In vitro and ex vivo activity of the fluoroquinolone DC-159a against mycobacteria.

Antimicrobial resistance is a global health problem. In 2021, it was estimated almost half a million of multidrug-resistant tuberculosis (MDR-TB) cases. Besides, non-tuberculous mycobacteria (NTM) are highly resistant to several drugs and the emergence of fluoroquinolone (FQ) resistant M. tuberculosis (Mtb) is also a global concern making treatments difficult and with variable outcome. The aim of this study was to evaluate the activity of the FQ, DC-159a, against Mtb and NTM and to explore the cross-resistance with the currently used FQs.A total of 12 pre-extensively drug-resistant (XDR) Mtb, 2 XDR, 36 fully drug susceptible strains and 41 NTM isolates were included to estimate the in vitro activity of DC-159a, moxifloxacin (MOX) and levofloxacin (LX), using minimal inhibitory and bactericidal concentration (MIC and MBC). The activity inside the human macrophages and pulmonary epithelial cells were also determined.DC-159a was active in vitro and ex vivo against mycobacteria. Besides, it was more active than MOX/LX. Moreover, no cross-resistance was evidenced between DC-159a and LX/MOX as DC-159a could inhibit Mtb and MAC strains that were already resistant to LX/MOX.DC-159a could be a possible candidate in new therapeutic regimens for MDR/ XDR-TB and mycobacterioses cases.

Virtuous and Vicious Cycles of Arm Use and Function Post-stroke.

Large doses of movement practice have been shown to restore upper extremities' motor function in a significant subset of individuals post-stroke. However, such large doses are both difficult to implement in the clinic and highly inefficient. In addition, an important reduction in upper extremity function and use is commonly seen following rehabilitation-induced gains, resulting in "rehabilitation in vain". For those with mild to moderate sensorimotor impairment, the limited spontaneous use of the more affected limb during activities of daily living has been previously proposed to cause a decline of motor function, initiating a vicious cycle of recovery, in which non-use and poor performance reinforce each other. Here, we review computational, experimental, and clinical studies that support the view that if arm use is raised above an effective threshold, one enters a virtuous cycle in which arm use and function can reinforce each other via self-practice in the wild. If not, one enters a vicious cycle of declining arm use and function. In turn, and in line with best practice therapy recommendations, this virtuous/vicious cycle model advocates for a paradigm shift in neurorehabilitation whereby rehabilitation be embedded in activities of daily living such that self-practice with the aid of wearable technology that reminds and motivates can enhance paretic limb use of those who possess adequate residual sensorimotor capacity. Altogether, this model points to a user-centered approach to recovery post-stroke that is tailored to the participant's level of arm use and designed to motivate and engage in self-practice through progressive success in accomplishing meaningful activities in the wild.

Levels and trends of perfluoroalkyl acids (PFAAs) in water (2013-2020) and fish from selected riverine basins in Spain.

The aim of the present study was to assess the presence of perfluoroalkyl acids (PFAAs), namely perfluoroalkane sulfonates and perfluoroalkyl carboxylic acids, in Spanish river basins in order to: identify potential spatiotemporal variations; evaluate the effectiveness of the measures implemented for the reduction/elimination of these pollutants; verify the fulfillment of the Environmental Quality Standards (EQSs) in the European Union. PFOS and PFOA were determined in 116 water samples from four sites in the Duero basin, the largest in the Iberian Peninsula, collected seasonally from 2013 to 2020. In addition, 30 fish sample composites from the sample banks of Duero, Tagus, Ebro, Eastern Cantabrian and Catalonian basins were analyzed for 15 PFAAs. Median PFOS and PFOA concentrations were 0.72 and 0.42 ng/L, ranging from values below the limit of quantification (LOQ) to 81 and 22 ng/L, respectively. During the studied period, 51% of water samples were above the EQS of 0.65 ng/L for PFOS. In the case of fish, the PFOS range was

In vitro anti-tuberculosis activity of azole drugs against Mycobacterium tuberculosis clinical isolates / Actividad in vitro de azoles contra aislamientos clínicos de Mycobacterium tuberculosis

