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00437.pdf

Memorias II Congreso Latinoamericano de Ingeniería Biomédica, Habana 2001, Mayo 23 al 25, 2001, La Habana, Cuba RADIATION ABSORBED DOSES AND SAFETY OF THE
HUMANIZED MONOCLONAL ANTIBODY H-R3 LABELED
WITH 99MTC
Leonel A. Torres, Marco A. Coca, Maria E. Solano, Alejandro Perera, Juan F. Batista, Abel Hernandez, Tania Crombet, Marilyn Perez, Mayra Ramos, Elvia L Sanchez, Susana Romero, Vicente Aguilar, Normando Iznaga Calle 34 #4501 esq. 45. Reparto Kohly, Playa, Havana, Cuba AP 11300 Telf.: (537) 23 0087 Fax: (537) 24 3298 ABSTRACT
(MAbs) could be useful for immunoscintigraphic diagnosis Introduction. Labeled humanized monoclonal antibodies (Mab) for the diagnosis and treatment of tumors using In order to decrease the immunogenicity and HAMA Nuclear Medicine procedures have contributed to decrease response to the murine MAbs, and increase the half-live of the immunogenicity and HAMA response of murine the molecule in the patient, it was developed a reshaped MAbs, increasing the effectiveness and usefulness of humanized monoclonal antibody H-R3. This study include radioimmunoscintigraphic procedures. The objective of the preliminary results of the internal radiation dosimetry, this study was to evaluate the internal radiation dosimetry biodistribution and safety of the 99mTc-labelled H-R3 used and safety of the 99mTc-labeled H-R3 Mab, in humans.
for radioimmunodiagnosis of tumors of epithelial origin.
Material and methods: Ten patients with suspicions ofepithelial tumors were included on this study. They 2. MATERIAL AND METHODS
received 1110 MBq of 99mTc H-R3, intravenously.
Multiple blood, urine samples and sequential whole-body Antibody Labelling and Quality Control.
images were collected at different times. The internal Freeze-dried kits with 3 mg of humanised MAb H-R3 radiation dosimetry was estimated using the Medical (Centre of Molecular Immunology, Havana, Cuba) were Internal Radiation Dosimetry (MIRD) Committee reconstituted with 1295 MBq of pertechnetate from a methodology. Biodistribution was computed from the 99Mo/99mTc generator (CIS-BioInternational, France).
scintigraphic images and regression analysis of the time- Labelled product was subjected to ascending paper activity curves was employed to compute the residence chromatography. Radiochemical purity higher 90% was times of the source organs. Hematological toxicity and considered satisfactory for patient’s administration.
adverse effects were evaluated using the WHOclassification. Results: Liver received the higher absorbed Studied Patients
dose (4,90E-02mGy/MBq), followed by the kidneys Ten adult patients (6 females and 4 males) were enrolled (1,60E-02mGy/MBq) and gallbladder wall (1,53E- on this study. They received 3 mg of MAb labeled with 02mGy/MBq). The effective dose and the effective 1110 MBq of 99mTc. All patients were suspected of equivalent dose were 0,0143 mSv/MBq and 0,0190 having either primary or recurrent cancer of epithelial mSv/MBq, respectively. The main source organs of this origin and had undergone CT scanning, x-ray and other radiopharmaceutical were liver, spleen, kidneys and heart.
determinations. Histopathological findings were considered No significant changes were observed in the hematological as a confirmation criterion (gold standard). All studied parameters and only two patients showed mild and patients gave their written informed consent previously to moderate adverse effects, solved during the studies and not Biodistribution studies and internal radiation dosimetry
Palabras clave: absorbed doses, monoclonal antibody,
analysis.
Anterior and posterior whole body images were acquired at5 min, 1 h, 3 h, 5 h and 24 h after administration of 1. INTRODUCTION.
radiopharmaceutical, using a gamma camera(SOPHYCAMMERA DS7, France). The gamma camerahead was fitted with a diverging parallel-hole collimator to Almost 90% of malignancies are epithelial derived tumors.
increase the lateral viewing aspect. A 20% window Cancer of epithelial origin constitutes one of the first centered on the 140 keV emission peak of the 99mTc was causes of death world-wide [1]. Tumors of epithelial employed. On the gantry movements were used a speed of origin, like cancer of lung, digestive tract, breast and 20 cm/min. All whole-body images were stored in the others, have an overexpression of the epidermal growth computer in 2048x512 byte mode matrix.
