Risk of a Second Skin Cancer in a Cohort of Patients With Nonmelanoma Skin Cancer -Basal Cell Carcinoma or Squamous Cell Carcinoma-Treated With Mohs Micrographic Surgery: A National Prospective Cohort Study. / Riesgo de aparición de segundas neoplasias cutáneas en una cohorte de pacientes diagnosticados de carcinoma queratinocítico (carcinoma basocelular y carcinoma epidermoide) tratados con cirugía de Mohs. Estudio de cohortes prospectivo nacional.

OBJECTIVE: Patients with nonmelanoma skin cancer (NMSC)-ie, basal cell carcinoma (BCC) or squamous cell carcinoma (SCC)-have an increased risk of developing a second skin cancer. The aim of this study was to describe the frequency, incidence per 1000 person-years, and predictors of a second skin cancer in a cohort of patients with NMSC treated with Mohs micrographic surgery (MMS). MATERIAL AND METHODS: Prospective study of a national cohort of patients with NMSC who underwent MMS at 22 Spanish hospitals between July 2013 and February 2020; case data were recorded in the REGESMOHS registry. The study variables included demographic characteristics, frequency and incidence per 1000 person-years of second skin cancers diagnosed during the study period, and risk factors identified using mixed-effects logistic regression. RESULTS: We analyzed data for 4768 patients who underwent MMS; 4397 (92%) had BCC and 371 (8%) had SCC. Mean follow-up was 2.4 years. Overall, 1201 patients (25%) developed a second skin cancer during follow-up; 1013 of the tumors were BCCs (21%), 154 were SCCs (3%), and 20 were melanomas (0.4%). The incidence was 107 per 1000 person-years (95% CI, 101-113) for any cancer, 90 per 1000 person-years (95% CI, 85-96) for BCC, 14 (95% CI, 12-16) per 1000 person-years for SCC, and 2 (95% CI, 1-3) per 1000 person-years for melanoma. More men than women developed a subsequent skin cancer (738 [61%] vs 463 [39%]). The main risk factors were a history of multiple tumors before diagnosis (relative risk [RR], 4.6; 95% CI, 2.9-7.1), immunosuppression (RR, 2.1; 95% CI, 1.4-3.1), and male sex (RR, 1.6; 95% CI, 1.4-1.9). CONCLUSION: Patients have an increased risk of developing a second tumor after MMS treatment of NMSC. Risk factors are a history of multiple tumors at diagnosis, immunosuppression, and male sex.

Mohs micrographic surgery in the elderly: comparison of tumours, surgery and first-year follow-up in patients younger and older than 80 years old in REGESMOHS.

BACKGROUND: The elderly population is increasing and more patients in this group undergo Mohs micrographic surgery (MMS). The few publications investigating MMS in elderly people conclude that it is a safe procedure; however, these are single-centre studies without a comparison group. OBJECTIVE: To compare the characteristics of patients, tumours, MMS and 1-year follow-up in patients younger than 80 years, with patients older than 80 years at the time of surgery. METHODS: Data was analysed from REGESMOHS, a prospective cohort study of patients treated with MMS. The participating centres were 19 Spanish hospitals where at least one MMS is performed per week. Data on characteristics of the patient, tumour and surgery were recorded. Follow-up data were collected from two visits; the first within 1 month postsurgery and the second within the first year. RESULTS: From July 2013 to October 2016, 2575 patients that underwent MMS were included in the registry. Of them, 1942 (75.4%) were aged <80 years and 633 (24.6%) were ≥80 years old. In the elderly, the tumour size was significantly higher with a higher proportion of squamous cell carcinoma. Regarding surgery, elderly more commonly had tumours with deeper invasion and required a higher number of Mohs surgery stages, leaving larger defects and requiring more time in the operating room. Despite this, the incidence of postoperative complications was the same in both groups (7%) and there were no significant differences in proportion of relapses in the first-year follow-up. CONCLUSION: The risk of short-term complications and relapses were similar in elderly and younger groups. MMS is a safe procedure in the elderly.