Background: Latent tuberculosis has been associated with the persistence of dormant Mycobacterium tuberculosis in the organism of infected individuals, who are reservoirs of the bacilli and the source for spreading the disease in the community. New active anti-TB drugs exerting their metabolic action at different stages and on latent/dormant bacilli are urgently required to avoid endogenous reactivations and to be part of treatments of multi- and extensively-drug resistant tuberculosis (M/XDR-TB). It was previously reported that azole drugs are active against M. tuberculosis. For that reason, the aims of this study were to determine the in vitro activity of azole drugs, imidazole (clotrimazole, CLO and econazole, ECO) and nitroimidazole (metronidazole, MZ and ipronidazole, IPZ), against a collection of MDR M. tuberculosis clinical isolates; and to analyze their potential use in both the LTB and the active forms of M/XDR-TB treatments. Methods: A total of 55 MDR M. tuberculosis isolates and H37Rv were included. MZ and IPZ activity against M. tuberculosis isolates were tested using anaerobic culture conditions. The activity of ECO and CLO was measured by the minimal inhibitory concentration (MIC) using a microdilution colorimetric method. Results: MZ and IPZ showed bacteriostatic activity against M. tuberculosis strains. MIC5o and MIC90 to ECO was 4.0 µg/ml, while MIC50 to CLO was 4.0 µg/ml and MIC90 was 8.0 µg/ml respectively. Conclusion: All azole compounds tested in the study showed inhibitory activity against MDR M. tuberculosis clinical isolates.

Introducción: La tuberculosis (TB) latente ha sido asociada a la persistencia de Mycobacterium tuberculosis durmientes en el organismo de las personas infectadas, las cuales constituyen un reservorio del bacilo y una fuente de diseminación de la enfermedad en la comunidad. Urge la necesidad de contar con nuevos fármacos antituberculosos con acción sobre el bacilo en estado latente/durmiente, a fin de evitar reactivaciones endógenas y para ser incluidas en el tratamiento de la TB multirresistente y extensivamente resistente (M/XDR-TB). Se ha reportado que los azoles son activos contra M. tuberculosis. Por esta razón, los objetivos del presente estudio fueron determinar la actividad in vitro sobre aislamientos clínicos de M/XDR-TB de distintos azoles, incluyendo los imidazoles econazol (ECO) y clotrimazol (CLO) y los 5-nitro-imidazoles ipronidazol (IPZ) y metronidazol (MZ), así como analizar su potencial uso contra las formas latente y activa de esta enfermedad. Métodos: Fueron incluidos 55 aislamientos clínicos de M. tuberculosis MDR y la cepa de referencia H37Rv. Se evaluó la actividad del MZ y el IPZ sobre los aislamientos en condiciones de cultivo anaeróbico, mientras que la actividad del ECO y el CLO fue estimada determinando la concentración inhibitoria mínima (CIM) mediante el método colorimétrico de microdilución en placa. Resultados: El MZ y el IPZ presentaron actividad bacteriostática frente a las cepas de M. tuberculosis. La CIM50 y CIM90 del ECO fue de 4 µg/ml, mientras que el CLO presentó una CIM50 de 4 µg/ml y una CIM90 de 8 µg/ml. Conclusión: Todos los compuestos azólicos evaluados presentaron actividad inhibitoria frente a aislamientos clínicos de M. tuberculosis.

In vitro anti-tuberculosis activity of azole drugs against Mycobacterium tuberculosis clinical isolates.

