Current sensors employed in medicine are used to detect chemical and biochemical abnormalities. Their applications range from biopsy (brain), enzyme-linked immunosorbent assay (ELISA) (spinal fluid), blood (bio-barcode), and sweat and urine bio-diagnostics where the primary focus is the selection of biomarkers that can pinpoint the occurrence of the disease. Emerging sensors for cholesterol detection are based on enzymatic functions, which degrade these molecules, where the signal can be visualized optically by using a transducer. Cholesterol is a steroid metabolite that is employed for the synthesis of steroid hormones, and the establishment of proper membrane permeability and fluidity. Since cholesterol is insoluble in blood, it is transported in the circulatory system within lipoproteins, complex spherical particles which have an exterior comprising of amphiphilic proteins and lipids with outward-facing surfaces that are water-soluble and inward-facing surfaces that are lipid-soluble. Low-density lipoprotein (LDL) is known as ‘bad’ cholesterol. High-density lipoprotein (HDL) is known as ‘good’ cholesterol. LDL is linked to cardiovascular conditions such as atherosclerosis and hypertension, which ultimately lead to coronary heart disease, myocardial and cerebral infarction (stroke). An appropriate therapeutic response to a sensor system for cholesterol, specifically LDL, detection implicates the design of an implantable system for stimuli-responsive drug release. The proposed system was designed to detect specific biochemical changes by employing nanoparticles made of glyceryl behenate, polyoxyethylene-polyoxypropylene block copolymer, avidin, biotin and anti-beta lipoprotein antibodies as sensors. This was achieved by coating nanoparticles with antibodies specific to the antigen (i.e. LDL) to create an antibody-conjugated antibody conjugated solid lipid nanoparticles (henceforth known as ‘antibody conjugated SLN). Fenofibrate was used as a model drug due to its low water solubility and to its lipophilic properties similar to statins. The antibody conjugated SLNs were of 150nm in size and had a zeta potential of -28mV. Their drug entrapment efficacy was 86%, with a drug release of 16mg/day due to Fickian diffusion and erosion mechanism. The slow release was due semi-crystalline structure determined by XRD and DSC. Antigen responsive hydrogel was designed by incorporation of thiolated antibody conjugated SLN via Traut’s reagents, polyethylene glycol diacrylate, methyl acrylic acid and polyethylene glycol 200. The osmotic pump was designed from polyethylene oxide, ethyl cellulose and mannitol. The drug reservoir was synthesized from ethyl cellulose coated gelatin capsule via coacervation phase separation method. The polymeric tube synthesized from ethyl cellulose, methyl cellulose and castor oil was coated with antigen responsive hydrogel. Ex vivo studies evaluating intravascular stability of the implant in correlation with mechanical analysis indicated the polymeric tube unstable. An 18-gauge catheter was used for forming an infusion tube as a substitute for the polymeric tube. The implant showed a correlation of Korsmeyer-Peppas drug release during in vivo and in vitro studies. A constant drug release of 881μg/day was observed during in vivo. This played a role in reduction of total cholesterol by means of reduction in LDL sub-fractions by 30%; in correlation with LDL particle enhance clearance from the plasma due to SLN-LDL uptake. An increase by 46% in HDL was observed, which correlated to fenofibrate therapeutic effect. Pharmacokinetic analysis indicated improved mean residence time and efficacy. This indicated that the device could be used for delivery of lipophilic drugs and detection of circulating biomarkers.

Dissertation--Upsala, 1855. Issued also as thesis, Stockholm.

