Association of Polymorphism rs17576 of the Metalloproteinase 9 Gene with Gestational Diabetes in Euro-Brazilian Pregnant Women.

BACKGROUND: Metalloproteinase 9 (MMP9) is involved in the degradation of extracellular matrix molecules, and its polymorphism rs17576 (Gln279Arg) has been associated with diabetes. We investigated the association of rs17576 in a case-control study with Euro-Brazilian women with gestational diabetes. METHODS: The study group consisted of a total of 262 Euro-Brazilian pregnant women classified as either healthy (n = 131, control) or with GDM (n = 131). Fluorescent probes with real time PCR (TaqMan system) were applied for genotyping. RESULTS: All groups were in Hardy-Weinberg equilibrium. The minor allele frequencies (G-allele) for rs17567 in healthy and GDM women were 27.1% [95% CI, 22 - 32] and 37.4% [95% CI, 32 - 43], p = 0.011, respectively. Genotypic comparison showed a significant difference (p < 0.05) between the groups. CONCLUSIONS: Polymorphism rs17567 was associated with GDM in the studied population and carriers of the G-allele showed an increased risk for gestational diabetes (Odds ratio 1.61; 95% CI, 1.1 - 2.3).

Herbaspirillum rubrisubalbicans, a mild pathogen impairs growth of rice by augmenting ethylene levels.

KEY MESSAGE: Herbaspirillum rubrisubalbicans decreases growth of rice. Inoculation of rice with H. rubrisubalbicans increased the ACCO mRNA levels and ethylene production. The H. rubrisubalbicans rice interactions were further characterized by proteomic approach. Herbaspirillum rubrisubalbicans is a well-known growth-promoting rhizobacteria that can also act as a mild phyto-pathogen. During colonisation of rice, RT-qPCR analyses showed that H. rubrisubalbicans up-regulates the methionine recycling pathway as well as phyto-siderophore synthesis genes. mRNA levels of ACC oxidase and ethylene levels also increased in rice roots but inoculation with H. rubrisubalbicans impaired growth of the rice plant. A proteomic approach was used to identify proteins specifically modulated by H. rubrisubalbicans in rice and amongst the differentially expressed proteins a V-ATPase and a 14-3-3 protein were down-regulated. Several proteins of H. rubrisubalbicans were identified, including the type VI secretion system effector Hcp1, suggesting that protein secretion play a role colonisation in rice. Finally, the alkyl hydroperoxide reductase, a primary scavenger of endogenous hydrogen peroxide was also identified. Monitoring the levels of reactive oxygen species in the epiphytic bacteria by flow cytometry revealed that H. rubrisubalbicans is subjected to oxidative stress, suggesting that the alkyl hydroperoxide reductase is an important regulator of redox homeostasis in plant-bacteria interactions.

Utilization of carbon sources by clinical isolates of Aeromonas.

Bacteria in the genus Aeromonas are primarily aquatic organisms; however, some species can cause diseases in humans, ranging from wound infections to septicemia, of which diarrhea is the most common condition. The ability to use a variety of carbon substrates is advantageous for pathogenic bacteria. Therefore, we used Biolog GN2 microplates to analyze the ability of 103 clinical, predominantly diarrheal, isolates of Aeromonas to use various carbon sources, and we verified whether, among the substrates metabolized by these strains, there were some endogenous to the human intestine. The results indicate that Aeromonas present great diversity in the utilization of carbon sources, and that they preferentially use carbohydrates and amino acids as carbon sources. Among the carbon sources metabolized by Aeromonas in vitro, some were found to be components of intestinal mucin, including aspartic acid, glutamic acid, l-serine, galactose, N-acetyl-glucosamine, and glucose, which were used by all strains tested. Additionally, mannose, d-serine, proline, threonine, and N-acetyl-galactosamine were used by several strains. The potential to metabolize substrates endogenous to the intestine may contribute to Aeromonas' capacity to grow in and colonize the intestine. We speculate that this may help explain the ability of Aeromonas to cause diarrhea.

The GCKR Gene Polymorphism rs780094 is a Risk Factor for Gestational Diabetes in a Brazilian Population.

