In doing so, a window of STI vulnerability is created during which potential pathogens including HIV enter the reproductive tract to infect host targets. “
“An expanding spectrum of acute and chronic non-infectious inflammatory diseases is uniquely responsive to IL-1β neutralization. TSA HDAC purchase IL-1β-mediated diseases are often called “auto-inflammatory” and the dominant finding is the release of the active form of IL-1β driven by endogenous molecules acting on the monocyte/macrophage. IL-1β activity is
tightly controlled and requires the conversion of the primary transcript, the inactive IL-1β precursor, to the click here active cytokine by limited proteolysis. Limited proteolysis can take place extracellularly by serine proteases, released in particular by infiltrating neutrophils or intracellularly by the cysteine protease caspase-1. Therefore, blocking IL-1β resolves inflammation regardless of how the cytokine is released from the cell or how the precursor is cleaved. Endogenous stimulants such as oxidized fatty acids and lipoproteins, high glucose
concentrations, uric acid crystals, activated complement, contents of necrotic cells, and cytokines, particularly IL-1 itself, induce the synthesis of the inactive IL-1β precursor, which awaits processing to the active form. Although bursts of IL-1β precipitate acute attacks of systemic or local inflammation, IL-1β also contributes to several SSR128129E chronic diseases. For example,
ischemic injury, such as myocardial infarction or stroke, causes acute and extensive damage, and slowly progressive inflammatory processes take place in atherosclerosis, type 2 diabetes, osteoarthritis and smoldering myeloma. Evidence for the involvement of IL-1β and the clinical results of reducing IL-1β activity in this broad spectrum of inflammatory diseases are the focus of this review. IL-1 has a long history 1; it begins with interest in the most salient manifestation of inflammation, fever. Indeed, the discovery of IL-1 as the quintessential inflammatory cytokine can be traced to the purification of the endogenous fever-producing molecule, leukocytic pyrogen, in 1977 2. Interest in this molecule increased when we reported that leukocytic pyrogen was the same molecule as lymphocyte activating factor 3, thus necessitating invention of the IL nomenclature. The term for IL-1 was assigned to the macrophage product and IL-2 for the T-cell product, even though there was no N-terminal amino acid sequence at that time that these were indeed different molecules.
1F). To analyze the interaction of LPL and calmodulin in more detail, we first analyzed the subcellular localization of calmodulin in T cells. In unstimulated cells that did not form a contact with APC, calmodulin and LPL were both equally distributed throughout the cytoplasm (Fig. 3A). Upon T-cell stimulation via superantigen-loaded APC for 45 min, in 48.09±0.16% of the T-cell/APC couples calmodulin translocated to the contact zone between T cells and APC where it colocalized with LPL. We reinforced this quantification by calculating the area corrected calmodulin
content within the contact zone of T cells and APC and subtracted an area corrected protein content within T-cell/T-cell and APC/APC contact zones 26. This analysis confirmed this website that calmodulin and LPL accumulated in the T-cell/APC contact zone (Supporting
Information Fig. 2). The interaction of calmodulin and LPL was shown by calmodulin pull-down experiments (Fig. 3B). A binding of LPL to calmodulin could only be seen in the presence of EGTA. Note that the calcium/calmodulin dependent ZD1839 purchase kinase type IV (CamKIV) was efficiently precipitated with calmodulin in the presence of calcium, whereas EGTA inhibited this interaction (Fig. 3C). These experiments explain at the same time the interaction of LPL and calmodulin in unstimulated cells, in which no calcium signal was induced (Fig. 3B). Although binding of LPL to calmodulin in the absence of calcium was Erastin cost unexpected, such interactions to calcium-free calmodulin (Apocalmodulin/ApoCam) were described for several proteins (reviewed in 27). We next analyzed whether inhibition
of calmodulin through the calmodulin antagonist W7 would lead to reduced LPL accumulation in the IS. MIFC analysis demonstrates that LPL recruitment was indeed diminished in the presence of W7 (Fig. 4A and B). The degree of inhibition is reminiscence of that observed for ΔCBD-LPL. Importantly, W7 also inhibited recruitment of the pSMAC-marker LFA-1, but not of the cSMAC-marker CD3 in the contact zone. The selective effects of W7 on the accumulation of pSMAC-markers in the IS was independently confirmed using LSM and EGFP-tagged LPL, F-actin or PKCΘ and staining of endogenous LFA-1 (Supporting Information Fig. 3). Also in these experiments the enrichment of LPL and the pSMAC-markers actin and LFA–1 were inhibited by W7, whereas it had no effect on the accumulation of the cSMAC-marker PKCθ in the IS. The reduced accumulation of ΔCBD-LPL (Fig. 1F), or of wt-LPL in the presence of calmodulin antagonists (Supporting Information Fig. 3) may be explained either by a diminished initial relocalization or a reduced maintenance of LPL in the contact zone. To discriminate between the two possibilities, we analyzed the relocalization kinetics and mean duration of wt-LPL and ΔCBD-LPL in the contact zone using time-lapse video-microscopy (TLV).
