allows reliable identification of microanastomotic complications with the described parameters.

Autonomization of free flaps in the oral cavity: A prospective clinical study.

Controversy exists over how long a free flap is dependent on its pedicle and if neovascularization is different between flap types, recipient sites, and irradiated and nonirradiated patients. An understanding of the timing of this process should optimize the safety of secondary procedures involving the flap. In a prospective clinical study, hemoglobin oxygenation and capillary flow were measured in 50 flaps (25 forearm flaps, 15 osteocutaneous fibula flaps, and 10 anterolateral thigh flaps) 4 and 12 weeks postoperatively. The flaps were located at the floor of the mouth, cheek, or tongue (n = 39) or at the hard or soft palate (n = 11). Measurements were carried out using the O2C monitoring system under temporary digital occlusion of the pedicle. After 4 weeks, 17 free flaps were found to be autonomized indicated by the O2C measurements comparing both values before and after digital compression of the vascular pedicle. After 12 weeks, 41 patients had completion of free flap autonomization, as indicated by the HbO(2) and CF before and after pedicle compression. The location of free flap in the lower jaw (P < 0.0001 after 4 weeks, P = 0.013 after 12 weeks), fasciocutaneous radial forearm flaps after 4 weeks (P < 0.0001), and not irradiated recipient site after 4 weeks (P = 0.014) were found to be positive factors significantly influencing autonomization. In conclusion, free flap autonomization depends on several variables which should be considered before further surgery after free flap reconstruction as the transferred tissue can be still dependent on its pedicle.

Real-time magnetic resonance imaging: mechanics of oral and facial function.

We report a summary of developmental work to explore, develop, and establish clinical applications of real-time magnetic resonance imaging (rtMRI) with a temporal resolution of 70 frames/second in oral and maxillofacial surgery (OMFS). Real-time MRI can contribute to procedure planning, diagnostics, rehabilitation, monitoring, and patient education. At present, conventional MRI is used extensively in the diagnosis, staging, and follow up of head and neck cancer patients, with scanning durations typically of several minutes and temporal resolution of up to 0.5 frames/second. The potential for rtMRI, where function can be assessed, could go far beyond the established clinical application of conventional MRI. Preliminary prototyping is a first stage in the establishment of rtMRI in OMFS. We follow best-practice approaches in co-creation across multiple disciplines, an indispensable aspect in the development of new methodologies and diagnostic tools.

Polyoxovanadates as new P-glycoprotein inhibitors: insights into the mechanism of inhibition.

A promising strategy to overcome multidrug resistance is the use of inhibitors of ABC drug transporters. For this reason, we evaluated the polyoxovanadates (POVs) [V10 O28 ]6- (V10 ), [H6 V14 O38 (PO4 )]5- (V14 ), [V15 O36 Cl]6- (V15 ) and [V18 O42 I]7- (V18 ) as inhibitors of three major multidrug resistance-linked ABC transporters: P-glycoprotein (P-gp), ABCG2 and MRP1. All of the POVs selectively inhibited P-gp. V10 and V18 were the two most promising compounds, with IC50 values of transport inhibition of 25.4 and 22.7 µm, respectively. Both compounds inhibited P-gp ATPase activity, with the same IC50 value of 1.26 µm. V10 and V18 triggered different conformational changes in the P-gp protein with time-dependent inhibition, which was confirmed using the synthesized salt of V10 with rhodamine B, RhoB-V10 . The hydrophilic nature of POVs supports the hypothesis that these compounds target an unusual ligand-binding site, opening new possibilities in the development of potent modulators of ABC transporters.

Mutational analysis of Saccharomyces cerevisiae Erf2 reveals a two-step reaction mechanism for protein palmitoylation by DHHC enzymes.

DHHC protein acyltransferases (PATs) catalyze the palmitoylation of eukaryotic proteins through an enzymatic mechanism that remains largely unexplored. In this study we have combined genetic and biochemical approaches to examine the molecular mechanism of palmitate transfer of the yeast Ras PAT, which is composed of Erf2 and Erf4. The palmitoylation reaction consists of two steps; they are autopalmitoylation of the enzyme to create a palmitoyl-Erf2 intermediate followed by the transfer of the palmitoyl moiety to the Ras substrate. Palmitoyl-CoA serves as the palmitate donor. To elucidate the kinetic properties of the Erf2·Erf4 PAT, we have developed a coupled enzyme assay that monitors the turnover of the palmitoyl-enzyme species indirectly by measuring the rate of CoASH release. Mutational analysis indicates that the DHHC motif constitutes the catalytic core of the enzyme required for autopalmitoylation and palmitoyl transfer to the Ras2 substrate. In the absence of Ras2, the palmitoyl-Erf2·Erf4 complex undergoes a cycle of hydrolysis and re-palmitoylation, implying that in the presence of palmitoyl-CoA, the complex is autopalmitoylated and competent to transfer palmitate to a protein substrate.

