Pharmacokinetics and safety of LEAD-452, an EGFR-specific 4-1BB-agonistic trimerbody in non-human primates.

LEAD-452 is a humanized bispecific EGFR-targeted 4-1BB-agonistic trimerbody with a unique trimeric configuration compared to other 4-1BB-specific antibodies that are currently in development. Indeed, enhanced tumor-specific costimulation and very remarkable safety and efficacy profiles have been observed in mouse models. Here, we conducted for the first time a preclinical pharmacokinetic and toxicity study in non-human primates (NHP) (Macaca fascicularis). LEAD-452 exhibits comparable binding affinity for human and macaque targets, indicating its pharmacological significance for safety testing across species. The NHP were administered LEAD-452 in a series of ascending doses, ranging from 0.1 mg/kg to 10 mg/kg, and repeated doses up to 20 mg/kg. The administration of LEAD-452 was found to be clinically well tolerated, with no major related adverse effects observed. Furthermore, there have been no reported cases of liver toxicity, thrombocytopenia, and neutropenia, which are commonly associated with treatments using conventional anti-4-1BB IgG-based antibodies. In addition, neither IgM nor IgG-based anti-drug antibodies were detected in serum samples from NHP during the study, regardless of the dose of LEAD-452 administered. These results support the clinical development of LEAD-452 for the treatment of solid tumors.

Characterization of a Trispecific PD-L1 Blocking Antibody That Exhibits EGFR-Conditional 4-1BB Agonist Activity.

Immune checkpoint blockade has changed the treatment paradigm for advanced solid tumors, but the overall response rates are still limited. The combination of checkpoint blockade with anti-4-1BB antibodies to stimulate tumor-infiltrating T cells has shown anti-tumor activity in human trials. However, the further clinical development of these antibodies has been hampered by significant off-tumor toxicities. Here, we generated an anti-4-1BB/EGFR/PD-L1 trispecific antibody consisting of a triple-targeting tandem trimerbody (TT) fused to an engineered silent Fc region. This antibody (IgTT-4E1-S) was designed to combine the blockade of the PD-L1/PD-1 axis with conditional 4-1BB costimulation specifically confined to the tumor microenvironment (TME). The antibody demonstrated simultaneous binding to purified EGFR, PD-L1, and 4-1BB in solution, effective blockade of the PD-L1/PD1 interaction, and potent 4-1BB-mediated costimulation, but only in the presence of EGFR-expressing cells. These results demonstrate the feasibility of IgTT-4E1-S specifically blocking the PD-L1/PD-1 axis and inducing EGFR-conditional 4-1BB agonist activity.

Evaluación del Rendimiento y Producción de Humedad de los Humidificadores de Burbuja usados con Mascarillas Oxígeno de Concentración Variable / Evaluation of the Performance and Humidity Production of Bubble Humidifiers used with Variable Concentration Oxygen Masks

Introducción. Introducción. El uso de humidificadores de burbuja asociados a equipos de oxigenoterapia es una práctica común en los centros hospitalarios de nuestro país. Sin embargo, no se ha evaluado el aporte real de humedad que entregan estos equipos cuando son usados con sistemas de concentración variable a distintas concentraciones de oxígeno. Además, se han descrito problemas de funcionamiento en la entrega de oxígeno cuando se han ocupado este tipo de humidificador. Objetivo. Determinar el aporte real de humedad que entregan los humidificadores de burbuja al ser usados con sistemas de oxigenoterapia de concentración variable. Métodos. Se evaluaron 10 humidificadores de burbuja, marca Hudson RCI®, con válvula liberadora de presión, los cuales se conectaron a un sistema de oxigenoterapia de concentración variable (Marca Hudson RCI®, modelo MULTI-VENT). La medición de la humedad relativa (HR) se realizó en una cámara de acrílico, donde se conectó el inyector del sistema de oxigenoterapia. Para medir H.R. y temperatura se utilizó un higrómetro digital (Veto®, Italia) y un higrómetro análogo (Hygromat®, Alemania). Cada medición requirió un tiempo de 5 minutos para lograr un valor estable. Se realizaron mediciones de la humedad relativa y temperatura entregada por este sistema, con y sin humidificador, a concentraciones de oxígeno de 0.24, 0.26, 0.28, 0.3, 0.35, 0.4 y 0.5. Resultados. Para las la temperatura, no hubo diferencias entre las mediciones realizadas con y sin humidificador de burbuja. Para la humedad relativa, sólo hubo diferencias estadísticamente significativas a concentraciones de oxígeno altas (> 0.35), pero con escaso cambio en la entrega de humedad absoluta. Cuando se utilizó flujos operativos ≥ 6 L/min se activó la válvula liberadora de presión del humidificador. Conclusión. El uso de humidificadores de burbuja asociados a sistemas de oxigenoterapia de concentración variable no genera un aumento significativo de la humedad entregada a concentraciones bajas de oxigeno, por lo cual se podría prescindir de su uso.

