Francisco Vera-Sempere

CD3+-Mediated Rejection and C4d Deposition in Two Composite Tissue (Bilateral Hand) Allograft Recipients After Induction With Alemtuzumab

Background. New therapies are being introduced in reconstructive transplant surgery to enhance composite tissue allograft survival. Methods. Alemtuzumab was used in two bilateral hand allograft recipients to cause lymphocyte depletion. Results. Although profound leukopenia and lymphopenia developed, several episodes of acute rejection occurred both in the early and late posttransplant period. Cell-mediated rejection was diagnosed during acute rejection episodes. Intraluminal C4d deposits were found in the capillaries not only accompanying cellular rejection, but also in the absence of clinical rejection. However, their significance is unclear because donor-specific antibodies were absent, there were no pathological signs of injury, allograft function was not impaired, and clinical signs of rejection resolved. Conclusion. These findings suggest that alemtuzumab may not prevent cell-mediated rejection of a hand allograft transplant. Furthermore, C4d deposition warrants attention in clinical composite tissue allotransplants.

Perioperative ischemic injury and fibrotic degeneration of muscle in a forearm allograft: functional follow-up at 32 months post transplantation.

Allografts of the forearm are still uncommon in the field of composite tissue allograft transplantation. In November 2007, a right-hand allograft and a left-hand/full-length forearm allograft were transplanted in a 30-year-old man who lost both hands and the vision in his left eye due to an explosion. The patient underwent alemtuzumab and steroid induction therapy. Tacrolimus, mycophenolate mofetil, and low doses of steroids were given to prevent rejection. The allografts were rejected 3 times, but these episodes were controlled successfully. The immunosuppressive regimen was switched from tacrolimus to sirolimus because of increased serum creatinine. The left allograft showed a flexion contracture due to muscle fibrosis that was conjectured to be associated with a perioperative ischemic injury and permitted only a Moberg-type key pinch. In contrast, an excellent grade of function was observed in the right allograft. The Disabilities of the Shoulder, Arm, and Hand score improved from 70.83 to 36.6 and intrinsic musculature returned to both allografts. The patient was able to work 2 years after transplantation. This is the first report of an ischemic injury related to the successful allotransplantation of a composite tissue.

Improvement in Renal Function After Late Conversion to Sirolimus-Based Immunosuppression in Composite Tissue Allotransplantation

1. Abraham G, Shroff S, Matcha J, et al. Deceased donor renal transplantation program in India. Kidney Int 2010; 77: 378. 2. Li R, Chen G, Guo H, et al. Prolonged cardiac allograft survival in presensitized rats after a high activity Yunnan-cobra venom factor therapy. Transplant Proc 2006; 38: 3263. 3. Mansy H, Filobbos P, Aly TF, et al. Massive haemorrhage and rupture of renal transplant from a donor who died of snake bite. Nephrol Dialysis Transplant 1998; 13: 1018.

Prefabrication of a free flap for tracheal reconstruction: an experimental study. Preliminary report.

&NA; The development of a prefabricated free flap that could have potential for tracheal reconstruction has been investigated in the goat model. Through a staged procedure, a composite cutaneous‐chondromucosal premolded, prevascularized flap was obtained by prefabrication techniques. The procedure comprised three surgical stages. In the first stage, on day 0, the cartilaginous framework was constructed, along with the vascular pedicle (implantation of an arteriovenous fistula as a vascular carrier). In the second stage, on day 50, the inner surface of the neotrachea was lined with nasal mucosa. In the third stage, on day 60, the flap was elevated and free transferred to reconstruct a 15‐cm circumferential defect in the cervical trachea. Ten animals were operated on, and the results were one infection, three early deaths, one freeflap failure with early tracheal stenosis, and five long‐term survivors without significant stenosis. The structure of the neotracheal flap closely resembled that of the native trachea: internal respiratory epithelial lining, cartilage rings, and fibrovascular tissue. Fiberoptic bronchoscopy was done to all the animals at 10 and 60 days, revealing no significant stenosis in the long‐term survivors.

In vivo microarterial freezing: Experimental study

In vivo freezing of microarteries with liquid nitrogen is known to relieve spasm without inducing thrombosis. In the present study the patency rates of microanastomoses in pre‐ and postfrozen vessels were investigated in the rat model. Freezing of the femoral artery was done with ethyl chloride. The artery was frozen before the anastomosis was performed on the left side (prefrozen), and after on the right side (postfrozen); patency rates were recorded at 2, 10, and 30 days. Patency rates were 100% in prefrozen vessels and 100% in posffrozen ones at all three time intervals. Histologic examination showed depopulation of all vessel walls with loss of the intima layer and fragmentation of the inner elastic lamina. Progressive cellular repopulation and regeneration of the endothelium occurred later, with no differences between pre‐ and postfrozen arteries. It is concluded that freezing of vessels with ethyl chloride reverts arterial spasm without inducing thrombosis and that frozen arteries can be repaired by microsurgical anastomosis with patency rates comparable to those of virgin arteries.

Cytological changes in the oral mucosa after use of a mouth rinse with alcohol: A prospective double blind control study

Aim: The aim of this preliminary study was to detect cytological changes in the oral mucosa after using a mouth wash with alcohol. Material and Methods: A prospective double-blind, controlled study was performed, for 6 months. Group 1 consisted of 30 subjects who used a mouth rinse with 26.9% of alcohol [Listerine®] and Group 2 consisted of 30 subjects who used a mouth rinse with the same ingredients but with no alcohol. We obtained three cytological samples from the oral mucosa. The presence of cytological atypia, binucleation and karyorrhesis, and type of cells were studied. We also used a fluorescent in situ hybridization technique (FISH) in 15 samples in each group, for the micronucleus. Results: We found no clinical mucosal alteration after using the mouth wash at the end of the study in either group. We observed no cytological differences between the groups at the end of the study (p>0.05). Regarding the study of the micronucleus by FISH, we observed no significant difference between the groups (p>0.05). Conclusions: Our results showed no cytological alteration in patients using a mouth rinse with alcohol, but these findings should be considered preliminary results, to be confirmed in a greater sample of patients. Key words:Mouth wash, oral mucosa, cytological change, alcohol.