Characterization of Surgical Procedures in the Spanish Mohs Surgery Registry (REGESMOHS) for 2013-2015. / Descripción de las intervenciones quirúrgicas recogidas en el registro español de cirugía de Mohs (REGESMOHS) (2013-2015).

INTRODUCTION: The Spanish Mohs Surgery Registry is used to collect data on the use and outcomes of Mohs micrographic surgery (MMS) in Spain. The aim of this study was to describe perioperative and intraoperative data recorded for MMS procedures performed between July 2013 (when the registry started) and January 2016. MATERIAL AND METHODS: Prospective cohort study of data from 18 hospitals. The data collected included type of anesthesia, surgical technique, hospital admission, number of Mohs stages, management of preoperative risk factors, additional treatments, previous treatments, type of tumor, operating time, and complications. RESULTS: Data were available for 1796 operations. The most common tumor treated by MMS was basal cell carcinoma (85.96%), followed by squamous cell carcinoma (6.18%), lentigo maligna (2.81%), and dermatofibrosarcoma protuberans (1.97%). Primary tumors accounted for 66.9% of all tumors operated on; 19.2% of tumors were recurrent and 13.9% were persistent. The most common previous treatment was surgical. MMS was mostly performed under local anesthesia (86.7% of cases) and as an outpatient procedure (71.8%). The frozen section technique was used in 89.5% of cases. One stage was needed to achieve tumor-free margins in 56.45% of patients; 2 stages were required in 32.1% of patients, 3 in 7.1%%, 4 in 2.7%, and 5 or more in 1.8%. The defect was reconstructed by the dermatologist in 98% of patients and the most common technique was flap closure (47.2%). Intraoperative complications were recorded for just 1.62% of patients and the median (interquartile range) duration of surgery was 75 (60-100) minutes. CONCLUSION: The characteristics of the patients and tumors treated by MMS are similar to those reported for similar studies in other geographic areas. Lentigo maligna and dermatofibrosarcoma protuberans accounted for a higher proportion of cases in our series, and repair of the surgical defect by a dermatologist was also more common. Operating times in MMS are not much longer than those reported for other procedures and the rate of intraoperative complications is very low.

Hip arthroscopy complications regarding surgery and early postoperative care: retrospective study and review of literature.

PURPOSE: The purpose is to describe the rate of complications in a series of hip arthroscopies performed at our center, as well as perform a systematic review of the current literature in order to compare our outcomes. METHODS: Two hundred and fifty-eight patients affected of femoroacetabular impingement and treated with hip arthroscopy have been studied. All minor and major complications were studied during the first postsurgery year. Furthermore, a systematic review was performed comparing major and minor complications with our series. Two attending orthopedic surgeons selected the different studies with the same inclusion and exclusion criteria, remaining with 48 studies that have been reviewed and included in the present study. RESULTS: Mean age was 36.6 years old (SD 17.45), and the ratio men:women was 137:121. The mean complication rate observed was 14.34% (37/258) of global complications. Only three patients showed major complications: femoral neck fracture, septic arthritis and avascular necrosis of the femoral head. Any of these patients had permanent side effects. CONCLUSIONS: Hip arthroscopy has a low rate of major complications, but a higher number of minor complications that could be avoided with certain preventive measures.

Autoantibodies against TIF-1-γ and CADM-140 in Spanish patients with clinically amyopathic dermatomyositis (CADM): clinical significance and diagnostic utility.

