An analytical strategy for designer benzodiazepines and Z-hypnotics determination in plasma samples using ultra-high performance liquid chromatography/tandem mass spectrometry after microextraction by packed sorbent.

The illicit market for new psychoactive substances (NPS) is continuously growing. Designer benzodiazepines (DBZD) and Z-hypnotics are increasingly being used for self-medication or recreational purposes. The limited regulation and little biological information available about NPS have raised the need for analytical methods capable of extracting and quantifying them in human biological fluids. In this work, a procedure based on microextraction by packed sorbent (MEPS) in combination with ultra-high performance liquid chromatography and tandem mass spectrometry (UHPLC-MS/MS) has been developed to determine the designer benzodiazepines (clonazolam, deschloroetizolam, nifoxipam, flubromazolam and meclonazepam), and the Z-hypnotics (zolpidem, zaleplon and zopiclone) in plasma. A 3342//16 asymmetric screening design was used to study extraction variables such as the type and volume of eluent, pH, number of extraction cycles, volume of washing solvent and type of sorbent. The ensuing analytical method was validated in terms of linearity by standard addition calibration curves at eight different analyte concentration levels from 0.5-500 ng mL-1. R2 values, limits of detection (LOD) and limits of quantification (LOQ) fell in the ranges 0.9900-0.9988, 0.5-5 ng mL-1 and 1-10 ng mL-1. Intra and interday precision expressed as relative standard deviations, were < 10.6 % and process efficiency ranged from 63 to 117 % for the quality control samples. The proposed method detected zolpidem and various other benzodiazepines in plasma samples from overdoses cases.

[Mid-term complications of the monopolar, modular radial head prosthesis]. / Complicaciones a mediano plazo de la prótesis monopolar modular de cabeza radial.

INTRODUCTION: The objective of this study is to assess the type, frequency and severity of complications after the implantation of the modular monopolar radial head prosthesis. MATERIAL AND METHODS: Forty-seven patients with 48 radial head prostheses implanted between 2009 and 2017 were reviewed retrospectively. Patients were evaluated clinical and radiographically for a mean follow-up of 43.55 months (range: 12-89). RESULTS: The same type of prosthesis was implanted in every patient (Ascension Modular Radial Head). The average score in the Mayo Elbow Performance Score was 88.29 ± 9.9 points. During the follow-up, three patients (6.25%) suffered from continuous pain. Twelve cases (25.5%) showed radiological oversizing, though only five were symptomatic. Heterotopic ossification was detected in twenty-seven cases (57.4%). Eleven patients (23.4%) developed postoperative stiffness. Nineteen cases (40.42%) showed periprosthetic osteolysis, from which seven were symptomatic. Thirteen patients (27%) developed surgery-related complications: three cases of infection, four cases of symptomatic loosening, two neurapraxies, one instability and three cases of oversizing with associated stiffness. Nine patients (18.75%) required reintervention. DISCUSSION: Our study obtains a 27% of overall complications, mostly related to oversizing and prosthetic loosening, and 19% of reinterventions. These results are similar to those presented in previous studies, with variations depending on the time of follow-up. Further research is also required to evaluate long-term results and the potential progression of the radiographic findings. CONCLUSION: Taken together, these data stress the need for improvement in both the surgical technique and the design of the implants.