Background. The use of bubble humidifiers associated with oxygen therapy equipment is a common practice in hospitals in our country. However, the real contribution of humidity delivered by these equipments when used with variable concentration systems at different oxygen concentrations has not been evaluated. In addition, operating problems have been described in the delivery of oxygen when this type of humidifier has been used. Objetive. Determine the real contribution of humidity delivered by bubble humidifiers when used with variable concentration oxygen therapy systems. Methods. 10 Hudson RCI® brand bubble humidifiers with a pressure release valve were evaluated, which were connected to a variable concentration oxygen therapy system (Hudson RCI® brand, MULTI-VENT model). The relative humidity (RH) was measured in an acrylic chamber, where the injector of the oxygen therapy system was connected. To measure R.H. and temperature, a digital hygrometer (Veto®, Italy) and an analog hygrometer (Hygromat®, Germany) were used. Each measurement required a time of 5 minutes to achieve a stable value. Measurements of the relative humidity and temperature delivered by this system were made, with and without a humidifier, at oxygen concentrations of 0.24, 0.26, 0.28, 0.3, 0.35, 0.4 and 0.5. Results. For temperature, there were no differences between the measurements made with and without a bubble humidifier. For relative humidity, there were only statistically significant differences at high oxygen concentrations (> 0.35), but with little change in absolute humidity delivery. When operating flows ≥ 6 L/min were used, the humidifier pressure relief valve was activated. Conclusion. The use of bubble humidifiers associated with variable concentration oxygen therapy systems does not generate a significant increase in the humidity delivered at low oxygen concentrations, so their use could be dispensed with.

A PD-L1/EGFR bispecific antibody combines immune checkpoint blockade and direct anti-cancer action for an enhanced anti-tumor response.

Immune checkpoint blockade (ICB) with antibodies has shown durable clinical responses in a wide range of cancer types, but the overall response rate is still limited. Other effective therapeutic modalities to increase the ICB response rates are urgently needed. New bispecific antibody (bsAb) formats combining the ICB effect and a direct action on cancer cells could improve the efficacy of current immunotherapies. Here, we report the development of a PD-L1/EGFR symmetric bsAb by fusing a dual-targeting tandem trimmer body with the human IgG1 hinge and Fc regions. The bsAb was characterized in vitro and the antitumor efficacy was evaluated in humanized mice bearing xenografts of aggressive triple-negative breast cancer and lung cancer. The IgG-like hexavalent bsAb, designated IgTT-1E, was able to simultaneously bind both EGFR and PD-L1 antigens, inhibit EGF-mediated proliferation, effectively block PD-1/PD-L1 interaction, and induce strong antigen-specific antibody-dependent cellular cytotoxicity activity in vitro. Potent therapeutic efficacies of IgTT-1E in two different humanized mouse models were observed, where tumor growth control was associated with a significantly increased proportion of CD8+ T cells. These results support the development of IgTT-1E for the treatment of EGFR+ cancers.

Tumor targeted 4-1BB agonist antibody-albumin fusions with high affinity to FcRn induce anti-tumor immunity without toxicity.

