Marital Status and Survival in Patients Diagnosed with Melanoma.

INTRODUCTION: Previous research suggests the presence of a spouse may considerably affect melanoma detection rates through more frequent examinations, better access to healthcare, and improved social support. Yet, the role of marital status on melanoma survival is currently unknown. The aim of this study is to assess whether marital status is associated with survival following melanoma diagnosis. METHODS: We performed secondary analysis of data from all participants of the Florida Cancer Data System (FCDS) and included adult melanoma patients diagnosed between 2001 and 2009 with follow-up information available until 2015. Marital status was categorized as single, married, divorced, or widowed. The primary outcome was survival interval after melanoma diagnosis, which was assessed according to the time from the date of diagnosis to the time of death or last contact. Cox proportional hazard models were used to assess the independent association between marital status and survival. RESULTS: We assessed data from 36,578 melanoma patients. Married patients were significantly more likely to survive than single patients (Hazard ratio (HR) = 0.65; 99% Confidence Interval (CI): 0.57-0.74; P < 0.001) after adjusting for age, sex, race, ethnicity, geographic location, insurance status, tobacco use, primary site, stage, and histology. There was no evidence of effect modification by gender (P=0.189). CONCLUSIONS: Married patients, including both men and women, had a 35% reduction in the risk of death after melanoma diagnosis compared with single patients, and mechanisms independent of earlier detection, such as social support, may play a role in survival in patients with melanoma.

Simplified risk stratification criteria for identification of patients with MRSA bacteremia at low risk of infective endocarditis: implications for avoiding routine transesophageal echocardiography in MRSA bacteremia.

The aim of this study was to identify patients with methicillin-resistant Staphylococcus aureus (MRSA) bacteremia with low risk of infective endocarditis (IE) who might not require routine trans-esophageal echocardiography (TEE). We retrospectively evaluated 398 patients presenting with MRSA bacteremia for the presence of the following clinical criteria: intravenous drug abuse (IVDA), long-term catheter, prolonged bacteremia, intra-cardiac device, prosthetic valve, hemodialysis dependency, vertebral/nonvertebral osteomyelitis, cardio-structural abnormality. IE was diagnosed using the modified Duke criteria. Of 398 patients with MRSA bacteremia, 26.4 % of cases were community-acquired, 56.3 % were health-care-associated, and 17.3 % were hospital-acquired. Of the group, 44 patients had definite IE, 119 had possible IE, and 235 had a rejected diagnosis. Out of 398 patients, 231 were evaluated with transthoracic echocardiography (TTE) or TEE. All 44 patients with definite IE fulfilled at least one criterion (sensitivity 100 %). Finally, a receiver operator characteristic (ROC) curve was obtained to evaluate the total risk score of our proposed criteria as a predictor of the presence of IE, and this was compared to the ROC curve of a previously proposed criteria. The area under the ROC curve for our criteria was 0.710, while the area under the ROC curve for the criteria previously proposed was 0.537 (p < 0.001). The p-value for comparing those 2 areas was less than 0.001, indicating statistical significance. Patients with MRSA bacteremia without any of our proposed clinical criteria have very low risk of developing IE and may not require routine TEE.

New contributions to the ORP & DO time profile characterization to improve biological nutrient removal.

Changes in the oxidation-reduction potential (ORP), pH and dissolved oxygen (DO), along with organic load and nutrient removal in a municipal wastewater treatment plant (WWTP) have been monitored throughout one year. The "nitrate knee" and the "nitrate break point" in ORP profiles, the "nitrate apex" and the "ammonia valley" in pH profiles and the "DO elbow" in DO profiles have been identified. Furthermore, these bending points have been correlated with the oxygen uptake rate (OUR), the temperature in the vessel and the aeration and non-aeration time profiles by using Principal Component Analysis (PCA). The data have been previously split up into wet and dry weather cycles by means of a K-means clustering algorithm. Finally, two new parameters have been defined: the "ORP Arrow", which is closely related to the inhibition of the denitrification process, and the "Oxygen Rise Average Slope" (ORAS), which shows the oxygen transfer rate.

Two-step chromatographic purification of recombinant Plasmodium falciparum circumsporozoite protein from Escherichia coli.