INTRODUCCIÓN: El objetivo de este estudio es evaluar el tipo, la frecuencia y la gravedad de las complicaciones después de la implantación de la prótesis monopolar modular de cabeza radial. MATERIAL Y MÉTODOS: Se revisaron retrospectivamente 47 pacientes con 48 prótesis de cabeza radial implantadas entre 2009 y 2017 durante una media de 43.55 meses (rango: 12-89). RESULTADOS: Se implantó el mismo tipo de prótesis en cada paciente (Ascension Modular Radial Head). La puntuación media obtenida en la clasificación Mayo Elbow Performance Score fue de 88.29 ± 9.9 puntos. Durante el seguimiento tres pacientes (6.25%) sufrieron dolor continuo. Doce casos (25.5%) mostraron sobredimensión radiológica, aunque sólo cinco fueron sintomáticos. Se detectó osificación heterotópica en 27 casos (57.4%), 11 pacientes (23.4%) desarrollaron rigidez postoperatoria, 19 casos (40.42%) mostraron osteólisis periprotésica, de los cuales siete fueron sintomáticos, 13 pacientes (27%) presentaron complicaciones: tres casos de infección, cuatro casos de aflojamiento sintomático, dos neuroapraxias, una inestabilidad y tres casos de sobredimensionamiento con rigidez asociada. Nueve pacientes (18.75%) fueron reintervenidos. DISCUSIÓN: Presentamos 27% de complicaciones globales, principalmente relacionadas con la sobredimensión y el aflojamiento protésico y 19% de reintervenciones. Estos resultados son similares a los descritos en estudios previos con variaciones en función del tiempo de seguimiento. Asimismo, se requieren nuevos estudios para evaluar los resultados a largo plazo y la posible progresión de los hallazgos radiográficos. CONCLUSIÓN: En conjunto, estos datos ponen de manifiesto la necesidad de mejoría tanto de la técnica quirúrgica como del diseño de los implantes.

[Medium term results of unipolar modular radial head arthroplasty]. / Resultados a medio plazo de la artroplastia monopolar modular de cabeza radial.

OBJECTIVE: Analyze the results of unipolar modular radial head arthroplasty in the treatment of complex fractures associated with elbow joint instability. MATERIAL AND METHOD: Retrospective design study of 44 patients, 17 men and 27 women with a mean age of 51 years (17-78) who suffered radial head fractures (Mason III) in the context of an unstable elbow injury. Mean follow-up was 21.2 months (12-60). Radial head replacement was performed with a unipolar modular prosthesis with a fixed stem (Ascension(®) Modular Radial Head [MRH]), associating repair of concomitant lesions. The Mayo scale Elbow Performance Score (MEPS) was used to perform the functional assessment. A radiological evaluation was performed at the last follow-up and the complications were recorded. RESULTS: Final arc of flexion-extension was 108° with an average of 135° of prono-Supination. At final follow-up, 82% of results were satisfactory according to the MEPS. The 40% of patients (17) had radiographic signs of lucencies around the stem, although most of them were asymptomatic. Two of them needed a second surgical procedure because of painful prosthetic loosening and further surgery was required for a prosthesis dislocation. DISCUSSION: Radial head implants are an adequate treatment option for restoring stability in complex radial head fractures. Modularity allows a wide range of size combination and helps to avoid overstuffing. Periprosthetic osteolysis could be associated with the presence of pain, so it is necessary to perform long-term studies to test the potential complications of this finding.

Resultados a medio plazo de la artroplastia monopolar modular de cabeza radial / Medium term results of unipolar modular radial head arthroplasty

Objetivo. Analizar los resultados de la artroplastia monopolar modular de la cabeza radial como tratamiento de fracturas complejas del codo asociadas a inestabilidad articular. Material y método. Estudio retrospectivo de 44 pacientes, 17 varones y 27 mujeres con una edad media de 51 años (17-78), que fueron intervenidos de fracturas de la cabeza radial tipo iii de Mason en el contexto de lesiones inestables del codo. El seguimiento medio fue de 21,2 meses (12-60). La sustitución de la cabeza radial se realizó con una prótesis monopolar modular de vástago fijo, denominada Ascension® Modular Radial Head (MRH), asociando la reparación de las lesiones concomitantes en el mismo acto quirúrgico. Para la evaluación funcional se utilizó la escala Mayo Elbow Performance Score (MEPS). También se registraron los hallazgos radiográficos y complicaciones durante el seguimiento. Resultados. El arco final de flexo-extension fue de 108° con una prono-supinación media de 135°. Al final del seguimiento, el 82% de los resultados fueron satisfactorios de acuerdo con la MEPS. El 40% de los pacientes (17) presentaron signos radiográficos de radiolucencia alrededor del vástago que resultaron asintomáticos. Dos de estos 17 pacientes precisaron reintervención por aflojamiento protésico doloroso y una nueva cirugía fue necesaria por luxación de la prótesis. Discusión. Los implantes monopolares son una opción válida de tratamiento para restaurar la estabilidad en las fracturas complejas de la cabeza radial. Su modularidad permite ampliar la selección de tamaños y evitar el pinzamiento radio-capitelar. La osteólisis periprotésica no es siempre sinónimo de ausencia de sintomatología, siendo necesarios estudios con seguimiento más prolongado para valorar la asociación de este hallazgo con el desarrollo de complicaciones potenciales (AU)

