Transplantation of NSCs to replace degenerated neurons or genetically modified NSCs producing neurotrophic factors have been used to protect striatal neurons against excitotoxic insults.[62] At present, little is known regarding whether implantation of NSCs prior to neuropathological Pritelivir cost damage could alter the progressive degeneration of striatal neurons and motor

deficits that occur in HD. This question is important since the genetic study of HD gene mutation[63] and neuroimaging can provide details on factors involved in the progression of HD,[64, 65] suggesting early intervention using brain transplantation could be effective in “pre-clinical” HD patients carrying the mutant HD gene. We have investigated the effectiveness of proactive transplantation of human NSCs into rat striatum of an HD rat model prior to lesion Rapamycin nmr formation and.demonstrated significantly improved motor performance and increased resistance to striatal neuron damage compared with control sham injections.[66] The neuroprotection provided by the proactive transplantation of human NSCs in the rat model of HD appears to be contributed by brain-derived neurotrophic factor (BDNF) secreted by the transplanted human NSCs. Rodents and primates with lesions

of the striatum induced by excitotoxic kainic acid (KA), or quinolinic acid (QA) have been used to simulate HD in animals and to test efficacy of experimental therapeutics on neural transplantation.[67] Excitotoxic animal models induced by QA, which stimulates glutamate receptors, and resembles the histopathologic characteristics of HD patients, cAMP were utilized for cell therapy with mouse embryonic

stem cells, mouse neural stem cells, mouse bone marrow mesenchymal stem cells and primary human neural precursor cells, and resulted in varying degrees of clinical improvement.[68-73] We have recently injected human NSCs intravenously in QA-HD model rats and demonstrated functional recovery in HD animals.[72, 73] The systemic transplantation of NSCs via an intravascular route is probably the least invasive method of cell administration.[73] Neural cell transplantation into striatum requires an invasive surgical technique using a stereotaxic frame. Non-invasive transplantation via intravenous routes, if effective in humans, is much more attractive. Systemic administration of 3-nitropropionic acid (3-NP) in rodents leads to metabolic impairment and gradual neurodegeneration of the basal ganglia with behavioral deficits similar to those associated with HD,[74, 75] and murine and human NSCs have been transplanted in the brain of 3-NP-HD animal models.[66, 76] The compound 3-NP is a toxin which inhibits the mitochondrial enzyme succinate dehydrogenase (SDH) and tricarboxylic acid (TCA) cycle, thereby interfering with the synthesis of ATP.[77] We have investigated the effectiveness of transplantation of human NSCs into adult rat striatum prior to striatal damage induced by 3-NP toxin.

T cells were purified with negative magnetic bead selection using the “Pan T cell isolation Kit” (Miltenyi Biotech, Bergisch Gladbach, Germany). Antibodies that were used in this study were specific for following markers: CD3ε,

LFA-1, CD2 (BD-Bioscience, Heidelberg, Germany), calmodulin (Zymed, Munich, Germany) and LPL 17. W7 was from Calbiochem (Darmstadt, Germany), Hoechst 33342 from Invitrogen (Karlsruhe, Germany), and BPB and Cytochalasin D from Staurosporine Sigma-Aldrich (Taufkirchen, Germany). For cDNA transfection into T cells, the “Human T Cell Nucleofector™ Kit” (Amaxa Biosystems, Cologne, Germany) was used. For the siRNA approaches, cells were electroporated with LPL-specific or control siRNA (Dharmacon, Lafayette, IN, USA). Thereafter, cells were stimulated with 2 μg/mL PHA for 16 h. The PHA was removed and the cells were transferred in medium containing 25 U/mL IL-2. After 2 days incubation cells were electroporated again and incubated for another 2 days at 37°C. The IL-2 was Doxorubicin datasheet removed and the cells were incubated in medium without IL-2 for 24 h prior to further experiments. Conjugates were formed between T cells and superantigen-loaded Raji B cells as described 17. Subcellular localization of proteins was determined by immunofluorescence and subsequent analysis with confocal LSM, TLV 17 or MIFC. Cells were stimulated and stained with