3A), and the PMNs still caused dyshesion of the cell layer (Table 1). The PMN-mediated dyshesion was greatly reduced in the presence of the protease inhibitor α1-antitrypsin, or peptide substrates of the PMN elastase, or a selective elastase inhibitor, indicating a major contribution of elastase (data summarized in Table 2). As expected, purified PMN elastase also caused a dyshesion of the tumor cells (data summarized in Table 1), which again was reversible, and could be inhibited by high serum concentrations
(data not shown). Pancreas elastase when used in comparable concentrations did not cause a dyshesion of cells after 2 h; only concentrations of more than 5 μg/mL and prolonged Idasanutlin order incubation (up to 4 h) resulted in some dyshesion. A likely target for elastase is the adhesion molecule E-cadherin that is expressed by T3M4 cells as shown by indirect immunofluorescence of confluent cell layers and by flow cytometry of dispersed cells (Figs 3B and C and Fig. 4A and D). Following exposure of T3M4 to PMNs or to PFA-fixed PMMs,
surface expression of E-cadherin was reduced (Fig. 3B). The loss of E-cadherin amounted to 45.9 ± 17.7% (mean ± SD of n = 5). Alpha-1-antitrypsin prevented the PMN-induced loss of E-cadherin, as did the elastase inhibitor or the respective substrate (examples in Fig. 3C). Also isolated PMN elastase caused a reduction of E-cadherin surface expression (example in Fig. 4C). By using an Ab that binds to LDK378 nmr the N-terminus of E-cadherin, the surface expression
was reduced, on average by 33.55 ± 19.2% within 2 h (mean ± SD of n = 7) (example in Fig. 4F). A mAb to E-cadherin that binds to a domain near the membrane showed no differences in binding to T3M4 compared with that of elastase-treated T3M4 cells (data not shown). The flow cytometry forward-sideward scatter image revealed that the majority of T3M4 cells were viable after the elastase Staurosporine solubility dmso treatment. The data so far implied an elastase-mediated loss of E-cadherin from the surface. Indeed, when T3M4 were treated with elastase for 2 h, E-cadherin within the membrane fraction was greatly reduced, but was conserved in the cytoplasm, as shown by western blotting (Fig. 4G). Since the Ab is directed to N-terminal region of the molecule, the data indicate cleavage of E-cadherin. Furthermore, a cleavage product of E-cadherin was detected in cell culture supernatants by ELISA. In untreated cells, a cleavage product concentration of 18.7 pg/mL was detected compared with one of 198.3 pg/mL in the elastase-treated cells (mean of three experiments performed in duplicates; p = 0.017 calculated by ANOVA). E-cadherin was not detectable in supernatants of MiaPaCa-2. Transfection of T3M4 with specific siRNA reduced the E-cadherin surface expression by more than 90% when measured after 48 h (Fig. 5A and B).
