A Phase I/II Clinical Trial to evaluate the efficacy of baricitinib to prevent respiratory insufficiency progression in onco-hematological patients affected with COVID19: A structured summary of a study protocol for a randomised controlled trial.

OBJECTIVES: Baricitinib is supposed to have a double effect on SARS-CoV2 infection. Firstly, it reduces the inflammatory response through the inhibition of the Januse-Kinase signalling transducer and activator of transcription (JAK-STAT) pathway. Moreover, it reduces the receptor mediated viral endocytosis by AP2-associated protein kinase 1 (AAK1) inhibition. We propose the use of baricinitib to prevent the progression of the respiratory insufficiency in SARS-CoV2 pneumonia in onco-haematological patients. In this phase Ib/II study, the primary objective in the safety cohort is to describe the incidence of severe adverse events associated with baricitinib administration. The primary objective of the randomized phase (baricitinib cohort versus standard of care cohort) is to evaluate the number of patients who did not require mechanical oxygen support since start of therapy until day +14 or discharge (whichever it comes first). The secondary objectives of the study (only randomized phase of the study) are represented by the comparison between the two arms of the study in terms of mortality and toxicity at day+30. Moreover, a description of the immunological related changes between the two arms of the study will be reported. TRIAL DESIGN: The trial is a phase I/II study with a safety run-in cohort (phase 1) followed by an open label phase II randomized controlled trial with an experimental arm compared to a standard of care arm. PARTICIPANTS: The study will be performed at the Institut Català d'Oncologia, a tertiary level oncological referral center in the Catalonia region (Spain). The eligibility criteria are: patients > 18 years affected by oncological diseases; ECOG performance status < 2 (Karnofsky score > 60%); a laboratory confirmed infection with SARS-CoV-2 by means of real -time PCR; radiological signs of low respiratory tract disease; absence of organ dysfunction (a total bilirubin within normal institutional limits, AST/ALT≤2.5 X institutional upper limit of normal, alkaline phosphatase ≤2.5 X institutional upper limit of normal, coagulation within normal institutional limits, creatinine clearance >30 mL/min/1.73 m2 for patients with creatinine levels above institutional normal); absence of HIV infection; no active or latent HBV or HCV infection. The exclusion criteria are: patients with oncological diseases who are not candidates to receive any active oncological treatment; hemodynamic instability at time of study enrollment; impossibility to receive oral medication; medical history of recent or active pulmonary embolism or deep venous thrombosis or patients at high-risk of suffering them (surgical intervention, immobilization); multi organ failure, rapid worsening of respiratory function with requirement of fraction of inspired oxygen (FiO2) > 50% or high-flow nasal cannula before initiation of study treatment; uncontrolled intercurrent illness (ongoing or severe active infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac arrhythmia, or psychiatric illness/social situations that would limit compliance with study requirements); allergy to one or more of study treatments; pregnant or breastfeeding women; positive pregnancy test in a pre-dose examination. Patients should have the ability to understand, and the willingness to sign, a written informed consent document; the willingness to accept randomization to any assigned treatment arm; and must agree not to enroll in another study of an investigational agent prior to completion of Day +28 of study. An electronic Case Report Form in the Research Electronic Data Capture (REDCap) platform will be used to collect the data of the trial. Removal from the study will apply in case of unacceptable adverse event(s), development of an intercurrent illness, condition or procedural complication, which could interfere with the patient's continued participation and voluntary patient withdrawal from study treatment (all patients are free to withdraw from participation in this study at any time, for any reasons, specified or unspecified, and without prejudice). INTERVENTION AND COMPARATOR: Treatment will be administered on an inpatient basis. We will compare the experimental treatment with baricitinib plus the institutional standard of care compared with the standard of care alone. During the phase I, we will define the dose-limiting toxicity of baricitinib and the dose to be used in the phase 2 part of the study. The starting baricitinib dose will be an oral tablet 4 mg-once daily which can be reduced to 2 mg depending on the observed toxicity. The minimum duration of therapy will be 5 days and it can be