Objective. Analyze the results of unipolar modular radial head arthroplasty in the treatment of complex fractures associated with elbow joint instability. Material and method. Retrospective design study of 44 patients, 17 men and 27 women with a mean age of 51 years (17-78) who suffered radial head fractures (Mason III) in the context of an unstable elbow injury. Mean follow-up was 21.2 months (12-60). Radial head replacement was performed with a unipolar modular prosthesis with a fixed stem (Ascension® Modular Radial Head [MRH]), associating repair of concomitant lesions. The Mayo scale Elbow Performance Score (MEPS) was used to perform the functional assessment. A radiological evaluation was performed at the last follow-up and the complications were recorded. Results. Final arc of flexion-extension was 108° with an average of 135° of prono-Supination. At final follow-up, 82% of results were satisfactory according to the MEPS. The 40% of patients (17) had radiographic signs of lucencies around the stem, although most of them were asymptomatic. Two of them needed a second surgical procedure because of painful prosthetic loosening and further surgery was required for a prosthesis dislocation. Discussion. Radial head implants are an adequate treatment option for restoring stability in complex radial head fractures. Modularity allows a wide range of size combination and helps to avoid overstuffing. Periprosthetic osteolysis could be associated with the presence of pain, so it is necessary to perform long-term studies to test the potential complications of this finding (AU)

Medición de 25-hidroxivitamina D sérica: comparación de dos inmunoensayos / Measurement of 25-hydroxyvitamin D in serum: comparison of two immunoassays

La concentración sérica de 25-hidroxivitamina D (25-OHD) es utilizada como indicador del estado nutricional de Vitamina D (VD). El método más utilizado para medirla es el RIA. El desarrollo reciente de métodos automatizados no radiactivos facilitaría la práctica diaria de laboratorio y el diagnóstico de necesidad de suplementación. Objetivos: Comparar los datos de 25-OHD obtenidos usando un RIA y un método de quimioluminiscencia (QLIA) automatizado disponible en nuestro medio. Materiales y métodos: Concentraciones de 25-OHD se midieron en suero de 45 pacientes: 8 hombres y 37 mujeres; 18 no suplementados y 27 suplementados con VD (n=5 con VD2 y n=22 con VD3). Las mediciones de 25-OHD se realizaron con un RIA y un QLIA automatizado (LIAISON), ambos DiaSorin. Se calcularon los coeficientes de variación intraensayo (CV intra) e interensayo (CV inter) para ambos métodos. Análisis estadístico: la comparación entre métodos se realizó con los programas Analyse-it y Med Calc Se consideró significativa una p<0.05. Resultados: Los CV% intra e inter fueron: para RIA menores de 10,6 y 19,9 vs QLIA menores de 8,0 y 13.2, respectivamente. En la población total y en el subgrupo no suplementado con VD los datos de RIA vs QLIA fueron: coeficiente de correlación de Pearson (0,9259 vs 0,9412), Bias%: (6.1 vs 2.7), coeficiente de concordancia (0,9244 vs 0,9329). Conclusiones: 1) Ambas metodologías son adecuadas para mediciones de 25OHD, especialmente en casos no medicados con VD, 2) La tendencia hacia un mayor bias% observado en pacientes suplementados con VD no parecería ser atribuible a variabilidad metodológica, y sugeriría que la VD exógena o alguno de sus metabolitos interactuaría en forma diferente en la medición de 25-OHD por cada una de las metodologías utilizadas. Mayor número de casos es necesario a fin de confirmar esta hipótesis.