Costimulation of tumor-infiltrating T lymphocytes by anti-4-1BB monoclonal antibodies (mAbs) has shown anti-tumor activity in human trials, but can be associated with significant off-tumor toxicities involving FcγR interactions. Here, we introduce albumin-fused mouse and human bispecific antibodies with clinically favorable pharmacokinetics designed to confine 4-1BB costimulation to the tumor microenvironment. These Fc-free 4-1BB agonists consist of an EGFR-specific VHH antibody, a 4-1BB-specific scFv, and a human albumin sequence engineered for high FcRn binding connected in tandem (LiTCo-Albu). We demonstrate in vitro cognate target engagement, EGFR-specific costimulatory activity, and FcRn-driven cellular recycling similar to non-fused FcRn high-binding albumin. The mouse LiTCo-Albu exhibited a prolonged circulatory half-life and in vivo tumor inhibition, with no indication of 4-1BB mAb-associated toxicity. Furthermore, we show a greater therapeutic effect when used in combination with PD-1-blocking mAbs. These findings demonstrate the feasibility of tumor-specific LiTCo-Albu antibodies for safe and effective costimulatory strategies in cancer immunotherapy.

Two-Minute Step Test as a Complement to Six-Minute Walk Test in Subjects With Treated Coronary Artery Disease.

The 2-Minute Step Test (2MST) has been presented as an alternative to the 6-Minute Walk Test (6MWT) based on the association between the two tests in older adults; however, some authors propose that it should not be a substitute but rather a complement to the latter in the fitness evaluation. Specifically, in coronary disease, despite the potential and clinical utility of 2MST, the relationship of both tests in this population is unknown. This study aimed to determine the relationship between 6MWT and 2MST and to explore the relationship of biodemographic factors for both tests in subjects with treated coronary artery disease. For this, the 6MWT and the 2MST were applied to patients with coronary artery disease treated in 6 hospitals in Chile between May 2019 and February 2020. Additionally, lower limb strength was assessed by a chair-stand test, grip strength was assessed by a dynamometer, and physical measurements were applied. In total, 163 participants underwent both tests (average age = 58.7 ± 9.8 years; 73.6% men; 64.4% revascularized by angioplasty; 28.2% revascularized by surgery, and 7.4% treated by drugs or thrombolysis). Heart rate was higher at the end of the 6MWT, while the perception of effort was greater at the end of the 2MST. There was a weak positive correlation between the 6MWT and the 2MST in subjects with treated coronary disease (r = 0.28, p = 0.0003). While age (r = -0.27), weight (r = 0.25), height (r = 0.49), and strength of both lower limbs (r = 0.41) and grip strength (r = 0.53) correlated weakly or moderately to the covered distance in 6MWT, the number of steps by the 2MST correlated only weakly to height (r = 0.23), lower limb strength (r = 0.34), and grip strength (r = 0.34). Age, weight, height, lower limb strength, and grip strength would explain better the meters walked in the 6MWT than the steps achieved in the 2MST. With these findings, we can conclude that, in patients with treated coronary artery disease, it does not seem advisable to replace 6MWT with 2MST when it is possible to do so. Additionally, the 2MST may provide additional information in the fitness evaluation. However, the usefulness of 2MST in this population needs to be further studied.

Trispecific T-cell engagers for dual tumor-targeting of colorectal cancer.

Retargeting of T lymphocytes toward cancer cells by bispecific antibodies has demonstrated its therapeutic potential, with one such antibody approved for the treatment of acute lymphoblastic leukemia (blinatumomab) and several other in clinical trials. However, improvement of their efficacy and selectivity for solid tumors is still required. Here, we describe a novel tandem T-cell recruiting trispecific antibody for the treatment of colorectal cancer (CRC). This construct, termed trispecific T-cell engager (TriTE), consists of a CD3-specific single-chain Fv (scFv) flanked by anti-epidermal growth factor receptor (EGFR) and anti-epithelial cell adhesion molecule (EpCAM) single-domain VHH antibodies. The TriTE was well expressed in mammalian and yeast cells, bound the cognate antigens of the three parental antibodies, and enabled the specific cytolysis of EGFR- and/or EpCAM-expressing cancer cells, without inducing T cell activation and cytoxicity against double-negative (EGFR-EpCAM-) cancer cells. Bivalent bispecific targeting of double-positive HCT116 cells by TriTE improved in vitro potency up to 100-fold compared to single-positive cells and significantly prolonged survival in vivo. In addition, it was less efficient at killing single-positive target cells than the corresponding bispecific controls, leading to potentially enhanced tumor specificity. Moreover, dual targeting of two tumor-associated antigens may contribute toward preventing the tumor escape by antigen loss caused by selective pressures from conventional single-targeting T-cell engagers, and may help to overcome antigenic heterogeneity.

