Duocarmycin SA Reduces Proliferation and Increases Apoptosis in Acute Myeloid Leukemia Cells In Vitro.

Acute myeloid leukemia (AML) is a hematological malignancy that is characterized by an expansion of immature myeloid precursors. Despite therapeutic advances, the prognosis of AML patients remains poor and there is a need for the evaluation of promising therapeutic candidates to treat the disease. The objective of this study was to evaluate the efficacy of duocarmycin Stable A (DSA) in AML cells in vitro. We hypothesized that DSA would induce DNA damage in the form of DNA double-strand breaks (DSBs) and exert cytotoxic effects on AML cells within the picomolar range. Human AML cell lines Molm-14 and HL-60 were used to perform 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), DNA DSBs, cell cycle, 5-ethynyl-2-deoxyuridine (EdU), colony formation unit (CFU), Annexin V, RNA sequencing and other assays described in this study. Our results showed that DSA induced DNA DSBs, induced cell cycle arrest at the G2M phase, reduced proliferation and increased apoptosis in AML cells. Additionally, RNA sequencing results showed that DSA regulates genes that are associated with cellular processes such as DNA repair, G2M checkpoint and apoptosis. These results suggest that DSA is efficacious in AML cells and is therefore a promising potential therapeutic candidate that can be further evaluated for the treatment of AML.

Forecasting of Airborne Conidia Quantities and Potential Insect Associations of Cryphonectria parasitica, the Causal Agent of Chestnut Blight, in England.

Sweet chestnut, an Asiatic tree introduced in many parts of Europe including the United Kingdom, is planted for nut production, timber, and amenity. Its major threat is the disease called blight, caused by the fungus Cryphonectria parasitica, which infects through wounds by airborne spores. Field trapping using sticky rods rotating traps was performed in an infected area in Devon (between May 2021 and April 2023). An improved dual hydrolysis Taqman probes real-time PCR was used. The number of spores was calculated by comparing the cycle threshold to the Ct of standards with known amounts of conidia or known target fragment copies cloned into a plasmid. Weekly spore counts were in the range of around 60 to approximately 8.5 × 103, with fluctuations of peaks (mainly in late summer-autumn 2021) and troughs. The effects of weather parameters were modelled, finding correlations between spore numbers and temperature, humidity, dewpoint, rainfall, wind speed, and wind duration. Additionally, an insect trapping was performed to confirm the presence/absence and quantity of C. parasitica conidia potentially phoretic on some insects by using the same molecular approach. None of the ten collected insect species harboured spores of this fungus.

Cryphonectria parasitica Detections in England, Jersey, and Guernsey during 2020-2023 Reveal Newly Affected Areas and Infections by the CHV1 Mycovirus.

In England, Cryphonectria parasitica was detected for the first time in 2011 in a nursery and in 2016 in the wider environment. Surveys between 2017 and 2020 identified the disease at different sites in Berkshire, Buckinghamshire, Cornwall, Derbyshire, Devon, Dorset, London, West Sussex, and the island of Jersey, while the present study comprises the results of the 2020-2023 survey with findings in Derbyshire, Devon, Kent, Nottinghamshire, Herefordshire, Leicestershire, London, West Sussex, and the islands of Jersey and Guernsey. A total of 226 suspected samples were collected from 72 surveyed sites, as far north as Edinburgh and as far west as Plymouth (both of which were negative), and 112 samples tested positive by real-time PCR and isolation from 35 sites. The 112 isolates were tested for the vegetative compatibility group (VCG), mating type, and Cryphonectria hypovirus 1 (CHV1). Twelve VCGs were identified, with two of them (EU-5 and EU-22) being the first records in the UK. Both mating types were present (37% MAT-1 and 63% MAT-2), but only one mating type was present per site and VCG, and perithecia were never observed. Cryphonectria hypovirus 1 (CHV1), consistently subtype-I haplotype E-5, was detected in three isolates at a low concentration (5.9, 21.1, and 33.0 ng/µL) from locations in London, Nottinghamshire, and Devon.

Glucocorticoid Receptor Regulates and Interacts with LEDGF/p75 to Promote Docetaxel Resistance in Prostate Cancer Cells.