Identification and characterization of a new true lipase isolated through metagenomic approach.

BACKGROUND: Metagenomics, the application of molecular genomics to consortia of non-cultivated microbes, has the potential to have a substantial impact on the search for novel industrial enzymes such as esterases (carboxyl ester hydrolases, EC 3.1.1.1) and lipases (triacylglycerol lipases, EC 3.1.1.3). In the current work, a novel lipase gene was identified from a fosmid metagenomic library constructed with the "prokaryotic-enriched" DNA from a fat-contaminated soil collected from a wastewater treatment plant. RESULTS: In preliminary screening on agar containing 1% tributyrin, 2661 of the approximately 500,000 clones in the metagenomic library showed activity. Of these, 127 showed activity on agar containing 1% tricaprylin, while 32 were shown to be true lipase producers through screening on agar containing 1% triolein. The clone with the largest halo was further characterized. Its lipase gene showed 72% identity to a putative lipase of Yersinia enterocolitica subsp. palearctica Y11. The lipase, named LipC12, belongs to family I.1 of bacterial lipases, has a chaperone-independent folding, does not possess disulfide bridges and is calcium ion dependent. It is stable from pH 6 to 11 and has activity from pH 4.5 to 10, with higher activities at alkaline pH values. LipC12 is stable up to 3.7 M NaCl and from 20 to 50°C, with maximum activity at 30°C over a 1 h incubation. The pure enzyme has specific activities of 1722 U/mg and 1767 U/mg against olive oil and pig fat, respectively. Moreover, it is highly stable in organic solvents at 15% and 30% (v/v). CONCLUSIONS: The combination of the use of a fat-contaminated soil, enrichment of prokaryotic DNA and a three-step screening strategy led to a high number of lipase-producing clones in the metagenomic library. The most notable properties of the new lipase that was isolated and characterized were a high specific activity against long chain triacylglycerols, activity and stability over a wide range of pH values, good thermal stability and stability in water-miscible organic solvents and at high salt concentrations. These characteristics suggest that this lipase has potential to perform well in biocatalytic processes, such as for hydrolysis and synthesis reactions involving long-chain triacylglycerols and fatty acid esters.

Immediate microsurgical reconstruction after tumor ablation predicts survival among patients with head and neck carcinoma.

BACKGROUND: The purpose of this study was to examine whether or not immediate microvascular free flap reconstruction influences survival among treated patients suffering from oral squamous cell carcinoma (OSCC), controlling for demographic, clinical, and tumor characteristics. METHODS: 773 patients with OSCC were treated with curative intent. Two hundred and seventy-four patients were immediately reconstructed using free microsurgical flaps. All measures were collected by chart abstraction. Kaplan-Meier plots and univariate log-rank test and multivariate Cox proportional hazards regression models were used to determine the association between possible predictor variables and survival time. In addition, a randomized matched-pair analysis was conducted to compensate different patient cohorts. RESULTS: After controlling for age, tumor stage, nodal stage, and tumor grade, which were independent and dependent predictors of survival, microsurgical free flap reconstruction was associated with survival [hazard ratio (HR) = 0.66, P < 0.001, 95% confidence interval (CI), 0.52 to 0.83]. In the randomized matched-pair analysis this effect became more evident (HR = 0.58, P < 0.001, 95% CI, 0.44 to 0.75). Controlling for extent of tumor, only T3 and T4 tumors were significantly associated with survival (P < 0.001, HR = 0.46, 95% CI, 0.31 to 0.69) when the area of defect was reconstructed, whereas in T1 and T2 tumors no significance was noted (P = 0.08, HR = 0.74, 95% CI, 0.53 to 1.04). CONCLUSIONS: Reconstruction of defects, especially in patients presenting with higher tumor stages, is not associated with shorter overall survival rates, as revealed in this study, and should be considered when functional deficits due to tumor ablation can be expected postoperatively.