The Plasmodium falciparum circumsporozoite (PfCS) protein (aa 19-405) has been cloned and expressed in E. coli. The protein was purified in a two-step process that was rapid and reproducible. E. coli cells were grown to a high density before induction for 1 h. Cells were disrupted by high pressure microfluidization and the total bacterial protein solubilized in 6 M Gu-HCl. The protein was refolded while bound to Ni-NTA agarose by exchange of 6 M Gu-HCl for 8 M urea and then slow removal of the urea. The eluted protein was further purified on Q Sepharose Fast Flow using conditions developed to remove E. coli proteins and reduce endotoxin (to 10 EU/50 microg). Yield was 20 mg of PfCS protein from 10 g of wet cell paste. The final protein product bound to HepG2 liver cells in culture and inhibited the invasion of those cells by sporozoites in an ISI assay greater than 80% over control cultures when used at 10 microg/ml.

Immunogenicity of Plasmodium falciparum circumsporozoite protein multiple antigen peptide vaccine formulated with different adjuvants.

Only low antibody levels were obtained from vaccinating human volunteers with single-chain peptide from the Plasmodium falciparum circumsporozoite protein (PfCSP). This resulted in modest protection against sporozoite challenge. In addition, HLA restriction limits the probability of synthesis of a vaccine effective for a diverse population. We report immunization studies with a multiple antigen peptide (MAP) system consisting of multiple copies of a B-cell epitope from the central repeat region of the PfCSP in combination with a universal T-cell epitope, the P2P30 portion of tetanus toxin. This MAP4(NANP)6P2P30 vaccine was highly immunogenic in four different strains of mice when used with various safe and nontoxic adjuvants. When this MAP vaccine was encapsulated in liposomes with lipid A and adsorbed to aluminium hydroxide and given three times at 4-week intervals, the resultant antibody prevented 100% of sporozoites from invading and developing into liver stage infection. This high degree of immunogenicity of MAP4(NANP)6P2P30 vaccine formulated in liposomes, lipid A and aluminum hydroxide provides the foundation for consideration of human trials with this formulation.

Heterogeneity of macrophage and T cell subpopulations in peripheral nerves from HIV infected individuals: A preliminary study.

GOAL: To determine the heterogeneity of surface marker expression of macrophages in peripheral nerve of patients who died with AIDS. BACKGROUND: Peripheral neuropathy occurs in 20%-40% of AIDS patients. There is evidence that activated macrophages may be involved in the neural damage associated with HIV-1 infection. We studied the expression of macrophage surface markers CD14, CD11c, CD68, and HLA-DR and also T cell surface markers CD3, CD4, and CD8 in peripheral nerves of AIDS patients. METHODS: Three levels of peripheral nerves (sciatic, tibial, or sural) were examined from a limited number of subjects consisting of 4 HIV-seropositive and 5 HIV-seronegative individuals. Standard immunohistochemical technique utilized alkaline phosphatase conjugate and fuchsin substrate. RESULTS: Surface antigen expression was significantly (p < .0025 increased in HIV-positive tissues compared with HIV-negative controls for CD14 and CD4 in sciatic nerves, CD68 and CD4 in tibial nerves, and CD68 in sural nerves. There were trends for increased expression of HLA-DR, CD3, and CD8 in sciatic nerves, CD11c and CD14 in tibial nerves, and CD14, HLA-DR, and CD4 in sural nerves in HIV-positive tissues compared with HIV-negative controls. CONCLUSION: During the course of AIDS there may be an involvement of all three levels of peripheral nerves suggesting that HIV-related neuropathy is a multifocal process.

Development of two monoclonal antibodies against Plasmodium falciparum sporozoite surface protein 2 and mapping of B-cell epitopes.