Patients with advanced prostate cancer (PCa) invariably develop resistance to anti-androgen therapy and taxane-based chemotherapy. Glucocorticoid receptor (GR) has been implicated in PCa therapy resistance; however, the mechanisms underlying GR-mediated chemoresistance remain unclear. Lens epithelium-derived growth factor p75 (LEDGF/p75, also known as PSIP1 and DFS70) is a glucocorticoid-induced transcription co-activator implicated in cancer chemoresistance. We investigated the contribution of the GR-LEDGF/p75 axis to docetaxel (DTX)-resistance in PCa cells. GR silencing in DTX-sensitive and -resistant PCa cells decreased LEDGF/p75 expression, and GR upregulation in enzalutamide-resistant cells correlated with increased LEDGF/p75 expression. ChIP-sequencing revealed GR binding sites in the LEDGF/p75 promoter. STRING protein-protein interaction analysis indicated that GR and LEDGF/p75 belong to the same transcriptional network, and immunochemical studies demonstrated their co-immunoprecipitation and co-localization in DTX-resistant cells. The GR modulators exicorilant and relacorilant increased the sensitivity of chemoresistant PCa cells to DTX-induced cell death, and this effect was more pronounced upon LEDGF/p75 silencing. RNA-sequencing of DTX-resistant cells with GR or LEDGF/p75 knockdown revealed a transcriptomic overlap targeting signaling pathways associated with cell survival and proliferation, cancer, and therapy resistance. These studies implicate the GR-LEDGF/p75 axis in PCa therapy resistance and provide a pre-clinical rationale for developing novel therapeutic strategies for advanced PCa.

Temperature Effects on the Cryphonectria hypovirus 1 Accumulation and Recovery within Its Fungal Host, the Chestnut Blight Pathogen Cryphonectria parasitica.

Biological control of Cryphonectria parasitica fungus, the causal agent of chestnut blight, by virus infection (hypovirulence) is an effective control strategy against chestnut blight in Europe and some parts of North America. The most studied mycovirus is the Cryphonectria hypovirus 1 (CHV1) type species of the Hypoviridae family. In this study, the CHV1 virus was studied within some highly infected British isolates of Cryphonectria parasitica, gained in the past through co-culture transmissions. The effects of six temperatures (5-30 °C, in 5 °C steps) on six infected isolates (three with viral strain E-5, and other three with viral strain L-18) and their respective negative non-infected controls, three isogenic virulent fungal isolates, were examined. Experiments were performed with the nine isolate types with three replicates on potato dextrose agar (PDA) with cellophane sheets per isolate and temperature. A recently developed rapid, specific, quantitative reverse transcription PCR (RT-qPCR) screening method was used. This enabled quantifying the concentration (nanograms per microliter or copy numbers) of the virus within each isolate repetition. The presence of the virus had a significant negative effect between 20 and 25 °C on the C. parasitica growth rate, which was anyway highly influenced by and positively correlated with the temperature. The temperature clearly determined the virus accumulation and its recovery from cold or heat, and the virus optimum temperature was estimated at 15-25 °C.

Glucocorticoid Receptor and ß-Catenin Interact in Prostate Cancer Cells and Their Co-Inhibition Attenuates Tumorsphere Formation, Stemness, and Docetaxel Resistance.

Therapy resistance hinders the efficacy of anti-androgen therapies and taxane-based chemotherapy for advanced prostate cancer (PCa). Glucocorticoid receptor (GR) signaling mediates resistance to androgen receptor signaling inhibitors (ARSI) and has also been recently implicated in PCa resistance to docetaxel (DTX), suggesting a role in therapy cross-resistance. Like GR, ß-catenin is upregulated in metastatic and therapy-resistant tumors and is a crucial regulator of cancer stemness and ARSI resistance. ß-catenin interacts with AR to promote PCa progression. Given the structural and functional similarities between AR and GR, we hypothesized that ß-catenin also interacts with GR to influence PCa stemness and chemoresistance. As expected, we observed that treatment with the glucocorticoid dexamethasone promotednuclear accumulation of GR and active ß-catenin in PCa cells. Co-immunoprecipitation studies showed that GR and ß-catenin interact in DTX-resistant and DTX-sensitive PCa cells. Pharmacological co-inhibition of GR and ß-catenin, using the GR modulator CORT-108297 and the selective ß-catenin inhibitor MSAB, enhanced cytotoxicity in DTX-resistant PCa cells grown in adherent and spheroid cultures and decreased CD44+/CD24- cell populations in tumorspheres. These results indicate that GR and ß-catenin influence cell survival, stemness, and tumorsphere formation in DTX-resistant cells. Their co-inhibition could be a promising therapeutic strategy to overcome PCa therapy cross-resistance.