Intraosseus plexiform schwannoma of the mandible: immunohistochemical differential diagnosis.

Schwannomas and neurofibromas are the most common benign tumors derived from peripheral nerves, and whereas the head and neck region is the most common location for the occurrence of benign neural sheath neoplasms, origin within the oral cavity is uncommon, and occurrence centrally in the jaws is most unusual. Plexiform (multinodular) schwannoma is an anatomically unique variant of schwannoma characterized grossly and/or microscopically by intraneural plexiform and often multinodular growth. In current report, we present the first reported case of intraosseous plexiform schwannoma of the mandible, an extremely rare benign neurogenic tumor, diagnosed by optical and immunohistochemical procedures, showing the importance of differential diagnosis of these unusual intraosseous mandibular tumors.

Recomendación para la determinación de HER2 en cáncer de mama. Consenso nacional de la Sociedad Española de Anatomía Patológica (SEAP) y de la Sociedad Española de Oncología Médica (SEOM)

Breast cancers with HER2 alterations are critical to identify because such tumors require unique treatment, including the use of targeted therapies. HER2 alterations at the DNA (amplification) and protein (overexpression) level usually occur in concert, and both in situ hybridization and immunohistochemistry can be accurate methods to assess these alterations. However, recent studies including those conducted by the Association for Quality Assessment of the Spanish Society of Pathology and the experience of several national reference centres for HER2 testing have suggested that serious reproducibility issues exist with both techniques. To address this, a joint committee of both the Spanish Society of Pathology and the Spanish Society of Medical Oncology has met to review guidelines for HER2 testing. Consensus recommendation are based not only on panellist’s experience but also in those consensus guidelines previously reported in several countries, such as United Stated, United Kingdom and Canada. These guidelines include minimal requirements that Pathology Department must meet in order to guarantee appropriate HER2 testing in breast cancer. Pathology laboratories that do not meet these standards must put effort to reach them and, in the meantime, send clinical cases to reference centres.

Differential expression of Cyclin D1 in keratin-producing odontogenic cysts

Objetives: The aim of the present study was to analyze the expression levels of Cyclin D1 (CCD1), a nuclear protein that plays a crucial role in cell cycle progression, in a series of keratin-producing odontogenic cysts. Study Design: A total of 58 keratin-producing odontogenic cysts, diagnosed over ten years and classified according to the WHO 2005 criteria, were immunohistochemically analyzed in terms of CCD1 expression, which was quantified in the basal, suprabasal and intermediate/superficial epithelial compartments. The extent of immunostaining was measured as a proportion of total epithelial thickness. Quantified immunohistochemical data were correlated with clinicopathological features and clinical recurrence. Results: Keratin-producing odontogenic cysts were classified as 6 syndromic keratocystic odontogenic tumors (S-KCOT), 40 sporadic or non-syndromic KCOT (NS-KCOT) and 12 orthokeratinized odontogenic cysts (OOC). Immunohistochemically, CCD1 staining was evident predominantly in the parabasal region of all cystic lesions, but among-lesion differences were apparent, showing a clear expansion of parabasal compartment especially in the S-KCOT, followed to a lesser extent in the NS-KCOT, and being much more reduced in the OOC, which had the greatest average epithelial thickness. Conclusions: The differential expression of CCD1 noted in the present study suggests that dysregulation of cell cycle progression from G1 to the S phase contributes to the different aggressiveness of these lesions. However, CCD1 expression levels did not predict NS-KCOT recurrence, which is likely influenced by factors unrelated to lesion biology. Key words:Keratin-producing odontogenic cyst, keratocyst, keratocystic odontogenic tumor, nevoid basal cell carcinoma syndrome, orthokeratinized odontogenic cyst, cyclin D1, immunohistochemistry.

A multiplex real-time PCR method for quantification of BK and JC polyomaviruses in renal transplant patients.

BackgroundInfection of BK or JC human polyomavirus can lead to polyomavirus-associated nephropathy in renal transplant patients. Thus effective management of these patients requires early detection and quantification of these viruses in urine and blood. DesignThe aim of this study was to evaluate and validate a multiplex real-time PCR-based method for monitoring BK and/or JC viral loads in renal transplant patients. Analytic parameters such as limit of quantification, linear dynamic range, sensitivity and specificity, as well as reliability of the assays were determined. Seventy-six plasma or urine samples spiked with variable amounts of BK and JC viral DNA ranging from low (7.0×103 or 1.5×104), to medium (1.0×106) to high (1.0×108 or 1.0×109 copies/mL) levels of viruses were tested. In addition, 45 clinical urine or plasma samples with known copy numbers of BK or JC viruses, which were isolated from the renal transplant patients from 4 US medical centers, were also tested. ResultsBK and/or JC viruses can be detected with distinguishable melting temperature of 64°C or 68°C, respectively. On the basis of the need for clinical monitoring of different types of specimens, the low limit of quantification for plasma or urine was set at 7.0×103 or 1.5×104 copies/mL, respectively with the linear dynamic range ≥6 logs. The assay exhibits 100% specificity, 97.9% sensitivity with low intra-assay and inter-assay variability (coefficient of variation <4%). ConclusionsThis clinically validated method has the necessary utility to monitor BK and JC viremia and viruria in renal transplant patients.