1A and Supporting Information Fig. 1). Supernatants from PerC (high percentage of B-1 cells) had very low levels of IgM, as were those from PLNs (very low frequencies of B-1 cells), which had none. In contrast, natural IgM levels were high in cultures of spleen cells. We also measured strong IgM production in BM cultures, a site that has not previously
shown to contain any mature B-1 cells (Fig. 1A). ELISPOT analysis confirmed spleen and BM as major sites of spontaneous IgM-secreting B cells (Fig. 1C). ELISPOT of PerC showed an unusual pattern. While we could discern many, very small spots, also noted in 31, they were not sufficiently distinguishable from the background Panobinostat to allow accurate counting. This is in contrast to ELISPOT analysis of PLNs that completely
lacked spots of any size (Fig. 1B and data not shown). Given Daporinad cell line that PerC supernatants contained very low levels of secreted IgM (Fig. 1A), we conclude that PerC cells might produce extremely small amounts of IgM per cell, whereas PLNs are completely devoid of IgM-secreting cells. Correlating the IgM concentrations in the supernatants of BM and spleen cultures with the frequency-measurements done by ELISPOT indicated that IgM secretion per antibody forming cell (AFC) was 2- to 3-fold higher in BM than in the spleen (Fig. 1D). To assess whether spontaneous IgM secretion in BALB/c mice is “natural” non-infection-induced IgM, we studied a known specificity of natural IgM, i.e. its binding to influenza virus 26. Antiviral natural antibodies are produced by the B-1 cell subset in non-infected mice 5, 26. Spleen and BM both contained influenza virus-binding IgM-secreting cells at frequencies of 6 and buy Staurosporine 15% of total IgM AFCs, respectively (Fig. 1E). Thus, at least some of the spontaneous IgM measured in the spleen and BM of BALB/c mice is “natural” IgM, and thus likely generated by B-1 cells. Activated B-2 cells down-regulate surface IgM expression during plasma cell differentiation 40. To determine whether differentiation to non-isotype class-switched,
IgM-secreting cells also involves down-regulation of surface IgM and to further characterize spontaneous IgM-secreting cells, we FACS-separated B-cell subsets based on combinations of surface IgM expression and other surface molecules. Sorting splenic CD19+ B cells based on surface expression of IgM and CD43, a marker expressed by B-1 cells and plasma blasts 41, revealed that IgM+CD43+ B cells contain the highest frequencies IgM-secreting B cells (Fig. 2A). A small proportion of IgM+CD43− subset also generated IgM AFC (Fig. 2A), possibly due to contaminating IgM+CD43+ cells and/or cells expressing very low levels of CD43. Very few IgM-secreting cells were detected among CD19− cells (data not shown). Thus, spontaneous IgM-secreting B cells do not down-regulate surface IgM or CD19. Consistent with data from the spleen, surface IgM-expressing B cells comprised the most of IgM-secreting cells expressed in the BM (Fig. 2B, top).
Because TRECs are stable within the original T cell and do not duplicate during mitosis they are diluted out in the periphery with antigen-driven or homeostatic
cell division . However, in healthy individuals, only homeostatic proliferation of naive T cells is likely to affect peripheral T cell TREC content significantly, as antigen-induced T cell proliferation will, to the most extent, affect memory/antigen-primed T cells with very minute amounts of TRECs, and thus not the population of RTE. Nevertheless, to exclude that the reduced TREC concentrations in peripheral blood lymphocytes from several UC patients, as well as CD patients, were caused by an increased peripheral T cell turnover we determined the frequencies C59 wnt research buy of proliferating T lymphocytes, detected as Ki67+CD3+ T lymphocytes, and found the prevalence to be equivalent in IBD patients and healthy individuals. Supporting the notion that the reduced TREC levels in peripheral blood T cells in several IBD patients are not caused by extensive proliferation
was also the finding of comparable frequencies of CD45RA+ as well as CD62L+ T lymphocytes in peripheral blood from IBD patients and healthy individuals. Thus, a likely explanation to the reduced TREC levels in peripheral blood from IBD patients could be see more enhanced migration of RTE from the blood to the inflamed mucosa, purging the peripheral blood of this population. The purpose of separating the integrin β7+ lymphocytes in peripheral blood was to analyse if there was a direct recruitment of gut homing T cells from the thymus.
The fact that the integrin β7+ population did not differ from unseparated lymphocytes regarding TREC content indicate that the majority of peripheral blood lymphocytes have divided, irrespective of integrin of β7+ expression. Although the frequency SPTLC1 of proliferating T lymphocytes was not estimated in the intestinal mucosa, the proliferation rate in UC patients would be increased rather than decreased compared to controls, due to the chronic inflammation. Thus, if anything, we are underestimating the amount of TRECs in mucosal lymphocytes of IBD patients by not expressing it relative to the proliferation rate of the T lymphocytes. Splitting the patient group into those with active disease versus those with inactive disease demonstrated that this recruitment was not limited to the actively inflamed mucosa, indicating a constant influx of RTE to the intestinal mucosa in UC patients also during remission. It would be very interesting to investigate the role of these RTE for the disease course, e.g.