fluorescently labeled antibodies and nuclear dyes (Hoechst) as indicated. Data acquisition was performed with an ImageStream (IS100) and data were analyzed with IDEAS 3.0 (Amnis, Seattle, WA, USA). To find the contact zone between T cells and APC, a Hoechst-dependent

valley mask was defined between T-cell/APC couples and combined with a T-cell mask (Supporting Information Fig. 1). Thereafter, protein accumulation was calculated as ratio between the fluorescence intensities Lck of the respective protein in the IS- and T-cell mask. The data were controlled by manually evaluation of 100 cells per sample. The size of the IS was calculated with the major axis feature and the size of T cells with the diameter feature on the T-cell mask. Both algorithms return the results in microns. The F-actin content in the cells was calculated as mean fluorescence intensity of the phalloidin staining within the T-cell mask. The plasmid pEGFP containing the wt-LPL cDNA was generated in our own laboratory 17. The plasmid was used to create mutants of LPL as follows: the two EF-hand calcium-binding domains of LPL at positions 22–27 (ΔEF1-LPL) and 62–73 (ΔEF2-LPL) or both calcium-binding domains (ΔEF1/2-LPL) 36, 37 were deleted using the QuickChange site-directed Mutagenesis XL Kit (Stratagene, La Jolla, CA, USA) according to the manufacturer’s instructions. The actin-binding domains at position 120–627 were removed by PCR amplification of the first 120 aa, which were subsequently introduced in pEGFP via EcoRI and XhoI.

Following lipopolysaccharide overnight treatment, BMDCs treated had a mature BMDC phenotype based on MHC class II high, CD40 and CD86 expression (P<0.05). To evaluate how HK or IR Brucella affected DC maturation, immature BMDCs were stimulated with either HK or IR rough vaccine strain RB51 or smooth pathogenic strain 2308 at 1 : 10 (DC : Brucella) or 1 : 100 CFU equivalents. Additional controls included media-only and lipopolysaccharide-treated BMDCs as well as live strain RB51- and 2308-infected (at MOI 1 : 10 or 1 : 100) BMDCs. Immature BMDCs treated overnight with media alone retained their immature phenotype with a reduced surface expression of MHC

class II and CD40, CD86 costimulatory markers MK-1775 clinical trial compared with lipopolysaccharide (Fig. 1a). Immature BMDCs stimulated with HK strain RB51 (HKRB51) at both 1 : 10 (P=0.0542) (not shown) and 1 : 100 (P=0.0018) CFU equivalents showed significant upregulation of MHC class II high expression compared with the media control (Fig. 1b). In addition, at corresponding doses of 1 : 10 and 1 : 100, HKRB51 had a higher mean (not statistically significant) MHC class II high expression than selleck HK strain 2308 (HK2308)-stimulated BMDCs (Fig. 1b). HK strain 2308 1 : 100 did not induce significant upregulation of MHC class II expression.

Furthermore, both HKRB51- and HK2308-stimulated DCs showed a nonsignificant dose-related increase in MHC class II high expression at 1 : 100 compared with 1 : 10. However, live strain RB51-infected BMDCs had greater MHC class II high expression than HKRB51 (not significant) and HK2308 (P≤0.05) at the corresponding doses (Fig. 1b). IR strain

RB51 (IRRB51) induced a relatively higher, but not significantly MHC class II high expression than IR strain 2308 (IR2308)-stimulated BMDCs at the corresponding doses. At 1 : 100, IRRB51 induced significantly (P≤0.05) higher MHC class II high expression than media (Fig. 1b). Moreover, IRRB51-induced mean DC–MHC class II high expression level was lower (not DOK2 significant) than that induced by HKRB51 at the respective doses (Fig. 1b). At both MOIs, live strain RB51 induced a higher MHC class II high expression on BMDCs compared with IRRB5,1 with significant differences (P≤0.05) at MOI 1 : 100 (Fig. 1b). Live strain RB51 at 1 : 100 also induced a significantly higher (P<0.05) MHC class II high expression than live strain 2308 at the same dose (Fig 1b). The expression levels of costimulatory molecules CD40 and CD86 (independent and coexpression) were also analyzed to assess the effect of live vs. HK or IR Brucella on DC maturation. Figure 1c shows CD40 expression on live, HK and IR Brucella-infected BMDCs. Only live, but not HK or IR, strain RB51-infected BMDCs at MOI 1 : 100 induced a significantly higher CD40 expression than the media control (P≤0.05). On comparing CD40 and CD86 expression, the results were similar.

