[Reconstruction of the medial patellofemoral ligament with quadricipital tendon autograft]. / Reconstrucción del ligamento patelofemoral medial con autoinjerto de tendón cuadricipital.

INTRODUCTION: Alterations of the patellofemoral joint are one of the most common causes of pain and inflammation as well as joint damage and instability of the knee. Patellofemoral instability is a common multifactorial entity, requiring realignment by proximal, distal, or a combination of techniques. Within the proximal procedures in soft tissues, is the medial patellofemoral ligament plasty (MPFL), which aims to realign the patella medially and restore the anatomy between the quadricipital tendon, patella and tubercle of the tibia. OBJECTIVE: To demonstrate that the plasty of the LPFM with quadriceps autograft is an effective technique with a low level of complications. MATERIAL AND METHODS: Observational, longitudinal, retrospective, descriptive, basic and clinical study. We present a series of 15 patients operated with this technique between October 2014 and September 2019. RESULTS: LPFM plasty with autograft quadricipital is a safe technique, which does not use implants, which reduces the risk of complications and restores the anatomy of the extensor apparatus of the knee. CONCLUSION: Our technique of repair of the medial patellofemoral ligament, with quadriceps autograft is a safe, reproducible technique, with good results in the medium term, as well as a low incidence of complications. Patients have returned to their previous sports activities without episodes of re-dislocation.

INTRODUCCIÓN: Las alteraciones de la articulación patelofemoral son una de las causas más comunes de dolor e inflamación así como de daño articular e inestabilidad de la rodilla. La inestabilidad patelofemoral es una entidad común multifactorial que requiere de una realineación mediante técnicas proximales, distales o una combinación de ellas. Entre los procedimientos proximales en tejidos blandos se encuentra la plastía del ligamento patelofemoral medial (LPFM), la cual tiene como objetivo realinear la patela hacia medial y restaurar la anatomía entre el tendón cuadricipital, patela y tubérculo de la tibia. OBJETIVO: Demostrar que la plastía del LPFM con autoinjerto de cuádriceps es una técnica efectiva y con bajo nivel de complicaciones. MATERIAL Y MÉTODOS: Estudio observacional, longitudinal, retrospectivo, descriptivo, básico y clínico. Se presenta una serie de 15 pacientes operados con esta técnica entre Octubre de 2014 y Septiembre de 2019. RESULTADOS: La plastía del LPFM con autoinjerto del cuadricipital es una técnica segura que no utiliza implantes, lo cual reduce el riesgo de complicaciones y restaura la anatomía del aparato extensor de la rodilla. CONCLUSIÓN: Nuestra técnica con autoinjerto de cuádriceps es segura, con buenos resultados a mediano plazo y baja incidencia de complicaciones. Los pacientes han regresado a sus actividades deportivas previas sin episodios de reluxación.

Implication of MT1-MMP in the maturation steps of benign melanocytic nevi.

BACKGROUND: Metalloproteinases (MMPs) are proteins involved in extracellular matrix breakdown and have been implicated in stages of migration and metastasis. MT1-MMP is an MMP anchored to the cell membrane. During maturation, melanocytic nevi penetrate the extracellular matrix and express MMPs. METHODS: We studied 10 junctional, 10 compound, and 10 intradermal nevi diagnosed by clinical and histological studies and by performing immunohistochemical study to assess MT1-MMP activity. RESULTS: We found evidence of MT1-MMP expression in melanocytic nevus cells, particularly around the entire border of cell nests. Expression was more intense in junctional nevi and gradually decreased with acquisition of intradermal component and became nonexistent in nevi in the deep dermis. CONCLUSIONS: MT1-MMP is expressed in the membrane of nevus cells, with expression greater in nest cells in contact with the extracellular matrix. The intensity of expression correlated inversely with the maturation phase of the nevus, being very high in junctional nevi and low in intradermal nevi.

In vitro capacitating effect of gamma-aminobutyric acid in ram spermatozoa.

We have evaluated the capacitating effect of gamma-aminobutyric acid (GABA) in ram spermatozoa in vitro, in a chemically defined medium, by means of the chlortetracycline (CTC) binding assay. Semen from adult Australian Merino rams was collected in an artificial vagina; spermatozoa were washed once in modified Biggers, Whitten, and Wittingham medium (m-BWW), without BSA or serum, and incubated in m-BWW alone or in m-BWW containing GABA, GABA agonists, or antagonists for 2 h at 38.5 degrees C under 5% CO2 in air. Samples were taken for assessment of CTC binding pattern or were further incubated for 15 min in the presence of 5 microM calcium ionophore A23187. Acrosomal exocytosis was evaluated by Pisum sativum agglutinin binding. Addition of GABA to the incubation medium resulted in a concentration-dependent increase in the percentage of CTC forms II and III, corresponding to mid-capacitated and capacitated spermatozoa, respectively. The effect was marginally significant at 1 microM and maximal at 20 microM. The action of 20 microM GABA was mimicked by the GABAB-receptor agonist, muscimol, but not by the GABAA-receptor agonist, baclofen, and completely blocked by the GABAA-receptor antagonists, bicuculline and picrotoxin, which lacked effect per se. In a separate set of experiments, incubation of spermatozoa with GABA at a concentration of 1 microM, which was insufficient to stimulate sperm capacitation, together with the neuroactive steroid allopregnanolone (1 microM) provoked a capacitating effect similar to that achieved by 20 microM GABA alone. These results show that GABA has a capacitating action on ram spermatozoa through a GABAA receptor-mediated mechanism.