Vinte e nove pacientes submetidos ao Transplante de Medula Óssea Alogênico foram analisados num estudo dirigido às transfusões de hemocomponentes, desde o período pré-transplante até o momento da alta hospitalar, que na maioria das vezes coincidiu com a independência às transfusões. Não houve um padrão único de requerimento transfusional e os fatores que determinaram este fato foram múltiplos. O tipo de doença de base, intercorrências clínicas e a presença da Doença do Enxerto Versus o Hospedeiro estiveram relacionados. As técnicas laboratoriais empregadas na rotina de avaliação dos pacientes ofereceram subsídio adequado, para em conjunto com o exame clínico, proporcionarem parâmetros decisivos na indicação das transfusões sangüíneas. Alguns pacientes apresentaram incompatibilidade ao sistema de grupo sangüíneo ABO com os seus respectivos doadores de medula, fato que não impediu a realização do transplante, devido ao emprego de métodos que permitiram superar esta dificuldade. Foi observada refratariedade às transfusões de concentrados de plaquetas em poucos pacientes da casuística, atribuída à presença de anticorpos contra antígenos do sistema de histocompatibilidade leucocitária humana (HLA). As transfusões de plaquetas obtidas de doadores com o sistema HLA compatível, membros familiares, foram eficazes nestes casos. O tempo de recuperação da função hematopoética da medula transplantada foi similar ao descrito na literatura. / 29 patients who underwent Allogeneic Bone Marrow Transplantation (Allo BMT) were analyzed regarding the need for transfusion of blood components from admission the date of discharge. Most of the time the discharge coincided with blood components transfusion independence. There was no specific pattern of blood products transfusion among these patients. The need for transfusion could be ascribed to the presence of several factors like baseline disease, clinical complications and graf versus host disease. Routine clinical parameters and laboratory techiniques were adequate to indicate the need for transfusion of blood components. Some of the patients were ABO incompatible with their respective marrow donors. We observed no adverse consequence for the outocome of these BMT procedures after the use of routine techiniques to overcome this incompatibility. Few patients were refractory to platelet transfusions that was ascribed to HLA directed antibodies. The use of HLA compatible platelet donors (family members) provided adequate platelet support in these cases. The time to hematopoietic recovery was compatible to the found in the international literature.

Thesis (M.S.P.H.)--University of Michigan, 1941.

Thesis (M.S.P.H.)--University of Michigan, 1941.

Proefschrift - Utrecht. / Summary in English.

Proefschrift--Nijmegen. / "Stellingen" [3] p. inserted. Bibliography: p. 119-122.

Thesis (Ph. D.)--University of Washington, 2005. / Vita. Includes bibliographical references (p. 173-178).

Vinte e nove pacientes submetidos ao Transplante de Medula Óssea Alogênico foram analisados num estudo dirigido às transfusões de hemocomponentes, desde o período pré-transplante até o momento da alta hospitalar, que na maioria das vezes coincidiu com a independência às transfusões. Não houve um padrão único de requerimento transfusional e os fatores que determinaram este fato foram múltiplos. O tipo de doença de base, intercorrências clínicas e a presença da Doença do Enxerto Versus o Hospedeiro estiveram relacionados. As técnicas laboratoriais empregadas na rotina de avaliação dos pacientes ofereceram subsídio adequado, para em conjunto com o exame clínico, proporcionarem parâmetros decisivos na indicação das transfusões sangüíneas. Alguns pacientes apresentaram incompatibilidade ao sistema de grupo sangüíneo ABO com os seus respectivos doadores de medula, fato que não impediu a realização do transplante, devido ao emprego de métodos que permitiram superar esta dificuldade. Foi observada refratariedade às transfusões de concentrados de plaquetas em poucos pacientes da casuística, atribuída à presença de anticorpos contra antígenos do sistema de histocompatibilidade leucocitária humana (HLA). As transfusões de plaquetas obtidas de doadores com o sistema HLA compatível, membros familiares, foram eficazes nestes casos. O tempo de recuperação da função hematopoética da medula transplantada foi similar ao descrito na literatura. / 29 patients who underwent Allogeneic Bone Marrow Transplantation (Allo BMT) were analyzed regarding the need for transfusion of blood components from admission the date of discharge. Most of the time the discharge coincided with blood components transfusion independence. There was no specific pattern of blood products transfusion among these patients. The need for transfusion could be ascribed to the presence of several factors like baseline disease, clinical complications and graf versus host disease. Routine clinical parameters and laboratory techiniques were adequate to indicate the need for transfusion of blood components. Some of the patients were ABO incompatible with their respective marrow donors. We observed no adverse consequence for the outocome of these BMT procedures after the use of routine techiniques to overcome this incompatibility. Few patients were refractory to platelet transfusions that was ascribed to HLA directed antibodies. The use of HLA compatible platelet donors (family members) provided adequate platelet support in these cases. The time to hematopoietic recovery was compatible to the found in the international literature.

