RESUMENES

Design and Development of Novel Specific Inhibitors for Plasmodium Falciparum Dihydroorotate Dehydrogenase. M. Solivan,C.Vlaar.
UPR- Medical Sciences Campus, School of Pharmacy, Department of Pharmaceutical Sciences, San Juan, PR.

Malaria is a serious endemic disease, which affects approximately 273 million people, with a reported annual death rate of 1.09 million, most of them children under five years. The responsible parasite, Plasmodium falciparum has developed widespread resistance against available chemotherapeutic agents and therefore there is an urgent need for new drugs. Dihydroorotate Dehydrogenase (DHODH) is a key enzyme in the biosynthesis of pyrimidines, and it has been shown that its inhibition can halt cell proliferation. Therefore, specific inhibition of P. falciparum DHODH, but not human DHODH could provide selective and relatively non-toxic drugs as a potential new treatment for malaria. In the present research, new derivatives of the class of pyrazoles (known to inhibit H Pylori-DHODH) are being synthesized. These new derivatives potentially bind covalently to two cysteine residues that are present in the active binding site of PfDHODH, but not in human DHODH. As potential groups that can react with the cysteine residues, pyrazoles with either disulfide or aldehydic groups have been synthesized. The development and optimization of a six-step synthesis for these new pyrazole derivatives is being presented. Supported by NIH Grant 3-S06-GM08224-21, MBRS-SCORE program.

Quality Assurance and Stability Sudies on Lupinus termis seeds and ointment formulation. L. Flores, M. Antoun. UPR-Medical Sciences Campus, School of Pharmacy, Department of Pharmaceutical Sciences, San Juan, PR.

Atopic dermatitis or Ezcema, as it is sometimes called, is a condition that can affect all age groups. Currently, there is no cure for atopic dermatitis, and corticosteroids are the primary drugs used to relieve the symptoms of the disease. Based on what is known in traditional medicine, and the published liture, the alcoholic extract of Lupinus seeds appears to have potential as an alternative treatment for this condition. The present thesis describes a preliminary Quality Assurance study done on the seeds of Lupinus termis, as well as a study done on the stability of a formulation of the alcoholic extract in an ointment base. Stability of the alcoholic extract in an ointment base evaluated during a period months under accelerated conditions and a period of 12 months for Long Term Conditions (i.e. Control Temperature, CRT). Two isolation methods to monitor the stability of a marker compound were used, Methanol extraction and Dichloromethanol/Isopropanol (3:1) extraction. There appears to be no significant different between both methods. This work has been sponsored by a grant from the National Center for Complementary and Alternative Medicine (1R21 AT 00961-01) and by a Research Resources, National Institutes of Health.

Inhibitory Effect of Simvastatin on Calcium Release in Smooth Muscle Cells. J. Bernstein. UPR- Medical Sciences Campus, School Medicine, Department of Physiology, San Juan, PR.

Ca++ mobilizing hormones transiently increase cytosolic calcium. Data by Escobales et al (J Cardiovasc Physiol 27: 383, 1996) show that Angiotensin II (ANG) and Simvastatin (SV) increased cytosolic calcium in rat vascular smooth muscle cells (SMC). SV also blocked the effect of ANG. Ca++ increases have been implicated in the development of atherosclerosis. Peaks were transformed into data points by UnScan (Silk) and analyzed by PeakFit v3 (Systat) using an Asymmetric Double Sigmoid model (ADS). 100 nM ANG produced a peak which is the result of 2 component peaks: amplitudes of 322.25 and 220.53 nM Ca and rates of entry of 81.75 and 51.21 nM Ca/sec, respectively. Addition of 20 uM SV resulted in 1 peak with amplitude of 51.09 nM and rate of rise of 6.83 nM Ca/sec. In the presence of ANG rates of Ca exit were 38.74 and 5.52 nM Ca/sec for the 1st and 2nd peak respectively and SV reduced this rate to 1.34 nM/sec. Because sigmoidal rates are influenced by peak size, data were normalized with respect to the SV peak. Rates of Ca entry into the cytosol were then 255.5 and 247.5 nM Ca/sec for the 1st and 2nd components, whereas exit rates were 185.8 and 26.52 nM Ca/sec, respectively. This shows that after ANG the 2 components are different. Furthermore, since rates in the presence of SV also remain different, this indicates that although both ANG and SV act on the same intracellular Ca++ pool, they do so by different mechanisms, a fact also concluded by Escobales et al. These facts demonstrate that Peak Analysis can be used to obtain information that is not apparent from a simple analysis of data in the form of peaks.

Cerebrospinal Fluid Biomarker Proteins Predict Cognitive Impairment in Human Immunodeficiency Virus-1 Infected Hispanic Women
J. Pérez Laspiur1; E. R. Anderson1; P. Ciborowski5,6; H. E. Gendelman5,6,7,8; V. Wojna1,4; J. Rodríguez-Orengo1,3; E. Rodríguez1; J. Anderson9; and L. M. Meléndez-Guerrero1,2
1Specialized Neurosciences Program, 2Departments of Microbiology, 3Biochemistry, and 4Neurology, University of Puerto Rico Medical Sciences Campus, San Juan, Puerto Rico 00935; 5Departments of Pharmacology and Experimental Neuroscience, 6Biochemistry, 7Pathology and Microbiology, and 8Internal Medicine, 9Statistics, University of Nebraska Medical Center, Omaha, NE 68198-5800