BACKGROUND: The glucokinase regulatory protein (GCKR) regulates the activity of the glucokinase (GCK), which plays a key role in glucose homeostasis. Genetic variants in GCK have been associated with diabetes and gestational diabetes (GDM). Due to the relationship between GCKRP and GCK, polymorphisms in GCKR are also candidates for genetic association with GDM. The aim of this study was to evaluate the association between the GCKR rs780094 polymorphism and GDM in a Brazilian population. METHODS: 252 unrelated Euro-Brazilian pregnant women were classified as control (healthy pregnant women, n = 125) and GDM (pregnant women with GDM, n = 127) age-matched groups. Clinical and anthropometric data were obtained from all subjects. The GCKR rs780094 polymorphism was genotyped using fluorescent probes (TaqMan® , code C_2862873_10). RESULTS: Both groups were in Hardy-Weinberg equilibrium. The GCKR rs780094 polymorphism was associated with GDM in codominant and dominant models (P = 0.022 and P = 0.010, respectively). The minor allele (T) frequency for the control group in the study was 38.4% (95% CI: 32-44%), similar to frequencies reported for other Caucasian populations. CONCLUSION: Carriers of the C allele of rs780094 were 1.41 (odds ratio, 95% CI, 0.97-2.03) times more likely to develop GDM.

Functional promoter polymorphisms of the receptor for advanced glycation end products in children and adolescents with type 1 diabetes.

RAGE promoter polymorphisms are associated with increases in RAGE expression. A case-control association study was conducted involving a Euro-Brazilian population of children and adolescents with type 1 diabetes (n = 90) and healthy controls (n = 105), which were matched by sex and age. Genotyping by PCR-RFLP the -429T>C (rs1800625), -374T>A (rs1800624), and 63 bp deletion/insertion (-407 to -345 bp) showed no significant differences (P > 0.05) between the groups.

Comparative proteomics analysis of the rice roots colonized by Herbaspirillum seropedicae strain SmR1 reveals induction of the methionine recycling in the plant host.

Although the use of plant growth-promoting bacteria in agriculture is a reality, the molecular basis of plant-bacterial interaction is still poorly understood. We used a proteomic approach to study the mechanisms of interaction of Herbaspirillum seropedicae SmR1 with rice. Root proteins of rice seedlings inoculated or noninoculated with H. seropedicae were separated by 2-D electrophoresis. Differentially expressed proteins were identified by MALDI-TOF/TOF and MASCOT program. Among the identified proteins of H. seropedicae, the dinitrogenase reductase NifH and glutamine synthetase GlnA, which participate in nitrogen fixation and ammonium assimilation, respectively, were the most abundant. The rice proteins up-regulated included the S-adenosylmethionine synthetase, methylthioribose kinase, and acireductone dioxygenase 1, all of which are involved in the methionine recycling. S-Adenosylmethionine synthetase catalyzes the synthesis of S-adenosylmethionine, an intermediate used in transmethylation reactions and in ethylene, polyamine, and phytosiderophore biosynthesis. RT-qPCR analysis also confirmed that the methionine recycling and phytosiderophore biosynthesis genes were up-regulated, while ACC oxidase mRNA level was down-regulated in rice roots colonized by bacteria. In agreement with these results, ethylene production was reduced approximately three-fold in rice roots colonized by H. seropedicae. The results suggest that H. seropedicae stimulates methionine recycling and phytosiderophore synthesis and diminishes ethylene synthesis in rice roots.

Prospection of plasma proteins as biomarkers for diabetes mellitus monitoring.