factor receptor (EGF-R) [2]. This fact is often related to All images were processed in a SOPHY-20P system using malignancy and poor prognosis of the disease [3]. Clinical the software BioDose v1.0, developed at the Centre for trials have shown that anti-EGF-R monoclonal antibodies Clinical Research. Geometric mean images were computed 950-7132-57-5 (c) 2001, Sociedad Cubana de Bioingeniería, artículo 00437 from the anterior and posterior whole-body scans. They were reviewed to determine the subjective biodistribution 99mTc-H-R3 Human biodistribution. Data corrected by of 99mTc-H-R3 and to select which organs and tissues were clearly visible in the total image series. Such organs were Regions of interest (ROI) were drawn over heart, liver,spleen, small intestine, kidneys, upper large intestine (ULI), lower large intestine (LLI) and urinary bladder. Total counts were computed for each one, at the different time intervals. For each source organ count values were converted to activity, corrected for decay and expressed as percentage of the total administered activity. Whole-body activity was initially 100 % following a exponential clearance by biological removal and physical decay of The MIRD Committee method was used to assess the internal radiation dosimetry. Time-activity curves for each source organ and the remainder of the body tissues were obtained and fitted to exponential disappearance curves.
Cumulative activities and residence times for each source The assessed absorbed dose of 8 target organs (mGy/MBq organ were estimated integrating the time-activity curves.
The absorbed doses to whole body and organs were or rad/mCi) after the injection of 99mTc- MAb ior H-R3 estimated using the residence times and the modified ”S” are presented in Table No II. The absorbed dose for the values reported on MIRD Pamphlets [6,7]. Based on the whole body and the effective dose are also reported. The results of the absorbed dose estimated to various target highest absorbed dose was received by the liver with a value of approximately 4,90E-02 mGy/MBq and thekidneys 1,60E-02mGy/MBq). The effective dose and the Clinical laboratory analysis and safety studies.
equivalent effective dose were 0,0143 mSv/MBq and Complete blood cell counts with differential and platelet 0,0190 mSv/MBq respectively. The main contributor counts, chemistry panel evaluation, liver function test and organs to the absorbed dose per organ were the hepatic urinary analysis were performed 1-7 days before and 24 h after antibody administration. Patients did not have any food for 12 hours before blood collection. Fifteen millilitres Normal organ dosimetry for the labelled MAb of blood were withdrawn from a patient’s vein 1 week before and 24 h after 99mTc-H-R3 administration. Five millilitres of blood were carefully added to a brand collection tube with 50 µL of 15% EDTA (K (Vacutainer, Becton Dickinson, USA) for haematology.
One drop of blood was extended in a microscope slide (Shanghai Company, China) for differential leukocyte counting. Five millilitres of blood were added to a centrifuge tube and were incubated at room temperature for 1 h. After that, sample was centrifuged at 3000 rpm for 10 min and two 1mL-specimens of serum were added in Eppendorf tubes for analytical chemistry tests. On the other hand one hundred millilitre urine sample of the first morning urination was collected in a sterile container to All patients were monitored up-to 24 h after injection in order to detect any adverse reaction.
3. RESULTS.
Biodistribution and Internal Radiation Dosimetry
Only two patients (20%) showed adverse reactions.
The computed biodistribution of the 99mTc-MAb H-R3 is Reactions were clasified as follows: Patient No.7, acuterhinitis at 30 minutes after injection. It was maintained for 6 hours, responding partially to the treatment with intramuscle Benadryl. According to the WHO criteria itwas considered as moderated reaction. On this patient thehematological analysis showed normal results. PatientNo.9, Chills that no required medication and disappearedat a few minutes. It was considered as mild reaction (grade1). The complementary blood examinations had alterationsin two parameters: glucose=21 milimols/L andalcalinephosphatase=312 u/L. These blood samples wereobtained a day after the injection of the product. We cannotestablish the causes of such abnormalities because no otherpatient injected had such abnormal parameters. On theother hand, no significant changes of the vital signs weredetected in any of the studied cases after the injection ofthe radiopharmaceutical and anyone patient showedadverse events at the administration site.
Clinical laboratory parameters before and 24 h after intravenous injection of the 99mTc-h R3 were assessed andno significant changes were observed. Additionally did notexist any abnormality repeated in more than one patient.
There were not hematologic and biochemical abnormalitieswith clinical significance that could be produced by theMAb or by radiation exposure.
