Nonmyeloablative Conditioning Generates Autoantigen‐Encoding Bone Marrow That Prevents and Cures an Experimental Autoimmune Disease

Autoimmune diseases result from chronic targeted immune responses that lead to tissue pathology and disease. The potential of autologous hematopoietic stem cells transplantation as a treatment for autoimmunity is currently being trialled but disease relapse is an issue. We have previously shown in a mouse model of experimental autoimmune encephalomyelitis (EAE) that the transplantation of bone marrow (BM) transduced to encode the autoantigen myelin oligodendrocyte glycoprotein (MOG) can prevent disease induction. However these studies were performed using lethal irradiation to generate BM chimeras and a critical factor for translation to humans would be the ability to utilize low toxic preconditioning regimes. In this study, treosulfan was used as a nonmyeloablative agent to generate BM chimeras encoding MOG and assessed in models of EAE induction and reversal. We find that treosulfan conditioning can promote a low degree of chimerism that is sufficient to promote antigen specific tolerance and protect mice from EAE. When incorporated into a curative protocol for treating mice with established EAE, nonmyeloablative conditioning and low chimerism was equally efficient in maintaining disease resistance. These studies further underpin the potential and feasibility of utilizing a gene therapy approach to treat autoimmune disease.     

Current Clinical Practice DIC 2002: A Review of Disseminated Intravascular Coagulation

The turn of the millennium has seen clear advances in the understanding and management of Disseminated Intravascular Coagulation (DIC). The recognition that its pathogenesis stems from sustained thrombin generation in fuelling the cycle between inflammation and coagulation has seen the first successful treatment in severe sepsis through targeting this activity. An advance in treatment brings heightened relevance to laboratory testing, which now emphasises earlier detection and better monitoring to facilitate improved risk-identification and assessment of therapeutic efficacy. This review article also provides insights into future strategies that might build on the foundation of improving prognosis for the patient with DIC.     

Modulation of metallothionein isoforms is associated with collagen deposition in proliferating keloid fibroblasts in vitro

Please cite this paper as: Modulation of metallothionein isoforms is associated with collagen deposition in proliferating keloid fibroblasts in vitro. Experimental Dermatology 2010; 19 : 987–993. Abstract: The keloid fibroblast (KF) is known to have higher proliferative capacity than normal dermal fibroblast (NF). Metallothionein (MT), a metal‐binding protein, has been reported to promote cell proliferation. In this study, we evaluated the expression of MT isoforms at the mRNA level in fetal bovine serum (FBS)‐stimulated proliferating KF. Although the morphological appearance of NF and KF was similar when viewed under light, confocal and transmission electron microscopy, there was surprisingly a generally lower expression of MT isoforms in KF when compared with NF and also reduced MT staining in dermal fibroblasts of keloids as opposed to normal skin. Primary cultures of KF grown in 5% FBS or 10% FBS compared to without FBS demonstrated significantly higher proliferative activity and more abundant deposition of collagen. Contrary to expectation, MT‐1A, ‐1F, ‐1G, ‐1X and ‐2A isoforms were significantly down‐regulated in proliferating KF. Moreover, stimulating KF with TGF β1, which is known to promote collagen synthesis and keloid formation, increased expression of Collagen 1A and 3A genes accompanied by reduction in MT‐2A gene expression. Furthermore, down‐regulation of the MT‐2A gene in proliferating KF by siRNA‐mediated silencing enhanced cell proliferation with concomitant up‐regulation of the NF‐κB gene and 10 of 13 other NF‐κB pathway–related genes analysed but no alteration of the Collagen 1 and Collagen 3 gene expression. It would appear that down‐regulation of MT isoforms in proliferating KF, in particular MT‐2A, enhances keloidogenesis with the possible involvement of the NF‐κB signalling pathway.     