Actin localization in ram spermatozoa: effect of freezing/thawing, capacitation and calcium ionophore-induced acrosomal exocytosis.

We have analyzed, by immunofluorescence, the localization of actin in ram spermatozoa, its colocalization with the actin-binding protein, gelsolin, and the effect of freeze/thawing, in vitro capacitation, and induced acrosomal exocytosis on its distribution. The monoclonal anti-actin and anti-gelsolin antibodies used recognized single bands at 43,000 and 90,000 kDa, respectively. In all spermatozoa, intense actin staining was observed in the whole length of the flagellum and, depending on the protocol used, in the neck and postacrosomal region of the head. Comparison of three staining methods, together with the use of NBD-phallacidin, allowed us to characterize ram sperm actin as a monomeric, intracellular, membrane-associated protein. Gelsolin was also present in ram spermatozoa and precisely colocalized with actin. Processes involving alterations in membrane structure such as freezing/thawing, in vitro capacitation, and calcium ionophore-induced acrosomal exocytosis provoked changes in the exposure of actin to the antibody. This strongly suggests a physical association of this protein to the plasma membrane, most likely by its intracellular side. The possible role of actin in sperm function is discussed.

The storage of pure ram semen at room temperature results in capacitation of a subpopulation of spermatozoa.

We investigated whether storage of pure ram semen at room temperature would facilitate the sperm capacitation process, as assessed by means of the chlortetracycline method. Objective motility, membrane integrity and ability of spermatozoa to undergo acrosome reaction induced by A23187 for 15 min were simultaneously evaluated to gain further insight into this process. Storage for 4 h at room temperature had a clear capacitating effect in approximately 50% of spermatozoa and increased their ability to respond to A23187. Beyond that time, the percentage of motility and membrane integrity remained unchanged. Moreover, storage did not alter the ability of those spermatozoa that remained noncapacitated under these conditions to become capacitated in SOF-m medium. Storage for 4 h increased the percentage of spermatozoa showing swelling of the apical ridge from 3 to 13%. In conclusion, storage of ram semen at room temperature for 4 h in the dark has a marked capacitating effect on a subpopulation of spermatozoa, without changes in motility or membrane integrity, and a low effect on the appearance of the acrosome. Since semen storage is generally included in different IVF protocols, the results presented here contribute toward a clearer understanding of its role in these procedures.

In vitro capacitation and induction of acrosomal exocytosis in ram spermatozoa as assessed by the chlortetracycline assay.

We have described the different patterns of chlortetracycline (CTC) binding to ram spermatozoa, immediately after ejaculation and upon in vitro capacitation and calcium ionophore-induced acrosomal exocytosis. Four different forms of CTC distribution were found. Form I showed an even distribution of fluorescence over the entire head, with a brighter band in the equatorial region. In Form II, uniform fluorescence was observed without equatorial band. Form III consisted of fluorescence in the anterior portion of the head. Form IV showed no fluorescence over the head. In all cases, fluorescence in the middle piece of the flagellum was observed as well. Immediately after ejaculation, Form I was the most abundant one (78%) in fresh semen with Forms II and III being relatively scarce (less than 15%). Form IV was virtually absent or appeared only occasionally. Incubation under in vitro capacitating conditions led to a significant decrease in Form I and to a significant increase in Forms II and III. Form II was mainly associated to intact acrosomes, while most spermatozoa in Form III showed intermediate forms of acrosomal status. Incubation of spermatozoa with the calcium ionophore A23187 resulted in 55% of spermatozoa showing Form IV, suggesting that it represents the acrosome-reacted stage. Form I was abruptly decreased at 30 min of incubation and was neglectible after 60 min. In contrast, Forms II and III increased at 30 min but decreased later on, suggesting that both forms represent intermediate stages before the acrosomal exocytosis. Analysis of acrosomal status in spermatozoa from individual CTC forms revealed that all spermatozoa that remained in Form II after incubation had intact acrosomes. Intermediate stages were predominant in Form III-spermatozoa, while most Form IV-spermatozoa underwent full acrosomal exocytosis. These results show that CTC binding can be used to monitor changes in ram spermatozoa during capacitation and acrosome-reaction.

[Genetic studies of panic disorder]. / Estudios genéticos del trastorno de pánico.

We review the literature on genetic factors in anxiety disorders and present data from a study in which we used the HLA system to evaluate distribution of different antigens in the members of a family with high morbidity of Panic Disorder. Our aim was to look for HLA haplotype sharing among the affected subjects. All members of the family were interviewed with the SCID interview to detect any psychiatric disorder. Third generation members under 18 were evaluated with a structured interview especially designed to identify separation anxiety disorder. In all cases we assessed 11 HLA-A, 16 HLA-B and 5 HLA-Cw antigens. The results suggest a genetic component for Panic Disorder, based on the presence of the same haplotype (A3B18) in the six members of the family suffering from Panic Disorder and Agoraphobia, compared with its absence in the others who were free of such disorders.