We found that both T conventional (Tconv; defined as FACS-sorted CD4+CD25−) and Tregs produced CXCL8 at similar concentrations (Fig. 1B and C) even in the absence

of TCR activation, suggesting that like endothelial cells, T cells may have preformed stores of CXCL8 15 that are released upon the shear stress of cell sorting. Notably, CXCL8 production by CD25− and CD25hi T cells was not restricted to cells with a naïve (CD45RA+) or memory (CD45RA−) phenotype. Similar results were obtained when cells were stimulated in the presence of exogenous IL-2 (data not shown). In parallel, we analyzed production of IFN-γ or IL-17 and confirmed that the CD25hiCD45RA− Tregs produce a significant amount of IL-17, and that neither CD25hiCD45RA− nor CD25hiCD45RA+ Tregs produced IFN-γ (Fig. 1B). These findings indicate that CD4+CD25hi Tregs produce CXCL8 irrespective of whether they are naïve or memory Panobinostat purchase cells and that this finding is not the result of contaminating IL-17-secreting cells. Isolation of cells on the basis of CD25, even in conjunction with other markers such as CD45, does not necessarily result in a homogeneous population of FOXP3+ cells. Therefore, to further confirm

that Tregs produce this chemokine, CXCL8 production was analyzed by intracellular staining. Ex vivo CD4+ T cells were stimulated with PMA/ionomycin for 6 h and CXCL8 producing cells were detected in both the FOXP3+ and FOXP3− populations (Fig. 1D and E). On average, 28.1%±1.0 (n=4, average±SEM) of stimulated CD4+FOXP3− T cells and 25.3%±4.1 (n=4) of stimulated CD4+FOXP3+ T cells were CXCL8+ (Fig. selleck kinase inhibitor 1E). To further confirm these data, as well as to determine the cytokine profile of these CXCL8+ T cells, naïve and memory Tconv and Tregs were sorted, expanded, and analyzed by intracellular staining. As shown in Fig. 1F and Supporting Information Fig. 1A and B, on average 12.8%±1.6 of FOXP3+CD45RA+ Tregs and 19.8%±2.6 of FOXP3+CD45RA− Tregs expressed CXCL8. Neither

the CD45RA+CXCL8+ nor the CD45RA−CXCL8+ Treg populations co-expressed significant levels of IFN-γ or IL-17, further confirming that Docetaxel mouse it is indeed the naturally occurring FOXP3+ Tregs that express CXCL8. A summary of CXCL8, IFN-γ, and IL-17 expression from expanded populations is seen in Supporting Information Table 2. To confirm whether FOXP3 directly regulates CXCL8 production, we investigated whether ectopic expression of FOXP3, which is known to reprogram Tconv cells into Tregs 16, modulates CXCL8 production. CD4+ T cells transduced with FOXP3 produced significantly more CXCL8 compared to control transduced cells, with the expected parallel suppression of IFN-γ production (Fig. 1G). Furthermore, FOXP3 directly transactivated the CXCL8 promoter, as evidenced by transient transfections using a CXCL8-promoter reporter construct (Fig. 1H). Together, these data conclusively demonstrate that FOXP3+ cells produce CXCL8 and indicate that FOXP3 directly regulates CXCL8 gene expression.