extended to 7 days. The standard of care will include the following therapies. Antibiotics will be individualized based on clinical suspicion, including the management of febrile neutropenia. Prophylaxis of thromboembolic disease will be administered to all participants. Remdesivir administration will be considered only in patients with severe pneumonia (SatO2 <94%) with less than 7 days of onset of symptoms and with supplemental oxygen requirements but not using high-flow nasal cannula, non-invasive or invasive mechanical ventilation or extracorporeal membrane oxygenation (ECMO). In the randomized phase, tocilizumab or interferon will not be allowed in the experimental arm. Tocilizumab can be used in patients in the standard of care arm at the discretion of the investigator. If it is prescribed it will be used according to the following criteria: patients who, according to his baseline clinical condition, would be an ICU tributary, interstitial pneumonia with severe respiratory failure, patients who are not on mechanical ventilation or ECMO and who are still progressing with corticoid treatment or if they are not candidates for corticosteroids. Mild ARDS (PAFI <300 mmHg) with radiological or blood gases deterioration that meets at least one of the following criteria: CRP >100mg/L D-Dimer >1,000µg/L LDH >400U/L Ferritin >700ng/ml Interleukin 6 ≥40ng/L. The use of tocilizumab is not recommended if there are AST/ALT values greater than 10 times the upper limit of normal, neutrophils <500 cells/mm3, sepsis due to other pathogens other than SARS-CoV-2, presence of comorbidity that can lead to a poor prognosis, complicated diverticulitis or intestinal perforation, ongoing skin infection. The dose will be that recommended by the Spanish Medicine Agency in patients ≥75Kg: 600mg dose whereas in patients <75kg: 400mg dose. Exceptionally, a second infusion can be assessed 12 hours after the first in those patients who experience a worsening of laboratory parameters after a first favourable response. The use of corticosteroids will be recommended in patients who have had symptoms for more than 7 days and who meet all the following criteria: need for oxygen support, non-invasive or invasive mechanical ventilation, acute respiratory failure or rapid deterioration of gas exchange, appearance or worsening of bilateral alveolar-interstitial infiltrates at the radiological level. In case of indication, it is recommended: dexamethasone 6mg/d p.o. or iv for 10 days or methylprednisolone 32mg/d orally or 30mg iv for 10 days or prednisone 40mg day p.o. for 10 days. MAIN OUTCOMES: Phase 1 part: to describe the toxicity profile of baricitinib in COVID19 oncological patients during the 5-7 day treatment period and until day +14 or discharge (whichever it comes first). Phase 2 part: to describe the number of patients in the experimental arm that will not require mechanical oxygen support compared to the standard of care arm until day +14 or discharge (whichever it comes first). RANDOMISATION: For the phase 2 of the study, the allocation ratio will be 1:1. Randomization process will be carried out electronically through the REDcap platform ( https://www.project-redcap.org/ ) BLINDING (MASKING): This is an open label study. No blinding will be performed. NUMBERS TO BE RANDOMISED (SAMPLE SIZE): The first part of the study (safety run-in cohort) will consist in the enrollment of 6 to 12 patients. In this population, we will test the toxicity of the experimental treatment. An incidence of severe adverse events grade 3-4 (graded by Common Terminology Criteria for Adverse Events v.5.0) inferior than 33% will be considered sufficient to follow with the next part of the study. The second part of the study we will perform an interim analysis of efficacy at first 64 assessed patients and a definitive one will analyze 128 assessed patients. Interim and definitive tests will be performed considering in both cases an alpha error of 0.05. We consider for the control arm this rate is expected to be 0.60 and for the experimental arm of 0.80. Considering this data, a superiority test to prove a difference of 0.20 with an overall alpha error of 0.10 and a beta error of 0.2 will be performed. Considering a 5% of dropout rate, it is expected that a total of 136 patients, 68 for each study arm, will be required to complete study accrual. TRIAL STATUS: Version 5.0. 14th October 2020 Recruitment started on the 16th of December 2020. Expected end of recruitment is June 2021. TRIAL REGISTRATION: AEMPs: 20-0356 EudraCT: 2020-001789-12, https://www.clinicaltrialsregister.eu/ctr-search/search (Not publically available as Phase I trial) Clinical trials: BARCOVID19, https://www.clinicaltrials.gov/ (In progress) FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol."

Presence of bacterial DNA in thrombotic material of patients with myocardial infarction.