o. and i.p. challenge regarding the cross-allergens (peanut, soy and fenugreek). click here Mice challenged p.o. with fenugreek and i.p. with soy in the fenugreek model (Fig. 1D) showed significantly higher MMCP-1 levels than controls and peanut challenged mice, while fenugreek-sensitized mice challenged with lupin showed higher levels than the controls only. Peanut challenged mice and unchallenged mice did not show significantly higher levels than control mice. In summary, mice challenged with the primary allergen displayed significantly higher levels of MMCP-1 than the other groups. Mice challenged with a potentially cross-reactive allergen showed higher levels of MMCP-1 than control mice, however,
the levels were comparable with mice that were only immunized and not challenged. There was a significant correlation between the anaphylaxis score and MMCP-1 with a Spearman’s ρ rank correlation coefficient of 0.417 for the lupin model, 0.448 for the fenugreek model and 0.409 for both models combined,
P ≤ 0.001. The involvement of IgE in the cross-allergic reactions was studied with different methods in the two models. In the lupin model, we used the PCA-test to investigate possible cross-reactions by injecting legumes other than lupin i.v. but no reactions could be observed in this test. In the fenugreek model, total IgE was measured in all mice both before and after challenge (Fig. 2A). Comparing total IgE levels before and after challenge in each group according to allergen challenge (t-test) revealed significant MK-1775 clinical trial differences in fenugreek challenged mice (P = 0.002), peanut challenged mice (P = 0.039) and lupin challenged mice (P = 0.047), but
not in soy challenged mice. Correspondingly, in the analysis of the groups before challenge, all groups had higher IgE levels than control mice, while total IgE levels after challenge with fenugreek, peanut or lupin were not significantly different from the controls. In Western Florfenicol blotting (Fig. 2B), we were only able to detect IgE binding to lupin in sera from mice immunized with lupin, where several IgE binding bands were revealed in the range from about 50 kDa to about 70 kDa. These sera also showed binding to a fenugreek band of approximately 50 kDa (Fig. 2B, arrow) and a band of approximately 60 kDa. As the latter band also could be seen with sera from naïve mice, this is presumably unspecific binding that might be due to the presence of lectin in the extract. Mice immunized with fenugreek showed IgE binding to fenugreek only, with several bands revealed between 50 kDa and about 150 kDa. No binding to peanut, soy or OVA was detected in any of the blots (not shown). Preincubation with the primary allergen inhibited all IgE binding, while potentially cross-reacting allergens did not inhibit the IgE binding substantially (not shown). Immunized mice showed high levels of IgG1 that were completely inhibited by preincubation with the primary allergen in both models (Fig. 3). In the lupin model (Fig.
9,15–18 Further studies are needed to increase our understanding of the roles of eosinophils and IL-5 in inflammatory responses and other diseases in which hypereosinophilia occurs. The differential migration of eosinophils versus neutrophils to thyroids of IFN-γ−/− and WT mice during the development of G-EAT offers a unique opportunity to examine the role of eosinophil trafficking to sites of inflammation and to investigate the potential role of these
cells in the induction and resolution of inflammation. Neutralization of IL-5 markedly inhibited migration of eosinophils to thyroids of IFN-γ−/− mice during development of G-EAT. However, IL-5 neutralization had no effect on the severity or rate of resolution of inflammation in G-EAT, suggesting that eosinophil migration has no apparent pathogenic role in G-EAT. WT and IFN-γ−/− DBA/1 mice were produced Metabolism inhibitor in our animal facilities at the University of Missouri as previously described.6–8 Both male and female mice (6–10 weeks old) were used. G-EAT was induced as previously described.1,5 Briefly, mice were injected intravenously
(i.v.) twice at 10-day intervals with 150 μg of MTg3 and 15 μg of lipopolysaccharide (LPS) (Escherichia coli 011:B4; Sigma Chemical Co., St Louis, MO). Seven days later, donor spleen cells were re-stimulated in vitro MLN8237 purchase with 25 μg/ml MTg and 5 ng/ml IL-12.1 Cells were harvested after 72 hr and washed twice, and 3·5 × 107 cells were transferred i.v. to 500-Rad irradiated
syngeneic recipients. Anti-IL-5 was purified from culture supernatants of the anti-IL-5-producing hybridoma TRFK-5 (provided by Dr Robert Coffman, DNAX Research Institute, Palo Alto, CA, USA) using protein G. IFN-γ−/− recipients of IFN-γ−/− donor cells were given 300 μg of anti-IL-5 intraperitoneally (i.p.) or rat immunoglobulin G (control IgG) every 4 days beginning on the Calpain day of cell transfer until euthanasia. WT recipients of WT donor cells were used for comparison. Thyroids were removed from groups of five or six recipient mice 20 days (peak of disease) or 40–50 days (fibrosis versus resolution) after cell transfer.1–6 Thyroids were fixed in formalin, sectioned and stained with haematoxylin and eosin (H&E), and scored quantitatively for G-EAT severity (the extent of inflammatory cell infiltration and thyroid follicle destruction) using a scale of 1+ to 5+, as described previously.6 1+ thyroiditis is defined as an infiltrate of at least 125 cells in one or several foci; 2+ is 10–20 foci of cellular infiltration involving up to 25% of the gland; 3+ indicates that 25–50% of the gland is infiltrated; 4+ indicates that > 50% of the gland is destroyed by infiltrating inflammatory cells; and 5+ indicates virtually complete destruction of the thyroid with few or no remaining follicles. Thyroid lesions were also evaluated qualitatively.