PURPOSE: This prospective study aimed to detect and identify plasma proteins differentially expressed between groups of Brazilian diagnosed with type 1 (T1DM), type 2 (T2DM) diabetes with good and poor glycemic control and the non-diabetic group denominated control group (CG). METHODS: Patients with T1DM and T2DM were subdivided according to their glycated haemoglobin (HbA1c) level: ≥ 53 mmol/mol and < 53 mmol/mol. Each subgroup was composed of ten subjects (n = 10). The plasma from each subgroup was pooled and depleted of albumin and IgG. The reminiscent proteins were quantified and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The relative volume of protein bands was determined by densitometry analysis, and those with differential abundance were identified by MALDI-TOF mass spectrometry. RESULTS: Alpha 2 - Macroglobulin (AMG) was 1.3-fold more abundant in T1DM with HbA1c ≥ 53 mmol/mol and < 53 mmol/mol and 1.4-fold more abundant in T2DM with HbA1c ≥ 53 mmol/mol compared to CG. Ceruloplasmin (Cp) and Haptoglobin (Hp) were overexpressed above 1.5-fold in all DM subgroups. Cp in T1DM and Hp in both types of DM were more expressed in HbA1c ≥ 53 mmol/mol than <53 mmol/mol. Apolipoprotein A-I (Apo A-I) was upregulated only in T2DM subgroups. CONCLUSION: In summary, three positive acute-phase proteins, AMG, Cp and Hp were more abundant in diabetic individuals regardless of the diabetes type. The highest Hp abundance in both types of DM with HbA1c ≥ 53 mmol/mol, reinforces Hp as a possible biomarker associated with diabetic complications.

Virulence characteristics and antimicrobial resistance of Aeromonas veronii biovar sobria 312M, a clinical isolate.

Aeromonas are bacteria widely distributed in the environment, and some species are able to cause infections in humans, of which diarrhea is the most common. The objective of this study was to evaluate the presence of virulence and antimicrobial resistance associated characteristics in A. veronii biovar sobria strain 312M isolated from diarrheal stools. For this, the genome sequencing and phenotypical tests were performed. The draft genome annotation revealed several complete pathways associated with carbon metabolism and a mucin-desulfating sulfatase which may contribute to intestine colonization, and a large number of virulence-associated genes encoding structures associated with adhesion, toxins, and secretion systems. The strain exhibited swimming and swarming motility, biofilm formation, and hemolytic activity. It was resistant to ampicillin, ampicillin/sulbactam, and amoxicillin-clavulanic acid. Although a cphA gene encoding a narrow-spectrum carbapenase was identified in the strain genome, no carbapenemase activity was detected in the antimicrobial susceptibility test. When compared with other A. veronii with complete genomes, the main differences in virulence characteristics are related to lateral flagella and type III and VI secretion systems; the antimicrobial resistance spectrum also varied among strains. The results indicated that A. veronii biovar sobria 312M presents high virulence potential and resistance to limited classes of antimicrobials.

d-GDM: A mobile diagnostic decision support system for gestational diabetes.

OBJECTIVE: The aim of the study is to describe a portable and convenient software to facilitate the diagnostics of gestational (GDM) and pre-gestational diabetes (PGDM). MATERIALS AND METHODS: An open source software, d-GDM, was developed in Java. The integrated development environment Android Studio was used as the Android operational system. The software for GDM diagnosis uses the criteria endorsed by the International Association of Diabetes and Pregnancy Study Group, modified by the World Health Organization. RESULTS: GDM diagnosis criteria is not simple to follow, therefore, errors or inconsistencies in diagnosis are expected and could delay the appropriate treatment. The d-GDM, was developed to assist GDM diagnosis with precision and consistency diagnostic reports. The open source software can be manipulated conveniently. The operator requires information regarding the gestational period and selects the appropriate glycaemic marker options from the menu. During operation, pressing the button "diagnosticar" on the screen will present the diagnosis and information for the follow up. d-GDM is available in Portuguese or English and can be downloaded from the Google PlayStore. A responsive web version of d-GDM is also available. The usefulness and accuracy of d-GDM was verify by field tests involving 22 subjects and 5 mobile phone brands. The approval regards user-friendliness and efficiency were 95% or higher. The GDM diagnosis were 100% correct, in this pilot test. d-GDM is a user-friendly, free software for diagnosis that was developed for mobile devices. It has the potential to contribute and facilitate the diagnosis of gestational diabetes for healthcare professionals.

Modulation of defence and iron homeostasis genes in rice roots by the diazotrophic endophyte Herbaspirillum seropedicae.