The Plasmodium yoelii sporozoite surface protein 2 (PySSP2) is the target of protective cellular immunity. Cytotoxic T cells specific for the Plasmodium falciparum analog PfSSP2, also known as thrombospondin-related anonymous protein (TRAP), are induced in human volunteers immunized with irradiated sporozoites. PfSSP2 is an important candidate antigen for a multicomponent malaria vaccine. We generated and characterized three monoclonal antibodies (MAbs) specific for PfSSP2/TRAP. The MAbs PfSSP2.1 (immunoglobulin G1 [IgG1]), PfSSP2.2 (IgG2a), and PfSSP2.3 (IgM) were species specific and identified three distinct B-cell epitopes containing sequences DRYI, CHPSDGKC, and TRPHGR, respectively. PfSSP2.1 partially inhibited P. falciparum liver-stage parasite development in human hepatocyte cultures (42 and 86% in two experiments at 100 microg/ml). Mice immunized with vaccinia virus expressing full-length PfSSP2 protein produced antibodies to (DRYIPYSP)3, and humans living in malaria-endemic areas (Indonesia and Kenya), who have lifelong exposure and partial clinical immunity to malaria, had antibodies to both (DRYIPYSP)3 and (CHPSDGKCN)2. Mice immunized with multiple antigen peptides MAP4 (DRYIPYSP)3P2P30 and MAP4 (CHPSDGKCN)3P2P30 in TiterMax developed antibodies to sporozoites that partially inhibited sporozoite invasion of human hepatoma cells (39 to 71% at a serum dilution of 1:50 in three different experiments). The modest inhibitory activities of the MAbs and the polyclonal antibodies to PfSSP2/TRAP epitopes do not suggest that a single-component vaccine designed to induce antibodies against PfSSP2/TRAP will be protective. Nonetheless, the MAbs directed against PfSSP2, and the peptides recognized by these MAbs, will be essential reagents in the development of PfSSP2/TRAP as a component of a multivalent P. falciparum human malaria vaccine.

Protection against malaria by Plasmodium yoelii sporozoite surface protein 2 linear peptide induction of CD4+ T cell- and IFN-gamma-dependent elimination of infected hepatocytes.

Plasmodium falciparum sporozoite surface protein 2 (SSP2), also known as TRAP, is included in experimental human malaria vaccines because Plasmodium yoelii SSP2 is the target of protective CD8+ CTL that eliminate P. yoelii-infected hepatocytes in mice. We now report that immunization with a synthetic branched-chain peptide including four copies of a PySSP2 sequence, NPNEPS, and two tetanus toxin T helper epitopes in the adjuvant TiterMax, or with an 18 amino acid peptide (NPNEPS)3 in the adjuvant protects A/J, but not BALB/c or C57BL/6 mice. Transfer of T lymphocyte-enriched immune splenocytes protects naive mice; in vivo depletion of CD4+ T cells eliminates vaccine-induced protection; and in vivo treatment with anti-IFN-gamma reverses vaccine-induced activity against infected hepatocytes. Lymph node cells from immunized A/J, BALB/c, and C57BL/6 mice recognize the (NPNEPS)3 peptide in vitro. However, the protected A/J mice respond with a predominantly Th1 pattern of lymphocyte response, and the non-protected strains of mice respond with a Th2 pattern. There are many examples of CD4+ T cells transferring protection against infectious organisms. However, to our knowledge, this is the first formal demonstration that immunization with a linear synthetic peptide induces CD4+ T cell-dependent, IFN-gamma dependent, genetically restricted sterile protective immunity against an infectious agent.

Identification and characterization of the protective hepatocyte erythrocyte protein 17 kDa gene of Plasmodium yoelii, homolog of Plasmodium falciparum exported protein 1.

We recently reported the discovery of a 17-kDa Plasmodium yoelii protein expressed in infected hepatocytes and erythrocytes, P. yoelii hepatocyte erythrocyte protein 17 (PyHEP17), and have demonstrated that this protein is a target of protective antibodies and T cells. Here, we report the identification and characterization of the gene encoding this protein and reveal that it is composed of two exons. Immunization of mice with PyHEP17 plasmid DNA induces antibodies, cytotoxic T lymphocytes, and protective immunity directed against the infected hepatocyte. Based on extensive sequence homology, expression pattern, and antigenic cross-reactivity, the Plasmodium falciparum homolog of PyHEP17 is identified as the protein known as exported protein-1 (PfExp-1), also called antigen 5.1, circumsporozoite related antigen, or QF116. Identity between PyHEP17 and PfExp-1 is 37% at the amino acid level (60/161 residues), mapping primarily to two regions within the second exon of 73% (16/22 residues) and 71% (25/35 residues) identity. On this basis, PfExp-1 is proposed as an important component of pre-erythrocytic human malaria vaccines.

Plasmodium yoelii: 17-kDa hepatic and erythrocytic stage protein is the target of an inhibitory monoclonal antibody.

