Clinical application of decellularized and lyophilized human amnion/chorion membrane grafts for closing post‐laryngectomy pharyngocutaneous fistulas

Background and Objectives

Squamous cell carcinoma is the most common pathological type among the cancers of the larynx. Standard treatment for squamous cell carcinoma of the larynx is the combination of chemotherapy, radiotherapy, and laryngectomy. Pharyngocutaneous fistula is a common complication of laryngectomy. We hypothesized that decellularized and lyophilized human amnion/chorion membrane can be an effective, non‐invasive method of treating pharyngocutaneous fistula.

Methods

A total of 67 patients with laryngeal squamous cell carcinoma were retrospectively analyzed after treatment in a prospective trial. After preoperative chemotherapy, radiotherapy, and total or extended laryngectomy, primary wound healing occurred in 42 (62.7%) patients. Pharyngocutaneous fistula developed in 8 (11.9%) patients. Decellularized and lyophilized human amnion/chorion membrane grafts were used to reconstruct the fistulas.

Results

The average time for the full healing of the wound in all patients after transplantation of these grafts was 18 days.

Conclusion

The advantages of using these grafts over other existing methods of pharyngocutaneous fistula treatment are that they are non‐invasive, prevent donor morbidity, and enable management of the wound without using classical wound gauze. J. Surg. Oncol. 2016;113:538–543. © 2016 The Authors. Journal of Surgical Oncology Published by Wiley Periodicals, Inc.     

Tamoxifen induces resistance to activated protein C

Introduction

The estrogen antagonist tamoxifen (TAM) increases the thrombotic risk similar to estrogen containing oral contraceptives (OC). In OC users this risk is attributed to alterations of hemostasis resulting in acquired resistance to activated protein C (APC). TAM-induced APC resistance has not been reported yet.

Materials and Methods

Blood samples were collected prospectively from women with breast cancer before (n = 25) and monthly after start of adjuvant TAM treatment (n = 75). APC resistance was evaluated on basis of the effect of APC on the endogenous thrombin generation potential. To detect increased in vivo APC generation APC plasma levels were measured using a highly sensitive oligonucleotide-based enzyme capture assay. Routine hemostasis parameters were measured additionally.

Results

APC sensitivity decreased by 41% (p = 0.001) compared to baseline after one month of TAM application and remained significantly decreased during the study period. Free protein S increased (p = 0.008) while other analyzed procoagulant factors, inhibitors, and activation markers of coagulation decreased or did not change significantly. In five patients the APC concentration increased to non-physiological levels but an overall significant increase of APC was not observed.

Conclusions

This is the first study showing acquired APC resistance under TAM therapy. Acquired APC resistance might explain the increased thrombotic risk during TAM treatment. Observed changes of hemostasis parameters suggest different determinants of TAM-induced APC resistance than in OC-induced APC resistance. The presence of acquired APC resistance in TAM patients warrants further evaluation if these patients may benefit from antithrombotic prophylaxis in the presence of additional thrombotic risk factors.     

The immunosuppressants cyclosporin A and FK506 equally ameliorate brain damage due to 30-min middle cerebral artery occlusion in hyperglycemic rats

In the present experiments, we compared the anti-ischemic effects of the immunosuppressants cyclosporin A (CsA) and FK506 in hyperglycemic animals subjected to 30 min of middle cerebral artery (MCA) occlusion. Both immunosuppressants were given as pre-treatment, the effect of treatment being evaluated by 2,3, 5-triphenyltetrazolium (TTC) staining after 3 days of recovery. Both FK506 and CsA reduced the infarct volume to less than 1/3 of control. In spite of CsA's known effect as a blocker of the mitochondrial transition (MPT) pore, it failed to give a more robust effect than FK506. If anything, FK506, which lacks an effect on the MPT pore, had a more pronounced anti-ischemic effect. We conclude that, in this model of infarction, an MPT may not play a major pathogenetic role.     