Using the Synergy between HPLC-MS and MALDI-MS Imaging to Explore the Lipidomics of Clear Cell Renal Cell Carcinoma.

Lipid imaging mass spectrometry (LIMS) has been tested in several pathological contexts, demonstrating its ability to segregate and isolate lipid signatures in complex tissues, thanks to the technique's spatial resolution. However, it cannot yet compete with the superior identification power of high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS), and therefore, very often, the latter is used to refine the assignment of the species detected by LIMS. Also, it is not clear if the differences in sensitivity and spatial resolution between the two techniques lead to a similar panel of biomarkers for a given disease. Here, we explore the capabilities of LIMS and HPLC-MS to produce a panel of lipid biomarkers to screen nephrectomy samples from 40 clear cell renal cell carcinoma patients. The same set of samples was explored by both techniques, and despite the important differences between them in terms of the number of detected and identified species (148 by LIMS and 344 by HPLC-MS in negative-ion mode) and the presence/absence of image capabilities, similar conclusions were reached: using the lipid fingerprint, it is possible to set up classifiers that correctly identify the samples as either healthy or tumor samples. The spatial resolution of LIMS enables extraction of additional information, such as the existence of necrotic areas or the existence of different tumor cell populations, but such information does not seem determinant for the correct classification of the samples, or it may be somehow compensated by the higher analytical power of HPLC-MS. Similar conclusions were reached with two very different techniques, validating their use for the discovery of lipid biomarkers.

The Nuclear Dense Fine Speckled (DFS) Immunofluorescence Pattern: Not All Roads Lead to DFS70/LEDGFp75.

The monospecific dense fine speckled (DFS) immunofluorescence assay (IFA) pattern is considered a potential marker to aid in exclusion of antinuclear antibody (ANA)-associated rheumatic diseases (AARD). This pattern is typically produced by autoantibodies against transcription co-activator DFS70/LEDGFp75, which are frequently found in healthy individuals and patients with miscellaneous inflammatory conditions. In AARD patients, these antibodies usually co-exist with disease-associated ANAs. Previous studies reported the occurrence of monospecific autoantibodies that generate a DFS-like or pseudo-DFS IFA pattern but do not react with DFS70/LEDGFp75. We characterized this pattern using confocal microscopy and immunoblotting. The target antigen associated with this pattern partially co-localized with DFS70/LEDGFp75 and its interacting partners H3K36me2, an active chromatin marker, and MLL, a transcription factor, in HEp-2 cells, suggesting a role in transcription. Immunoblotting did not reveal a common protein band immunoreactive with antibodies producing the pseudo-DFS pattern, suggesting they may recognize diverse proteins or conformational epitopes. Given the subjectivity of the HEp-2 IFA test, the awareness of pseudo-DFS autoantibodies reinforces recommendations for confirmatory testing when reporting patient antibodies producing a putative DFS pattern in a clinical setting. Future studies should focus on defining the potential diagnostic utility of the pseudo-DFS pattern and its associated antigen(s).

Canker Development and Biocontrol Potential of CHV-1 Infected English Isolates of Cryphonectria parasitica Is Dependent on the Virus Concentration and the Compatibility of the Fungal Inoculums.

Biological control of Cryphonectria parasitica fungus, causal agent of chestnut blight, by virus infection (hypovirulence) has been shown to be an effective control strategy against chestnut blight in Europe and some parts of North America. The most studied mycovirus is the Cryphonectria hypovirus 1 (CHV-1) type species of the Hypoviridae family. To efficiently provide biocontrol, the virus must be able to induce hypovirulence in its fungal host in chestnut trees. Here, two different CHV-1 subtype I virus strains (E-5 and L-18), gained by transmissions, were tested for their hypovirulence induction, biocontrol potential, and transmission between vegetatively compatible (VCG) and incompatible fungal isolate groups in sweet chestnut seedlings and branches. Both strains of CHV-1 showed great biocontrol potential and could protect trees by efficiently transmitting CHV-1 by hyphal anastomosis between fungal isolates of the same VCG and converting virulent to hypovirulent cankers. The hypovirulent effect was positively correlated with the virus concentration, tested by four different reverse-transcription PCRs, two end-point and two real-time methods, one of which represents a newly developed real-time PCR for the detection and quantification of CHV-1.