Cognitive, motor and/or behavioral impairments are commonly a result of continuous human immunodeficiency virus (HIV) infection and immune activation of brain mononuclear phagocytes (MP). MP-induced inflammatory neurotoxic molecules present commonly in cerebrospinal fluid (CSF) can induce neurodegeneration and, in measure, predict the onset and progression of dementia in HIV infected patients. In the current report, a proteomics platform combining surface enhanced laser desorption ionization time of flight (SELDI-TOF), reverse-phase high performance liquid chromatography (RP-HPLC), one dimensional SDS-PAGE electrophoresis (1DE) and liquid chromatography tandem mass spectrometry (LC-MS/MS) was used to assess whether a unique biomarker signature is indicative of HIV-1-associated dementia (HAD). CSF was collected from 10 HIV-1 infected Hispanic women with HAD (n=6) or nondemented (ND; n=4). Differentially expressed proteins were identified by SELDI-TOF followed by RP-HPLC fractionation and rigorous statistical analysis. Subsequently, 1DE revealed bands corresponding to proteins of similar mass and these were identified by LC-MS/MS. Of most interest, three proteins unique to HAD were familial ALS mutant of Cu+2, Zn+2 superoxide dismutase, apolipoprotein E, and osteopontin. Normal superoxide dismutase was present in ND. Osteopontin and apolipoprotein E were present in HAD. These macrophage proteins contribute to inflammation and neurodegeneration. The results of this study demonstrate that important macrophage proteins potentially contribute to HAD development in Hispanic women.

Melatonin content determination using HPLC with UV/VIS detection. J. Vega, J. Morreno, V. Lopez, J. Bloom, Ph.D. UPR- Department of Chemical, Natural Sciences Faculty, Río Piedras, PR.

A new method for the determination and quantization of melatonin in dietary supplements was developed using HPLC with UV/VIS detection. The sample was extracted from the matrices (capsules, tablets, capsules with herbs) using methanol as an extraction solvent after sonication and centrifugation of the sample solution. The assay of the samples showed 87.7-108.5% concordance with that of the label claim in the different matrices studied. Results obtained from the method showed a LOD of 40ng/mL and a LOQ of 80ng/mL within the linearity range determined (0.040 µg/mL–25.0µg/mL). The method procedure was tested for roughness/robustness, with addition stability studies for standard solutions. The calibration curve range used during the analysis was between 1.60 - 25.00µg/mL. The spiked recovery results for the active ingredient from the different matrices were between 99.8-105.7% with an RSD of 2.1%. The standard solution showed an adequate stability for analysis one week after its preparation. No significant degradation in the melatonin peak at 4.4min was observed when the samples were submitted to different degrading conditions such as acid, base, and oxidative environment at room temperature. Minimum degradation was observed when sample containing 30% hydrogen peroxide is heated above 90ºC.

The Adjuvants Polyethyleneimine and GM-CSF Enhance the Immunogenicity of a Dengue-4 Env DNA Vaccine Candidate. M. Rodríguez-González; I. Martínez. Department of Microbiology and Medical Zoology, UPR School of Medicine, San Juan, PR.

Dengue (DEN) is an RNA virus that is endemic in about 100 countries in the tropical area. There is no licensed vaccine for prevention of illnesses caused by this virus. In our laboratory, we have developed a DEN-4 DNA vaccine candidate that is immunogenic and protective in mice. In the present studies, we tested two adjuvants to improve the immunogenicity of this vector. One is the polymer polyethyleneimine (PEI) that has the capacity to bind DNA through electrostatic interactions, and facilitates endosomal lysis. The other adjuvant tested, GM-CSF, is a cytokine that has the ability to recruit dendritic cells, which are professional antigen presenting cells. Western Blot and immunofluorescence assays data demonstrated that the DEN-4 Env vector complexed with PEI can be expressed in mouse myoblast cells. We also found that intramuscular injection of either the DEN-4 Env vector complexed with PEI, or coinjected with a plasmid that expresses GM-CSF, induced significant total antibodies in Balb/c mice. Lower, but significant, total antibody titers were induced by our vaccine candidate administered in combination with PEI and GM-CSF. The results obtained in these experiments will contribute to the knowledge and development of vaccines against DEN viruses.

Identification of Genes that Suppress Nikkomycin Z (NZ)–induced Lethality when Overexpressed in Saccharomyces cerevisiae Lacking the Myosin Type II Gene MYO1. N.L. Díaz-Blanco, José R. Rodríguez-Medina
UPR-Medical Sciences Campus, School of Medicine, Department of Biochemistry, San Juan, PR.

Cell wall biogenesis in type II myosin deficient (myo1) strains of the budding yeast Saccharomyces cerevisiae is significantly affected. These strains require chitin synthase III for survival and hence are hypersensitive to Nikkomycin Z (NZ), a competitive inhibitor of the enzyme catalytic subunit (Chs3p). myo1 strains are highly dependent on de novo synthesis of Chs3p suggesting that its normal recycling pathway may be deficient. To clarify the mechanism for NZ-hypersensitivity in myo1 strains and determine if Chs3p traffic is modified, a haploid myo1 strain was transformed with a yeast cDNA library and transformants were positively selected for growth on media containing an inhibitory dose of NZ (30 g/ml). Selected colonies were confirmed for NZ-resistance in a liquid culture assay format: resistant strains were scored as those with growth ratio greater than 40% relative to untreated cultures. We have sequenced cDNA clones that confer NZ-resistance between 40–100%, compared to untransformed myo1 cells which typically exhibit 10-15% NZ-resistance. To date, most genes are related to four major pathways: the ubiquitin mediated proteolytic pathway, transcription initiation and control, the lipid, fatty-acid and sterol metabolism pathway and protein synthesis. The restoration of cell wall function by the overexpression of specific genes supports the hypothesis that certain proteins involved in cell wall biogenesis may become depleted in myo1 cells. The cDNA clones are being analyzed to establish a detailed NZ-resistance pathway.

Induction of Male-Typical Sexual Behaviors in AAS-Exposed Females
J. Parrilla1;V. Rundle-González2; J.L. Barreto-Estrada3; J.C. Jorge3 Department of Sciences, Mathematics and Technology, UNE1; UPR-Río Piedras2; UPR-RCM Department of Anatomy3, San Juan, PR.