Subeffective doses of dexketoprofen trometamol enhance the potency and duration of fentanyl antinociception

The combination of classic non-steroidal antiinflammatory drugs (NSAIDs) with opiates induces more analgesia than the summed effect of each drug given separately. No studies have been performed using new generation NSAIDs and fentanyl nor on the duration of this effect. We have studied the analgesic effect of fentanyl alone and after the administration of subeffective doses of dexketoprofen trometamol in rat nociceptive responses. The responses were evoked by noxious mechanical stimulation and were recorded as single motor units in male Wistar rats anaesthetized with alpha-chloralose. The effective dose 50 (ED(50)) observed with fentanyl was 22.4 +/- 1.5 microg kg(-1) and full recovery was apparent 20 min later. The administration of a total dose of 40 microg kg(-1) of dexketoprofen trometamol did not induce any significant effect on the nociceptive responses. In the presence of dexketoprofen trometamol, the ED(50) for fentanyl was 5 fold lower than before: 3.8 +/- 1.1 microg kg(-1) and no significant recovery was observed 45 min later. The opioid antagonist naloxone (200 microg kg(-1)) did not reverse the effect, although in control experiments the same dose was able to prevent any action of fentanyl given alone. We conclude that the combination of fentanyl and subeffective doses of dexketoprofen trometamol induces a more potent and longer lasting analgesic effect than that observed with fentanyl alone, and that this is not an opioid mediated action.     

Peri-implant soft tissue integration of immediately loaded implants in the posterior macaque mandible: a histomorphometric study

BACKGROUND: Today, one critical goal in implant placement is the achievement of optimal soft tissue integration. Reports thus far have demonstrated successful soft tissue preservation in delayed loaded implants placed in anterior jaws. The aim of this study was to histomorphometrically examine the soft tissues around immediately loaded implants placed in the macaque posterior mandible. METHODS: Splinted crowns on screw-shaped titanium implants (8 mm length, 3.5 mm diameter) were utilized. Three implants each were placed in the premolar-molar edentulous mandibular segments of 6 adult monkeys (Macaca fascicularis); one side served as the control (delayed loading) and the other as the test sites (immediate loading). The animals were sacrificed after 3 months of loading. Histomorphometry of 6 soft tissue indices including the sulcus depth (SD), junctional epithelium (JE), connective tissue contact (CTC), biologic width (BW = SD + JE + CTC), DIM (distance between the implant top and coronal gingiva), and DIB (distance between the implant top and first implant-to-bone contact) was performed on non-decalcified sections. RESULTS: No significant differences in the mean soft tissue scores (mm) between the test (SD = 0.68 +/- 0.63; JE = 1.71 +/- 1.04; CTC = 1.51 +/- 1.14; DIM = 2.27 +/- 1.18; DIB = 1.32 +/- 1.21; BW = 3.9) and control (SD = 0.88 + 0.57; JE = 1.66 + 0.77; CTC = 1.24 +/- 0.92; DIM = 2.38 +/- 0.81; DIB = 1.19 +/- 0.91; BW = 3.78) groups were observed (P > 0.01). CONCLUSION: These findings suggest that the dimensions of the peri-implant soft tissues were within the biologic range and were not influenced by immediate functional loading or posterior location of the implants in the macaque mandible.     

Transcorporal artificial urinary sphincter cuff placement in cases requiring revision for erosion and urethral atrophy