Infectious agents have been suggested to be involved in etiopathogenesis of Acute Coronary Syndrome (ACS). However, the relationship between bacterial infection and acute myocardial infarction (AMI) has not yet been completely clarified. The objective of this study is to detect bacterial DNA in thrombotic material of patients with ACS with ST-segment elevation (STEMI) treated with Primary Percutaneous Coronary Intervention (PPCI). We studied 109 consecutive patients with STEMI, who underwent thrombus aspiration and arterial peripheral blood sampling. Testing for bacterial DNA was performed by probe-based real-time Polymerase Chain Reaction (PCR). 12 probes and primers were used for the detection of Aggregatibacter actinomycetemcomitans, Chlamydia pneumoniae, viridans group streptococci, Porphyromonas gingivalis, Fusobacterium nucleatum, Tannarella forsythia, Treponema denticola, Helycobacter pylori, Mycoplasma pneumoniae, Staphylococus aureus,  Prevotella intermedia and Streptococcus mutans. Thus, DNA of four species of bacteria was detected in 10 of the 109 patients studied. The most frequent species was viridans group streptococci (6 patients, 5.5%), followed by Staphylococus aureus (2 patients, 1.8%). Moreover, a patient had DNA of Porphyromonas gingivalis (0.9%); and another patient had DNA of Prevotella intermedia (0.9%). Bacterial DNA was not detected in peripheral blood of any of our patients. In conclusion, DNA of four species of endodontic and periodontal bacteria was detected in thrombotic material of 10 STEMI patients. Bacterial DNA was not detected in the peripheral blood of patients with bacterial DNA in their thrombotic material. Bacteria could be latently present in plaques and might play a role in plaque instability and thrombus formation leading to ACS.

[Effects of education and strength training on functional tests among older people with osteoarthritis]. / Programas de Educación en Salud y Entrenamiento de la Fuerza en adultos mayores con artrosis de cadera leve a moderada.

BACKGROUND: Hip and knee osteoarthritis are important causes of pain and disability among older people. Education and strength training can alleviate symptoms and avoid functional deterioration. AIM: To assess muscle strength, fall risk and quality of life of older people with osteoarthritis and the effects of physiotherapy education and strength training on these variables. MATERIAL AND METHODS: Thirty participants aged 78 ± 5 years (63% women) were randomly assigned to receive physiotherapy (Controls), physiotherapy plus education (Group 1) and physiotherapy plus strength training (group 2). At baseline and after 16 weeks of intervention, patients were evaluated with the Senior Fitness Test, Timed Up and Go and Quality of Life score short form (SF-36). RESULTS: During the intervention period, Senior Fitness Test and Timed Up and Go scores improved in all groups and SF-36 did not change. The improvement in Senior Fitness Test and Timed Up and Go was more marked in Groups 1 and 2 than in the control group. CONCLUSIONS: Education and strength training improve functional tests among older people with osteoarthritis.

Programas de Educación en Salud y Entrenamiento de la Fuerza en adultos mayores con artrosis de cadera leve a moderada / Effects of education and strength training on functional tests among older people with osteoarthritis

Background: Hip and knee osteoarthritis are important causes of pain and disability among older people. Education and strength training can alleviate symptoms and avoid functional deterioration. Aim: To assess muscle strength, fall risk and quality of life of older people with osteoarthritis and the effects of physiotherapy education and strength training on these variables. Material and Methods: Thirty participants aged 78 ± 5 years (63% women) were randomly assigned to receive physiotherapy (Controls), physiotherapy plus education (Group 1) and physiotherapy plus strength training (group 2). At baseline and after 16 weeks of intervention, patients were evaluated with the Senior Fitness Test, Timed Up and Go and Quality of Life score short form (SF-36). Results: During the intervention period, Senior Fitness Test and Timed Up and Go scores improved in all groups and SF-36 did not change. The improvement in Senior Fitness Test and Timed Up and Go was more marked in Groups 1 and 2 than in the control group. Conclusions: Education and strength training improve functional tests among older people with osteoarthritis.

T-box 3 negatively regulates osteoblast differentiation by inhibiting expression of osterix and runx2.