BACKGROUND: Latent tuberculosis has been associated with the persistence of dormant Mycobacterium tuberculosis in the organism of infected individuals, who are reservoirs of the bacilli and the source for spreading the disease in the community. New active anti-TB drugs exerting their metabolic action at different stages and on latent/dormant bacilli are urgently required to avoid endogenous reactivations and to be part of treatments of multi- and extensively-drug resistant tuberculosis (M/XDR-TB). It was previously reported that azole drugs are active against M. tuberculosis. For that reason, the aims of this study were to determine the in vitro activity of azole drugs, imidazole (clotrimazole, CLO and econazole, ECO) and nitroimidazole (metronidazole, MZ and ipronidazole, IPZ), against a collection of MDR M. tuberculosis clinical isolates; and to analyze their potential use in both the LTB and the active forms of M/XDR-TB treatments. METHODS: A total of 55 MDR M. tuberculosis isolates and H37Rv were included. MZ and IPZ activity against M. tuberculosis isolates were tested using anaerobic culture conditions. The activity of ECO and CLO was measured by the minimal inhibitory concentration (MIC) using a microdilution colorimetric method. RESULTS: MZ and IPZ showed bacteriostatic activity against M. tuberculosis strains. MIC50 and MIC90 to ECO was 4.0µg/ml, while MIC50 to CLO was 4.0µg/ml and MIC90 was 8.0µg/ml respectively. CONCLUSION: All azole compounds tested in the study showed inhibitory activity against MDR M. tuberculosis clinical isolates.

Fitness of drug resistant Mycobacterium tuberculosis and the impact on the transmission among household contacts.

There has been an on-going debate on whether the development of drug resistance in Mycobacterium tuberculosis reduces its relative fitness and its ability to cause disease. The aim of this study was to explore this relationship. For this purpose, we evaluated the in vitro growth of clinical isolates and the transmission of the strains within the patients' households. Clinical and epidemiological data from patients in households, drug-susceptibility and genetic patterns of the isolates were collected. BACTEC MGIT 960™ system with the Epicenter™ software was used to perform fitness experiments and calculate the relative fitness (RF) comparing with the H73Rv reference strain. From 39 households, 124 patients and 388 contacts were included. Concerning transmission, 20 Multi drug-resistant (MDR) and 16 drug sensitive (DS) index cases generated 23 and 28 secondary cases, respectively. An average RF drop of 16.7% was found for MDR strains, but only mutations in rpoB codons 531 were associated with reduced fitness. When the strains were transmitted, their RF tended to decrease, and strains with low RF were less frequently transmitted. Within the limitations of this study, the results showed that the decrease in RF was associated to a limited transmission among the households' contacts.

Predictive value of direct nitrate reductase assay and its clinical performance in the detection of multi- and extensively drug-resistant tuberculosis.

Conventional culture and drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are laborious and time consuming. For this reason alternative rapid culture and DST techniques are urgently needed to shorten the time for drug-resistance detection. A total of 222 smear-positive sputum samples were evaluated by the direct nitrate reductase assay (D-NRA) on Lowenstein-Jensen medium, for the rapid and simultaneous detection of resistance to isoniazid, rifampicin, kanamycin and ofloxacin. p-Nitrobenzoic acid was also included for identification of the M. tuberculosis complex. Results were compared with the BACTEC MGIT 960 as gold standard. The general performance of the D-NRA was very good, reaching a global value of 97 %. D-NRA had a turn-around time of 16.9 days to obtain results while that of the indirect MGIT 960 system was 29 days. D-NRA is a low-cost technology, easy to set up in clinical laboratories and suitable to be used for DST of M. tuberculosis in all smear-positive samples.

First evaluation in Argentina of the GenoType® MTBDRplus assay for multidrug-resistant Mycobacterium tuberculosis detection from clinical isolates and specimens.

Tuberculosis (tB) and multidrug and extensively drug-resistant (dR) tB are important public health problems that are spreading worldwide. The aims of this study were to determine the sensitivity and specificity of the genotype® mtBdRplus assay from smear-positive clinical specimens and isolates and to explore its possible application in routine work. Clinical samples were previously decontaminated using naoH-n-acetyl-l-cystein or naoH-Clna hypertonic solution for Ziehl-neelsen staining and cultures. The leftover sediments of smear-positive samples were stored at -20 °C, 70 of which were selected to be included in this study according to their dR profile. thirty dR Mycobacterium tuberculosis isolates were also assessed. Sequencing was used as gold standard to detect mutations conferring isoniazid (InH) and rifampicin (RIF) resistance. Valid results were obtained in 94.0 % of the samples and 85.5 % (53/62) of the InH-R samples were properly identified. mutations in the katGS315t gene and inhA C-15t gene promoter region were present in 59.7 % (37/62) and 25.8 % (16/62) of the InH-R samples, respectively. the system could also identify 97.7 % (41/42) of the RIF-R samples; the mutations found were rpoBS531l (66.7 %, 28/42), d516V (19.0 %, 8/42), H526Y and S531p/W (4.8 %, 2/42 each one), and S522l/Q (2.4 %, 1/42). a 98.8 % concordance between the genotype assay and sequencing was obtained. genotype® mtBdRplus has demonstrated to be easy to implement and to perform in clinical laboratories and useful for a rapid detection of dR M. tuberculosis from decontaminated sputa and clinical isolates. Therefore, this assay could be applied as a rapid tool to predict InH-R and/or RIF-R in dR risk cases.