PURPOSE: A distal cuff location is often required in patients undergoing artificial urinary sphincter reimplantation after previous erosion or in those requiring revision because of urethral atrophy at the original cuff site. Dissecting the urethra at a more distal site increases the risk of urethral injury and erosion, and often the urethral circumference is so small that a 4 cm. cuff is too large. We present a novel technique for distal cuff placement using transcorporal dissection that leaves corporal tunica albuginea on the dorsal surface of the urethra, allowing for its safer mobilization and adding to its bulk. MATERIALS AND METHODS: We reviewed the charts of 31 men who underwent this technique and contacted 26 by telephone. The indications for distal transcorporal cuff placement varied. In 7 men with inadequate urethral coaptation with a 4 cm. proximal cuff at initial implantation a primary transcorporal tandem cuff was implanted distal. In 8 men persistent or recurrent incontinence despite a 4 cm. proximal cuff led to secondary distal reimplantation. Previous artificial urinary sphincter erosion and/or infection in 10 cases, previous urethral surgery at the optimal cuff site in 5 and radiation changes at the optimal cuff site in 1 led to selection of the more distal site and technique. Of the transcorporally placed cuffs 18 were 4 cm. and 13 were 4.5 cm. Preoperatively 5.2 pads were used daily. Of the 31 patients 27 were impotent preoperatively, 1 had normal erections, 1 had partial erections with the MUSE drug delivery system (Vivus, Inc., Menlo Park, California) and 2 had a previously placed penile prosthesis. RESULTS: At a mean followup of 17 months 26 of the 31 patients (84%) had occasional or no stress incontinence requiring 0 to 1 pad daily, 2 with pure urge incontinence used 1 to 2 pads daily and 3 had mixed incontinence requiring 0 to 3 pads daily. Of the 26 men surveyed 25 were very satisfied with the postoperative level of incontinence. Postoperatively erectile function deteriorated in 1 patient and was unchanged in the remainder. There was no erosion or infection of the transcorporally placed cuffs, although 3 were replaced for malfunction. CONCLUSIONS: This technique offers significant advantages in cases of revision. The technique protects the urethra from intraoperative dissection injury and decreases the risk of erosion because the urethra is buttressed at its vulnerable location. In addition, bulk is added to the urethra, allowing for better cuff sizing, which is usually a problem at this location where the urethra is small, thereby, improving continence in revised cases. Our success has recently led us to abandon tandem cuff placement altogether. There is a potential for deteriorating erectile function in potent men who undergo implantation in this fashion.     

Locoregional adoptive immunotherapy resulted in regression in distant metastases of a recurrent esophageal cancer

 Esophageal cancer is one of the most common malignant diseases. However, postoperative recurrences are still resistant to currently available radiochemotherapy. We recently reported a study on the initial clinical efficacy of locoregional adoptive immunotherapy for advanced esophageal cancer. We report here our clinical experience of remarked responses in distant metastatic lesions in a patient with recurrent cancer after receiving this immunotherapy. A male patient underwent curative surgery, and presented with multiple recurrent metastases in the supraclavicular lymph nodes (LNs), liver, and abdominal aortic LNs. Autologous tumor-activated lymphocytes (AuTLs) generated ex vivo were regionally injected into supraclavicular LNs every 2 weeks 13 times. Mean numbers of the administrated cells were 0.8 × 10⁹ cells/injection. AuTLs established from peripheral blood lymphocytes stimulated by autologous tumor cells with interleukin-2 were tested for their cytotoxicity before every treatment. During immunotherapy, Grade 2 diarrhea and fever were observed. The clinical partial responses were obtained in all lesions and were sustained for 11 months. Because clinical toxicity was tolerable, this immunotherapy might be useful for patients with far-advanced esophageal cancers. Received: June 19, 2002 / Accepted: September 26, 2002 Correspondence to:U. Toh     

Intraarterial cellular immunotherapy for patients with inoperable liver metastases of esophageal cancer

PURPOSE: We report 2 patients with refractory liver metastatic tumor after esophagectomy for advanced esophageal cancer, who responded markedly to locoregional cellular immunotherapy by repeated intraarterial infusions of autologous tumor cell-activated T lymphocytes (AuTL), even after they failed the standard chemotherapy of cisplatin (CDDP) and 5-fluorouracil (5-FU). METHODS: AuTL administrations were made through the hepatic artery via a subcutaneous reservoir located at the right upper leg. Six injections were administered to both patients, repeated at 2-week intervals. The total number of administered T cells reached 2.4 x 10(9) and 3.1 x 10(9), respectively. RESULTS: A 39% and 51% regression in each infused field, compared with the size of liver tumor before treatment, was observed by computed tomography (CT) scan in patient 1 and 2, respectively. The responses continued up to the 10th and 11th month after the intraarterial infusion, confirmed by follow-up CT scan. The adverse effects of intraarterial immunotherapy were tolerable, with grade 1-2 fever and nausea in each patient. CONCLUSIONS: Clinical regression of liver metastases of esophageal cancer was observed in both patients who received this intraarterial cellular immunotherapy. Liver metastases of esophageal cancer may be controlled effectively and safely by repeating the intraarterial AuTL infusion as a locoregional immunotherapy over a long period.