Infected hepatocytes are important targets for malaria vaccines. To identify Plasmodium yoelii proteins expressed in infected hepatocytes, we immunized BALB/c ByJ mice with P. yoelii liver stage schizonts and produced a panel of monoclonal antibodies (Mabs). An IgG1 Mab, navy yoelii liver stage 3 (NYLS3), had the strongest reactivity against liver stage parasites and was selected for further characterization. The Mab does not recognize P. yoelii sporozoites, but recognizes liver stage parasites within 6 hr of invasion of mouse hepatocytes and throughout the hepatic and asexual erythrocytic stages of the parasite life cycle as determined by the immunofluorescent antibody test. This Mab is species-specific, and it reacts with liver stages of P. yoelii but does not react with liver stages of other Plasmodium species. The protein recognized by this Mab is present on the parasitophorous vacuole membrane of infected hepatocytes and erythrocytes as demonstrated by immunoelectron microscopy and has a relative molecular weight of 17 kDa as demonstrated by immunoblot of an extract of infected erythrocytes. It is therefore designated P. yoelii hepatic and erythrocytic stage protein, 17 kDa or PyHEP17. When added to primary cultures of mouse hepatocytes 24 hr after inoculation with P. yoelii sporozoites, when all sporozoites have invaded hepatocytes, NYLS3 eliminates up to 98% of liver-stage parasites. Intravenous injection of NYLS3 into mice delays the onset and reduces the density of blood-stage parasitemia after sporozoite or blood-stage challenge. The P. falciparum and P. vivax homologs of PyHEP17 may therefore be important targets for vaccines designed to attack the hepatic and erythrocytic stages of the parasite life cycle.

Nitric oxide-mediated antiplasmodial activity in human and murine hepatocytes induced by gamma interferon and the parasite itself: enhancement by exogenous tetrahydrobiopterin.

Expression of inducible nitric oxide (NO) synthase has been shown to inhibit the development of several pathogens, including fungi, bacteria, parasites, and viruses. However, there is still controversy as to whether this effector mechanism can inhibit the development of human pathogens. We now report that gamma interferon (IFN-gamma) induces the elimination of Plasmodium falciparum-infected primary human hepatocytes from cultures and that the antimalarial activity is dependent on NO. Infection with the parasite alone in the absence of added IFN-gamma caused a 10-fold increase in NO formation. Both spontaneous inhibition and IFN-gamma-induced inhibition of Plasmodium yoelii-infected murine hepatocytes were increased with the addition of the NO synthase cofactor tetrahydrobiopterin, or sepiapterin, which is converted to tetrahydrobiopterin. These results indicate that under in vitro conditions the parasite itself provides a signal that triggers induction of the NO pathway in human and murine hepatocytes and that NO formation in infected hepatocytes is limited by tetrahydrobiopterin availability.

[A low-level stress test, in the early phase of myocardial infarction and its correlation with coronary angiography]. / Prueba de esfuerzo de bajo nivel, en la fase temprana del infarto del miocardio y su correlación con la coronariografía.

In a prospective 24-month trial at the Instituto Nacional de Cardiologia, 56 patients were studied. All patients had acute myocardial infarction (AMI), diagnosed by clinical, electrocardiographic and enzymatic means. They were studied in two groups: Group A with single localized AMI (n = 30) and Group B with AMI at two locations (n = 26); a resting electrocardiogram (EKG) was analyzed in each case and a low level stress test was performed within the 2nd and 3rd postinfarction weeks; coronary angiography was done between the 8th and 9th postinfarction weeks. In Group A the low level stress test (LLST) was positive for ischemia at a distance from the infarction site in 21, and eighteen of them had multi-vessel injuries (MVI); in 9 the LLST was negative; of these 7 had single-vessel injury; only the remaining 2 had MVI (p less than 0.001) with 90% sensitivity and 78% specificity. In Group B there was no significant relationship between LLST and coronary angiography (64% sensitivity, and 62% specificity). Relating the ischemic change at a distance in the resting EKG with coronary angiography, we found 75% sensitivity and 55% specificity in Group A. In Group B, sensitivity and specificity were even lower. We conclude that LLST in the early postinfarction phase in Group A is a safe and reliable method to suspect MVI, allowing the early identification of patients with lesions that could be treated by surgical means.

[Exercise test with cold water intake. Preliminary results]. / Prueba de esfuerzo con ingestión de agua fría. Resultados preliminares.