Obstructive uropathy as the only manifestation of flare in a patient with systemic lupus erythematosus and anti-phospholipid syndrome.

Chronic interstitial cystitis and ureteral stenosis has occasionally been reported in systemic lupus erythematosus, mostly associated with gastrointestinal symptoms. We report a case of obstructive uropathy associated to chronic interstitial cystitis as the only manifestation of lupus flare in a patient with SLE and anti-phospholipid syndrome (APS) who had been in remission for many years. The development of chronic interstitial cystitis in patients with SLE and APS has not been previously reported. Histopathological study of her urinary bladder and ureteral meatus showed chronic inflammatory infiltrate in the subepithelium. Lack of significant lower urinary tract symptoms and gastrointestinal involvement were some of the factors that could have prevented an earlier diagnosis. Obstructive uropathy and renal insufficiency initially improved with immunosuppressive treatment and endoureteral protheses, but poor compliance to the therapy led to ominous ending.

Cell division in the unicellular microalga Dunaliella viridis depends on phosphorylation of extracellular signal-regulated kinases (ERKs).

In mammalian cells, MAPKs are involved in both stress response (JNK and p38 pathways) and cell proliferation and differentiation [extracellular signal-regulated kinase (ERK)] through protein kinase cascades. Exposure of Dunaliella viridis cell cultures to PD98059, a very specific inhibitor of the ERK signalling pathway, resulted in a total arrest of cell proliferation and a complete dephosphorylation of ERK. As shown by flow cytometry analysis of propidium iodide-stained cells, PD98059 stopped mitosis at the G(2) phase after the S phase has been completed. Multiple physiological parameters such as cell motility and reducing power generation (NADPH) clearly indicate that the treated cells are wholly viable. Exposure of D. viridis to environmental stresses that impair cell division, such as hyperosmotic shock, nitrogen starvation, or sublethal UV irradiation, caused a marked decrease in the phospho-ERK levels as detected by western blot. Two 400 bp polynucleotides from D. viridis with high homologies to published sequences of ERK1 and ERK2 were cloned, sequenced, and submitted to GenBank. Northern blot analysis revealed two mRNA bands of approximately 1.9 kb, consistent with the expected size of ERK proteins ( approximately 40 kDa). Sequence analysis showed that they contained several mitogen-activated protein kinase (MAPK) conserved domains, including II, III, VIb, VII, and the double phosphorylation motif. Interestingly, in D. viridis, this motif was T*DY* instead of the canonic T*EY*. Based on this finding, ERK plant sequences can be divided into two groups, one termed the T*DY* branch and the other termed the T*EY* branch. The molecular and functional data presented here suggest that ERK is a very ancient signalling pathway and that it was already present in the last common ancestor of all eukaryotic cells.