T-box (Tbx)3, a known transcriptional repressor, is a member of a family of transcription factors, which contain a highly homologous DNA binding domain known as the Tbx domain. Based on the knowledge that mutation of the Tbx3 gene results in limb malformation, Tbx3 regulates osteoblast proliferation and its expression increases during osteoblast differentiation, we predicted that Tbx3 is an important regulator of osteoblast cell functions. In this study, we evaluated the consequence of transgenic overexpression of Tbx3 on osteoblast differentiation. Retroviral overexpression increased Tbx3 expression >100-fold at the mRNA and protein level. Overexpression of Tbx3 blocked mineralized nodule formation (28 +/- 8 vs. 7 +/- 1%) in MC3T3-E1 cells. In support of these data, alkaline phosphatase (ALP) activity was reduced 33-70% (P < 0.05) in both MC3T3-E1 cells and primary calvaria osteoblasts overexpressing Tbx3. In contrast, Tbx3 overexpression did not alter ALP activity in bone marrow stromal cells. Tbx3 overexpression blocked the increase in expression of key osteoblast marker genes, ALP, bone sialoprotein, and osteocalcin that occurs during normal osteoblast differentiation, but had little or no effect on expression of proliferation genes p53 and Myc. In addition, Tbx3 overexpression abolished increased osterix and runx2 expression observed during normal osteoblast differentiation, but the change in Msx1 and Msx2 expression over time was similar between control and Tbx3 overexpressing cells. Interestingly, osterix and runx2, but not Msx1 and Msx2, contain Tbx binding site in the regulatory region. Based on these data and our previous findings, we conclude that Tbx3 promotes proliferation and suppresses differentiation of osteoblasts and may be involved in regulating expression of key transcription factors involved in osteoblast differentiation.

[Multifocal electroretinogram and age-related macular degeneration]. / Electrorretinograma multifocal y degeneración macular asociada a la edad.

PURPOSE: To assess the value of multifocal electroretinogram (mfERG) in clinical evaluation of the response of patients with choroidal neovascularization (CNV), secondary to age-related macular degeneration, to photodynamic therapy (PDT). METHODS: We designed a prospective non-comparative case series and enrolled 23 eyes of 22 patients with predominantly classic CNV. The initial evaluation included clinical examination with measurement of visual acuity (Colenbrander), biomicroscopic evaluation (BMC), retinography, fluorescein angiography (FA), Humphrey 24-2 visual field examination and mfERG. All patients were treated with PDT. Clinical follow-up was performed two and six months after treatment with visual acuity evaluation, BMC, visual field assessment and mfERG. FA was repeated after an interval of between four and six months in all patients. RESULTS: Visual acuity remained stable or improved in all patients included in the study. Visual field examination gave erratic and unreliable results because of patients' unstable fixation. N1 and P1 wave peak amplitudes in the central rings did not change significantly after treatment but they improved in rings 3 (p=0.000), 4 and 5 (p=0.008). CONCLUSIONS: Multifocal ERG is a useful tool for the clinical follow-up of CNV. It offers interesting non-subjective data of retinal sensitivity of patients with macular diseases treated with PDT. In addition we obtained a central retinal sensitivity map where we were able to evaluate the extent and depth of retinal damage.

Electrorretinograma multifocal y degeneración macular asociada a la edad / Multifocal electroretinogram and age-related macular degeneration

Objetivo: Determinar la utilidad del electrorretinograma multifocal (mfERG) en la evaluación clínica de la respuesta al tratamiento con terapia fotodinámica (TFD) en pacientes con neovascularización coroidea (NVC) secundaria a degeneración macular asociada a la edad (DMAE).Métodos: Se ha realizado un estudio prospectivo en 23 ojos de 22 pacientes con NVC predominantemente clásica. Al inicio se realizó una medición de la agudeza visual mediante optotipos de Colenbrander, exploración biomicroscópica (BMC), retinografía, angiografía fluoresceínica (AGF), campimetría de Humphrey 24-2 y mfERG. Todos los pacientes fueron tratados mediante TFD. Se ha repetido la exploración de la agudeza visual, BMC, campimetría y mfERG a los dos y a los seis meses del tratamiento y se ha realizado una AGF tras un intervalo entre los cuatro y seis meses del inicio del estudio.Resultados: La agudeza visual permanece estable o mejora en todos los pacientes incluidos en el estudio. El estudio campimétrico nos ofrece resultados erráticos y poco fiables debido a la mala fijación que presentan estos pacientes. La densidad de respuesta de las ondas N1 y P1 del mfERG en los anillos centrales no muestra variaciones significativas después del tratamiento, sin embargo mejora en los anillos 3 (p=0,000), 4 y 5 (p=0,008).Conclusiones: El mfERG es una prueba de utilidad en el seguimiento de la NVC puesto que nos permite medir objetivamente la sensibilidad retiniana en pacientes con maculopatías tratadas con terapia fotodinámica. Por otra parte obtenemos un mapa de sensibilidades de la retina central en el que podemos valorar la extensión y la profundidad de la lesión