Rice is staple food of nearly half the world's population. Rice yields must therefore increase to feed ever larger populations. By colonising rice and other plants, Herbaspirillum spp. stimulate plant growth and productivity. However the molecular factors involved are largely unknown. To further explore this interaction, the transcription profiles of Nipponbare rice roots inoculated with Herbaspirillum seropedicae were determined by RNA-seq. Mapping the 104 million reads against the Oryza sativa cv. Nipponbare genome produced 65 million unique mapped reads that represented 13,840 transcripts each with at least two-times coverage. About 7.4% (1,014) genes were differentially regulated and of these 255 changed expression levels more than two times. Several of the repressed genes encoded proteins related to plant defence (e.g. a putative probenazole inducible protein), plant disease resistance as well as enzymes involved in flavonoid and isoprenoid synthesis. Genes related to the synthesis and efflux of phytosiderophores (PS) and transport of PS-iron complexes were induced by the bacteria. These data suggest that the bacterium represses the rice defence system while concomitantly activating iron uptake. Transcripts of H. seropedicae were also detected amongst which transcripts of genes involved in nitrogen fixation, cell motility and cell wall synthesis were the most expressed.

[Light and hens as attraction factors of Nyssomyia whitmani in a rural area, Southern Brazil]. / Luz e galinhas como fatores de atração de Nyssomyia whitmani em ambiente rural, Paraná, Brasil.

OBJECTIVE: To verify the influence of traps with electric light and hens as factors that attract sandflies and compare results between capture methods. METHODS: The study was conducted in the Palmital Farm, Southern Brazil. Sandfly collections were conducted with Falcão traps and an electric aspirator, fortnightly, between 8 p.m. and 11 p.m. in the presence or absence of light and hens in peridomiciliary areas, from September 1998 to June 1999. RESULTS: A total of 43,767 specimens from eight species of sandflies were collected: Nyssomyia whitmani, N. neivai and Migonemyia migonei constituting 99.9% of the total collected, with predominance of N. whitmani. The number of this species collected inside the hen's shed in the presence of hens (21,045) was greater than in their absence (10,434). In the presence of hens, with distinct intensities of light, a larger number of N. whitmani samples were collected with 3W light. In the presence of hens and light (3W), the number of N. whitmani collected with the electric aspirator (5,141) was superior to that collected with the Falcão trap (1,675). In the absence of light, with or without the presence hens, there was no difference between the numbers of N. whitmani collected with the electric aspirator or the Falcão trap. CONCLUSIONS: Hens and electric light together attract more N. whitmani to peridomicilary areas. The number of N. whitmani collected with an electric aspirator inside a hen's shed with the presence of hens and light is greater than those collected with a Falcão trap in the same conditions.

Serum Fluorescent Advanced Glycation End (F-AGE) products in gestational diabetes patients.

OBJECTIVES: Advanced glycation end products (AGEs) are involved in the pathogenesis and complications of diabetes mellitus (DM). Gestational DM (GDM) is characterized by increased glycemia and oxidative stress, which are factors associated with high serum AGE concentrations. The aim of this study was to evaluate the utility of a serum fluorescence AGE (F-AGE) method as a screening tool for gestational diabetes. SUBJECTS AND METHODS: Serum samples from 225 GDM patients and 217 healthy pregnant women (healthy controls) were diluted 50-fold in phosphate-buffered saline, and the AGEs were estimated by fluorometric analysis (λEx 350 nm/ λEm 440 nm). RESULTS: No significant (P > 0.05) differences in AGE concentrations, expressed in Arbitrary Units (UA/mL × 104), were observed in the women with GDM or in the healthy controls. Furthermore, F-AGE concentrations did not change significantly during the pregnancy (12-32 weeks of gestation). Only the GDM group had a positive correlation (r = 0.421; P < 0.001) between F-AGEs and serum creatinine concentrations. CONCLUSION: It was not possible to distinguish women with gestational diabetes from the healthy controls on the basis of serum F-AGE concentrations.

Type 2 diabetes-associated genetic variants of FTO, LEPR, PPARg, and TCF7L2 in gestational diabetes in a Brazilian population.