Activation of purified allogeneic CD4(+) T cells by rat bone marrow-derived dendritic cells induces concurrent secretion of IFN-gamma, IL-4, and IL-10

Dendritic cells (DCs) are highly specialized antigen-presenting cells that play a key role in the initiation and regulation of immune responses. The ability of DCs to process antigens and the outcome of their interaction with T cells are largely dependent on phenotype as well as maturation state of DCs. In this study, we generated DCs from rat bone marrow precursors. Bone marrow cells cultured in the presence of granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4, and Flt-3 ligand (FL) produced immature DCs that expressed intermediate levels of major histocompatibility complex (MHC) class II, low levels of CD80 and CD86 molecules and displayed a high capacity of endocytosis. Bone marrow-derived DCs (BMDCs) matured in the presence of lipopolysaccharide (LPS) upregulated expression of MHC class II, CD80 and CD86, while their phagocytic capacity was dramatically reduced. Mature BMDCs stimulated vigorous proliferation of purified allogeneic CD4(+) T cells in a primary mixed leukocyte reaction (MLR) and elicited a mixed cytokine profile in allogeneic CD4(+) T cells: DCs activated CD4(+) T cells to produce interferon (IFN)-gamma, IL-4, and IL-10. Thus, rat BMDCs effectively internalize antigens and stimulate T cell proliferation but fail to induce an unidirectional polarization of T helper (T(H)) cells and in this respect differ from both human and mouse DCs.     

Attenuation correction reveals gender-related differences in the normal values of transient ischemic dilation index in rest-exercise stress sestamibi myocardial perfusion imaging

BACKGROUND: Transient ischemic dilation (TID) has been established as an important independent marker of severe and extensive coronary artery disease (CAD) in myocardial perfusion imaging (MPI). The accuracy of the TID index is dependent on a well-determined threshold (normal limits) between normal and abnormal values for each study protocol. To date, the effects of neither gender nor attenuation correction (AC) on TID normal limits have been established. Thus, the objectives of this study were to determine if AC processing changes the normal value of the TID index and if there were gender-related differences in the TID index of normal patients who had undergone rest/exercise-stress technetium-99m sestamibi MPI. METHODS AND RESULTS: Seventy-five patients (33 women, 42 men; mean age, 57.7 +/- 11.7 y and 55.9 +/- 10.0 y, respectively) with less than a 5% likelihood of CAD, who had undergone low-dose rest/high-dose exercise-stress Tc-99m sestamibi MPI, were studied. All studies were acquired using simultaneous emission/transmission scans and were corrected for attenuation, scatter, and resolution effects using the ExSPECT II method. Both the AC and non-AC studies were analyzed using the Emory Cardiac Toolbox (ECTb; Syntermed, Inc, Atlanta, Ga) quantitative software. The TID index was calculated automatically as the ratio of stress mean left ventricular volumes to rest mean left ventricular volumes by ECTb. Patients were grouped by gender and the TID indices from AC and non-AC studies were compared. Linear regressions of the TID index and body mass index were analyzed to exclude differences in body size between male and female patients as a confounding factor in gender-related differences in TID. The TID index upper normal limits were calculated as the mean value plus 2 standard deviations (SDs). AC processing did not change the TID index significantly whether the genders were combined or separated (AC TID = 0.97 +/- 0.14 vs non-AC TID = 0.98 +/- 0.12 for all patients). Female patients showed higher mean TID indices than male patients in both AC (1.01 +/- 0.15 vs 0.95 +/- 0.12) and non-AC studies (1.00 +/- 0.15 vs. 0.97 +/- 0.10), but this difference was statistically significant only in AC studies (p = .03). TID indices remained constant across the range of body mass index studied. The TID index upper normal limit was 1.31 for female and 1.18 for male patients. CONCLUSION: TID normal values for rest/exercise-stress Tc-99m sestamibi MPI are gender-dependent and not affected by AC processing. Thus, diagnosticians should take into account these gender-related differences, as compared with the traditional value generated from mostly male populations, to ensure both men and women have the same overall accuracy of using the TID index in the diagnosis and prognosis of CAD.