Purpose: To assess the value of multifocal electroretinogram (mfERG) in clinical evaluation of the response of patients with choroidal neovascularization (CNV), secondary to age-related macular degeneration, to photodynamic therapy (PDT) Methods: We designed a prospective non-comparative case series and enrolled 23 eyes of 22 patients with predominantly classic CNV. The initial evaluation included clinical examination with measurement of visual acuity (Colenbrander), biomicroscopic evaluation (BMC), retinography, fluorescein angiography (FA), Humphrey 24-2 visual field examination and mfERG. All patients were treated with PDT. Clinical follow-up was performed two and six months after treatment with visual acuity evaluation, BMC, visual field assessment and mfERG. FA was repeated after an interval of between four and six months in all patients. Results: Visual acuity remained stable or improved in all patients included in the study. Visual field examination gave erratic and unreliable results because of patients’ unstable fixation. N1 and P1 wave peak amplitudes in the central rings did not change significantly after treatment but they improved in rings 3 (p=0.000), 4 and 5 (p=0.008). Conclusions: Multifocal ERG is a useful tool for the clinical follow-up of CNV. It offers interesting non-subjective data of retinal sensitivity of patients with macular diseases treated with PDT. In addition we obtained a central retinal sensitivity map where we were able to evaluate the extent and depth of retinal damage

Eficacia de un programa de capacitación en medidas básicas de prevención de infecciones intrahospitalarias / Effectiveness of a program of qualification in basic measures of prevention of intrahospitable infections

Objetivos: Determinar la eficacia de un programa de capacitación en prevención de infecciones intrahospitalarias (IIH) para modificar conocimientos, actitudes y prácticas (CAP) del personal de salud hospitalario. Materiales y métodos: Estudio prospectivo de intervención, desarrollado en el Hospital Hipólito Unanue de Tacna, Perú, en el año 2000. Antes y después de la intervención se evaluó el nivel CAP en el personal asistencial de los servicios de hospitalización a través de encuestas. La intervención consistió en capacitaciones sobre medidas básicas para prevención de IIH y observaciones periódicas de las prácticas. Se comparó las proporciones según niveles CAP usando la prueba de McNemar. Resultados: Se incluyó al 73,7 por ciento (129/175) del personal; solo 22,9 por ciento (11/48) de médicos completaron el estudio. En general, >50 por ciento mostró niveles adecuados de CAP desde el inicio. Solamente se halló mejoría significativa en conocimientos (p<0,004) y prácticas (<0,001) del grupo enfermeras/ obstetrices/ técnicos. Si bien los servicios de hospitalización especializados tuvieron mayor nivel CAP que los básicos, sólo en estos últimos se mostró mejoras significativas en el nivel de prácticas (p<0,001). El cumplimiento rutinario de las medidas de bioseguridad pasó de 1 por ciento a 89,8 por ciento. Conclusiones: La implementación de un programa hospitalario de capacitación y supervisión permanente para la prevención de IIH mostró mejorar el nivel de conocimientos y prácticas en el personal no médico.

Objectives: To determine the efficacy of a qualification program on nosocomial infection (NI) prevention, to modify knowledge, attitudes and practices (KAP) of the hospital health care providers. Materials and methods: Prospective intervention study, performed in the Hipólito Unanue Hospital in Tacna, Peru, 2000. Before and after the intervention, KAP level was evaluated in hospitalization services using questionnaires. The intervention consisted of training on basic measures for NI prevention and periodic practice observations. Proportions according to KAP levels were compared using McNemar‘s test, considering p<0,05 as statistically significant. Results: 73,7% (129/175) of the hospital staff was included; only 22,9% (11/48) physicians completed the study. In general, >50% showed good KAP levels since the initial stages. We only found significant improvement in knowledge (p<0,004) and practices (<0,001) of the nurses & technicians group. Although special hospitalization services had a higher KAP level than the basic ones, only the practices in the basic services showed significant improvements (p<0,001). The routine fulfillment in the biosafety measures shifted from 1% to 89,8%. Conclusions: The implementation of a hospital program of qualification and permanent supervision for NI prevention was shown to improve the level of knowledge and practices in the nurses & technicians group.