Thirty healthy individuals with no history of cardiovascular disease were studied to determine the electrocardiographic effects of maximal exercise immediately followed by ingestion of ice water. The subjects were subgrouped according to their training into (A) high (N = 5), (B) moderate (N = 14) and (C) low (N = 11) levels. Electrocardiograms (ECGs) were taken at rest and at rest with ingestion of ice water followed by maximal stress tests. Maximal stress tests were repeated followed by ingestion of ice water at the beginning of and at 2, 3, 6 and 9 minutes of recuperation. The stress test combining maximal effort and ice water ingestion was positive in all members of Group A, in 4 from Group B and in 1 from Group C. A stress test associating maximal effort with ice water ingestion is a useful method of detecting subjects susceptible to changes in ECG which appear to be secondary to coronary spasm. It has a low cost it is simple to perform and represents minimal risk.

[Effects of aerobic physical conditioning on the profile of plasma lipoproteins in a group of healthy volunteers]. / Efectos del acondicionamiento físico aeróbico sobre el perfil de lipoproteinas plasmáticas en un grupo de voluntarios sanos.

To evaluate the effects of aerobic physical conditioning on plasma lipoproteins, we studied 26 previously untrained, apparently healthy, non obese volunteers. All participants underwent a treadmill test performed according to the protocol of Bruce with the direct measurement of maximal oxygen consumption (VO2max). A program of aerobic exercise was prescribed for each volunteer at 70% of their corresponding VO2max. At baseline and at the end of weeks 4, 8 and 12 of the exercise program, cholesterol and triglycerides were measured by enzymatic analysis in total plasma and in the lipoprotein fractions separated by preparative ultracentrifugation and precipitation methods. At the end of week 12, the VO2max measurement was repeated. At the end of the protocol, mean VO2max increased from the value of 39.9 observed at baseline to 94.4 ml/kg/min (p less than 0.01). There were no variations in mean body weight, diet or smoking status of the participants during the exercise program. Cholesterol associated with High-density lipoproteins (C-HDL) increased from 42.5 to 46.1 mg/dl (p less than 0.05). This effect was first noticeable at week 8. We didn't observe significant changes in Total Cholesterol nor the Cholesterol fraction associated with Low-density lipoproteins (C-LDL). Total triglycerides decreased at weeks 4 and 8 but returned to near baseline values at week 12. The C-LDL/C-HDL ratio considered as an index of a high coronary risk decreased from 2.32 at baseline to 2.02 (p less than 0.05) at week 12. Thirteen of the twenty six initial volunteers completed the physical conditioning program as planned, the rest were eliminated at different stages of the protocol due to incomplete adherence to their exercise schedules.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunofluorescent and immunoelectron microscopic localization of protein antigens in red cells infected with the human malaria Plasmodium falciparum.

Erythrocytes infected with the human malaria Plasmodium falciparum produce elevations of the surface membrane of the red cell called knobs. Through the use of transmission electron microscopy and a post-embedding protein A-immunogold technique, it was possible to show changes in the distribution of band 3, glycophorin A and spectrin in the region of the knob. These proteins appeared to be aggregated or condensed in the area of the knob, whereas the remainder of the red cell surface showed no such dense clusters; haemoglobin and the histidine-rich protein of P. lophurae could not be localized to the knobby protuberances. It was not possible to detect any changes in protein distribution using the light microscope and indirect immunofluorescence.

[Response of normal children to the treadmill exercise test using the Bruce protocol]. / Respuesta de los niños sanos a la prueba de esfuerzo en banda sinfin con el protocolo de Bruce.

Fifty nine boys and 41 girls underwent exercise stress testing (ETT), utilizing the Bruce protocol. Their mean age was 10 years. They were grouped by sex, age and body surface area. Blood pressure (BP), heart rate (HR) at rest, during exercise and after were monitored as well as the duration of the test and the energy cost. The HR and-BP had a similar linear relationship in both groups during the different stages of the test. The duration of the test expressed in minutes was 11.8 +/- 1.2 in boys and 10.7 +/- 1.2 in girls (P = 0.001). The oxygen consumption (ML/kg/min) was 45.2 +/- 4.9 and 41.9 +/- 4.5 that is equivalent to 12.9 +/- 1.4 and 11.9 +/- 1.2 mets for each group respectively. The group of boys of 6 (9.8) and 14 years of age (13.6) (P = 0.002) and in the girls in the 7 (9.5) and 10 years age group (11.8) P = 0.05. We conclude that 1) The ETT can be done in children safely but was have to take in consideration their age, sex, and body surface area in evaluating the results. 2) This study gives a reference to evaluate children with an without heart disease.