OBJECTIVE: Gestational diabetes mellitus (GDM) is a metabolic disorder that shares pathophysiologic features with type 2 diabetes mellitus. The aim of this study was to investigate the association of the polymorphisms fat mass and obesity-associated (FTO) rs1421085, leptin receptor (LEPR) rs1137100, rs1137101, peroxisome proliferator-activated receptor gamma (PPARg) rs1801282, and transcription factor 7-like 2 (TCF7L2) rs7901695 with GDM. SUBJECTS AND METHODS: 252 unrelated Euro-Brazilian pregnant women were classified into two groups according to the 2015 criteria of the American and Brazilian Diabetes Association: healthy pregnant women (n = 125) and pregnant women with GDM (n = 127), matched by age. The polymorphisms were genotyped using fluorescent probes (TaqMan®). RESULTS: All groups were in Hardy-Weinberg equilibrium. The genotype and allele frequencies of the studied polymorphisms did not show significant differences between the groups (P > 0.05). In the healthy and GDM groups, the C allele frequencies (95% CI) of the FTO rs1421085 polymorphism were 36.8% [31-43%] and 35.0% [29-41%]; the G allele frequencies (95% CI) of the LEPR rs1137100 polymorphism were 24.8% [19-30%] and 22.8% [18-28%]; the G allele frequencies (95% CI) of the LEPR rs1137101 polymorphism were 43.6% [37-50%] and 42.9% [37-49%]; the G allele frequencies (95% CI) of the PPARg rs1801282 polymorphism were 7.6% [4-11%] and 8.3% [5-12%]; and the C allele frequencies (95% CI) of the TCF7L2 rs7901695 polymorphism were 33.6% [28-39%] and 39.0% [33-45%], respectively. CONCLUSION: The studied polymorphisms were not associated with GDM in a Brazilian population.

The rs10885122 polymorphism of the adrenoceptor alpha 2A (ADRA2A) gene in Euro-Brazilians with type 2 diabetes mellitus.

OBJECTIVE: To investigate the association of the rs10885122G>T polymorphism in the ADRA2A gene in a Euro-Brazilian sample of healthy (controls) and type 2 diabetic (T2D) subjects. SUBJECTS AND METHODS: We used fluorescent probes (TaqMan) to genotype 241 subjects, that is, 121 healthy and 120 T2D subjects, who were classified based on the Brazilian Diabetes Association (2013) and American Diabetes Association (2014) criteria. RESULTS: The genotype and allele frequencies showed no significant (P > 0.05) difference between the two studied groups. The minor allele (T) frequencies (95%CI) for rs10885122 were 19% (14-24%) and 20% (15-26%) for healthy and T2D groups, respectively. Carriers of the T allele (genotypes GT+TT) were significantly associated (P = 0.016) with approximately a 7-kg body weight reduction compared with the genotype GG, which was only found in the T2D group. CONCLUSION: The rs10885122G>T polymorphism of the ADRA2A gene was not associated with T2D in Euro-Brazilians, and carriers of the T allele had lower body weight in the presence of T2D.

Polymorphisms in FTO and TCF7L2 genes of Euro-Brazilian women with gestational diabetes.

OBJECTIVE: To investigate the association between fat mass and obesity-associated (FTO) gene polymorphisms rs8050136C>A and rs9939609T>A, and transcription factor 7-like 2 (TCF7L2) gene polymorphisms rs12255372G>T and rs7903146C>T, in a sample group of pregnant Euro-Brazilian women with or without gestational diabetes mellitus (GDM). METHODS: Subjects were classified as either healthy pregnant control (n=200) or GDM (n=200) according to the 2010 criteria of the American Diabetes Association. The polymorphisms were genotyped using fluorescent probes (TaqMan®). RESULTS: All groups were in the Hardy-Weinberg equilibrium. The genotype and allele frequencies of the examined polymorphisms did not exhibit significant difference (P>0.05) between the groups. In the healthy and GDM pregnant women groups, the A-allele frequencies (95% CI) of FTO polymorphisms rs8050136 and rs9939609 were 39% (34-44%); 38% (33-43%) and 40% (35-45%); 41% (36-46%), respectively; and the T-allele frequencies of TCF7L2 polymorphisms rs12255372 and rs7903146 were 30% (26-35%), 32% (27-37%) and 29% (25-34%), 36% (31-41%), respectively. CONCLUSION: The examined polymorphisms were not associated with GDM in the Euro-Brazilian population studied.

[Influence of rearrangement and cleaning of the peridomiciliary area and building disinsectization on sandfly population density in the municipality of Doutor Camargo, Paraná State, Brazil]. / Influência da reorganização, da limpeza do peridomicílio e a da desinsetização de edificações na densidade populacional de flebotomíneos no Município de Doutor Camargo, Estado do Paraná, Brasil.