[Granulocyte-apheresis in ocular Behçet's disease resistant to medical treatment]. / Granulocito-aferesis en Behçet ocular refractario al tratamiento médico.

OBJECTIVE: To assess the feasibility of granulocyte apheresis (GCAP) in patients with Ocular Behçet's Disease (BD) resistant to immunosuppressive therapy (prednisone, cyclosporin, azathyoprine or mycophenolate mofetil). METHODS: Prospective observational study of five cases, carried out in a university centre. Four patients were resistant to medical treatment and one refused immunosuppressive drugs. The intervention procedure consisted of an extracorporeal GCAP using a column filled with cellulose acetate beads (Adacolumn, JIMRO, Takasaki, Japan). All patients received underwent a schedule of therapy of five sessions, once a week for five consecutive weeks. Visual acuity (Snellen lines), the degree of intraocular inflammation and doses of immunosuppressive therapy were measured and observed every week. RESULTS: Visual acuity improved in the five cases. Intraocular inflammation was measured and observed in every case, relapses were avoided, and treatment with prednisone was reduced by more than half of the initial dose (average reduction 52.7%, SD 14). CONCLUSIONS: GCAP has been shown to be safe and effective as a new therapy in five cases of Ocular Behçet's Disease refractory to medical treatment. Further research is needed in order to confirm the promising results of these initial investigations.

Granulocito-aferesis en Behçet ocular refractario al tratamiento médico / Granulocyte-Apheresis in ocular Behçets disease resistant to medical treatment

Objetivo: El objetivo del estudio fue evaluar la viabilidad de la granulocitoaferesis (GCAP) en pacientes con enfermedad de Behçet (EB) ocular resistente al tratamiento con inmunosupresores (prednisona, ciclosporina, azatioprina o micofenolato mofetilo). Métodos: Estudio prospectivo de cinco pacientes, realizado en un hospital universitario de tercer nivel. Cuatro pacientes eran resistentes al tratamiento médico y uno rechazaba los fármacos inmunosupresores. El procedimiento consistió en realizar una GCAP extracorpórea mediante una columna rellena con gránulos de acetato de celulosa (Adacolumn®, JIMRO, Takasaki, Japón). Todos los pacientes fueron tratados con cinco sesiones, una semanal durante cinco semanas consecutivas. En cada sesión se controlaba la agudeza visual (AV) mediante las lineas de Snellen, el grado de inflamación intraocular y las dosis de tratamiento inmunosupresor. Resultados: La agudeza visual mejoró en los cinco casos. La inflamación intraocular se controló, no hubo recidivas y el tratamiento con prednisona se redujo en más de la mitad de la dosis inicial (reducción media de 52,7%, DE 14 ). Conclusiones: La GCAP ha demostrado ser un nuevo tratamiento seguro y eficaz en cinco casos de EB ocular resistente al tratamiento médico. Es preciso seguir investigando para confirmar los esperanzadores resultados de estos estudios iniciales(AU)

Objective: To assess the feasibility of granulocyte apheresis (GCAP) in patients with Ocular Behçets Disease (BD) resistant to immunosuppressive therapy (prednisone, cyclosporin, azathyoprine or mycophenolate mofetil). Methods: Prospective observational study of five cases, carried out in a university centre. Four patients were resistant to medical treatment and one refused immunosuppressive drugs. The intervention procedure consisted of an extracorporeal GCAP using a column filled with cellulose acetate beads (Adacolumn, JIMRO, Takasaki, Japan). All patients received underwent a schedule of therapy of five sessions, once a week for five consecutive weeks. Visual acuity (Snellen lines), the degree of intraocular inflammation and doses of immunosuppressive therapy were measured and observed every week. Results: Visual acuity improved in the five cases. Intraocular inflammation was measured and observed in every case, relapses were avoided, and treatment with prednisone was reduced by more than half of the initial dose (average reduction 52.7%, SD 14). Conclusions: GCAP has been shown to be safe and effective as a new therapy in five cases of Ocular Behçets Disease refractory to medical treatment. Further research is needed in order to confirm the promising results of these initial investigations(AU)

Clinical pharmacokinetics of toremifene.