Purified B cells were cultured for 3 days and stained with 5 μg/mL propidium iodide (PI; Invitrogen) or TUNEL (Roche, Switzerland) according to the manufacturer’s recommendations. The cells were analyzed on a FACS Calibur (BD). Supernatants were collected from naive and memory B cells grown for 7 days

(0.2×106 cells/500 μL in 48-well plates). Secreted Igs were measured by Human IgA, IgM and IgG ELISA Quantitation Sets from Bethyl Laboratories (TX, USA). Absorbance was measured MAPK Inhibitor Library concentration by a Sunrise Plate Reader (Tecan, Switzerland) set at 450 nm. All labeling reactions were performed by incubating cells with Abs for 30 min at 4°C. When an unconjugated primary Ab was used, the cells were washed twice before incubation with the secondary Ab. The cells were analyzed on a FACS Calibur Flow Cytometer, LSR II or FACS Canto (all from BD). Data were collected using FACS Diva software whereas analysis was performed using FlowJo (Tree Star, OR, USA) or Cytobank software (www.cytobank.org). CD19+CD27− naive and CD19+CD27+ memory B cells were obtained by staining CD19+ selected B cells from peripheral blood with anti-CD19 PECy5 and anti-CD27 PE mAbs and sorting on a FACS DiVa or FACS Aria Flow Cytometer (BD). We did not divide between Selleck Roxadustat switched memory and IgM-memory B cells, but grouped them together as one population. Different subpopulations

from tonsils were obtained by staining the single-cell suspension with anti-CD38 FITC, anti-CD19 PE, anti-IgD PerCPCy5.5, anti-CD5 PECy7 and CD27 allophycocyanin and sorting the following Mirabegron populations: naive B cells (CD19+IgD+CD38−CD27−CD5−), memory B cells (CD19+IgD−CD38−CD27+CD5−), GC B cells (CD19+IgD−CD38+CD27−CD5−) and non-B cells (CD19−). Cells were stimulated for 1 h or as indicated, before they were lysed in Tris lysis buffer as described previously 55. Cell lysates were electrophoresed using 10% SDS-polyacrylamide gels (Pierce, IL, USA) and transferred to PVDF membranes

(Millipore, MA, USA). The membranes were blocked for 60 min with 5% BSA (Sigma-Aldrich) with TBST before they were incubated with primary Abs overnight. After washing in TBST, the membranes were incubated for 60 min with HRP-conjugated anti-rabbit or anti-goat IgG Ab (Dako). Protein bands were visualized by the ECL or ECL-plus detection system (GE Healthcare, NJ, USA). Protein expression was quantified by scanning hyperfilms and using Quantity One Software (Bio-Rad, CA, USA). Cells were fixed in 4% PFA (Electron Microscopy Sciences, PA, USA) in PBS, washed in PBS and permeabilized in 90% methanol in PBS at −20°C. After washing in PBS, cells were incubated in blocking buffer (1 mg/mL human γ-globulin (Sigma) in 0.9% NaCl) at room temperature for 30 min, followed by incubation with primary Abs (diluted in blocking buffer) in a humid chamber at 4°C overnight.

Microsurgery, 2011. “
“Introduction: Microsurgical lower extremity flap reconstruction provides a valuable option for soft tissue reconstruction in comorbid patients. Limb salvage with flap reconstruction can result in limb length preservation. Despite this, few Selleckchem MK-2206 studies have examined the impact of salvage on patient-centered metrics in this cohort of patients. Therefore, we investigated quality of life and patient satisfaction following microsurgical

lower extremity reconstruction in this high-risk patient population. Factors that resulted in improved patient-centered outcomes were also identified. Methods: A retrospective review was conducted of all patients who had lower PLX-4720 supplier extremity free flap reconstruction (FFR) following lower extremity wounds. High-risk patients were identified as having multiple comorbidities and chronic wounds. Patients with traumatic wounds were excluded from analysis. Quality of life was evaluated with the Short Form-12 (SF-12) validated survey. Phone interviews were conducted for survey evaluations. Results: From 2005 to 2010, 57 patients had lower extremity flap reconstruction that met the inclusion criteria. Average follow-up was 236.6 weeks (range, 111–461). Comorbidities included diabetes (36%),