Anabolic androgenic steroids (AAS) have been misused by athletes for more than four decades. Unfortunately, a growing number of females and adolescents are becoming AAS users at alarming rates. Previous immunohistochemistry data have shown that modulation of GABA-IR in female mice was observed in endocrine-related regions (mPOA and VMN) after AAS exposure. For this reason, we have investigated the effects of anabolic steroids in sexual behaviors between control and AAS treated females. The AAS, 17&#945;-methyltestosterone (17&#945;-meT; 7.5 mg/kg) or saline were administered for a two-week period through an osmotic pump. Parameters such as frequency of mounts, fights and escapes, in addition to pelvic thrust and lordosis were measured. Interestingly, we have found that AAS-treated females (F/AAS) showed a low number of mounts when paired with an intact male (P<0.005). This behavior was accompanied by an increased number of fights with the male (P<0.01) and escapes from the male (P<0.05). However, F/AAS showed a strong significant tendency to mount a female control (F/C) (P<0.001) in a male-like pattern. When compared mounts and pelvic thrust to F/C, there was no difference between a male and F/AAS. Finally, lordosis strength (LS) was greater when a male and a F/C where paired (P<0.05). The behavioral results presented in this study correlates with the GABAergic modulation observed in endocrine-related regions of the brain, suggesting that it mediates some of the effects induced upon androgen exposure. Supported by MBRS-RISE to UNE (1R25-GM066250-01A1) and to MSC-UPR (GM61838), NIH COBRE (RR15565), RCMI (G12RR03051)

Differential Expression of Biogenic Amines in the CNS of Macrobrachium rosenbergii and cDNA Library Construction. N. Vázquez-Acevedo1,2; D. Reyes-Colón1,2; A.B. Kohn3; N.M. Rivera1; M.A. Sosa1,2. UPR 1Institute of Neurobiology; UPR 2 School of Medicine, Department of Anatomy; and 3Whitney Laboratory, University of Florida, St. Augustine, Florida, USA

The giant prawn Macrobrachium rosenbergii is a freshwater shrimp whose adult males develop through three morphological types (small [SC], yellow [YC], and blue [BC] claws) that differ in their claw-to-body-length ratio, claw color, and morphology. Each morphotype represents a level in a dominance hierarchy, BC males being most dominant. We are using this model to study the mechanisms involved in modulation of aggression, focusing on the role played by biogenic amines. One possible mechanism is differences in expression or function of aminergic receptors. We cloned one tyramine (Tyrcrust1) and two serotonin (5-HTcrust1 and 5-HTcrust 2) receptors in the prawn and have preliminary evidence suggesting they may be differentially expressed in different areas of the prawn’s central nervous system (CNS). Results from quantitative RT-PCR experiments indicate that the 5-HTcrust1 receptor is expressed in lower amounts in the brain and thoracic ganglia, and 5-HTcrust2 receptor is expressed in lower amounts in the abdominal ganglia of the SC prawn. Tyrcrust1 is expressed in higher amounts in most areas of the ventral nerve cord of the SC prawn. Real-time PCR experiments are being conducted to confirm these results. We have constructed an amplified cDNA library for expressed sequence tags (ESTs) collection from the CNS of the SC male. We plan to also construct cDNA libraries from the CNS of the other prawn morphotypes, hoping to identify other aminergic receptors and other members of the pathway involved in aggressive behavior in the prawn. Supported by: NIH/MBRS S06GM008224, NIMH/MRISP MH48190, NIH/RCMI G12RR03051; NIH R01NS39103.

Characterization of HIV-1 Primary Isolates and Effects on Macrophage Protein Expresión. D. Toro-Nieves1; M. Plaud1, 2; J. Pérez1; V. Wojna2, H. Gendelman3; L. Meléndez-Guerrero1, 2. 1UPR Medical Sciences Campus Department of Microbiology, San Juan PR. 2UPR Medical Sciences Campus NeuroAIDS Program. 3University of Nebraska Medical Center, Omaha NE.

HIV invades the central nervous system early after viral infection and induces cognitive damage known as HIV associated dementia (HAD). HIV infected monocytes and lymphocytes cross the blood-brain barrier releasing virus and toxins into the brain that impact disease neuropathogenesis. We hypothesize that replication of HIV variants will influence macrophage proteome and HAD. HIV primary isolates from peripheral blood of women with HAD were compared for tropism to isolates from women with normal cognition. For macrophage tropism, HIV isolates were inoculated and propagated on monocyte-derived-macrophages (MDM) and tested for HIV p24 antigen. For lymphocyte tropism, the T cell line, MT2 was inoculated with isolates and monitored for syncytia formation. HIV inoculated MDM were lysed and analyzed by SELDI-TOF proteomics studies. Some MDM showed initial abortive infections with isolates. Three isolates showed increased but low viral replication and two showed high levels of infection. Protein profiles of MDM lysates from uninfected and abortive infections showed intensity differences at 11, 13, and 15kD. The peak at 11kD was decreased in lysates from MDM inoculated with HIV from demented patients as compared with isolates from patients with normal cognition and uninfected (p<0.05). HIV primary isolates showed different replication kinetics on MDM. The MDM profiles changed with HIV replication levels. The decrease of the peak at 11kD in MDM inoculated with isolates from patients with HAD that showed abortive infections may be produced in response to endocyted HIV particles.

Inhibitory Activity and Protein Profile Characterization of Supernatant from Placental Macrophages. K. Garcia-Crespo 1; V. Garcia 1; L. Meléndez-Guerrero1. 1UPR, School of Medicine, Department of Microbiology, San Juan, PR.

It is well documented that soluble factors secreted in the placenta are able to inhibit viral replication. Studies done recently in our laboratory have shown that supernatant from placental macrophages (PM) is capable of reducing viral replication in monocyte derived macrophages (MDM). In this study we further characterize the antiviral effects and protein content of this supernatant. To determine if the factors found in the supernatant are produced constitutively PM were isolated from term placenta and cultured for collection of supernatants at days 3,6,9 and 12. MDM were infected with HIV-BAL and cultured with the collected supernatant or with medium alone as a positive control. Viral replication was determined by p24 antigen assays. We also characterized the protein profiles of PM supernatant by comparing them to protein profiles from MDM. PM and MDM were cultured and supernatant was collected at days 6 and 12. This supernatant was then, analyzed using SELDI-TOF technology from Ciphergen to obtain a protein profile. We found a reduction in HIV-1 replication in MDM cultured with PM supernatant from days 6, 9 and 12 as compared with the positive control. We compared the protein profiles from MDM and PM supernatants and found that there were four significantly different peaks between the two profiles. Two of the peaks were found to be over-expressed in MDM and two of the peaks were over-expressed in PM. We hypothesize that at least one of the proteins secreted by PM could be a factor associated with the antiviral activity of the supernatant.