Epidemiología de la vejiga hiperactiva sobre una población de 3.692 consultas urológicas y 2.030 estudios urodinámicos / Overactive bladder epidemiology in a population of 3.692 urology visits and 2.030 urodynamic studies

Introducción: La sociedad Internacional de Continencia ha definido: 1. El Síndrome de vejiga hiperactiva (VHA) como un "diagnóstico sintomático" basado en la presencia de urgencia miccional, con o sin incontinencia , usualmente acompañadas de polaquiuria diurna y nocturna, en ausencia obvia de enfermedad metabólica o neurológica. 2. Vejiga Hiperactiva ("detrusor hiperactivo") como una "observación urodinámica" que se caracteriza por contracciones involuntarias del detrusor, durante la fase del llenado que pueden ser espontáneas o provocadas. Sobre la base de estudios poblacionales realizados en Europa y EE. UU. la prevalencia de vejiga hiperactiva (VHA) ha sido reportada en alrededor de 16 por ciento. En la Argentina no se conoce la epidemiología de la VHA y no hemos encontrado publicaciones nacionales en nuestra revisión bibliográfica. Objetivos: Establecer la tasa de prevalencia de VHA por sexo y edad en una población de 3.692 consultas urológicas y 2.030 estudios urodinámicos a fin de conocer la frecuencia e importancia de esta patología en los pacientes que consultaron en nuestro Servicio. Material y Métodos: Estudio de carácter epidemiológico, retrospectivo, sobre una población de 3.692 pacientes que consultaron consecutivamente, de los cuales 2.030 fueron estudiados mediante urodinamia de presión/flujo, estimandose la tasa de prevalencia de VHA. Se recolectó información adicional sobre edad y sexo de los pacientes con el objeto de ajustar la prevalencia de VHA a los diferentes estratos etarios y de género. Conclusiones: La prevalencia de VHA en nuestra población fue del 23,4 por ciento: dos o tres de cada 10 consultas. El 56,5 por ciento de los pacientes con VHA fueron de sexo masculino y el 43,5 por ciento del femenino. El 54 por ciento de todas las VHA ocurrieron en mayores de 60 años, 65 por ciento de las masculinas y 40 por ciento de las femeninas, siendo el riesgo de VHA en nuestra comunidad en ese grupo etario hasta 7 veces superior respecto de la gente más joven. La manifestación clínica prevalente de VHA fue la incontinencia de orina, seguida por la urgencia y la polaquiuria. La incontinencia fue 5 veces más frecuente en mujeres que en hombres y la polaquiuria fue 13 veces más frecuente en hombres que en mujeres

Epidemiología de la vejiga hiperactiva sobre una población de 3.692 consultas urológicas y 2.030 estudios urodinámicos / Overactive bladder epidemiology in a population of 3.692 urology visits and 2.030 urodynamic studies

Introducción: La sociedad Internacional de Continencia ha definido: 1. El Síndrome de vejiga hiperactiva (VHA) como un "diagnóstico sintomático" basado en la presencia de urgencia miccional, con o sin incontinencia , usualmente acompañadas de polaquiuria diurna y nocturna, en ausencia obvia de enfermedad metabólica o neurológica. 2. Vejiga Hiperactiva ("detrusor hiperactivo") como una "observación urodinámica" que se caracteriza por contracciones involuntarias del detrusor, durante la fase del llenado que pueden ser espontáneas o provocadas. Sobre la base de estudios poblacionales realizados en Europa y EE. UU. la prevalencia de vejiga hiperactiva (VHA) ha sido reportada en alrededor de 16 por ciento. En la Argentina no se conoce la epidemiología de la VHA y no hemos encontrado publicaciones nacionales en nuestra revisión bibliográfica. Objetivos: Establecer la tasa de prevalencia de VHA por sexo y edad en una población de 3.692 consultas urológicas y 2.030 estudios urodinámicos a fin de conocer la frecuencia e importancia de esta patología en los pacientes que consultaron en nuestro Servicio. Material y Métodos: Estudio de carácter epidemiológico, retrospectivo, sobre una población de 3.692 pacientes que consultaron consecutivamente, de los cuales 2.030 fueron estudiados mediante urodinamia de presión/flujo, estimandose la tasa de prevalencia de VHA. Se recolectó información adicional sobre edad y sexo de los pacientes con el objeto de ajustar la prevalencia de VHA a los diferentes estratos etarios y de género. Conclusiones: La prevalencia de VHA en nuestra población fue del 23,4 por ciento: dos o tres de cada 10 consultas. El 56,5 por ciento de los pacientes con VHA fueron de sexo masculino y el 43,5 por ciento del femenino. El 54 por ciento de todas las VHA ocurrieron en mayores de 60 años, 65 por ciento de las masculinas y 40 por ciento de las femeninas, siendo el riesgo de VHA en nuestra comunidad en ese grupo etario hasta 7 veces superior respecto de la gente más joven. La manifestación clínica prevalente de VHA fue la incontinencia de orina, seguida por la urgencia y la polaquiuria. La incontinencia fue 5 veces más frecuente en mujeres que en hombres y la polaquiuria fue 13 veces más frecuente en hombres que en mujeres