Prevalencia de edema cerebral en pacientes con cetoacidosis diabética / Prevalence of cerebral edema among diabetic ketoacidosis patients

Introducción. El edema cerebral (EC) es la complicación más grave de la cetoacidosis diabética (CAD) en niños. La patogénesis del EC no se conoce con exactitud y su aparición ha sido relacionada con la terapia de rehidratación endovenosa en el tratamiento inicial.Objetivos. Estimar la prevalencia de EC en pacientes con CAD tratados en el Hospital General de Niños Pedro de Elizalde mediante rehidratación endovenosa y analizar potenciales factores de riesgo para el desarrollo de EC.Materiales y método. Estudio de diseño transversal para prevalencia y un análisis exploratorio para comparar las características clínicas y de laboratorio entre los pacientes con y sin EC. Se incluyeron pacientes de 1 a 18 años hospitalizados con diagnóstico de CAD desde el 1 de enero de 2005 hasta el 31 de diciembre de 2014.Resultados. Se analizaron 693 episodios de CAD en 561 historias clínicas. En 10 pacientes, se evidenció EC (el 1,44 %; intervalo de confianza del 95 %: 0,8-2,6). Los pacientes con EC presentaron mayor uremia (p < 0,001), menor presión de dióxido de carbono (p < 0,001) y menor natremia (p < 0,001) que aquellos pacientes sin EC.Conclusión. La prevalencia de EC en pacientes con CAD fue del 1,44 %, menor que la reportada en nuestro país (del 1,8 %). Los factores de riesgo al ingresar asociados a su desarrollo fueron la presencia de uremia elevada, hiponatremia e hipocapnia.

Introduction. Cerebral edema (CE) is the most severe complication of diabetic ketoacidosis (DKA) in children. There is no accurate knowledge of CE pathogenesis and its onset has been related to intravenous rehydration therapy during the initial treatment.Objectives. To estimate the prevalence of CE among DKA patients treated at Hospital General de Niños Pedro de Elizalde with intravenous rehydration and analyze potential risk factors for the development of CE.Materials and methods. Cross-sectional prevalence study and exploratory analysis to compare clinical and laboratory characteristics between patients with and without CE. Patients aged 1-18 years hospitalized with the diagnosis of DKA between January 1st, 2005 and December 31st, 2014 were included.Results. A total of 693 DKA events from 561 medical records were analyzed. Ten patients had evidence of CE (1.44 %; 95 % confidence interval: 0.8-2.6). Patients with CE had higher serum urea levels (p < 0.001), lower carbon dioxide pressure (p < 0.001), and lower serum sodium levels (p < 0.001) than those without CE.Conclusion. The prevalence of CE among DKA patients was 1.44 %, smaller than that reported in our country (1.8 %). The risk factors at admission associated with CE development were high serum urea levels, hyponatremia, and hypocapnia.

TGF-ß-induced IGFBP-3 is a key paracrine factor from activated pericytes that promotes colorectal cancer cell migration and invasion.

The crosstalk between cancer cells and the tumor microenvironment has been implicated in cancer progression and metastasis. Fibroblasts and immune cells are widely known to be attracted to and modified by cancer cells. However, the role of pericytes in the tumor microenvironment beyond endothelium stabilization is poorly understood. Here, we report that pericytes promoted colorectal cancer (CRC) cell proliferation, migration, invasion, stemness, and chemoresistance in vitro, as well as tumor growth in a xenograft CRC model. We demonstrate that coculture with human CRC cells induced broad transcriptomic changes in pericytes, mostly associated with TGF-ß receptor activation. The prognostic value of a TGF-ß response signature in pericytes was analyzed in CRC patient data sets. This signature was found to be a good predictor of CRC relapse. Moreover, in response to stimulation by CRC cells, pericytes expressed high levels of TGF-ß1, initiating an autocrine activation loop. Investigation of secreted mediators and underlying molecular mechanisms revealed that IGFBP-3 is a key paracrine factor from activated pericytes affecting CRC cell migration and invasion. In summary, we demonstrate that the interplay between pericytes and CRC cells triggers a vicious cycle that stimulates pericyte cytokine secretion, in turn increasing CRC cell tumorigenic properties. Overall, we provide another example of how cancer cells can manipulate the tumor microenvironment.