Serum concentration of 25-hydroxyvitamin D (25OHD) is used as an indicator of nutritional status of Vitamin D (VD). The methodolgy more frequently used for its measurement is RIA. The recent development of automated non-radioactive methodologies would help the laboratory daily practice to diagnose the need for supplementation. Objectives: To compare the data of 25-OHD obtained using a RIA and an automated chemiluminescence method (CLIA) automated available in our country. Materials and methods: Concentrations of 25-OHD were measured in serum of 45 patients: 8 men and 37 women, 18 unsupplemented and 27 supplemented with VD (n=5 with VD2 and n=22 with VD3). For 25-OHD measurements we used a RIA and a QLIA under an automated platform (LIAISON), both DiaSorin. We calculated intra-assay (intra) and interassay (inter) coefficients of variation (CV%) for both methods. Statistical analysis: comparison between methods was conducted with Analyse-it and Med Calc softwares; p <0.05 was considered significant. Results: The intra and inter CV% were below 19.9 and 10.6 for RIA vs 8.0 and 13.2 for CLIA, respectively. In the overall population and in the subgroup never supplemented with VD, data for RIA vs CLIA were: Pearson correlation coefficient (0.9259 vs 0.9412), Bias% (6.1 vs. 2.7), concordance coefficient (0.9244 vs 0.9329). Conclusions: 1) Both methods are suitable for measurements of 25OHD, particularly in cases not medicated with VD, 2) The trend toward greater bias% observed in patients supplemented with VD does not appear to be attributable to methodological variability, and suggests that exogenous VD or its metabolites interact differently in the measurement of 25-OHD by each of the methodologies used. A higher number of cases is needed to confirm this hypothesis.

Economic contribution of draught animals to Mazahua smallholder Campesino farming systems in the highlands of Central Mexico.

The economic contribution of draught animals to smallholder Mazahua campesino systems in two mountain villages of San Felipe del Progreso, in the central highlands of Mexico, was assessed. Campesinos rely on draught animals for cultivation tasks, as pack animals, and as transport for agricultural and domestic activities. The villages were San Pablo Tlalchichilpa (SPT) and La Concepción Mayorazgo (LCM). Twelve households that possessed draught animals were monitored from July 1999 to June 2000, nine in SPT and three in LCM, in terms of animal inventories and income from their draught animals, in cash and opportunity values. Equines in SPT have substituted bulls, and are recognized for their multipurpose contribution, while in LCM bulls are still used for ploughing the land. Overall total mean gross income was US dollar 490.78 per farm per year, plus US dollar 56 as opportunity value of the fertilizer value of manure for both villages. Deducting estimated costs, owning draught animals leaves a mean net margin of US dollar 412.50/year in SPT and of US dollar 285.64/year in LCM. There is a significant correlation (p < 0.05) between ownership of draught animals and incomes, with a regression coefficient of US dollar 279.16 per year per draught animal. Besides positive economic returns, having work animals alleviates drudgery for the campesino families.

[Reaction surveillance: organization in public care/hospitals in Marseille]. / Réactovigilance: quelle organisation à l'Assistance publique-Hôpitaux de Marseille ?

For the French Drug Agency (AFSSAPS), the aim of the reactovigilance is to monitor the adverse effects resulting from the use of IVD-MD. More precisely, the goal is to evaluate all incidents and risks that can result in inadequate results or negative effects to the user and therefore for the patient. According to the French Law and before the transposition in the French Law of the European Guideline 98/79/CE, the University Hospital of Marseille has decided to organize its own reactovigilance network. Since 2001, an institutional organization has been set up. For each of the 33 labs of the University hospital one reactovigilant has been nominated as well as one coordinating reactovigilant. Specialized structures have been created: one central committee and one experts group. Standardized operating procedures have been established. The reactovigilance system is also integrated to the vigilance coordination network of the Marseille University Hospital. This organization allows to achieve tracability of all alerts, as well as information and training for the professionals. Four missions are defined: collection and management of all incidents IVD-MD-related; diffusion of AFFSAPS and industrials alerts to all labs in order to take appropriate measures; Tracability of alerts and incidents in all labs and in central committee, tracability of reactives according to French guidelines (GBEA). For the first 10 months of 2002, 46 alerts have been forwarded to all lab's correspondants. In the same period, one real adverse event has been locally notified and after analyse will be forwarded to the AFSSAPS. All these factors should contribute to the health professionals interest and participation in reactovigilance activities.