Social Inequities in the Impact of COVID-19 Lockdown Measures on the Mental Health of a Large Sample of the Colombian Population (PSY-COVID Study).

The identification of general population groups particularly vulnerable to the impact of COVID-19 lockdown measures on mental health and the development of healthcare policies are priority challenges in the current and future pandemics. This study aimed to identify the personal and social determinants of the impact of COVID-19 lockdown measures on mental health in a large sample of the Colombian population. In this cross-sectional study, an anonymous online survey was answered by 18,061 participants from the general population residing in Colombia during the first wave of the COVID-19 outbreak (from 20 May to 20 June 2020). The risk of depression, anxiety, and somatization disorders were measured using the Patient Health Questionnaire (PHQ-2), Generalized Anxiety Disorder Scale (GAD-2), and Somatic Symptom Questionnaire (SSQ-5), respectively. Overall, 35% of participants showed risk of depression, 29% of anxiety, and 31% of somatization. According to the analysis of social determinants of health, the most affected groups were people with low incomes, students, and young adults (18-29 years). Specifically, low-income young females were the most at-risk population group. These findings show how the lockdown measures affected the general population's mental health in Colombia and highlight some social risk factors in health.

The LEDGF/p75 Integrase Binding Domain Interactome Contributes to the Survival, Clonogenicity, and Tumorsphere Formation of Docetaxel-Resistant Prostate Cancer Cells.

Patients with prostate cancer (PCa) receiving docetaxel chemotherapy invariably develop chemoresistance. The transcription co-activator lens epithelium-derived growth factor p75 (LEDGF/p75), also known as DFS70 and PSIP1, is upregulated in several human cancers, including PCa and promotes resistance to docetaxel and other drugs. The C-terminal region of LEDGF/p75 contains an integrase binding domain (IBD) that tethers nuclear proteins, including the HIV-1 integrase and transcription factors, to active chromatin to promote viral integration and transcription of cellular survival genes. Here, we investigated the contribution of the LEDGF/p75 IBD interactome to PCa chemoresistance. Quantitative immunoblotting revealed that LEDGF/p75 and its IBD-interacting partners are endogenously upregulated in docetaxel-resistant PCa cell lines compared to docetaxel-sensitive parental cells. Using specific human autoantibodies, we co-immunoprecipitated LEDGF/p75 with its endogenous IBD-interacting partners JPO2, menin, MLL, IWS1, ASK1, and PogZ, as well as transcription factors c-MYC and HRP2, in docetaxel-resistant cells, and confirmed their nuclear co-localization by confocal microscopy. Depletion of LEDGF/p75 and selected interacting partners robustly decreased the survival, clonogenicity, and tumorsphere formation capacity of docetaxel-resistant cells. These results implicate the LEDGF/p75 IBD interactome in PCa chemoresistance and could lead to novel therapeutic strategies targeting this protein complex for the treatment of docetaxel-resistant tumors.

[Multidisciplinary pelvic floor team: 5 year experience.] / Unidad funcional multidisciplinar de suelo pélvico: experiencia de 5 años.

INTRODUCTION: Pelvic floor dysfunction (PFD) is expressed by many different symptoms that can affect pelvic floor organs. These symptoms can appear alone or associated. The involvement of different organs and the wide variety of symptoms we have to deal with, has become teamwork essential for diagnosis and treatment of PFD. Currently, multidisciplinary pelvic floor teams are essential when dealing with PFD. OBJECTIVE: The main objective of the article is to analyze characteristics and pathologies of the patients discussed during these multidisciplinary meetings. MATERIAL AND METHODS: This is a descriptive study. Patients followed in the multidisciplinary pelvic floorteam at Galdakao-Usansolo Hospital were recruited. We analyzed the patients discussed at the meetings from January 2014 until March 2019. RESULTS: We recruited 55 patients. 89.09% were female and the mean age of the analyzed patients was 55.36 years. The most frequent reasons for medicalconsultation were pelvic floor prolapses in female (22.45%) and pelvic floor pain in male (33.33%). Association between symptoms occur in 78.18% of the patients.41.82% of patients were referred to more than 2 specialists and 21.82% needed associated treatments. CONCLUSIONS: Pelvic floor dysfunction is manifested by a combination of  symptoms which need different specialists, not only for diagnosis, but also for treatment. This is why multidisciplinary teams have become important for the management of pelvic floor disfunction.

