A real-time polymerase chain reaction assay for the identification and quantification of American Leishmania species on the basis of glucose-6-phosphate dehydrogenase

A real-time polymerase chain reaction (PCR) test was developed on the basis of the Leishmania glucose-6-phosphate dehydrogenase locus that enables identification and quantification of parasites. Using two independent pairs of primers in SYBR-Green assays, the test identified etiologic agents of cutaneous leishmaniasis belonging to both subgenera, Leishmania (Viannia) and Leishmania (Leishmania) in the Americas. Furthermore, use of TaqMan probes enables distinction between L. (V.) braziliensis or L. (V.) peruviania from the other L. (Viannia) species. All assays were negative with DNA of related trypanosomatids, humans, and mice. The parasite burden was estimated by normalizing the number of organisms per total amount of DNA in the sample or per host glyceraldehyde-3-phosphate dehydrogenase copies. The real-time PCR assay for L. (Leishmania) subgenus showed a good linear correlation with quantification on the basis of a limiting dilution assay in experimentally infected mice. The test successfully identifies and quantifies Leishmania in human biopsy specimens and represents a new tool to study leishmaniasis.

Association of soluble tumor necrosis factor receptors 1 and 2 with nephropathy, cardiovascular events, and total mortality in type 2 diabetes

AIMS/HYPOTHESIS: Soluble tumor necrosis factor receptors 1 and 2 (sTNFR1 and sTNFR2) contribute to experimental diabetic kidney disease, a condition with substantially increased cardiovascular risk when present in patients. Therefore, we aimed to explore the levels of sTNFRs, and their association with prevalent kidney disease, incident cardiovascular disease, and risk of mortality independently of baseline kidney function and microalbuminuria in a cohort of patients with type 2 diabetes. In pre-defined secondary analyses we also investigated whether the sTNFRs predict adverse outcome in the absence of diabetic kidney disease. METHODS: The CARDIPP study, a cohort study of 607 diabetes patients [mean age 61 years, 44 % women, 45 cardiovascular events (fatal/non-fatal myocardial infarction or stroke) and 44 deaths during follow-up (mean 7.6 years)] was used. RESULTS: Higher sTNFR1 and sTNFR2 were associated with higher odds of prevalent kidney disease [odd ratio (OR) per standard deviation (SD) increase 1.60, 95 % confidence interval (CI) 1.32-1.93, p < 0.001 and OR 1.54, 95 % CI 1.21-1.97, p = 0.001, respectively]. In Cox regression models adjusting for age, sex, glomerular filtration rate and urinary albumin/creatinine ratio, higher sTNFR1 and sTNFR2 predicted incident cardiovascular events [hazard ratio (HR) per SD increase, 1.66, 95 % CI 1.29-2.174, p < 0.001 and HR 1.47, 95 % CI 1.13-1.91, p = 0.004, respectively]. Results were similar in separate models with adjustments for inflammatory markers, HbA1c, or established cardiovascular risk factors, or when participants with diabetic kidney disease at baseline were excluded (p < 0.01 for all). Both sTNFRs were associated with mortality. CONCLUSIONS/INTERPRETATIONS: Higher circulating sTNFR1 and sTNFR2 are associated with diabetic kidney disease, and predicts incident cardiovascular disease and mortality independently of microalbuminuria and kidney function, even in those without kidney disease. Our findings support the clinical utility of sTNFRs as prognostic markers in type 2 diabetes.

Mixed-signal analog-digital circuits design on the pre-diffused digital array using trapezoidal association of transistors

The mixed-signal and analog design on a pre-diffused array is a challenging task, given that the digital array is a linear matrix arrangement of minimum-length transistors. To surmount this drawback a specific discipline for designing analog circuits over such array is required. An important novel technique proposed is the use of TAT (Trapezoidal Associations of Transistors) composite transistors on the semi-custom Sea-Of-Transistors (SOT) array. The analysis and advantages of TAT arrangement are extensively analyzed and demonstrated, with simulation and measurement comparisons to equivalent single transistors. Basic analog cells were also designed as well in full-custom and TAT versions in 1.0mm and 0.5mm digital CMOS technologies. Most of the circuits were prototyped in full-custom and TAT-based on pre-diffused SOT arrays. An innovative demonstration of the TAT technique is shown with the design and implementation of a mixed-signal analog system, i. e., a fully differential 2nd order Sigma-Delta Analog-to-Digital (A/D) modulator, fabricated in both full-custom and SOT array methodologies in 0.5mm CMOS technology from MOSIS foundry. Three test-chips were designed and fabricated in 0.5mm. Two of them are IC chips containing the full-custom and SOT array versions of a 2nd-Order Sigma-Delta A/D modulator. The third IC contains a transistors-structure (TAT and single) and analog cells placed side-by-side, block components (Comparator and Folded-cascode OTA) of the Sigma-Delta modulator.