BACKGROUND: Patients with clinically amyopathic dermatomyositis (CADM) appear to be at risk for developing cancer and interstitial lung diseases, but population data to confirm this hypothesis are limited. Moreover, CADM presents cutaneous and histological findings that may overlap with subacute cutaneous lupus erythematosus (SCLE). OBJECTIVES: To determine the association between myositis-specific autoantibodies, myositis-associated autoantibodies and CADM in Spanish patients. In addition, to study the usefulness of these autoantibodies in the differential diagnosis between CADM and SCLE. METHODS: Serum samples were tested for myositis-specific autoantibodies and myositis-associated autoantibodies through immunoprecipitation and other standardized methods. RESULTS: Anti-CADM-p140 and anti-p155 antibodies were the only myositis-specific autoantibodies found and were associated with interstitial lung diseases and cancer respectively. No myositis-associated autoantibodies were found in CADM. Moreover, clinical subsets and proportions seemed to differ from Asian cohorts, where anti-CADM-p140 is considered a CADM hallmark antibody and a risk factor for the development of interstitial lung disease. Interestingly, anti-SSA was highly associated with SCLE, whereas no myositis-specific autoantibodies were found in this entity. LIMITATIONS OF THE STUDY: Association between CADM and myositis-specific autoantibodies and differences between CADM and SCLE were tested on a relatively small cohort of patients. CONCLUSION: There is an association between cancer-associated myositis and interstitial lung diseases and their hallmark autoantibodies in our cohort. In addition, the combined determination of myositis-specific autoantibodies and SSA autoantibodies may help to accurately discriminate SCLE from CADM.

[Treatment of lymphatic malformations with OK-432 sclerosis. Our experience]. / Tratamiento de las malformaciones linfáticas mediante esclerosis con OK-432. Nuestra experiencia.

INTRODUCTION: The elective treatment of lymphatic malformations has always been surgical resection. However sclerotherapy with a variety of substances is being used as an alternative to surgery specially in the last years. The aim of this essay is to present our results using sclerosis with OK-432. MATERIALS AND METHODS: We have reviewed all patients affected by lymphatic malformation that have been treated by sclerosis with OK-432. Each procedure was performed under general anesthesia by a radiologist and a pediatric surgeon. RESULTS: Fifteen patients have been treated. On the whole, 36 procedures have been performed, with a mean of 2.4 for each patient. The mean amount of OK-432 injected was 1.5 milligrams (mg) in each procedure. 73% of the patients showed nearly complete or complete response. There were two complications; one of them was a limited bleeding and the other one, a case of high fever resolved by antipyretics and antibiotics. Microcystic or mixed malformations required a higher number of procedures and presented poorer response. CONCLUSION: Sclerotherapy with OK-432 is an effective and safe treatment for lymphatic malformation specially the macrocystic ones, therefore it should be considered as first line therapeutic option.

[Ten years of laparoscope hernia repairs. Indications and results]. / Diez años de herniorrafias laparoscópicas. Indicaciones y resultados.

INTRODUCTION: The inguinal hernia surgery is common in the pediatric population although laparoscopic management today, is not considered the standard technique. The aim of this study is to review the results of our study and evaluate the contralateral side in patients with unilateral hernia. MATERIAL AND METHODS: Retrospective study of laparoscopic herniorraphy at our hospital since the beginning of the art in 1999 with a total of 348 patients. The indications of laparoscopy have been the availability of umbilical hernia associated, doubt diagnosis, recurrence of open surgery, bilateral tumor and casual diagnosis. We analyzed the following parameters: sex, age, weight, associated diagnosis, intra and postoperative complications, recurrence and follow-up. RESULTS: 474 hernias were repaired laparoscopically in 348 patients. A predominance of males against females (30%). In 37 patients (11%) the diagnosis was made during another operation. 39 patients with recidive open surgery were operated by laparoscopy. We had only 2% intraoperative complications and 2% of recurrences. The follow-up period is 10 years. CONCLUSIONS: Laparoscopic herniorraphy may be a routine procedure with results comparable to open surgery. Laparoscopy offers a good overview of the anatomy, easily identifying the defect, with a technique increasingly used in the field of pediatric surgery.

[Indications, technique and outcomes of the one-time laparoscopic orchidopexy]. / Indicaciones, técnica y resultados de la orquidopexia laparoscópica en un solo tiempo.