PVD (24.6%), and ESRD (7%). Limb length preservation and ambulation occurred in 82.5% (47/57). Revisional surgery occurred in 33.3% (19/57). Survey response rate was 63%. Average SF-12 PCS and MCS scores were 44.9 and 59.8 for patients able to achieve ambulation and 27.6 and 61.2 for nonambulatory patients. Conclusions: Microsurgical flap reconstruction is a valuable reconstructive

option in high-risk patients. Quality of life is comparable with 4��8C a normalized population if limb salvage is successful. Quality of life is decreased significantly when failure to ambulate occurs in this patient cohort. © 2013 Wiley Periodicals, Inc. Microsurgery 34:1–4, 2014. Lower extremity reconstruction with the aim toward limb salvage in the co-morbid patient population is a difficult undertaking for the reconstructive surgeon. Co-morbidities such as diabetes mellitus, peripheral vascular disease, and renal failure add complexity to microsurgical reconstruction. Systemic vascular changes such as recipient vessel disease, recipient site scarring, and donor vessel disease may pose a technical challenge. However, successful outcomes in lower extremity reconstruction are well demonstrated in this patient population and provide patients with the option of limb salvage.[1, 2] Early successful outcomes are predicated by overcoming compromised vascular inflow and by controlling infection. Following the early postoperative period, achieving successful long-term outcomes becomes more challenging. Traditionally flap survival was the marker for defining a successful outcome.

To the best of our knowledge, the present study is the first to identify in humans the ability of α-defensins, endogenous antimicrobial peptides from PMNs, to induce the expression of epithelial MxA, a potent antiviral protein against both RNA and DNA viruses. This innate antiviral immune mechanism could play an important role in maintaining healthy periodontal tissues. α-defensin-induced

MxA is an additional Galunisertib pathway to the well-recognized type I IFN induction [[35, 36]]. This function seems to be unique to α-defensin, because other antimicrobial peptides in healthy periodontal tissue (β-defensins and LL-37) induced only negligible MxA expression. It should be noted that α-defensins are known to upregulate co-stimulatory molecule and CD91 expression on antigen presenting check details dendritic cells [[37]]. There is little available information regarding innate antiviral immunity in the oral cavity. The human mouth harbors millions of microbes; however, we rarely develop serious infections [[38]]. Our previous research demonstrated TLRs and RLRs, key microbial sensors, in cells of periodontal tissues, which are critical for innate immune activation and local defense [[7-9]]. In the present study, we observed expression of MxA, PKR, OAS, and SLPI in healthy periodontal tissues, thus highlighting the role of innate antiviral immunity in periodontal tissue. MxA proteins are key mediators of innate antiviral

resistance induced in cells by type I (α/β) and type III (λ) IFNs [[29]]. The human MxA gene belongs to the class of IFN-stimulated genes (ISGs) and it is used as a surrogate marker N-acetylglucosamine-1-phosphate transferase for type I IFN activity in various experimental and clinical settings. Santoro et al. [[39]] used MxA to identify type I IFN in oral lichen planus. They found large numbers of MxA-positive cells in the lesion; therefore, a role of type I IFN in the pathology of oral lichen planus was postulated. We are unaware of any previous study of MxA in periodontal disease. Our consistent finding of positive immunostaining of MxA protein

in epithelium of healthy periodontal tissues (n = 9) was somewhat unexpected, since real-time PCR detected only negligible expression of type I IFN or type III IFN in healthy tissue specimens. Interestingly, the level of MxA proteins in the epithelial layer was significantly higher in healthy periodontal tissues than in periodontitis (Table 1). While searching for candidate MxA inducers, we treated primary HGECs with a variety of antimicrobial molecules, which are constitutively expressed in gingival epithelium. We clearly observed MxA protein expression after treatment with α-defensin-1, -2, or -3, but not with the other antimicrobial peptides β-defensin-1, -2, -3, or LL-37. At present, it is not clear how α-defensins induce MxA expression. Our data strongly suggest that induction of MxA expression by α-defensin-1 is not dependent on type I IFN as neutralizing antibodies against type I IFN had no effect on the MxA expression.