Three DNA Regions Required for Cointegration during Conjugal Transfer of the Gonococcal 5.2 kb Beta-Lactamase Plasmid pSJ790. R. Scharbaai; A. González; L.J. Torres-Bauzá. UPR- Medical Sciences Campus, Department of Microbiology, San Juan, PR.

The gonococcal 5.2 kb beta-lactamase plasmid pSJ790 is mobilized to Escherichia coli by conjugative plasmids R64, N3 and tetM. Previous restriction map and Southern Blot analysis of randomly selected transconjugants revealed that pSJ790 fused (cointegrate) with the conjugative plasmids by more than one mechanism. In order to locate the DNA regions required for pSJ790 mobilization, the R-plasmid was digested with BamH1 and HindIII into three fragments of 2.4 kb, 1.8 kb and 1.0 kb and cloned to pACYC184 vector. The clones (pACYC2.4, pACYC1.8 and pACYC1.0) were then subjected to mobilization assays in the presence of conjugative tetM to recipient E. coli HMS174. Mating experiments showed that the 2.4 kb BamH1-BamH1 fragment (pACYC2.4) is required for mobilization with the help of tetM (mobilization frequency of 2.2 x 10-2 transconjugants / donor cell). Mating assays of the recombinants were repeated using the conjugative plasmids R64 and N3. However, since pACYC184 was observed to mobilize at a low frequency; the three BamH1-HindIII fragments were cloned to pUC19 vector. Mobilization assays of the resultant clones (pUC2.4, pUC1.8 and pUC1.0) showed that both R64 and N3 were able to mobilize pUC1.0 at almost the same frequency (1.9 x 10-7 and 2.2 x 10-7 transconjugants / donor cells; respectively). The clone pUC1.8 was mobilized at a moderate to low frequency by N3 and R64 (1.4 x 10-4 and 4.9 x 10-8 transconjugants / donor cells; respectively). Conclusions: The three BamH1-HindIII fragments of 2.4 kb 1.8 kb and 1.0 kb of pSJ790 contain active DNA sites for pSJ790 mobilization by cointegration with conjugative plasmids.

Partial Characterization of the Insulin Receptor Gene From Pink Convict Cichlids. L. Del Valle; Y. Gerena; L. Serra; A.Schwartz , E. Fernandez-Repollet. University of Puerto Rico, Medical Sciences Campus, School of Medicine, Pharmacology Department.

The insulin receptor (IR) is a cell surface integral membrane protein which plays an essential physiological role in glucose metabolism. The origin of the glucose regulation system is believed to have accompanied early evolution of vertebrates. Although the IR sequence has been identified in most organisms including a partial cDNA sequence of a nonmammalian vertebrate zebrafish (Danio rerio), there have been no reports of its identification and characterization in the vertebrate pink convict cichlid (Cryptoheros nigrofasciatus). In an attempt to characterize the IR sequence from pink convict cichlids, we have compared the IR gene sequences of several organisms including zebrafish. We also isolated total RNA and amplify a partial cDNA sequence of the IR gene from pink convict cichlids. IR sequences of several species including, human, turbot fish, and Danio rerio were identified and aligned using GeneDoc Software. Total RNA was isolated from pink convict cichlids using RNAStat 60 reagent. cDNA synthesis was performed from total RNA in a reaction primed with oligo(dT) primers. Bioinformatic analysis revealed a highly conservative pattern (>89%) of the IR sequence among D. rerio, H. sapiens, M. musculus and R. norvergicus. Total RNA was successfully isolated from pink convict cichlids and RNA gel electrophoresis revealed a 4.7kb and 1.9kb fragments corresponding to the 28S and 18S rRNA. Studies are in progress to amplify a 399bp fragment, corresponding to the partial cDNA sequence of the pink convict cichlid IR gene and generate an IR-EGFP fusion protein to visualize the localization of this receptor during development.

Senger's Disease: An Association of Bilateral Cataracts, Mitochondrial Myopathy of the Heart and Skeletal Muscle and Lactic Acidosis. Case Report with Electron Microscopy Findings. M. Torrellas, M. S. Correa-Rivas MD, A. González-Pesante MD; L. Colón-Castillo MD. Department of Pathology and Laboratory Medicine, Medical Sciences Campus, UPR, San Juan, PR.

Senger’s disease is a rare inherited disorder characterized by congenital cataracts, hypertrophic cardiomyopathy, mitochondrial myopathy of the heart and skeletal muscle, and lactic acidosis. The pattern of inheritance is autosomal recessive, although an X-linked dominant inheritance with variable expression has also been described. The disease was first described by Senger et al. in 1975, and since then very few cases have been reported in the medical literature. We present a 5 month old male patient with bilateral congenital cataracts, hypertrophic cardiomyopathy, and lactic acidosis who during an evaluation at the Emergency Room rapidly deteriorated developing cardiac failure and death. Senger’s disease was clinically considered. The autopsy revealed an infant with anasarca, bilateral congenital cataracts, hypertrophic cardiomyopathy, and skeletal myopathy. Microscopy of the heart muscle revealed clearing of the myocytes, and sections of the skeletal muscle revealed chronic infiltrates including lymphocytes, histiocytes and plasma cells, degenerated fibers, and basophilic fibers with nuclei of regeneration. At the ultrastructural level, skeletal muscle fibers were well preserved with increased number of normal and abnormal mitochondria with bizarre shapes, varying sizes, and with the presence of longitudinal electron dense inclusions. The autopsy findings of bilateral cataracts, hypertrophic cardiomyopathy, skeletal myopathy and the electron microscopy findings along with lactic acidosis and metabolic work up supported the clinical diagnosis of Senger's disease.