Anti-tumoral potential of a human granulysin-based, CEA-targeted cytolytic immunotoxin.

Granulysin is a protein present in the granules of human cytotoxic T lymphocytes (CTL) and natural killer (NK) cells, with cytolytic activity against microbes and tumors. Previous work demonstrated the therapeutic effect of intratumoral injection of recombinant granulysin using in vivo models of breast cancer and multiple myeloma. In the present work we have developed a granulysin gene fusion to the anti-carcinoembryonic antigen (CEA/CEACAM5) single chain Fv antibody fragment MFE23. Both granulysin and the granulysin-based immunotoxin were expressed in Pichia pastoris. The immunotoxin specifically recognized CEA, purified or expressed on the cell surface. Moreover, the bioactivity of the immunotoxin against several CEA+ cell lines was higher than that of granulysin alone. Granulysin and the immunotoxin were tested as a treatment in in vivo xenograft models in athymic mice. When injected intratumorally, both granulysin and the immunotoxin were able to inhibit tumor growth. Furthermore, systemic administration of the immunotoxin demonstrated a decrease in tumor growth in a CEA+ tumor-bearing mouse model, whereas granulysin did not exhibit a therapeutic effect. This is the first granulysin-based immunotoxin and the present work constitutes the proof of concept of its therapeutic potential.

A tumor-targeted trimeric 4-1BB-agonistic antibody induces potent anti-tumor immunity without systemic toxicity.

The costimulation of immune cells using first-generation anti-4-1BB monoclonal antibodies (mAbs) has demonstrated anti-tumor activity in human trials. Further clinical development, however, is restricted by significant off-tumor toxicities associated with FcγR interactions. Here, we have designed an Fc-free tumor-targeted 4-1BB-agonistic trimerbody, 1D8N/CEGa1, consisting of three anti-4-1BB single-chain variable fragments and three anti-EGFR single-domain antibodies positioned in an extended hexagonal conformation around the collagen XVIII homotrimerization domain. The1D8N/CEGa1 trimerbody demonstrated high-avidity binding to 4-1BB and EGFR and a potent in vitro costimulatory capacity in the presence of EGFR. The trimerbody rapidly accumulates in EGFR-positive tumors and exhibits anti-tumor activity similar to IgG-based 4-1BB-agonistic mAbs. Importantly, treatment with 1D8N/CEGa1 does not induce systemic inflammatory cytokine production or hepatotoxicity associated with IgG-based 4-1BB agonists. These results implicate FcγR interactions in the 4-1BB-agonist-associated immune abnormalities, and promote the use of the non-canonical antibody presented in this work for safe and effective costimulatory strategies in cancer immunotherapy.

Tratamiento laparoscópico de hernia incisional con malla biocompatible de ptfe intraperitoneal / Laparoscopic treatment of incisional hernia with ptfe biological mesh

Una hernia Incisional (HI) puede definirse como cualquier defecto de la pared abdominal, con o sin aumento de volumen, en el área de una cicatriz postoperatoria, perceptible o palpable por el examen clínico o imagenológico(1). La hernioplastía intraperitoneal laparoscópica constituye una opción segura para el tratamiento de hernias incisionales de la pared abdominal, asociada a una baja tasa de recurrencia que alcanza a 6,25% a largo plazo(1). Se presenta un caso de Hernioplastia laparoscópica con colocación de malla de PTFE intraperitoneal, como una alternativa válida para la reparación de hernias incisionales.

An incisional hernia can be defined a s a ny d efect o f t he abdominal wall, with or without an increase in volume, in the area of a postoperative scar, perceptible or palpable by clinical or imaging examination. The l aparoscopic i ntraperitoneal h ernioplasty is a safe option for the treatment of incisional hernias of the abdominal wall, associated with a low recurrence rate that reaches 6.25% in the long term. In this time, a case of laparoscopic hernioplasty with placement of a intraperitoneal PTFE mesh is presented as a valid alternative for the repair of incisional hernias.

ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy.

The redirection of T cell activity using bispecific antibodies is one of the most promising cancer immunotherapy approaches currently in development, but it is limited by cytokine storm-related toxicities, as well as the pharmacokinetics and tumor-penetrating capabilities of current bispecific antibody formats. Here, we have engineered the ATTACK (Asymmetric Tandem Trimerbody for T cell Activation and Cancer Killing), a novel T cell-recruiting bispecific antibody which combines three EGFR-binding single-domain antibodies (VHH; clone EgA1) with a single CD3-binding single-chain variable fragment (scFv; clone OKT3) in an intermediate molecular weight package. The two specificities are oriented in opposite directions in order to simultaneously engage cancer cells and T cell effectors, and thereby promote immunological synapse formation. EgA1 ATTACK was expressed as a homogenous, non-aggregating, soluble protein by mammalian cells and demonstrated an enhanced binding to EGFR, but not CD3, when compared to the previously characterized tandem bispecific antibody which has one EgA1 VHH and one OKT3 scFv per molecule. EgA1 ATTACK induced synapse formation and early signaling pathways downstream of TCR engagement at lower concentrations than the tandem VHH-scFv bispecific antibody. Furthermore, it demonstrated extremely potent, dose-dependent cytotoxicity when retargeting human T cells towards EGFR-expressing cells, with an efficacy over 15-fold higher than that of the tandem VHH-scFv bispecific antibody. These results suggest that the ATTACK is an ideal format for the development of the next-generation of T cell-redirecting bispecific antibodies.

COMPORTAMIENTOS DE RIESGO Y NIVEL DE CONOCIMIENTO DE INFECCIONES DE TRANSMISIÓN SEXUAL EN ESTUDIANTES UNIVERSITARIOS / RISK BEHAVIORS AND LEVEL OF KNOWLEDGE ABOUT SEXUALLY TRANSMITTED INFECTIONS IN UNIVERSITY STUDENTS

Las infecciones de transmisión sexual constituyen un problema epidemiológico y clínico de primer orden en todo el mundo por las secuelas que pueden ocasionar y por sus consecuencias económicas. Los adolescentes y jóvenes tienen mayor riesgo de presentarlas debido a factores como el inicio temprano de relaciones sexuales, la promiscuidad, entre otros. El objetivo de esta investigación fue determinar los comportamientos de riesgo y nivel de conocimiento de infecciones de transmisión sexual en estudiantes de Bacteriología de la ciudad de Cartagena. Estudio correlacional de corte transversal, en donde se aplicó un cuestionario para determinar comportamientos de riesgo y nivel de conocimiento a 128 estudiantes de Bacteriología. El 78,1 % de los estudiantes encuestados sí ha tenido relaciones sexuales. El 55 % ingiere bebidas alcohólicas antes de una relación sexual. A través del análisis estadístico se estableció una asociación entre la presencia de ITS con el consumo de drogas (p=0,042). Los estudiantes presentaron un nivel de conocimiento regular. No se encontró asociación estadísticamente significativa entre los comportamientos de riesgo y el nivel de conocimiento (p> 0,05). Los resultados encontrados en esta investigación evidencian la necesidad de realizar actividades de prevención para motivar cambios de comportamiento que permitan disminuir el riesgo de contagio de estas enfermedades.

Sexually transmitted infections are an epidemiologic and clinical problem of first order in all the world by effect that can produce and their economic consequences. Adolescents and young are in most risk to have these diseases by facts such as premature sexual relationships and promiscuity. The objective of this investigation was to determine risk behaviors and level of knowledge about sexually transmitted infections in students' community. Correlational cross-sectional study. A questionnaire was applied to determine risk behaviors and level of knowledge about sexually transmitted infections to 128 students' community. In this study was found 78.1% of students had sexual relationships. 55% of students drink alcohol before a sexual relationship. By means of statistical analysis was found association between sexually transmitted infections and drug use (p=0.042). Students had a regular level of knowledge. It not was found association between risk behaviors and level of knowledge (p>0.05). Results found in this investigation demonstrate the necessity of making activities for prevention of these infections and motivating changes of behaviors for reducing risk of contagion of these infections.

Immune Regulation by Pericytes: Modulating Innate and Adaptive Immunity.