ELISPOT analysis of antigen-specific SB203580 research buy IFN-γ production by CD8+ T cells has been previously described 44. Lymphocytes were harvested from the spleens of WT BALB/c mice and sorted for B220+Thy-1.2−120G8− cells on a FACSAria. 3×106 purified (>98% purity by FACS) B cells were adoptively transferred by intravenous injection into naïve BALB/c mice prior to adoptive transfer of TCR-Tg cells and immunization. Data
analysis and presentation were performed using Prism (GraphPad Software). This work was supported by NIH grant AI44375. M. G. O. was supported by a fellowship from the Malaria Research Institute. The authors are grateful for the support of the Bloomberg Family Foundation. PDL-1 blocking antibodies were kindly provided by Lieping Chen. Conflict of interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available
as submitted by the authors. “
“An association study of a cohort of 177 Sudanese patients infected with Schistosoma mansoni [82 (46%) males and 95 (54%) females] was conducted to evaluate the factors controlling the regression of liver fibrosis 39 months after treatment with praziquantel using ultrasound evaluation. Periportal fibrosis (PPF) was regressed in 63 (35.6%) patients, while the disease progressed to higher grades in 24 (13.6%) patients. The grade of PPF did not change in 90 (50.8%) patients.
The mean values of portal vein diameter, splenic vein Torin 1 in vivo diameter and index liver size in subjects in whom PPF regressed after treatment were significantly lower than in subjects in whom the disease Mannose-binding protein-associated serine protease was progressed (P<0.0001, P=0.031 and P=0.003, respectively). The progression of hepatic fibrosis in males (15, 8.5%) was greater than that in females (9, 5.1%). Patients with regression or progression phenotypes tend to cluster in certain families. Our study indicated that regression, progression and stabilization of PPF after praziquantel therapy is controlled by gender, age, grade of fibrosis and possibly inherited factors. Human schistosomiasis is a major health problem in many countries including Sudan. The disease is chronic and debilitating, and remains one of the most prevalent parasitic infections in tropical and subtropical environments (WHO, 1993). Despite control efforts in a number of countries, 200 millions of people are still infected, and 10% develop severe disease with Symmers fibrosis (WHO, 1998). Mortality due to Schistosoma mansoni infections is mainly the consequence of portal hypertension that is caused by hepatic periportal fibrosis (PPF) (Dessein et al., 1999a). In PPF, varying degrees of inflammation and collagen surrounding the portal vein and its tributaries are observed (Homeida et al., 1991).