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Ruptured Abdominal Aortic Aneurysm in an Infant: Case Report. M. García-Maldonado 1, María S. Correa-Rivas, E. Márquez-Grau 2. 1Department of Pathology and Laboratory Medicine, and 2Department of Surgery, Medical Sciences Campus, School of Medicine, UPR, San Juan, PR.

Aneurysms of the aorta are rare in children and young adults. Among the described conditions associated with aneurysm are coarctation of the aorta, cardiac valvular disease, umbilical artery catheterization, trauma, infection, arteritis, tuberous sclerosis, and connective tissue diseases. Some cases have no distinguishable cause and are thus idiopathic. We present a one month old infant who during a hospitalization due to a Staphylococcus aureus infection at two weeks old, was found with an abdominal aortic aneurysm on radiologic images. The patient was transferred to our Institution for further management, but died before any surgical intervention. Autopsy revealed a 6 cm abdominal aortic aneurysm located at the diaphragmatic hiatus above the celiac trunk, displaced superiorly to the right hemithorax, and ruptured to the right pleural cavity with a thrombus formation. Another 1.5 cm aneurysm was noted at the bifurcation of the iliac artery; this one not ruptured. Microscopic sections of the aorta stained with hematoxylin and eosin, and elastin stains revealed disruption of the elastic layer and calcifications within the thrombus, aortic wall, and diaphragm, adjacent to the aneurysmal sac. The presence of these calcificatons and well organized thrombus with fibrin deposition suggested a chronic ongoing process, probably congenital, though in our case, the predisposing factor to the aneurysm development remained undetermined. We discuss this unusual disease in children and the different proposed causes of abdominal aorta aneurysm recently reviewed in the medical literature.

Acute Myelogenous leukemia/ Transient Myeloproliferative disorder in Down syndrome. Report of a case. M. Rosa-Gallardo; R.Velez-Rosario. Department of Pathology and Laboratory Medicine. University of Puerto Rico. Medical Sciences Campus.

Individuals with Down Syndrome (DS) have an increased predisposition to develop acute leukemia. In approximately 50% of cases it appears to be acute megakaryoblastic leukemia. In some cases the disease undergoes spontaneous remission during the first one to three months of life. This process has been referred to as transient myeloproliferative disorder or transient leukemia.
We report the case of a female newborn with DS associated with acute myelogenous leukemia consistent with Megakaryoblastic origin. This patient also had severe cardiac malformations and suffered other perinatal complications that caused her death at 19 days after birth. We will review the clinical and pathological characteristics of this interesting entity

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Association of Extensive Brain Calcifications, Myelofibrosis and Retinopathy in a 12 Year Old Child
D. Negrón, L. Colón-Castillo, I. Morales-Melecio, M. S. Correa-Rivas. Department of Pathology and Laboratory Medicine, Medical Sciences Campus, School of Medicine, UPR, San Juan, PR.

Intracerebral calcifications associated with retinopathy have been described in association with several rare syndromes as well as familial cases of idiopathic brain calcifications with autosomal dominant inheritance. In 1992, T. Revesz published a case report describing a 6 month old male infant who was found to have bilateral exudative retinopathy and subsequently developed severe aplastic anemia, leading to his death. Additional clinical findings included intrauterine growth retardation, cerebral calcifications, cerebellar hypoplasia, and progressive psychomotor retardation. We report a 12 year old male child with similar findings. The developmental history of this boy was otherwise unremarkable with an above average intelligence. He was well at home, but unexpectedly presented acute neurological deterioration and death. The postmortem neuropathologic examination revealed multiple intraventricular and intraparenchymal coarse calcifications as well as intraparenchymal hemorrhages, cerebral edema and bilateral subarachnoid hemorrhages in the occipital lobes and cerebellum. We discuss several conditions associated with intracerebral calcifications such as dyskeratosis congenita, Coat´s disease, Fanconi anemia and Revesz syndrome,which were considered in the differential diagnosis of this difficult case. The presence of exudative retinopathy and bone marrow abnormalities should alert clinicians to pursue further findings associated with these conditions.

Metanephric Adenoma in a 16 Year Old Child: Case Report. L. Rivera1, M. S. Correa-Rivas1, S. Gutierrez1, A. Ramírez2, P. Collazo3. 1 Department of Pathology, 2Department of Urology and 3Department of Radiology, UPR, Medical Sciences Campus, School of Medicine, Sam Juan, PR

Metanephric adenoma is a rare epithelial renal neoplasm with characteristic histology and no malignant potential. It occurs at all ages most commonly in the fifth and sixth decades, with a 2:1 female to male ratio. They ranged widely in size and are located mostly in the renal cortex. Etiology is unknown, but they may be associated with smoke, tubular nephrosclerosis, and dyalisis. Approximately 50 % are incidental findings while others present with polycythemia, abdominal or flank pain, mass, hematuria or hypertension. We report a 16 year old girl found with a metanephric adenoma of the left kidney, which showed typical clinical, microscopic and immunohistochemical findings. It is important to differentiate metanephric adenoma from other more aggressive renal tumors such as renal cell carcinoma and Wilm’s tumor, since metanephric adenoma has a benign course and should be treated in a conservative manner.

Exposure to Television in a Group of Children living in San Juan. I. Matías1; I. Ancalle1; L. García2. San Juan City Hospital, Department of Pediatrics1; and University of Puerto Rico School of Medicine, Department of Pediatrics2, San Juan, Puerto Rico.