Phlebotomine sandfly captures were performed in the domicile, domestic animal shelters, and peridomicile before and after rearrangement, cleaning, and disinsectization. Phlebotomine sandflies were captured with Falcão traps from October 1998 to April 2000 in the Recanto Marista district, Doutor Camargo municipality, Paraná State. The results were compared with those from October 1996 to September 1997. In this survey, 122,616 phlebotomine sandflies were captured using 10 traps, as compared to 157,983 in the previous period, with 6 traps. Lutzomyia neivai predominated in both periods. During the former period an hourly average of 1,641.9 phlebotomine sandflies were captured as compared to 806.7 in the latter. The results with four traps installed in the same locations shows that the hourly average was 1,318.8 in the former period and 156.0 in the latter. The large reduction in the number of phlebotomine sandflies captured in the second period may be due to the impact of alterations in the environment and the disinsectization of the buildings after the first period.

[Ecology of Lutzomyia (Nyssomyia) whitmani in an urban area in Maringá, Paraná, Brazil]. / Ecologia de Lutzomyia (Nyssomyia) whitmani em área urbana do município de Maringá, Paraná.

OBJECTIVES: To study the fauna, hours of activity and seasonality of phlebotomines in forest animal habitats in the Ingá Park, on the urban perimeter of the municipality of Maringá, Paraná State, Brazil. METHODS: Phlebotomines were collected on the urban perimeter using Shannon traps from 6:00 p.m. to 6:00 a.m. and Falcão traps from 8:00 p.m. to 12:00 midnight, on two nights per month from October 1998 to September 2000. The Shannon traps were installed close to forest animal habitats and on the banks of the Moscados stream; the Falcão traps were placed in shelters for captive birds, forest mammals and reptiles. RESULTS: The Shannon traps yielded 13,656 phlebotomines, with predominance of Lutzomyia whitmani (98.7%). The Falcão traps yielded 4,040 phlebotomines, with L. whitmani again predominating (96.3%). L. whitmani was most frequently found between 12:00 midnight and 2:00 a.m. in the Shannon traps. This species was most frequently collected in the months of August (55.1%) in the Shannon traps, and in March 1999 (19.2%), November 1999 (15.9%) and August 2000 (20.6%) in the Falcão traps. More phlebotomines were collected in Falcão traps installed in shelters for captive mammals (84.0%). CONCLUSION: There was a clear predominance of L. whitmani in the Ing Park. The greatest frequency of L. whitmani was in August from Shannon traps, and in March and November from Falcão traps. The peak activity of L. whitmani took place between 12:00 midnight and 2:00 a.m.

d-GDM: A mobile diagnostic decision support system for gestational diabetes

ABSTRACT Objective The aim of the study is to describe a portable and convenient software to facilitate the diagnostics of gestational (GDM) and pre-gestational diabetes (PGDM). Materials and methods An open source software, d-GDM, was developed in Java. The integrated development environment Android Studio was used as the Android operational system. The software for GDM diagnosis uses the criteria endorsed by the International Association of Diabetes and Pregnancy Study Group, modified by the World Health Organization. Results GDM diagnosis criteria is not simple to follow, therefore, errors or inconsistencies in diagnosis are expected and could delay the appropriate treatment. The d-GDM, was developed to assist GDM diagnosis with precision and consistency diagnostic reports. The open source software can be manipulated conveniently. The operator requires information regarding the gestational period and selects the appropriate glycaemic marker options from the menu. During operation, pressing the button "diagnosticar" on the screen will present the diagnosis and information for the follow up. d-GDM is available in Portuguese or English and can be downloaded from the Google PlayStore. A responsive web version of d-GDM is also available. The usefulness and accuracy of d-GDM was verify by field tests involving 22 subjects and 5 mobile phone brands. The approval regards user-friendliness and efficiency were 95% or higher. The GDM diagnosis were 100% correct, in this pilot test. d-GDM is a user-friendly, free software for diagnosis that was developed for mobile devices. It has the potential to contribute and facilitate the diagnosis of gestational diabetes for healthcare professionals.