A colorimetric assay for determination of cell viability in algal cultures.

In this work, we propose the determination of cell viability in algal cultures by using a colorimetric assay widely used for estimation of cell proliferation in animal cell cultures. The method is based on in vivo reduction by metabolically active cells of a tetrazolium compound (MTS=3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenil)-2H-tetrazolium, inner salt) to a colored formazan, with maximal absorbance at 490 nm, that is released to the culture medium. For this purpose, we have tested two microalgae with high commercial value (Dunaliella and Spirulina) and two seaweeds with different morphology (Ulva and Gracilaria). Color development in this assay is directly proportional to the number of viable cells, to the incubation time in the presence of the assay solution, and to the incubation temperature. A direct significant correlation was found between algal photosynthesis rate and color development in all species used through this work. Moreover, the intensity of absorbance at 490 nm was significantly lower in stressed cells (e.g. in nutrient-limited cultures, in the presence of toxic substances, and in osmotically-stressed cultures). We conclude that cell viability of algal cultures can be rapidly and easily estimated through colorimetric determination of the reduction of MTS to formazan.

Aqueous phase disinfection with power ultrasound: process kinetics and effect of solid catalysts.

The effectiveness of power ultrasound as a viable alternative for destroying pathogenic organisms in homogeneous and heterogeneous mixtures of aqueous solutions was investigated. The method involved monitoring of total coliform bacteria during sonication of E. coli suspensions in the absence and presence of equivalent mass concentrations of ceramic, metallic zinc, and activated carbon. It was found that disinfection by ultrasound is accelerated with solids in the order activated carbon > ceramic > metallic zinc. Process kinetics for each test system were assessed by nonlinear regression analysis of bacterial density vs time data, and the predicted model was found to resemble a well-known expression describing chlorination kinetics. The model denoted that in the presence of activated carbon the process rate was pseudo first order, and the required contact time to accomplish 50% kill was 2.8, 2.4, and 4 times shorter than it was in the zinc-catalyzed, ceramic-catalyzed, and noncatalyzed systems, respectively. It was further found that catalytic effects faded away with increased sonication time and/or reduced number of bacteria, denoting (i) decreased probability of bacterial contact with the solid-liquid interface; (ii) erosion of solid surfaces by vibrational effects; and (iii) reduced cavity formation due to degassing effects of ultrasound.

[Prevalence of male sexual dysfunctions]. / Prevalencia de las disfunciones sexuales masculinas.

OBJECTIVE: To determine the prevalence rate of male sexual dysfunction in 5323 urologic consultations. MATERIALS AND METHODS: In order to establish which age groups show an important consultation rate due to male sexual dysfunction, the number of consultations and the theoretical distribution according to age in the province of Córdoba were contrasted, estimating the risk of suffering from sexual dysfunction, and determining the age distribution and tendency analysis. RESULTS: From 5323 urologic consultations in a period of 30 months, 779 were due to sexual dysfunction--14.63% (IC 95% 12-17%); 54.3% were due to erectile sexual dysfunction; 33% were due to ejaculation disorders; and 12.7% were because of other because of other dysfunctions. CONCLUSIONS: The consultation rate increases significantly with age. In patients younger than 20-years-old the most frequent complaint is ejaculation disorder while in patients younger than 40-years-old the risk of erectile sexual dysfunction is low, increasing twice as much at 40 be considered an important health problem.