Pericytes (PC) are mural cells that surround endothelial cells in small blood vessels. PC have traditionally been credited with structural functions, being essential for vessel maturation and stabilization. However, an accumulating body of evidence suggests that PC also display immune properties. They can respond to a series of pro-inflammatory stimuli and are able to sense different types of danger due to their expression of functional pattern-recognition receptors, contributing to the onset of innate immune responses. In this context, PC not only secrete a variety of chemokines but also overexpress adhesion molecules such as ICAM-1 and VCAM-1 involved in the control of immune cell trafficking across vessel walls. In addition to their role in innate immunity, PC are involved in adaptive immunity. It has been reported that interaction with PC anergizes T cells, which is attributed, at least in part, to the expression of PD-L1. As components of the tumor microenvironment, PC can also modulate the antitumor immune response. However, their role is complex, and further studies will be required to better understand the crosstalk of PC with immune cells in order to consider them as potential therapeutic targets. In any case, PC will be looked at with new eyes by immunologists from now on.

Intramolecular trimerization, a novel strategy for making multispecific antibodies with controlled orientation of the antigen binding domains.

Here, we describe a new strategy that allows the rapid and efficient engineering of mono and multispecific trivalent antibodies. By fusing single-domain antibodies from camelid heavy-chain-only immunoglobulins (VHHs) to the N-terminus of a human collagen XVIII trimerization domain (TIE(XVIII)) we produced monospecific trimerbodies that were efficiently secreted as soluble functional proteins by mammalian cells. The purified VHH-TIE(XVIII) trimerbodies were trimeric in solution and exhibited excellent antigen binding capacity. Furthermore, by connecting with two additional glycine-serine-based linkers three VHH-TIE(XVIII) modules on a single polypeptide chain, we present an approach for the rational design of multispecific tandem trimerbodies with defined stoichiometry and controlled orientation. Using this technology we report here the construction and characterization of a tandem VHH-based trimerbody capable of simultaneously binding to three different antigens: carcinoembryonic antigen (CEA), epidermal growth factor receptor (EGFR) and green fluorescence protein (GFP). Multispecific tandem VHH-based trimerbodies were well expressed in mammalian cells, had good biophysical properties and were capable of simultaneously binding their targeted antigens. Importantly, these antibodies were very effective in inhibiting the proliferation of human epidermoid carcinoma A431 cells. Multispecific VHH-based trimerbodies are therefore ideal candidates for future applications in various therapeutic areas.

Abordaje laparoscópico en peritonitis por perforación vesical / Laparoscopic in bladder perforation peritonitis

Las lesiones vesicales son causadas por traumatismos cerrados de alta energía que interrumpen la pelvis ósea, un golpe directo a una vejiga distendida, lesiones penetrantes o causas iatrogénicas varias. 1 Se presenta el caso de una paciente de sexo femenino, joven, con patología de base en estudio por el Servicio de Urología, quien en el contexto del diagnóstico de dicha patología fue sometida a un procedimiento urológico invasivo (RTU), con abdomen peritoneal 24hs después del procedimiento. En el acto quirúrgico se constató aproximadamente 1000cc de orina y perforación vesical de 1cm de diámetro con mucosa exteriorizada hacia cavidad peritoneal. Se realizó biopsia de la mucosa vesical, rafia de la perforación en un plano, lavado y aspirado de cavidad por videolaparoscopía, y colocación de drenaje tubular y sonda vesical en permanencia.

Bladder injuries are caused by high energy blunt trauma thatdisrupt the pelvis bone, a direct blow to a distended bladder, penetratinginjuries or some iatrogenic causes. The case of a youngfemale patient, underlying pathology study by the Department ofUrology, who in the context of diagnosis of this disease underwenta urological invasive procedure (TUR), with peritoneal abdomenpresents 24 hours after the procedure. During surgery it was foundabout 1000cc of urine and bladder perforation 1 cm in diameterwith externalized mucosa into peritoneal cavity. Biopsy of bladdermucosa, closure of defect with absorbable material and drainage ofthe cavity was performed entirely by videolaparoscopy.

Role of nucleotide-binding oligomerization domain 1 (NOD1) in pericyte-mediated vascular inflammation.