The investigation of a male with impalpable testes is one of the most frequent diagnostic indications of laparoscopy and it is accepted as a therapeutic tool for the treatment of the intra-abdominal testis. We have studied thirteen patients being surgically operated by a video-assisted orchidopexy without spermatic vessels section. The technique consists of wide mobilization of the spermatic vessels and the vas deferens from the posterior peritoneum, sectioning the gubernaculum and descending of the testis to the scrotum. In all the cases, the internal groin ring was later closed by means of a laparoscopy. There were no intraoperative complications. In 100% of the cases, the testis was descended to the scrotum. The surgical time oscillated between 40 and 80 minutes (60 minutes mean). At follow-up from 6 months to 4 years, the outcome was satisfactory in 11 patients, whereas in 2 cases the operated testis ascended.

Tetraspanins in intercellular adhesion of polarized epithelial cells: spatial and functional relationship to integrins and cadherins.

The subcellular distribution of tetraspanin molecules and their functional relationship with integrins in cell-cell adhesion was studied in detail in different polarized epithelial cell models. CD9, CD81 and CD151 tetraspanins were localized at lateral cell-cell contact sites in a similar distribution to E-cadherin. Interestingly, CD9 was partially localized at the apical microvillae of Madin-Darby canine kidney cells forming multimolecular complexes distinct from those found on the basolateral membrane, suggesting the coexistence of differential tetraspanin webs with different subcellular localization. We found that tetraspanin-associated beta1 integrins at cell-to-cell contacts were in a low-affinity conformational state, and that their localization at intercellular contacts was independent of cadherin expression and adhesion. Furthermore, integrin-tetraspanin complexes were functionally relevant in cell-cell adhesion in a cadherin-independent manner, without requiring a conformational change of the integrin moiety. Nevertheless, the integrin alpha3beta1 was ligand-binding competent and this binding did not disrupt association to tetraspanins. Moreover, Chinese hamster ovary cells treated with anti-tetraspanin mAbs or activatory anti-beta1 integrin mAbs were able to develop tubule-like structures. Together, these data support tetraspanin association as a new regulatory mechanism of integrin function and suggest a role for tetraspanins-integrin complexes in providing the cell with the spatial cues necessary for their proper polarization.

Functional analysis of ligand-binding and signal transduction domains of CD69 and CD23 C-type lectin leukocyte receptors.

CD69 and CD23 are leukocyte receptors with distinctive pattern of cell expression and functional features that belong to different C-type lectin receptor subfamilies. To assess the functional equivalence of different domains of these structurally related proteins, a series of CD69/CD23 chimeras exchanging the carbohydrate recognition domain, the neck region, and the transmembrane and cytoplasmic domains were generated. Biochemical analysis revealed the importance of the neck region (Cys68) in the dimerization of CD69. Functional analysis of these chimeras in RBL-2H3 mast cells and Jurkat T cell lines showed the interchangeability of structural domains of both proteins regarding Ca2+ fluxes, serotonin release, and TNF-alpha synthesis. The type of the signal transduced mainly relied on the cytoplasmic domain and was independent of receptor oligomerization. The cytoplasmic domain of CD69 transduced a Ca2+-mediated signaling that was dependent on the extracellular uptake of Ca2+. Furthermore, a significant production of TNF-alpha was induced through the cytoplasmic domain of CD69 in RBL-2H3 cells, which was additive to that promoted via FcepsilonRI, thus suggesting a role for CD69 in the late phase of reactions mediated by mast cells. Our results provide new important data on the functional equivalence of homologous domains of these two leukocyte receptors.

The tyrosine kinase PYK-2/RAFTK regulates natural killer (NK) cell cytotoxic response, and is translocated and activated upon specific target cell recognition and killing.