An exploratory study on how to improve the economy of American Samoa

Deep in the South Pacific region about 2,300 miles southwest of the Hawaiian islands1 lies a United States territory that many Americans have never heard of nor known anything about. However, some famous Americans such as Troy Polamalu of the Pittsburgh Steelers, semi retired professional wrestler Dwayne “The Rock” Johnson, and Hawaii Congresswoman Tulsi Gabbard have genealogical roots there. More importantly, many of the Territory’s sons and daughters have served and lost their lives for the United States flag and the cause of freedom around the world. This place is called American Samoa, a collection of seven islands that if glued together would have a total landmass of approximately 76 square miles, just a tad bigger than the capital city of the United States. According to the United States Census Bureau, there were 55,519 residents of American Samoa in 2010.1 The majority of them are ethnic Samoans, a Polynesian sect that traces its history back to early migrants from Southeast Asia who settled the islands around 1500 B.C.2 3 The climate is warm all year long and the forests along the mountains are ripe with vegetation. The main island is Tutuila with its beautiful and coveted landlocked harbor that was used as a coaling station by the United States naval ships during World War II. In fact, it was the Pago Pago Harbor that diminished the impact of the 2009 Tsunami that devastated the Samoan islands by channeling the waters of the Pacific Ocean towards the end of the harbor instead of flooding many other villages surrounding the Pago Pago Bay area. Lives and property were destroyed near the end of the Harbor but it could have been worse for the entire Bay area. Locally grown foods include coconut, taro, banana, guava, sugar cane, papaya, yam, pineapple, and breadfruit. It is completely surrounded by the Pacific Ocean from which the locals obtain a variety of seafood. There is a popular saying in Samoa that goes, “In Samoa, it is impossible to starve 1 American Samoa Department of Commerce, 2012 Statistical Yearbook, http://www.doc.as/wpcontent/uploads/2011/06/2012-Statistical-Yearbook-1.pdf 2 U.S. Census Bureau News, U.S. Census Bureau Releases 2010 Census Population Counts for American Samoa, http://www.census.gov/2010census/news/releases/operations/cb11-cn177.html (Aug. 24, 2011). 3 3 J. Robert Shaffer, American Samoa: 100 Years Under the United States Flag (Honolulu, Hawaii: Island Heritage Publishing, 2000), 34. 4 because people live off of the land’s and the ocean’s abundant resources.” To the west of American Samoa lies a larger group of four islands that make up the Sovereign State of Samoa, which became independent from New Zealand in 1962. Samoa and American Samoa share the same language, culture, and religion but are divided by government and political systems. The focus of this study will be on American Samoa, which became a United States territory in 1900 when the principal chiefs of Tutuila (the largest island in American Samoa) ceded the islands to the United States.

Physiological response of American bullfrog tadpoles to stressor conditions of capture and hypoxia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Association of an insulin-like growth factor 1 gene microsatellite with phenotypic variation and estimated breeding values of growth traits in Canchim cattle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Association of acepromazine with propofol in giant amazon turtles Podocnemis expansa reared in captivity

PURPOSE: To evaluate the effects of different concentrations of an anesthetic association in giant amazon turtles (Podocnemis expansa).METHODS: Twenty healthy P. expansa of both sexes weighing between 1.0 and 1.5kg commercially bred in the Araguaia River Valley, Goias, Brazil, were separated into two groups (G1 n=10 and G2 n=10). Each group received a respective protocol: P1=acepromazine (0.5 mg/kg IM) and propofol (5 mg/kg IV) and P2=acepromazine (0.5 mg/kg IM) and propofol (10 mg/kg IV). The acepromazine was administered in the left thoracic member and the propofol in the cervical vertebral sinus. Assessments were made of the anesthetic parameters of locomotion, muscle relaxation, response to pain stimuli in the right thoracic and pelvic members and heartbeat.RESULTS: The anesthetic induction time was the same for both protocols (P1 and P2); however the P2 effects were of a longer duration.CONCLUSION: The sedation achieved with both protocols (P1 and P2) were satisfactory for the biological sample collection, physical examinations and minor surgeries on this species.