CGD abscesses were consistently larger than in WT animals

with equivalent challenge (Fig. 1B) and flow cytometry of collagenase D-released abscess cells indicated that a majority of cells were Gr-1+neutrophils, F4/80+ macrophages, and CD11c+ DCs (Fig. 1C). The total number of cells within a WT abscess was 5.3×106 while the CGD abscess yielded 3.06×107 cells, a 5.7-fold increase that correlates well with the increased abscess size. Finally, H&E staining of abscess sections showed the difference in overall size and revealed distinct areas of increased neutrophilic infiltrate in Talazoparib chemical structure the CGD abscess (Fig. 1D). These data establish that the CGD mutation results in extreme sensitivity to abscess formation in response to GlyAg/SCC exposure characterized by more severe pathology and either increased neutrophil infiltrate or defective clearance (e.g. efferocytosis) following the initial insult. To discern the hyperresponsiveness mechanism, mice were challenged with 100 μg GlyAg and selleckchem 1:4 diluted SCC for analysis of cellular infiltration (Fig. 2). At the times indicated, a peritoneal lavage was performed. Recovered cells were analyzed by flow cytometry while lavage supernatants were tested for nitrate and nitrite levels as markers of NO synthesis. Unchallenged CGD animals showed elevated baseline NO levels compared with WT (p<0.03);

however, this difference increased dramatically over the first 24-h period upon challenge (Fig. 2A), demonstrating the hyperresponsiveness MTMR9 to the GlyAg+SCC stimulation despite the modestly increased baseline. Remarkably,

total cellular influx into the peritoneum was not significantly different at most time points (Fig. 2B) and no consistent proportional differences in neutrophil, macrophage, or CD4+ T-cell populations were seen between WT and CGD (Fig. 2C and D), although modest differences in neutrophils were seen at 24 h (Fig. 2D). These data suggest that the proportional increase in neutrophils visible by H&E within the abscess (Fig. 1C and D) was mostly likely due to defects in neutrophil clearance rather than increased peritoneal infiltration, which is consistent with previous reports 27–29. More importantly, these findings suggest that the >10-fold increase in NO detected in the peritoneal lavage (Fig. 2A) was not due to increased cell numbers, but was more likely the result of changes in per-cell production of NO. iNOS expression was examined in isolated WT and CGD cells to establish the source of increased NO levels. Lavage cells were collected from GlyAg challenged mice for mRNA isolation and detection of the iNOS transcript using RT-PCR. In vivo challenge induced CGD cells to transcribe iNOS mRNA to a remarkably greater extent compared with WT cells at 24 h (Fig. 3A).

48 ± 0.05) at 3 weeks after surgery (0.95 ± 0.04). In the PD group, FEZ1 protein levels then decreased but at 5 weeks after injury were still selleck chemicals higher compared with the sham control group (Figure 2E,F). Along with FEZ1 expression, GFAP expression in striatum and substantia nigra was enhanced at 2 weeks after injury, peaked (0.77 ± 0.04 compared with 0.64 ± 0.03 in striatum, and 0.47 ± 0.05 compared with 0.27 ± 0.04 in substantia nigra), and then decreased (Figure 2G–J). In striatum of PD rats, GFAP expression levels were markedly higher at 2–4 weeks compared with the sham group (Figure 2G,H). However, in substantia nigra, GFAP expression levels were

increased at 2–5 weeks in the PD group compared with the sham group (Figure 2I,J). Because we found increased expression of FEZ1 and GFAP using real-time PCR and Western blot analysis, we chose two time points, 2 and 5 weeks after surgery, to examine brain sections from the PD and sham groups for immunohistochemical staining (Figure 3). This immunohistochemical