Toremifene is a chlorinated triphenylethylene derivative of tamoxifen approved for use in the treatment of patients with metastatic breast cancer. Toremifene is well tolerated in patients, and common adverse effects of this drug include vasomotor symptoms such as hot flashes and vaginal discharge. This compound is administered to patients orally at a dose of 60 mg/day, although alternative methods of administration have been investigated. Oral bioavailability is estimated to be approximately 100%. At steady state, toremifene and its metabolites are highly protein bound (>95%). Toremifene is metabolised in the liver by cytochrome P450 enzymes, and it is eliminated primarily in the faeces following enterohepatic circulation. The half-life of toremifene is approximately 5 days, and steady state is reached by 6 weeks depending on the dose given. The pharmacokinetics of toremifene have been shown to be altered by certain liver conditions, but age and kidney function do not appear to be as significant.

Nueva técnica de reconstrucción en hemipelvectomía parcial interna. Informe de dos casos clínicos / A new tecnique for reconstruction after partial internal hemi-pelvectomy

La cirugía conservadora de tumores óseos localizados en pelvis, representa un desafío para el equipo médico quirúrgico, del cual exige una incuestionable experiencia. La resección innominada tipo II, II-A y la IIA + III presenta serios problemas de reconstrucción, debido a que el remanente óseo es un hueso plano delgado, con una anatomía irregular, que impide efectuar una osteosíntesis estable y suficiente. El objetivo del presente trabajo es dar a conocer una nueva técnica de reconstrucción para la hemipelvectomía parcial interna, mediante el uso de un clavo en "y", bloqueado con pernos a fémur y con cerclajes de alambre al iliaco. Se presenta el reporte preliminar en dos pacientes. Se pretende asegurar la artrodesis femoroiliaca o cuando menos una pseudoartrosis no dolorosa. No hubo acortamiento significativo de la extremidad y la reincorporación a la actividad fue rápida.

Long-term pharmacological kindling increases in vitro release of IR-Met and IR-Leu-enkephalin from amygdala.

Met-enkephalin release is increased from amygdala and striatum 1 and 15 days after pharmacological kindling with pentylenetetrazol, following potassium-induced depolarization in vitro via a Ca2+ dependent mechanism. Leu-enkephalin release was only enhanced in amygdala and striatum 1 day after the last seizure. IR-Met-enkephalin amygdala tissue content enhanced 1 and 15 days after seizure. In striatum, we found an IR-Met-enkephalin decrease 35 days after the last stimulus. IR-Leu-enkephalin amygdala tissue content enhanced 1 day after the last seizure, and no significant increases were found in striatum 1, 15 and 35 days after the last seizure. In this paper, we show that opioid peptides release is differentially enhanced in rat brain for several days after the last seizure, thus suggesting that opioid peptides may have a protective action against seizure activity.

Diurnal variations of opioid peptides and synenkephalin in vitro release in the amygdala of kindled rats.

Pentylenetetrazol (PTZ) kindling was induced in male Wistar rats (250-300 g) by daily intraperitoneal injections of 35 mg/kg of the convulsant agent. Immunoreactive (IR)-Met-enkephalin (IR-ME), IR-Leu-enkephalin (IR-LE), IR-heptapeptide (IR-HE), IR-octapeptide (IR-OC) and IR-synenkephalin (IR-Syn) in vitro release was measured from amygdala slices 24 h after the last stimulus, in groups of eight rats, every 4 h beginning at 08:00 h. Opioid peptides in vitro release displayed diurnal variations. IR-ME and IR-Syn showed maximal levels before the onset of darkness (16:00 h). IR-LE and IR-OC release was enhanced 4 h later (20:00 h), no changes were detected for IR-HE. These results show that endogenous opioid system (EOS) release displays diurnal variations. The peak for the analysed peptides was reached before and during the dark phase. It is suggested that EOS release enhancement in PTZ-kindled rats, seems to be due to a compensatory mechanism against the excitation induced by the blockade of the GABAergic transmission.