Prevalencia de las disfunciones sexuales masculinas. / [Prevalence of male sexual dysfunctions]

OBJECTIVE: To determine the prevalence rate of male sexual dysfunction in 5323 urologic consultations. MATERIALS AND METHODS: In order to establish which age groups show an important consultation rate due to male sexual dysfunction, the number of consultations and the theoretical distribution according to age in the province of Córdoba were contrasted, estimating the risk of suffering from sexual dysfunction, and determining the age distribution and tendency analysis. RESULTS: From 5323 urologic consultations in a period of 30 months, 779 were due to sexual dysfunction--14.63

(IC 95

12-17

); 54.3

were due to erectile sexual dysfunction; 33

were due to ejaculation disorders; and 12.7

were because of other because of other dysfunctions. CONCLUSIONS: The consultation rate increases significantly with age. In patients younger than 20-years-old the most frequent complaint is ejaculation disorder while in patients younger than 40-years-old the risk of erectile sexual dysfunction is low, increasing twice as much at 40 be considered an important health problem.

[Mitomycin C for the treatment of superficial tumors of the bladder (Tis-Ta-T1) with one or more risk factors. Argentinian Cooperative Group of Oncourology (GCAO)]. / Mitomicina-C para el tratamiento de los tumores superficiales de vejiga (Tis-Ta-T1) con uno o más factores de riesgo. Grupo Cooperativo Argentino de Oncourología (GCAO).

OBJECTIVES: To determine the efficacy and toxicity of intravesical mitomycin C in superficial bladder tumors (Tis Ta T1) completely resected with one or more risk factors. (Protocol U. 01/90). METHODS: The patients received six instillations weekly of 40 mg mitomycin C within 15 days post-TUR of a superficial bladder tumor with one or more risk factors: histological grade 3, tumor size more than 3 cm and/or multicentric lesion. The patients were evaluated by cystoscopy every three months for the first two years and every six months thereafter. Toxicity was evaluated according to Miller's score. Tumor recurrence, disease-free interval and survival were analyzed. RESULTS: 126 patients were entered into the study; of these, 110 were evaluable. At 18 months mean follow-up (range 6-36 months), 77 patients (70%) remain disease-free; the mean time to recurrence was 13.8 months. There were no differences between patients with one, two or three risk factors or those who received or did not receive previous treatments. The patients tolerated the treatment well; there were no dropouts or systemic toxicity. CONCLUSIONS: 1. At 18 months mean follow-up, 77 of 110 patients (70%) remain disease-free; 2. The mean time to recurrence was 13.8 months; 3. Local toxicity was minimal; 4. There were no dropouts due to toxicity; 5. Systemic toxicity was not observed.

Characterization of monoclonal antibodies against ovine prolactin: suitability for use in immunocytochemical analysis of rat prolactin.

The aim of this study was to identify a monoclonal antibody (MAb) suitable for use in the immunocytochemical localization of prolactin in rat tissues. We took advantage of the conservation of certain amino acid sequences in prolactin among species by examining the crossreactivity patterns of five MAb, originally generated to ovine prolactin, with rat prolactin by enzyme-linked immunoassay (ELISA), Western blot analysis, and immunocytochemistry. Two of five antibodies (17D9 and 6F11) showed reactivity with 100 ng of immobilized rat prolactin (NIH RP-3) by ELISA, 6F11 reacting more strongly than 17D9. Only 6F11 reacted with prolactin in lysates of GH4C1 rat pituitary tumor cells by Western blot analysis. When we examined the crossreactivity of the MAb with rat prolactin in monolayer cultures of GH4C1 cells by indirect immunofluorescence, we found that both 17D9 and 6F11 reacted strongly with the cultures. The distribution of staining with 17D9 or 6F11 was coincident with staining with a polyclonal antiserum to rat prolactin. Preabsorption of the antibodies with a 20-fold excess of purified rat prolactin abolished the staining of GH4C1 cell cultures with either antibody. Therefore, we have selected from a series of MAb raised to ovine prolactin two antibodies (17D9 and 6F11) that react specifically with rat prolactin in immunocytochemical studies, whereas 6F11 also reacts strongly with rat prolactin by ELISA and Western blot analysis.