analysis at 2 weeks after surgery indicated that FEZ1 protein expression in PD rats GPCR Compound Library was increased compared with the sham group. To determine the cellular localization and the temporal changes of FEZ1 immunoreactivity in brain of PD rats, we performed immunofluorescent staining on transverse cryosections. Because previous data have shown that FEZ1 was expressed in the cytoplasm of astrocytes and neurones and that FEZ1 may play important roles in human astrocytes [28, 29], we investigated whether FEZ1 colocalized with TH (a positive marker for dopamine neurones) or with GFAP (a positive marker for astrocytes). In sham-operated controls, FEZ1 mostly colocalized with TH (Figure 4A) but not with GFAP (Figure 4C). In contrast, at 2 weeks after injury, when FEZ1 had reached peak expression,

we found that FEZ1 was expressed in many TH-negative cells in PD group brain sections (Figure 4A). Double immunofluorescent staining demonstrated that these TH-negative cells were mostly selleckchem GFAP-positive astrocytes (Figure 4B,C). Cells morphologically looking like TH cells but only stained by FEZ1 might be other types of neurones. Furthermore, triple immunostaining was performed using FEZ1, TH and GFAP to better understand the redistribution of FEZ1 immunostaining in the PD group (Figure 5). Brain tissues from the sham and PD groups (2 weeks after 6-OHDA injection) were transversely sectioned and triple immunolabelled with FEZ1, TH and GFAP (Figure 5A–P). Next, we counted FEZ1-positive cells, FEZ1-positive astrocytes and FEZ1-positive dopamine neurones (Figure 5Q). FEZ1-positive dopaminergic neurones constituted the majority of FEZ1-positive cells in substantia nigra of the sham group. However, FEZ1-positive astrocytes composed the majority of FEZ1-positive cells in the PD group. The proportion of FEZ1-positive in other cell types was unchanged.

The donor site complication of abdominal hernia is well-addressed with mesh INK 128 in vitro placement at our

center. In this clinical scenario, we show successful microvascular flap coverage utilizing both the superior and inferior epigastric neurovascular bundles and the entire rectus muscle to create two flaps, thereby sparing our young trauma patient both a second operation for a second free flap, as well as a second donor site for another flap. Careful consideration should be given to the use of this flap as a double transfer in cases such as this with two medium-sized defects in which a large portion of the standard inferior-based flap will be discarded. However, it must be recognized that the size and quality of the superior vessels will ultimately determine feasibility and that other available free tissue transfer options may be required. “
“A neuroma is a collection of disorganized nerve sprouts emanating from an interruption of axonal continuity, forming within a collagen scar as the nerve attempts to regenerate. Lingual neuroma formation secondary to iatrogenic trauma to the Roxadustat mw tongue is likely not uncommon; however, we could not find a report in the literature of treatment of a distal tongue end-neuroma treated by resection and implantation into muscle. Here we describe a patient who experienced debilitating chronic tongue pain after excision of a benign mass. After failing conservative management, the patient

was taken to the operating room where an end-neuroma of the lingual nerve was identified and successfully treated by excision and burying of the free proximal stump in the mylohyoid muscle. At 17 months postoperatively, she remains pain free without dysesthesias. © 2013 Wiley Periodicals, Inc. Microsurgery 33:575–577, 2013. “
“With ZD1839 in vitro recent advances in free tissue transfer, soft tissue defects involving the knee can be covered perfectly utilizing various free flaps. Yet the success of this operation depends on a secure

nontraumatic recipient pedicle around the knee area. The purpose of this study is to introduce the descending branch (DB) of the lateral circumflex femoral artery (LCFA) as a new recipient pedicle for knee defect coverage. Through autopsies of eight cadavers and a total of 11 extremities involving the area 10- and 15-cm above the upper margin of the patella, the number and sizes of the artery and vein of the descending branch of the lateral circumflex femoral artery were investigated. In a clinical setting, two cases of soft tissue defects in the area of the knee were reconstructed utilizing the DB of the LCFA with an anterolateral thigh perforator (ALTP) free flap on the ipsilateral side. Anatomical: The descending branches of the lateral circumflex femoral vessels measuring 10- and 15-cm above the lateral aspect of the patella numbered 1 artery and about 1.5 veins. The diameters of these vessels ranged from 1.0 to 2.0 mm (1.4 ± 0.4 mm) for the artery at 10-cm site and 1.0 to 3.