4. CONCLUSIONS
The obtained results of Biodistribution and Dosimetry of99mTc-labeled Monoclonal Antibody H-R3 have shownthat liver is the target organ of this product and presentedan uptake peak at 1hr post-injection with a high retention.
This organ also received the higher absorbed dose. Theseresults should be taking into account for a future use of thiscompound for therapeutical purposes and for its current usein diagnostic procedures.
5. REFERENCES.
[1] Sheryl L. Parker Sl, Tong T, Bolden S, Wingo PA. Cancer Statistics. Ca.
A. Cancer Journal for Clinicians. CA 1996, 46: 15-27.
[2] Modjtahedi H, Eccles S, Sandle J, Box G, Titley J and Christopher D.
Differentiation or immune destruction. Two pathways for therapy ofsquamous cell carcinomas with antibodies to the epidermal growth factorreceptor. Cancer Res. 1994; 54: 1695-1701.
[3] Fontanini G, Vignati S, Bigini D, Mussi A, Lucchi H, et al. Epidermalgrowth factor receptor (HEGF-R) expresion in non-small cell lungcarcinomas correlates with metastatic involvement of hilar and mediastinallymph nodes in the squamous subtype. Eur. J. Cancer. 1995; 31A: 178-183.
[4] Ramos-Suzarte M, Rodriguez N, Oliva JP, Iznaga N, Perera N , MoralesA, et al. 99mTc-Labeled Antihuman Epidermal Growth Factor ReceptorAntibody in Patients with Tumors of Epithelial Origin: Part III. ClinicalTrials Safety and Diagnostic Efficacy. J Nucl Med 1999; 40 768-775.
[5] Iznaga-Escobar N, Torres LA, Morales A, Ramos M, Rodriguez N, PerezN, et al. 99mTc-Labeled-anti-human epidermal growth factor receptorantibody (ior egf/r3) in patients with tumor of epithelial origin: II.
Pharmacokinetics and clearances. J Nucl Med 1998; 39: 1918-1927.
[6] Sorenson J.A., Phelps M. E. Physics in Nuclear Medicine. SecondEdition, Orlando, Florida. Grune an Stratton, 1987.
[7] Snyder WS., Ford, MR. et al. "S" absorbed dose units cumulated activityfor selected radionuclides and organs. MIRD pamphlet no. 11. New York:Society of Nuclear Medicine, 1975.
DOSIS DE RADIACIÓN ABSORBIDAS Y SEGURIDAD DEL
ANTICUERPO MONOCLONAL HUMANIZADO H-R3
MARCADO CON 99MTC
RESÚMEN
Introducción. Los anticuerpos monoclonales humanizados (AcM) marcados para el diagnóstico y eltratamiento de tumores usados en la Medicina Nucleares han contribuido a disminuir la inmunogenicidad y larespuesta HAMA de los AcM murinos, aumentado la efectividad y utilidad de los procedimientos deradioinmunodiagnóstico. El objetivo de este estudio fue evaluar las dosis de de radiación interna y laseguridad del AcM H-R3 marcado con 99mTc, en humanos. Materiales y métodos: Diez pacientes consospechas de tumores epiteliales fueron incluidos en este estudio. Ellos recibieron 1110 MBq de 99mTc-H-R3, por vía intravenosa. Múltiples muestras de sangre y orina e imágenes secuenciales de cuerpo enterofueron colectadas en diferentes tiempos. Las dosis de radiación interna fueron estimadas usando lametodología del Comité de Dosimetría Interna de la Sociedad Americana de Medicina Nuclear (MIRD). Labiodistribución se calculó a partir de las imágenes gammagráficas. Se empleó análisis de regresión de lascurvas de actividad-tiempo para calcular los tiempos de residencia de los órganos fuente. Se evaluaron latoxicidad hematológica y los efectos adversos usando la clasificación de la OMS. Resultados: El hígadorecibió la dosis absorbida más alta (4,90E-02mGy/MBq), seguido por los riñones (1,60E-02mGy/MBq) y lapared de la vesícula (1,53E-02mGy/MBq). La dosis efectiva y la dosis efectiva equivalente fueron 0,0143mSv/MBq y 0,0190 mSv/MBq, respectivamente. Los principales órganos fuentes de este estudio fueronhígado, bazo, riñones y corazón. No se observo ningún cambio significativo en los parámetros hematológicosy sólo dos pacientes mostraron efectos adversos ligeros y moderados, no relacionados con las exposiciones ala radiación y resueltos durante los estudios.

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