Media communication influences children perception of their environment, interpersonal relations, and their behavior. The American Academy of Pediatrics (AAP) recommends that children TV exposure be limited and under supervision. Objective: To assess the time of exposure to TV among children and its supervision. Methods: A survey was answered by 42 caretakers of children between the ages of 1 to 10 years who visited a primary care clinic in San Juan or who were admitted at the San Juan City Hospital. The Institutional Review Board approved the study. Results: The mean age of children was 5 y/o, 40% were females and 60% males. All of them had exposure to TV for a mean of 4 hours during school days and 6 hours on weekends. Seventy-four had a TV in their bedroom. Thirty-three percent of children chose the programs to be watched. Ninety-eight percent of caretakers referred to supervise the children while watching TV at least most of the time with 67% acknowledging the TV programs’ classification. Eating in front of the TV was associated to a higher prevalence of obesity (54% vs 21%, p=0.0472). Conclusions: Children in this sample are exposed to TV for more hours than recommended by the AAP. Although most of the caretakers referred to supervise the children while watching TV, there is lack of knowledge about the chosen programs classification. As previously reported in the literature, children who ate in front of the TV had a higher prevalence of obesity. Education of caretakers is warranted in order to decrease the negative effects associated to excessive exposure to TV and to decrease the morbidity associated to obesity.

Serous Surface Papillary Carcinoma of Peritoneum: An Autopsy Case Report. M.L. Rivera Zengotita; C. Climent Peris. Department of Pathology and Laboratory Medicine, UPR School of Medicine, San Juan, PR.

Serous surface papillary carcinoma of peritoneum is a rare primary tumor of the peritoneum that diffusely involves the peritoneal surface but spares or superficially invades the ovaries. We report the case of a 69 year old female without history of systemic illness who died a few hours after hospital admission due to respiratory failure secondary to massive right pleural effusion. Post-mortem cytologic examination of the effusion fluid revealed adenocarcinoma cells. Autopsy demonstrated yellow-white small (0.3 cm) nodularities located on the right parietal pelvic peritoneum and massive right pleural effusion (1,350 ml of bloody fluid). Microscopic examination of the peritoneal nodularities revealed serous surface papillary carcinoma of the peritoneum. The tumor was also present on the surface of one ovary without cortical invasion and in one breast. This patient had an unusual and catastrophic clinical presentation of serous surface papillary carcinoma of the peritoneum.

Evidence Based Management of a Glioblastoma Multiforme in an Infant. M. Torres-Serrant, M.E. Echevarría, M.S. Correa-Rivas, L. Colón-Castillo, F. Montalvo. Department of Pathology and Laboratory Medicine, Medical Sciences Campus, School of Medicine, UPR, San Juan, PR.

Glioblastoma multiforme (GBM) is one of the most common and aggressive primary brain tumors. It is a high grade astrocytoma most commonly seen in adults. In children it is less common, accounting for approximately 10% of all pediatric central nervous system tumors. Despite multi-modality treatment with gross tumor resection, radiation therapy and chemotherapy, survival rates remain poor for children, ranging from 20-30% two years after diagnosis. We present the case of an infant girl born without perinatal complications that developed status epilepticus at 6 months old. Head CT scan showed a supratentorial cortical lesion within the right parieto-occipital lobe with both solid and cystic components. MRI confirmed these findings showing an enhancing lesion in the right parietal lobe. Patient underwent gross total resection of the tumor. On microscopy, the tumor revealed marked nuclear atypia, proliferation of hyperplastic vessels, and necrosis surrounded by a palisade of anaplastic cells; all features characteristic of glioblastoma. No radiation therapy was given at that time since she was less than 3 years old, but she received chemotherapy with carboplatin, vincristine and cyclophosphamide for a two year period. Despite the dismal prognosis of this neoplasm, especially without multi-modality treatment, our patient has survived with minimal neurologic deficits, and continues without evidence of tumor recurrence five years after diagnosis. This case supports the idea that glioblastomas in early childhood can be managed successfully with gross total resection and appropriate adjuvant chemotherapy.

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Treatment Response to Peg-Interferon and Ribavirin in Hispanics with Hepatitis C. J. Iturrino, C.J. Romero-Marrero, C.J. Sanchez, G. Nogueras, A. Ortiz, P.J. Costas, E.A. Torres. UPR- Medical Sciences Campus, Department of Medicine, San Juan, PR.

Response to hepatitis C (HCV) therapy is influenced by ethnic/racial factors. We performed a retrospective analysis to describe the end of treatment response (ETR) and sustained viral response (SVR) to Peg-interferon and Ribavirin in treatment-naïve chronic HCV Puerto Rican patients in a government-sponsored pilot clinic. 156 (106 M:50 F, mean age 41.9) patients were started in treatment. Genotype 1 was found in 72.4%. Treatment was discontinued in 47 (30%): 24 (51%) were non-responders, 16 (34%) had adverse effects, 5 (10.7%) developed co-morbidities during treatment, 2 (4.3%) had unrelated deaths. Another 46 patients (29.5%) were lost to follow-up after having started treatment. 63 patients (40.4%) completed treatment. When patients with an early discontinuation are excluded, 37/63 (58.7%) had ETR, while the ETR on those intended to treat was 23.7%. Recurrence of viremia was observed in 5/37 (13.5%) of those with ETR. SVR was seen in 16/63 (25.4%) patients who completed treatment, while the SVR on those intended to treat was 16/156 (10.3%). Sixteen of 37 (43.2%) patients were lost to follow up to assess SVR. No correlation was found between ETR and age, gender, viral genotype, initial HCV RNA, early virological response, or degree of fibrosis (p>0.05). Although insufficient patients completed follow-up to achieve statistical significance, SVR to pegylated interferon and ribavirin seems to be lower in Hispanics than reported in other populations. The high rate of discontinuation of treatment in our population surpasses the previously reported rates in U.S. Latinos and Caucasians.

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Obesity Risk Factors for Puerto Rican Type 2 diabetes Patients. J. Rodriguez, R. J. Rodríguez. UPR, Medical Sciences Campus, Department of Gerontology, School Graduate of Public Health, San Juan, PR.