INTRODUCCIÓN: La disfunción del suelo pélvico se presenta de múltiples formas y se manifiesta con síntomas que afectan a diferentes órganos del suelo pélvico. Estos síntomas pueden aparecer aislados o asociados entre sí, implicando así a diferentes especialidades a la hora del diagnóstico y tratamiento. Los comités multidisciplinares se han convertido en una herramienta imprescindible en el manejo de la patología del suelo pélvico. OBJETIVO: El objetivo principal es estudiar las características de la patología que presentan los pacientes abordados en el Comité Funcional Multidisciplinar de Suelo Pélvico (CFMSP).MATERIAL Y MÉTODOS: Estudio descriptivo transversal de los pacientes abordados en el CFMSP del Hospital de Galdakao-Usansolo desde 2014 hasta marzo de 2019. RESULTADOS: En el comité se abordaron 55 pacientes, de las cuales el 89,09% fueron mujeres. La edad media fue de 55,36 años. El motivo de consulta másfrecuente fue el prolapso de órganos pélvicos (22,45%) en la población femenina y el dolor (33,33%) en la población masculina. El 78,18% presentaba más de unsíntoma asociado. Los pacientes que requirieron ser derivados a más de 2 especialidades supuso el 41,82% y el 21,82% de los pacientes precisaron más de una modalidad de tratamiento. CONCLUSIONES: La patología del suelo pélvico se manifiesta como asociación de síntomas que requieren la implicación de diferentes especialistas para el diagnóstico y tratamiento. Es el motivo por el que las unidades multidisciplinares son cada vez más necesarias en el abordaje integral de la patología del suelo pélvico.

Hypovirulent effect of the Cryphonectria hypovirus 1 in British isolates of Cryphonectria parasitica.

BACKGROUND: Chestnut blight, caused by Cryphonectria parasitica, is controlled in many European countries by the naturally occurring mycovirus Cryphonectria hypovirus 1 (CHV-1). During surveys of recently identified chestnut blight outbreak in England, CHV-1 was detected in several individuals of the pathogen isolated from affected trees. We investigated two of these CHV-1-infected isolates (L-6 and Db-1) as potential biocontrol agents for deployment in the UK comparing their virulence against virus-free (M1275) and hypovirulent (M784) European isolates by inoculating sweet chestnut seedlings. RESULTS: Both the European CHV-1 M784 hypovirulent isolate and UK L-6 isolate formed significantly smaller lesions in sweet chestnut seedling bark than the other three isolates (Db-1, and virulent isolates FTC121 and M1275). The highest virus concentration was detected in isolate M784, followed by L-6, with the lowest concentration in isolate Db-1. White colony colouration indicative of hypovirulence was common in colonies re-isolated from smaller lesions, and the same isolates also tended to be slower growing in culture, have a higher virus concentration, and caused less epicormic growth and fewer stromata to be present in plants. L-6 and Db-1 virus sequences, respectively, matched the virus haplotype E-5 detected previously in Switzerland and a mutation of the same subtype I haplotype. CONCLUSION: Isolate L-6 could potentially act as biocontrol for chestnut blight outbreaks in the UK but further laboratory and field experiments are needed. © 2019 Crown copyright. Pest Management Science © 2019 Society of Chemical Industry.

[Elevation of Ca 125 in a patient with giant congenital hydronephrosis].

OBJECTIVE: To report a new case with elevation of Ca 125 and hydronephrotic kidney without neoplastic disease, having special reference to clinical aspects. METHOD: The clinical history, anatomopathological and immunohistochemical findings are described. We performed a bibliographic review. We report the case of a 27 year-old female with the diagnosis of giant hydronephrosis and Ca125 elevation (313 u/ml hydronephrotic kidney urine and 112.3 u/ml serum). RESULT: She underwent a radical nephrectomy and Ca125 levels decreased. After 9 years the patient did not have any neoplastic disease and Ca125 levels are normal. CONCLUSIONS: The Ca125 rise with hydronephrotic kidney may be usual although there are not enough studies. We think that in patients with hydronephrotic kidney and malignant neoplasm the Ca125 antigen can lose sensitivity for the early diagnosis, staging and follow up of the malignant diseases.