Hydrogen peroxide mediates vascular cell adhesion molecule-1 expression from interleukin-18-activated hepatic sinusoidal endothelium: implications for circulating cancer cell arrest in the murine liver.

The mechanism of intrasinusoidal arrest of circulating cancer cells, which is a critical step in liver metastasis, appears to be facilitated by tumor-derived proinflammatory factors that increase sinusoidal cell adhesion receptors for cancer cells. However, how this prometastatic microenvironment is up-regulated remains unknown. Using intrasplenically injected B16 melanoma (B16M) cells, we show that the expression of vascular cell adhesion molecule-1 (VCAM-1) significantly increased in hepatic sinusoidal endothelium (HSE) cells over physiologic baseline within the first 24 hours of metastatic cancer cell infiltration in the liver. This correlated with increased in vitro adhesion of B16M cells to HSE cells isolated from B16M cell-injected mice. In vivo VCAM-1 blockade with specific antibodies before B16M cell injection decreased sinusoidal retention of luciferase-transfected B16M cells by 85%, and metastasis development by 75%, indicating that VCAM-1 expression on tumor-activated HSE cells had a prometastatic contribution. Because VCAM-1 expression is oxidative stress-inducible, recombinant catalase was in vivo administered, resulting in a complete abrogation of both VCAM-1 expression and B16M cell adhesion increases in HSE cells isolated from B16M cell-injected mice. Catalase also abrogated the proadhesive response of HSE cells to B16M-conditioned medium (B16M-CM) in vitro, although this did not affect the concomitant release of major proinflammatory cytokines by HSE cells. HSE cells treated with B16M-CM released interleukin (IL)-18 via tumor necrosis factor-alpha (TNF-alpha)-dependent IL-1beta in vitro. In turn, H(2)O(2) production from B16M-CM-treated HSE cells was regulated by IL-18. Thus, liver-infiltrating B16M cells activated their adhesion to HSE through a sequential process involving TNF-alpha-dependent IL-1beta, which induced IL-18 to up-regulate VCAM-1 via H(2)O(2). The pivotal position of H(2)O(2) was further supported by the fact that incubation of HSE cells with nontoxic concentrations of H(2)O(2) directly enhanced VCAM-1-dependent B16M cell adhesion in vitro without proinflammatory cytokine mediation, which emphasizes the key role of oxidative stress in the pathogenesis of liver inflammation and metastasis.

Differential activities of the SoxR protein of Escherichia coli: SoxS is not required for gene activation under iron deprivation.

When Escherichia coli cells are under superoxide stress, proteins SoxR and SoxS, acting sequentially, control the expression of a set of repair and defense genes. One of these genes, fumC, encoding fumarase C, was reported to be also activated by iron deprivation in a soxRS-dependent manner. However, the same condition failed to induce the expression of a soxS'::lacZ fusion. The expression of acnA (aconitase A) is also activated by SoxR alone when under iron deprivation, but not of sodA (Mn-superoxide-dismutase). SoxR completely inhibited the migration of a DNA fragment containing the promoter region of fumC, in gel-shift experiments. SoxR might bind to a different region than SoxS within the fumC promoter, or an unknown intermediate other than SoxS might be acting. It is possible that the regulatory role of SoxR is more complex than previously considered.

The Neurospora am gene and NADP-specific glutamate dehydrogenase: mutational sequence changes and functional effects--more mutants and a summary.

A further series of mutant am alleles, encoding potentially active NADP-specific glutamate dehydrogenase (GDH) and capable of complementation in heterocaryons, have been characterized with respect to both GDH properties and DNA sequence changes. Several mutants previously studied, and some of their same-site or second-site revertants, have also been sequenced for the first time. We present a summary of what is known of the properties of all am mutants that have been defined at the sequence level.