The compartmentalization of plasma membrane proteins has a key role in regulation of lymphocyte activation and development of immunity. We found that the proline-rich tyrosine kinase-2 (PYK-2/RAFTK) colocalized with the microtubule-organizing center (MTOC) at the trailing edge of migrating natural killer (NK) cells. When polyclonal NK cells bound to K562 targets, PYK-2 translocated to the area of NK-target cell interaction. The specificity of this process was assessed with NK cell clones bearing activatory or inhibitory forms of CD94/NKG2. The translocation of PYK-2, MTOC, and paxillin to the area of NK-target cell contact was regulated upon specific recognition of target cells through NK cell receptors, controlling target cell killing. Furthermore, parallel in vitro kinase assays showed that PYK-2 was activated in response to signals that specifically triggered its translocation and NK cell mediated cytotoxicity. The overexpression of both the wt and a dominant-negative mutant of PYK-2, but not ZAP-70 wt, prevented the specific translocation of the MTOC and paxillin, and blocked the cytotoxic response of NK cells. Our data indicate that subcellular compartmentalization of PYK-2 correlates with effective signal transduction. Furthermore, they also suggest an important role for PYK-2 on the assembly of the signaling complexes that regulate the cytotoxic response.

Rho GTPases control migration and polarization of adhesion molecules and cytoskeletal ERM components in T lymphocytes.

Motile lymphocytes adopt a polarized morphology with different adhesion molecules (ICAM, CD43 and CD44) and ERM actin-binding proteins concentrated on the uropod, a slender posterior appendage with important functions in cell-cell interactions and lymphocyte recruitment. We have studied the role of Rho family of GTPases (Rho, Rac and Cdc42) in the control of lymphocyte polarity and migration by analyzing the effects of exogenously introduced Rho GTPase mutants. Transfection of T cell lines that constitutively display a polarized motile morphology with activated mutants of RhoA, Rac1 and Cdc42 impaired cell polarization. A guanosine nucleotide exchange factor for Rac, Tiam-1, induced the same effect as activated Rac1. Conversely, dominant negative forms of the three GTP-binding proteins induced a polarized phenotype in constitutively round-shaped T cells with redistribution of ICAM-3 and moesin to the uropod in an integrin-dependent manner. On the other hand, overexpression of dominant negative Cdc42 and activated mutants of all three Rho GTPases significantly inhibited SDF-1alpha-induced T cell chemotaxis. Together, these data demonstrate that Rho GTPases regulate lymphocyte polarization and chemokine-induced migration, and underscore the key role of Cdc42 in lymphocyte directional migration.

Activation of peripheral blood T cells by interaction and migration through endothelium: role of lymphocyte function antigen-1/intercellular adhesion molecule-1 and interleukin-15.

Cell adhesion molecules have a key role in the migration of T cells to inflammatory foci. However, the effect of the endothelial-lymphocyte interaction on the activation of the latter cells remains unresolved. We have studied the effect of resting and stimulated endothelial cells (ECs) on the activation of peripheral blood T cells (PBTLs), as assessed by the expression of CD69 and CD25 activation antigens. The incubation of PBTLs with tumor necrosis factor-alpha-activated EC monolayers, either alive or fixed, induced the expression of CD69 but not CD25, preferentially in the CD8(+) CD45RO+ cell subset. Furthermore, it induced the production of cytokines such as IFN-gamma, but not that of interleukin-2 (IL-2) and IL-4. EC treated with other stimuli such as IL-1beta, IFN-gamma, or lipopolysaccharide also showed the same proactivatory effect on T cells. Lymphocyte activation was almost completely inhibited by blocking anti-CD18 and anti-intercellular adhesion molecule-1 (anti-ICAM-1) monoclonal antibodies (MoAbs), but only slightly affected by MoAbs against CD49d, vascular cell adhesion molecule-1, and anti-IL-15. In addition, the interaction of PBTL with immobilized ICAM-1 induced CD69 expression in the same memory T-cell subset. IL-15 induced T-cell activation with expression of CD69 and CD25, and production of IFN-gamma, and its effect was additive with that triggered by cell adhesion to either EC or immobilized ICAM-1. The transmigration of PBTLs through either confluent EC monolayers or ICAM-1-coated membranes also induced efficiently the expression of CD69. When IL-15 was used as chemoattractant in these assays, a further enhancement in CD69 expression was observed in migrated cells. Together these results indicate that stimulated endothelium may have an important role in T-cell activation, through the lymphocyte function antigen-1/ICAM-1 pathway, and that IL-15 efficiently cooperates in this phenomenon. These observations could account for the abundance of CD69(+) cells in the lymphocytic infiltrates of several chronic inflammatory diseases.