Muscle response to the association of statin and physical exercise in rats

Physical exercise and statins, which are recommended for the treatment of dyslipidemia, are independently associated to the occurrence of muscle injury. The objective is analyze the effect of aerobic exercise associated to the use of simvastatin on the morphology of the gastrocnemius muscle. Thirty Wistar rats were divided into six groups, two of which received a standard diet (1 sedentary and 1 exercised) and four (1 sedentary with medication, 1 sedentary without medication, 1 exercised with medication, 1 exercised without medication) received a hypercholesterolemic diet (standard diet with the addition of cholesterol and coconut oil). Simvastatin (20 mg/Kg) was administered five days a week for eight weeks, together with aerobic training on a treadmill (9.75 m/min) for 60 minutes a day. The gastrocnemius muscle was removed, sliced, stained with Hematoxylin-Eosin and submitted to a histochemical reaction to determine mitochondrial activity. The data were analyzed using a paired t-test, analysis of variance and Scheffe's post hoc test (p<0.05). Greater histological alterations were found in the medicated and exercised animals, with a greater frequency of occurrence as well. The histochemical analysis revealed that the medicated groups had fibers with more intensive mitochondrial activity alongside fibers with an absence of reaction. The morphometric analysis revealed no significant differences between groups. It is suggested that simvastatin is a medication that leads to the occurrence of muscle injury and its administration in association with physical activity may exacerbate these injuries. This finding may be related to cellular respiration.

Association of ABO gene mutations resulting in a rare B subgroup

Background and Objectives B subgroups are rare and the genetic analysis reported to date has been limited.Materials and Methods Serological and molecular investigations were performed in blood from a B-subgroup donor.Results Red cells did not react with anti-B and anti-AB reagents. However, cells absorbed anti-B. Red cells presented positive reactions with anti-H, and saliva secreted H substance. The molecular study demonstrated a B allele with the substitutions 467C>T, 646T>A, 681G>A, 771C>T, 796C>A, 803G>C, 829G>A and an 0 allele with the sequence of 002.Conclusions It is probable that the presence in exon 7 of some of the 002 substitutions could have weakened the enzymatic activity of the encoded B transferase.

Analysis of the molecular association of rifampicin with hydroxypropyl-²-cyclodextrin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Association of diabetes and cigarette smoke exposure on the glycemia and liver glycogen of pregnant Wistar rats

Purpose: To evaluate cigarette smoke exposure and/or diabetes association effects on the glycemia and liver glycogen levels of pregnant Wistar rats. Methods: 60 adult rats were randomly distributed into (n= 10/group): non-diabetic exposed to filtered air (G1); non-diabetic exposed to cigarette smoke only before pregnancy (G2); non-diabetic exposed to cigarette smoke before and during pregnancy (G3); diabetic exposed to filtered air (G4); diabetic exposed to cigarette smoke only before pregnancy (G5), and diabetic exposed to cigarette smoke before and during pregnancy (G6). Glycemia was determined at days 0 and 21 of pregnancy. Liver samples were collected for liver glycogen determinations. Results: At day 21 of pregnancy, glycemia was higher in G5 and G6 compared to G4 group. G2 (2.43 +/- 0.43), G3 (3.20 +/- 0.49), G4 (2.62 +/- 0.34), G5 (2.65 +/- 0.27) and G6 groups (1.94 +/- 0.35) presented decreased liver glycogen concentrations compared to G1 (4.20 +/- 0.18 mg/100mg liver tissue) (p<0.05). G5 and G6 groups presented decreased maternal weight gain and litter weight. Conclusions: Severe diabetes and cigarette smoke exposure, alone or associated, caused impairment in liver glycogen storage at term pregnancy. Due to the fact that liver glycogen storages were considered determinant for glucose tolerance, it is relevant to point out a rigid clinical glycemic control and to stop smoking so earlier in pregnancy programming.