We have recently described the response of human brain pericytes to lipopolysaccharide (LPS) through toll-like receptor 4 (TLR4). However, Gram-negative pathogen-associated molecular patterns include not only LPS but also peptidoglycan (PGN). Given that the presence of co-purified PGN in the LPS preparation previously used could not be ruled out, we decided to analyse the expression of the intracellular PGN receptors NOD1 and NOD2 in HBP and compare the responses to their cognate agonists and ultrapure LPS. Our findings show for the first time that NOD1 is expressed in pericytes, whereas NOD2 expression is barely detectable. The NOD1 agonist C12-iE-DAP induced IL6 and IL8 gene expression by pericytes as well as release of cytokines into culture supernatant. Moreover, we demonstrated the synergistic effects of NOD1 and TLR4 agonists on the induction of IL8. Using NOD1 silencing in HBP, we showed a requirement for C12-iE-DAP-dependent signalling. Finally, we could discriminate NOD1 and TLR4 pathways in pericytes by pharmacological targeting of RIPK2, a kinase involved in NOD1 but not in TLR4 signalling cascade. p38 MAPK and NF-κB appear to be downstream mediators in the NOD1 pathway. In summary, these results indicate that pericytes can sense Gram-negative bacterial products by both NOD1 and TLR4 receptors, acting through distinct pathways. This provides new insight about how brain pericytes participate in the inflammatory response and may have implications for disease management.

Utilidad de la cpre en la pancreatitis aguda biliar e ictericia / Utility in ercp acute biliary pancreatitis amd jaundice

El uso de la Colangio-Pancreatografía Retrógrada Endoscópica (CPRE) en la Pancreatitis Aguda, fundamentalmente en la de origen Biliar (PAB) no ha estado exento de polémicas. Se han realizado varios estudios prospectivos randomizados comparando la desobstrucción endoscópica biliopancreática temprana versus el tratamiento conservador. Los cuales no presentan una diferencia significativa entre la morbilidad y la mortalidad de la pancreatitis aguda e ictericia entre ambos grupos. Otros estudios más actuales (Revisiones sistemáticas y Resúmenes estructurados) presentan resultados similares.Con respecto a la decisión de realizar o no una CPRE, la American Society for Gastrointestinal Endoscopy (ASGE), publicó una guía en el 2010, en la cual se emplean predictores clínicos para asignar tres grupos de riesgo de coledocolitiasis.La indicación de la CPRE en pacientes con pancreatitis aguda e ictericia que no presenten colangitis o alto riesgo de coledocolitiasis, aún merece una mejor evaluación de la misma ya que los factores indicadores de litiasis coledociana no siempre están presentes y si lo están, presentan falsos positivos que nos obliga a extremar recursos para evitar una CPRE innecesaria, de alto costo en nuestro medio y no exenta de complicaciones.

The use of endoscopic retrograde cholangiopancreatography-Pancreatography (ERCP) in Acute Pancreatitis, mainly in the home Billiards (PAB) has not been without controversy. There have been several prospective randomized studies comparing early unblocking biliopancreatic endoscopic versus conservative treatment. Which do not show a significant difference between the morbidity and mortality of acute pancreatitis and jaundice between the two groups. Other more recent studies show similar results (systematic and structured summaries Revisions). With respect to the decision to perform or not an ERCP, the American Society for Gastrointestinal Endoscopy (ASGE), published a guide in 2010, in which clinical predictors are used to assign three groups of risk of choledocholithiasis. The indication for ERCP in patients with acute pancreatitis and jaundice which do not cholangitis or high risk of choledocholithiasis, still deserves a better assessment of it as indicators of cholelithiasis factors are not always present and if they are, have false positives requiring us to take extra resources to avoid unnecessary ERCP, high cost in our midst and not without complications.

Relación odotólogo paciente / The dentist patient relationship

El objetivo de este trabajo es ampliar el campo de estudio de las relaciones odontólogo/paciente y médico/paciente, para analizar los antecedentes de la entrevista y sus consecuencias. Se analizan las diferencias entre estas relaciones profecionales y se concluye en la definitiva diferenciación entre ellas, que obliga a la odontología a elaborar el estudio particular de sus relaciones desde el nivel psicológico.