[Wrist arthroscopy for fractures of the distal end of the radius]. / Artroscopía de muñieca en fracturas del extremo distal del radio.

INTRODUCTION: Fractures of the distal end of the radius are very frequent in the orthopedist's practice. The anatomical reduction of the articular surface and the extra-articular alignment of the radius are extremely important to minimize complications. The purpose of the study is to make an arthroscopic diagnosis of the lesions associated to the distal radius and assist in the reduction of the distal radius fracture. MATERIAL AND METHODS: The study was conducted from March 2007 to February 2008. We performed 20 wrist arthroscopies in 20 patients with an intraarticular fracture of the distal end of the radius. They were classified according to Frykman's criteria. Patient age ranged from 17 to 67, with a mean of 38.7 +/- 15.9 years. 60% were males and 40% females; mean follow-up was 12 months; the chondral lesions were classified according to Outerbridge, the interosseous ligament lesions and the triangular fibrocartilage lesions, according to Palmer's criteria. We did articular debridement, radiofrequency chondroplasty, and assisted the fracture reduction. We used Henry's palmar approach and did open reduction and internal fixation with a T-shaped palmar plate and finally performed an arthroscopy to check the reduction. RESULTS: 8 cases had chondral lesions (grades II to IV); 7 cases had lesion of the triangular fibrocartilage (central isolated ruptures, radial detachment of the triangular fibrocartilage from the sigmoid notch), 5 cases had lesion of the scapholunate ligament; one case had a lesion of the luno-pyramidal ligament, and in 6 cases no associated lesions were found; 8 cases had a single lesion and 6 cases had 2 to 3 concomitant lesions. Two patients had been lost at the 3- and 6-month follow-up visits; 35% of those that were followed-up had pain at 3 months (7 patients) and 15% at 6 months. Only 2 cases had scapholunate instability and only one case had distal radioulnar instability. CONCLUSIONS: Arthroscopy has proven useful in identifying hidden lesions, assisting in fracture reduction and even treating the lesions found at the time of the diagnosis. Chondral lesions were the most frequent ones, followed by lesions of the triangular fibrocartilage; a close relation was found between the type of Frykman fracture and the associated lesion. Type 7 and 8 lesions were more frequently associated with chondral lesions and with a higher mean age. At the end of the follow-up period the scapholunate ligament lesions remained unstable. Chondral lesions were related with persistent pain at 3 months.

Determination of the effective dose of an experimental fasciolicide in naturally and experimentally infected cattle.

The aim of the present study was to determine the effective dose of an experimental fasciolicide called compound alpha or 5-chloro-2-methylthio-6-(1-napthyloxy)1H-benzimidazole in experimentally and naturally infected cattle. In the first experiment, 24 fluke-free heifers were each infected with 800 metacercariae of Fasciola hepatica and re-infected on day 45 with other 600 cysts per animal. On day 75, when the animals had 4- and 10-week-old flukes respectively, they were divided into four groups (G) of six animals each according to fluke egg counts. Groups 1-3 received compound alpha at 10, 12 and 14 mg/kg/p.o., respectively. G4 remained as an untreated control. Twenty days after treatment, the animals were sacrificed for the recovery of flukes. Efficacy was assessed as a percentage of egg or fluke reduction relative to the untreated control. In the second experiment (naturally infected cattle), 24-year-old steers positive to F. hepatica eggs were blocked into four groups of five animals each. Groups 1-3 received compound alpha at 10, 12 and 14 mg/kg/p.o., respectively. Group 4 served as a non-treated control. All procedures to determine efficacy were carried out as mentioned in the first experiment. The results in the first study showed a percentage on egg reduction of 97.3, 100 and 100 and overall fluke reduction of 94.3, 100 and 100 for Groups 1-3, respectively. In the second study, the percentage of egg reduction was of 87.5, 99.1 and 100 and overall efficacy regarding fluke reduction was of 84.2, 99.6, and 100 for Groups 1-3, respectively. It is concluded that the effective dose selected for compound alpha was of 12 mg/kg/p.o. in cattle having an induced or natural F. hepatica infection.