The prevalence of Diabetes mellitus type 2(DMT2) is high in the Puerto Rican population as compared to other metabolic diseases in the island (CDC, 2002; Perez, Perez, & Suarez, 2001). Diabetes have been identified as the third reason of mortality in PR (PR Health Department,1999-2001). Phenotypic components of human obesity, such as anormal body weight (BW) and body mass index (BMI) are related to a higher risk in the develop of DMT2. Research at the CDPR explored the relation between DMT2 patients an risk behaviors that affect their life-expectancy.Objective:To explore risk factors for DMT2(i.e., obesity)in a patients sample at the CDPR. Method:A valid clinical and socio-demographic health information questionnaire to screen patient's need, clinical variables, behaviors and attitudes related to DMT2 was used with 100 patients in a 6 months period at the CDPR.Descriptive statistics and ANOVA was performed.Results:Descriptive statistics included 66% female vs. 34% male with a mean age of 58.9 years. Of the total sample, 64.4% subjects report significant body weight changes,59.6% gaining weigth,and 50% reporting physical inactivity. Adjustment by age and gender was done, statistical differences was found between Diabetes knowledge and risk behaviors prevention (i.e., vaccines (r = .41, p= .000), avoid hypoglycemia (r =.34, p=.001), avoid smoking (r =.25, p=.012), and changes in body weigh (r =.30, p=.003).Conclusions:It is imperative to develop preventive lifestyle educational behavioral programs that help DMT2 Hispanic patients to improve their quality of life via the identification, and prevention, of risk behaviors.

Current Role of Protocols in Obstetrical Patient Care at University Hospitals. V. Tamayo Agrait, K. Adamsons. UPR- Medical Sciences Campus, Department of OB-GYN, School of Medicine, San Juan, PR.

In the past, protocols guiding patient care were mostly used at community hospitals, where a large portion of residents were foreign graduates. These tools served as education resources to ensure better compliance with local and national standards. In our specialty, this use of protocols has been largely replaced by educational materials, as the Compendium, an annual publication of the American College of Obstetricians and Gynecologists. This volume contains the most relevant topics to the practitioner and is often considered a source of present standards of care in patient management for medical and medico-legal purposes. The purpose of our study was to determine whether leading university hospitals in our specialty use protocols, and if they do, what are their purposes. We selected 34 institutions for mailing our questionnaires, including six of outstanding reputation (Harvard, Columbia, Johns Hopkins, Stanford, University of Pennsylvania and Yale). Thirty of them replied. Sixty percent of them were users, while 40 percent were non-users (although two of them mention use of "standarized approches or "guidelines"). Of particular interest was that only one institution of "outstanding" reputation did not use protocols. All users agreed that use of protocols improves patient care. Forty percent of them considered revision and updating of the protocols burdensome. We are proposing that protocols, usually derived from textbooks, are essential to promote patient management according to standards of care, as evidenced by its use on most of the leading University Hospitals covered by our study.

El Impacto del Practicum de Servicio y Aprendizaje de la Escuela de Farmacia en la Península de Cantera y Barrio Obrero. S. Rivera-Sárate, M. L. González-Cordero. Práctica en Farmacia, Escuela de Farmacia, UPR

Objetivos:
Exponer al estudiante a los asuntos de salud de la comunidad mediante su colaboración en cuatro organizaciones comunitarias.
Ofrecer un servicio a la comunidad de acuerdo a las necesidades identificadas por las organizaciones y contextualizadas a la profesión de farmacia.
Contribuir al desarrollo de las competencias de los estudiantes del Programa de Doctorado en Farmacia.

Estrategias:
Los estudiantes se asignan a una organización y se familiarizan con la misión, metas, objetivos y operación de ésta. La organización identifica la necesidad de salud de la comunidad y los estudiantes en colaboración con la organización desarrollan un plan de trabajo para atender la misma. Este plan se discute con los miembros de la organización, se revisa, se implanta y evalúa. Los estudiantes ofrecen recomendaciones a las organizaciones con el fin de mejorar su impacto en la comunidad. Este Practicum incluye la realización de un diario reflexivo donde se describen las experiencias de los estudiantes, una evaluación de la organización y una evaluación de los estudiantes. La población impactada incluye: niños, jóvenes, adultos y pacientes de edad avanzada. Algunas de las necesidades identificadas son: desconocimiento de la manufactura de medicamentos y educación sobre medicamentos u condiciones. Las actividades realizadas incluyen: desarrollo de material educativo, charlas, visitas al hogar y a la industria farmacéutica.

Logros:
La evaluación de este Practicum sugiere que los objetivos han sido alcanzados.

Proyecciones:
Continuar ofreciendo servicio a las organizaciones participantes y expandir el Practicum.

A novel teaching strategy in the UPR School of Dentistry, Graduate Postdoctoral Residency Program: Use of Objective Structured Clinical Examinations. N. Ortiz 1. Office for the Development of Clinical Skills, University of Puerto Rico School of Medicine. San Juan, Puerto Rico.

Current and projected approaches to dental education have created a wide interest in learning tools that imitate real life conditions. Standardized patients (SPs) are the gold standard to measure the competence of health professionals. SPs are used as part of the objective structured clinical examination (OSCE). The use of OSCEs in training dental professionals is core for history taking, physical exam and in decision making process to establish a treatment plan. It was not until three years ago that the General Practice Residency Program (GPR) in Dentistry School began to use them. Objective: Use OSCEs to assess competency in history, physical exam, and decision making skills in the Physical Diagnosis Course of the GPR Program. Strategies: Residents were taught to gather history and physical exam. Were encouraged to develop a diagnostic, treatment, and educational plan. Outcome was measured using OSCEs. Outcomes: The average in history taking skills was 90%; physical exam, 88%; in critical thinking, 86%. Projections: OSCEs will continue to be used in the Physical Diagnosis Course for dental students. Immediate feedback given by the faculty physician will be established as part of the examination. Supported in part by the GPR Program, UPR School of Dentistry and the Office of Clinical Skills, UPR School of Medicine.

Cesarean delivery as a barrier for breastfeeding initiation and duration: The Puerto Rican experience. N. Pérez-Ríos1, A.P. Ortiz1, and G. Ramos-Valencia1. UPR 1School of Public Health, Department of Biostatistics and Epidemiology, San Juan, PR.