EGF modulation of the ras-related rhoB gene expression in human breast-cancer cell lines.

The mRNA levels of the ras-related human rhoA, rhoB and rhoC genes were studied in human breast-cancer cell lines (HBCal), and in normal and immortalized mammary epithelial cells (HMEC) by Northern blot analysis and in situ hybridization. In contrast to the ubiquitous rhoA and rhoC gene expression, dramatic variations in the mRNA level of the rhoB gene were evidenced. The rhoB mRNA level appeared to be inversely correlated to the amounts of the epidermal-growth-factor(EGF) receptors in these cells. The rhoB transcripts were detected at high levels in ZR75-1, MCF7, HSL 53, HSL 59, HSL 90, T47D and SKBR3 HBCal, at hardly detectable levels in BT 20, MDA-MB 231 and H466B HBCal and at intermediate levels in normal and immortalized breast epithelial cells. Rapid and transient induction of the rhoB transcription was observed after EGF treatment in serum-deprived MDA-MB231, T47D and immortalized epithelial cells. In contrast, no modulation of rhoB expression by EGF could be objectified in the MCF7 and ZR75-1 cell lines. Yet a normal function of EGF receptors was evidenced, since the immediate early gene c-fos was rapidly induced, suggesting a constitutive expression of rhoB in these cell lines bypassing the regulation by EGF. In human mammary epithelial cells, rhoB mRNA is rapidly and transiently induced with EGF concentrations known to stimulate cell proliferation. This suggests that the rhoB product might be involved in a cascade that initiates or promotes cell proliferation, and plays an important role in EGF-stimulated growth of breast normal and cancer cells.

p53 mutations and overexpression in locally advanced breast cancers.

Alterations in the p53 gene were analysed in 39 patients with locally advanced breast cancers (LABCs) (stage III-IV) with inflammatory signs in most cases (UICC stage T4d = 32 patients) by molecular and immunohistochemical (IHC) approaches. All patients were included in the same therapy protocol. Using polymerase chain reaction (PCR) and a single-strand conformational polymorphism migration technique (SSCP), the presence of mutations in exons 2-11, covering the entire coding sequence of the p53 gene, was evaluated. Using the mouse specific anti-human p53 monoclonal antibody (PAb 1801), we also looked for overexpression of the p53 protein in tissue sections. In 16 cases shifted bands were reproducibly identified by PCR-SSCP, and all but one (localised to exon 10) were in exons 5-8, the usual mutational hotspots. Fifteen of these 16 samples were sequenced and 14 of the suspected mutations (36%) were confirmed. Most of them (12) were single nucleotide substitutions, and transitions were more frequent (eight cases) than transversions (four cases). Fourteen of the tumour samples were positively stained with the monoclonal antibody PAb 1801, 11 with nuclear staining only, two with mixed cytoplasmic and nuclear staining and one with cytoplasmic staining only. Staining patterns were very heterogeneous in terms of the percentage of positive cells (10-75%) and their distribution in the tissue section (isolated foci or dispersed cells). In 11 of the 14 mutated cases a positive immunostaining was observed. The presence of a p53 mutation was significantly associated with larger tumour diameter (chi 2 = 7.490, P = 0.0062) and the presence of clinical metastases (stage IV) (chi 2 = 10.113, P = 0.0015). A non-statistically significant trend of association was observed between p53 mutation, negative oestrogen receptors and lower response rate to therapy. Our results in this group of patients and the heterogeneity of the staining of tumour cells in tissue sections suggest that p53 mutations could be a late event in this non-familial form of breast cancer.

Cytogenetic characterization of a new human papillary thyroid carcinoma permanent cell line (GLAG-66).

A new permanent cell line (GLAG-66) has been established from the metastases of a papillary thyroid carcinoma in a male patient. Herein are reported the cytogenetic characteristics of this new cell line, which is tumorigenic in athymic mice. An aneuploid chromosomal pattern was observed (48 chromosomes) with various chromosomal abnormalities. The karyotype was: 48,XY,der(1)t(9;1;9), +der(8)t(1;8),der(9)t(1;9),der(9)t(1;9), +14. This cell line should prove to be of great value in the study of the biology of human papillary thyroid carcinomas.