Talassemias são as mais comuns desordens monogenéticas em humanos; são caracterizadas pela presença de anemia microcíticas e hipocrômicas que resultam da redução ou ausência na síntese de uma ou mais cadeias globínicas. No Brasil a beta-talassemia tem importante significado clínico e sua ocorrência pode ser explicada como conseqüência da grande mistura étnica. A beta-talassemia é extremamente heterogênea e mais de 200 mutações têm sido descritas causando diferentes graus de anemia. Essas mutações são regionalmente específicas e cada país possui seu grupo de alterações característico. No estado homozigoto são classificadas clinicamente como major e no estado heterozigoto como intermédia e minor. Os pacientes com talassemia intermédia podem não apresentar sintomas clínicos, mas constatam-se características laboratoriais específicas. A avaliação dos índices hematológicos, dosagens dos parâmetros bioquímicos e dos valores das hemoglobinas fetal e HbA2 em combinação com o entendimento das possíveis interações gênicas são os procedimentos recomendados para o diagnóstico laboratorial. O objetivo deste estudo foi determinar a presença de mutações em pacientes que utilizam a Divisão de Laboratório Central do Hospital das Clínicas da Faculdade de Medicina da Universidade São Paulo (DLC-HC-FMUSP) e que apresentavam perfil laboratorial sugestivo para beta-talassemia. Estudou-se a região do gene da globina em onde estão descritas as mutações mais freqüentes para nosso meio através da técnica de PCR, e posterior seqüenciamento para identificação das mutações. As mutações observadas na população avaliada (N=40), em ordem crescente de freqüência foram: Codon 39 (CT), IVS-I-110 (GA), IVS-I-1(GA), IVS-I-5 (GC), IVS-I-6 (TC), IVS-I-Exon-II (GA), Exon-II-IVS-II (GA), Stop Cd 6 (GT), Stop Cd15 (GA), Del C Cd 44 e 5UTR+20. Os primeiros quatro genótipos representaram 82,5% das mutações observadas neste trabalho. O uso combinado dos índices hematimétricos, parâmetros bioquímicos e avaliação das características morfológicas das hemácias demonstraram eficiência no rastreamento de pacientes portadores de beta-talassemia, assintomáticos A técnica do PCR e posterior processo de seqüenciamento demonstraram elevada eficiência na determinação das alterações moleculares no grupo de pacientes avaliados. / Thalassemias are the most common monogenic disorders in humans; are characterized by hypochromic and microcytic anemia arising from the reduction or absence in the synthesis of one or more chains globínicas. In Brazil, beta-thalassemia has important clinical significance and its occurrence can be explained as a result of the large ethnic mix. The beta-thalassemia is extremely heterogeneous and more than 200 mutations have been described causing varying degrees of anemia, these mutations are regionally specific and each country has its characteristic group of amendments. In the homozygous state are clinically classified as major and the heterozygous state as intermediate and minor. Patients with thalassemia can not present interim clinical symptoms, but have characteristics specific laboratory. The evaluation of hematologic indices, strengths and biochemical parameters of fetal hemoglobin and HbA2 values in combination with an understanding of possible interactions genes, are the recommended procedures for laboratory diagnosis. The purpose of this study was to determine the presence of mutations in patients who use the Division of Central Laboratory of the Hospital of the Faculty of Medicine of the University Sao Paulo (DLC-HCFMUSP) who presented suggestive profile laboratory for beta-thalassemia. It was studied the region of the gene of beta globin where they are frequently described the changes to our region through the PCR technique, and subsequent sequencing to identify the mutations. Of the 40 patients evaluated were found the following changes: Codon 39 (CT), IVS-I-110 (GA), IVS-I-1 (GA), IVS-I-5 (GC), IVS-I-6 (TC), IVS-I-Exon-II (GA), Exon-II-IVS-II (GA), Stop Cd 6 (GT), Stop Cd15 (G A), Del Cd 44 and 5UTR+20. The first four genotypes accounted for 82.5% of mutations observed in this study. The combined use of hematologic indices, biochemical parameters and evaluation of morphological characteristics of red blood cells demonstrated efficiency in the tracking of patients with beta- thalassemia, asymptomatic. The technique of PCR and subsequent process of sequencing demonstrated high efficiency in determining the molecular changes in the group of patients evaluated.