During 1990-1996, 30 to 37% of all births in Puerto Rico were performed through cesarean section. This represents a public health concern, as studies performed in other populations suggest that cesarean section is a barrier for breastfeeding initiation and duration. Our study pretend to examine the association between cesarean section delivery and the initiation and duration of breastfeeding in a representative sample of 1,958 Puerto Rican women aged 15-49 years. Bivariate and multivariate logistic regression analysis was used to examine the association between birth type and initiation of breastfeeding. We also used Survival analysis and Cox Regression to observe the effect of birth type on the duration of breastfeeding. The proportion of cesarean section was 36.1%. In covariate adjusted multivariable logistic regression models, cesarean section was negatively related to breastfeeding initiation (OR=.61; IC 95%; .49, .75), but no association was found with breastfeeding duration (HR=.99, IC 95%: 0.83-1.18). This is the first population based study to confirm that cesarean section is a barrier for initiation of breastfeeding in PR. Health education campaigns targeting hospital personnel and women who have cesarean sections should be developed, in order to increase breast feeding initiation among these women.

Colorectal Cancer Risk Index and Physical Activity in Hispanic Adults Living in a Rural Area of Puerto Rico M. Torres-Cintron1, F. A. Ramirez-Marrero, FACSM2, C. M. Nazario1, E. Suarez1. 1University of Puerto Rico - Medical Sciences Campus, San Juan, Puerto Rico. 2University of Puerto Rico - Rio Piedras Campus, San Juan, Puerto Rico.

Colorectal cancer is the second leading cause of cancer mortality in Puerto Rico. The purpose of this study was to estimate the association between physical activity and colorectal cancer risk in a group of Hispanic adults living in Barranquitas, Puerto Rico. A colorectal cancer risk index was developed based on the presence of known risk factors. Risk points were assigned according to the strength of the causal association of the risk factors and summed. Population average of colorectal cancer risk was obtained from the prevalence of risk factors using the PR-BRFSS and the prevalence of risk factors of the study population. Physical activity level was determined using the International Physical Activity Questionnaire. Participants were classified as highly active (&#8805;1500 MET-min/wk) and not highly active (<1500 MET-min/wk). Personal interviews were performed on 126 participants [females=83 (66%), males=43 (40%); mean age = 58.7 yrs] living in a rural municipality of Puerto Rico. A logistic regression model was used to estimate the magnitud of the association between the colorectal cancer risk index and the physical activity. The average risk index for men was 20.17, and for women 20.84. Forty-two percent had a high colorectal cancer risk index, and 71% were classified as highly active. When the model was adjusted by sex, education level, medical insurance and calcium use, the risk of colorectal cancer was twice for those with an activity level <1500 MET-min/wk compared with those with an activity level &#8805;1500 MET-min/wk (POR= 2.14; IC 95%: 0.84, 5.42), however, this association was not statistically significant.

Risk factors associated with youth violence among 5th and 6th grade students in Puerto Rico, 2001: a cross-sectional study. M. C. Mercado-Crespo1, C. N. Vélez-Santori1, G. Ramos-Valencia1, M. R. Moscoso2. 1UPR – Graduate School of Public Health, Medical Sciences Campus, San Juan, PR; 2UPR - Río Piedras Campus, San Juan, PR.

In Puerto Rico, violence is the fourth cause of death among children aged 10-14. Although there are many surveys of 7th-12th grade violence, data on younger children is scarce. The purpose of this investigation was to estimate the prevalence of youth violence risk factors among school-children ages 9 to 13.

A secondary analysis of data from an island-wide representative sample of all 5th and 6th grade students in public and private schools in PR was carried out, with approval of UPR-RCM's IRB. "Consulta Juvenil: V" project selected the sample through a multistage stratified cluster sampling design, sponsored by Puerto Rico's Mental Health and Addiction Services Administration. Risk factors were classified as individual, family, school, peer, situational or community related factors, and analyzed in relation to the reported violent behavior.

The prevalence of at least one violent incident among preadolescents in PR was 43%, school-violence being most common. Contrary to previous 7th-12th grade studies, individual, family and school risk factors were responsible of the most variability in violent behavior, whereas peer, situational and community risk factors contributed the least. Being male and of higher grade level was responsible of the greatest change of units in the violence index. Church-attendance was identified as a protective factor.

Findings were consistent in stating violent behavior varies according to the type of risk factors present in the individual. Further, longitudinal studies and the development of a violence surveillance system in PR are recommended.

El impacto de la diabetes y la obesidad en la población de adultos mayores en Puerto Rico. A. García, M. McEniry, M. Larriuz, H. Strmic-Pawl. Universidad de Puerto Rico, Recinto de Ciencias Médicas, Departamento de Ciencias Sociales, Escuela Graduada Salud Pública, Recinto de Ciencias Médicas, San Juan, PR.

Los adultos mayores en Puerto Rico tienden a tener una mayor prevalencia de diabetes, especialmente entre las mujeres. En este trabajo se destacan algunos hallazgos sobre diabetes y obesidad obtenidos en el estudio PREHCO que recogió información de 5,336 personas de 60 años o más seleccionadas en un muestra representativa de esta la población en Puerto Rico. La prevalencia global de diabetes fue de 28 por ciento, probablemente subestimada ya que es autoreportada. Se estima que los residentes en Puerto Rico tienen 1.8 veces más probabilidad de padecer de diabetes que los blancos no hispanos en los Estados Unidos. La diabetes es mayor entre las mujeres. Las personas diabéticas padecen más de enfermedades crónicas y tienen más limitaciones en las actividades de la vida diaria. Los diabéticos y las diabéticas consumen más medicamentos y usan más los servicios de salud que las personas que no son diabéticas.

La obesidad es un conocido factor de riesgo para la diabetes, razón por la cual no sea sorprendente que estos datos demuestren una relativamente alta prevalencia de obesidad. Las personas obesas tiene una mayor prevalencia de diabetes que las personas no obesas.