[New blood transfusion partners in France]. / Les nouveaux acteurs de la transfusion sanguine en France.

The French Blood Agency, created in 1993, controls all aspects of the blood transfusion system. It is responsible for restructuring the blood transfusion system and for Good Manufacturing Practices. All incidences are reported to the blood monitoring system which also obtains further descriptions of such incidents. Blood transfusion policies have been modified to include monitoring of blood-associated morbidity. Successive reforms have led to the establishment of a new public system for blood transfusion. New structures and more formal designation of responsibilities are needed. The function of these new structures is to better control transfusion activities and prevent unknown risk.

IL-18 regulates IL-1beta-dependent hepatic melanoma metastasis via vascular cell adhesion molecule-1.

Proinflammatory cytokines, including IL-1beta and tumor necrosis factor-alpha (TNF-alpha), promote cancer cell adhesion and liver metastases by up-regulating the expression of vascular cell adhesion molecule-1 (VCAM-1) on hepatic sinusoidal endothelium (HSE). In this study, hepatic metastasis after intrasplenically injected mouse B16 melanoma (B16M) cells was reduced 84-95% in mice with null mutations for either IL-1beta or the IL-1beta-converting enzyme (ICE, caspase-1) compared with wild-type mice. On day 12, 47% of wild-type mice were dead compared with 19% of either IL-1beta or ICE-deficient mice. In vitro, conditioned medium from B16M cells (B16M-CM) induced the release of TNF-alpha and IL-1beta from cultures of primary murine HSE. The effect of B16M-CM on HSE resulted in increased numbers of B16M cells adhering to HSE, which was completely abrogated by a specific inhibitor of ICE, anti-IL-18 or IL-18-binding protein. Exogenous IL-18 added to HSE also increased the number of adhering melanoma cells; however, this was not affected by IL-1 receptor blockade or TNF neutralization but rather by anti-VCAM-1. These results demonstrate a role for IL-1beta and IL-18 in the development of hepatic metastases of B16M in vivo. In vitro, soluble products from B16M cells stimulate HSE to sequentially release TNF-alpha, IL-1beta, and IL-18. The IL-18 cytokine increases expression of VCAM-1 and the adherence of melanoma cells.

[Serologic diagnosis of human leptospirosis with 3 variants of the passive hemagglutination technique]. / Diagnóstico serológico de la leptospirosis humana mediante 3 variantes de la técnica de hemaglutinación pasiva.

A serological study was conducted to evaluate 3 variants of the positive hemagglutination technique (HA) with the use of formalized erythrocytes of fresh lamb (HC) and of formalized Type O, Rh-negative human red blood cells (HH). No significant statistical difference were found between the results of HA and HC and between HC and HH. Unspecific reaction was only observed in 2 of the 200 sera studied by HH and HC. Heterophil antibodies were just observed in 2.5% of the sera studied by HH, and in 100% of the sera studied by HC. The qualitative parameters were determined for the variants used, and this way the repetition and reproducibility of the most useful variant was evaluated.

Mannose receptor-mediated endothelial cell activation contributes to B16 melanoma cell adhesion and metastasis in liver.

The role of mannose receptors from hepatic sinusoidal endothelium (HSE) in liver colonization by B16 melanoma (B16M) cells was studied. The expression of high mannose-type oligosaccharides on the surface of B16M cells was enhanced by in vitro treatment with 1-deoximannojirimycin (1-DMM). There was a significant (P < 0.01) enhancement of hepatic metastasis when B16M cells were 1-DMM-treated before being intrasplenically injected into C57BL/6J mice. Intraperitoneal administration of 5 mg/kg recombinant human interleukin-1 receptor antagonist (rHuIL-1Ra) inhibited the 1-DMM-induced enhancement of metastasis. Expression of high mannose-type oligosaccharides on the surface of 1-DMM-treated B16M cells and their in vitro adhesion to the HSE was significantly correlated (R = 0.82). The addition of either 100 microg/ml mannan or paraformaldehyde (PFA)-fixed 1-DMM-treated B16M cells to cultured HSE for a period of 12 h significantly (P < 0.01) increased the release of IL-1beta from the HSE compared to that liberated by the HSE incubated with either basal medium or PFA-fixed untreated B16M cells. The same HSE treatments also significantly (P < 0.01) increased the degree of adhesion of other B16M cells to HSE, being abrogated by anti-mouse vascular cell adhesion molecule-1 (VCAM-1) antibodies. The conditioned media from HSE cultures, activated by PFA-fixed, 1-DMM-treated B16M cells significantly (P < 0.01) increased B16M cell proliferation when compared to conditioned media from HSE cultures incubated with PFA-fixed, untreated B16M cells. Thus, 1-DMM treatment of B16M cells enhanced the development of hepatic metastasis by IL-1-dependent mechanisms. The mechanism is consistent with in vitro mannose receptor-mediated melanoma cell attachment to the HSE, which subsequently upregulates IL-1beta release, VCAM-1-dependent adherence, and melanoma growth factor(s) release by HSE.