Regulation of endothelial cell motility by complexes of tetraspan molecules CD81/TAPA-1 and CD151/PETA-3 with alpha3 beta1 integrin localized at endothelial lateral junctions.

Cell-to-cell junction structures play a key role in cell growth rate control and cell polarization. In endothelial cells (EC), these structures are also involved in regulation of vascular permeability and leukocyte extravasation. To identify novel components in EC intercellular junctions, mAbs against these cells were produced and selected using a morphological screening by immunofluorescence microscopy. Two novel mAbs, LIA1/1 and VJ1/16, specifically recognized a 25-kD protein that was selectively localized at cell-cell junctions of EC, both in the primary formation of cell monolayers and when EC reorganized in the process of wound healing. This antigen corresponded to the recently cloned platelet-endothelial tetraspan antigen CD151/PETA-3 (platelet-endothelial tetraspan antigen-3), and was consistently detected at EC cell-cell contact sites. In addition to CD151/PETA-3, two other members of the tetraspan superfamily, CD9 and CD81/ TAPA-1 (target of antiproliferative antibody-1), localized at endothelial cell-to-cell junctions. Biochemical analysis demonstrated molecular associations among tetraspan molecules themselves and those of CD151/ PETA-3 and CD9 with alpha3 beta1 integrin. Interestingly, mAbs directed to both CD151/PETA-3 and CD81/ TAPA-1 as well as mAb specific for alpha3 integrin, were able to inhibit the migration of ECs in the process of wound healing. The engagement of CD151/PETA-3 and CD81/TAPA-1 inhibited the movement of individual ECs, as determined by quantitative time-lapse video microscopy studies. Furthermore, mAbs against the CD151/PETA-3 molecule diminished the rate of EC invasion into collagen gels. In addition, these mAbs were able to increase the adhesion of EC to extracellular matrix proteins. Together these results indicate that CD81/TAPA-1 and CD151/PETA-3 tetraspan molecules are components of the endothelial lateral junctions implicated in the regulation of cell motility, either directly or by modulation of the function of the associated integrin heterodimers.

Utilización de células permeabilizadas de K.fragilis para la obtención de leche con bajo contenido de lactosa / Use of permeabilized K. fragilis cells in low-lactose milk production

Células de Kluyveromyces fragilis aisladas por el Instituto de Investigaciones de Derivados de la Cana de Azúcar (ICIDCA) fueron permeabilizadas con una mezcla de tolueno-etanol (1:4) y 0,02 % de tritón X-100 durante 20minutos. La lactasa de células permeabilizadas presentó una actividad máxima a la temperatura de 55-C y una buena estabilidad a 50-C por más de 6 horas, mientras que el pH óptimo de 6,5 es similar a la enzima libre. La afinidad (Km) de la enzima en las células permeabilizadas, para el sustrato ONPG, fue de 2,63 mM y de 30,52 mM para la lactosa. Los resultados de la hidrólisis de lactosa en leche descremada en polvo se corresponden con los parámetros cinéticos obtenidos, alcanzándose un grado de hidrólisis de 93,17 % a las 5 horas a 50 -C y un porcentaje de conversión de monosacáridos de 73,65 %, utilizando 2 gramos de células enteras permeabilizadas por litro de leche