It is possible that under different conditions CD8+CD28− T cells with regulatory properties are more prominent, and under these circumstances the use of MSC should be reconsidered. IL-15 is a cytokine that promotes CD8+CD28− T cell proliferation . Interestingly, IL-15, next to IL-7, is crucial for the homeostatic maintenance of T cells in the absence of antigenic stimuli and expedites the loss of CD28 expression . During normal exposure to antigen CD28 expression is transiently reduced but returns quickly to basal expression levels. Repeated NVP-BGJ398 chemical structure antigen exposure due to the natural ageing process, viral infections or viral reactivation
in immunocompromised patients causes a decline in CD28 expression, leading eventually to total loss of CD28. Surprisingly, we found that in our setting CD28+ T cells did not lose CD28 during allogeneic stimulation with PBMC, confirming that extended
rounds of antigen exposure are required to initiate reduction of CD28. Permanent decline of CD28 expression entails telomere AZD8055 nmr shortening and reduction of telomerase activity and is attributed to a defect in the CD28 promotor leading to transcriptional inactivation [50-54]. We, however, found that CD8+ T cells that were initially CD28− gained CD28 expression during allogeneic stimulation with PBMCs. Reinduction of CD28 expression in CD4+CD28− T cells is a known phenomenon and only possible until CD28− T cells have reached terminal differentiation. Warrington et al. described that combined stimulation of T cell receptor (TCR) and IL-12 receptor restored CD28 transcription and protein expression, Metalloexopeptidase while single stimulation of either the TCR or the IL-12 receptor was not sufficient . IL-12 is produced by phagocytic cells, B cells and other antigen-presenting cells  and therefore potentially contributes to the CD28 re-expression in originally CD8+CD28− T cells in MLR. Although CD28 expression can be influenced up to a certain stage during T cell differentiation, MSC did not affect the immunophenotypical changes of CD8+CD28− T cells, nor did they cause loss of CD28 expression
in CD8+CD28+ T cells. Further, we found that MSC did not induce apoptosis in CD8+CD28− T cells, despite their ability to express Fas ligand (FasL) or to initiate the programmed death (PD)-1/PD-ligand 1 (PD-L1) pathway [57, 58]. These observations indicate that MSC solely have an anti-proliferative effect on CD8+CD28− T cells. Co-administration of MSC with other immunosuppressive drugs is not always encouraged; agents such as tacrolimus, mammalian target of rapamycin (mTor) inhibitor rapamycin and rabbit anti-thymocyte globulin (rATG) negatively affect the suppressive capacity of MSC in vitro [59-61]. At same time, MSC are able to reduce the efficacy of tacrolimus and rapamycin [59, 60]. As MSC lack expression of the CTLA-4 ligands CD80 and CD86, it was not surprising that belatacept did not diminish MSC function .
These findings highlight the considerable variation in the number of SIEV trunks as well as their source of regional drainage, and show the importance of consideration of such variation. © 2011 Wiley-Liss, Inc. Microsurgery, 2011. “
“So far, predictive models with individualized estimates of prognosis for patients with peripheral nerve injuries are lacking.
Our group has previously shown the prognostic value of a standardized scoring system by examining the functional outcome after acute, sharp complete laceration and repair of median and/or ulnar nerves at various levels in the forearm. In the present study, we further explore the potential mathematical model in order to devise an effective prognostic scoring system. We retrospectively collected medical record data of SP600125 nmr 73 cases with a peripheral nerve injury in the upper extremity in order to estimate which patients would return to work, and what time was necessary to return to the pre-injury work. Postoperative assessment followed the protocol described by Rosén and Lundborg. We found that
return to pre-injury work can be predicted with high sensitivity (100%) and specificity (95%) using the total numerical score of the Rosén and Lundborg protocol at the third follow-up interval (TS3) as well as the difference between the TS3 and the total score at second follow-up interval (TS2). In addition, the factors age and type of injured nerve (median, ulnar, or combined) can determine the time of return to Fludarabine work based on a mathematical
model. This prognostic protocol can be a useful tool to provide information about the functional and social prospects of the patients with these types of injuries. © 2012 Wiley Periodicals, Inc. Microsurgery, 2013. “
“Introduction: The originally described distally based sural flap technique has a risk of partial or total flap necrosis as high as 25%. The purpose of this study was to compare the medicinal leech therapy (MLT) with venous catheterization (VC) for blood find more volume removal, infection, wound dehiscence, and flap necrosis in the distally based sural flap with venous congestion. Patients and methods: Fifty-six conventional distally based sural flaps with venous congestion during reconstructive surgeries were randomly divided into two groups, MLT group and VC group. The results of comparisons were analyzed using SPSS software (SPSS for Windows Ver.11.5). Results: There were significant differences in terms of the average volume of removed blood (53.6cc vs.172.2cc), infection (10.7% vs. 34.6%), wound dehiscence (10.7% vs. 42.3%), flap necrosis (3.6% vs. 19.2%), and nursing (7.8 vs. 5.19) and patient’s satisfaction (8.03 vs. 5.6) in the VC group and MLT group, respectively. Although local heparin irrigation was performed in the VC group, the catheter was exchanged in 10 patients due to obstruction by clot.