[The reactogenicity and immunogenicity of the first Cuban vaccine against human leptospirosis]. / Reactogenicidad e inmunogenicidad de la primera vacuna cubana contra la leptospirosis humana.

A controlled double-blind trial was conducted with the participation of 80 adult volunteers of both sexes, who were randomly divided into groups of 40 individuals each one. The case-base study received the vaccine and the control group was administered placebo to know the safety, the behaviour of reactogenicity, and to star the immunogenicity studies of the first Cuban vaccine against human leptospirosis. The vaccine used in the case-base study was an inactivated and trivalent vaccine containing strains of Leptospira canicola, icterohaemorrhagiae and pomona, since they have the highest circulation in the country. The results obtained showed the inocuity of the vaccine as no adverse severe reactions were detected. The general symptomatology observed was low, where as febricula was the most common general symptom. It appeared during the first 3 days of observation and there were no significant differences between the 2 group. Only a mild pain at the site of the injection was reported as a local symptom, which was more frequent in the vaccinated group than in the control group (7.8 against 1.5%, respectively). The seroconversion obtained was of 29% by microagglutination, and of 34.2% by ELISA. The final results allowed to conclude that this vaccine is safe for human adults at the ages under study, and give the possibility to continue other studies in more advanced stages to complete the requirements for obtaining its license.

Mercury induces multiple antibiotic resistance in Escherichia coli through activation of SoxR, a redox-sensing regulatory protein.

Sub-inhibitory mercury concentrations are capable of partially activating SoxR, as shown by the augmented expression of a soxS'::lacZ fusion, and a diminished sensitivity to antibiotics caused by mercury treatment. Mercury may elevate the intracellular concentration of superoxide or perhaps act as a putative metal ligand for SoxR.

Cloning of a novel neuronal nitric oxide synthase expressed in penis and lower urinary tract.

The neuronal nitric oxide synthase (nNOS) is the main NOS isoform in the urogenital tract catalyzing the synthesis of nitric oxide, the mediator of penile erection and presumably an important factor in the control of urinary voiding. We have cloned from the rat penile corpora cavernosa a cDNA coding for a novel nNOS differing from the cerebellar nNOS by the presence of a 102 nucleotides stretch and other features. This new species is the only nNOS mRNA expressed in the rat penis, urethra, prostate, and skeletal muscle, coexists with the cerebellar nNOS in the pelvic plexus and bladder, and is detectable in the cerebellum. The novel insert is present in human penile RNA and is transcribed from intron 16. The features and distribution of the penile nNOS suggest that it is may be regulated differentially from the cerebellar nNOS.

Crystallization and preliminary X-ray analysis of the NADP-specific glutamate dehydrogenase from Neurospora crassa.

The NADP-linked glutamate dehydrogenase from Neurospora crassa has been crystallized by the hanging-drop method of vapour diffusion in the presence of 0.1 M glutamate. The crystals are trigonal and are in space group P3(1)21 with unit-cell dimensions of a = b = 196.6, c = 102.0 A and with a trimer in the asymmetric unit. A full structure determination of this enzyme will lead to an understanding of the molecular basis of inter-allelic complementation observed with hybrid hexamers of naturally occurring mutants.