Relation between mast cells concentration and serotonin expression in chagasic megacolon development.

Chagas' disease is still reaching about 10 million people in the world. In South America, one of the most severe forms of this disease is the megacolon, characterized by severe constipation, dilated sigmoid colon and rectum and severe malnutrition. Previous data suggested that mast cells and serotonin (5-hydroxytryptamine [5-HT]) expression could be involved in intestinal homeostasis control, avoiding the chagasic megacolon development. The aim at this study was to characterize the presence of mast cells and expression of serotonin in chagasic patients with and without megacolon and evaluate the relation between mast cells, serotonin and megacolon development. Our results demonstrated that patients without megacolon feature a large amount of serotonin and few mast cells, while patients with megacolon feature low serotonin expression and a lot of mast cells. We believe that serotonin may be involved in the inflammatory process control, triggered by mast cells, and the presence of this substance in large quantities of the intestine could represent a mechanism of megacolon prevention.

Genomic and physiological characterization of Kitasatospora sp. nov., an actinobacterium with potential for biotechnological application isolated from Cerrado soil.

An Actinobacteria - Kitasatospora sp. K002 - was isolated from the soil of Cerrado, a savanna-like Brazilian biome. Herein, we conducted a phylogenetic, phenotypic and physiological characterization, revealing its potential for biotechnological applications. Kitasatospora sp. K002 is an aerobic, non-motile, Gram-positive bacteria that forms grayish-white mycelium on solid cultures and submerged spores with vegetative mycelia on liquid cultures. The strain showed antibacterial activity against Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli. Genomic analysis indicated that Kitasatospora xanthocidica JCM 4862 is the closest strain to K002, with a dDDH of 32.8-37.8% and an ANI of 86.86% and the pangenome investigations identified a high number of rare genes. A total of 60 gene clusters of 22 different types were detected by AntiSMASH, and 22 gene clusters showed low similarity (< 10%) with known compounds, which suggests the potential production of novel bioactive compounds. In addition, phylogenetic analysis and morphophysiological characterization clearly distinguished Kitasatospora sp. K002 from other related species. Therefore, we propose that Kitasatospora sp. K002 should be recognized as a new species of the genus Kitasatospora - Kitasatospora brasiliensis sp. nov. (type strains = K002).

Association between IGF2 and CYP21 gene polymorphisms and characteristics of economic interest in Nellore cattle.

We analyzed two single nucleotide polymorphisms (SNPs) of the IGF2 and CYP21 genes in Nellore cattle participating in the Brazilian Animal Breeding Program. The SNPs were found in exon 6 of the IGF2 (insulin-like growth factor 2) gene (RFLP/MboII) as well as in the promoter region of the CYP21 (steroid 21-hydroxylase) gene (RFLP/HpaII) of these animals. The TC heterozygotes were significantly more frequent than CC and TT homozygotes in the RFLP/MboII polymorphism. The T allele was significantly more frequent than the C allele in RFLP/HpaII polymorphism. This population was found to be in Hardy-Weinberg equilibrium for these SNPs. Association of these polymorphisms with expected progeny differences of reproductive and productive traits was investigated, but proved to be significant only for DP550 (expected progeny differenced for weight at 365 days - IGF2 - RFLP/MboII) and DP450 (expected progeny differenced for weight at 450 days - CYP21 - RFLP/HpaII). This is the first study on the occurrence of these two polymorphisms in this Zebu breed of cattle. A total of 147 Nellore animals participating in the Breeding Program of the Nellore Breed (PMGRN) under the management of the National Association of Breeders and Researchers (ANCP) in the city of Ribeirão Preto were analyzed.

Identification of a DNA methylation point in the promoter region of the bovine CYP21 gene.

The CYP21 (steroid 21-hydroxylase) gene is involved in the synthesis of steroid hormones. Bov-A2 is a retroposon that is common in ruminant genomes. The promoter region of bovine CYP21 contains a short interspersed nucleotide element of Bov-A2, which overlaps a putative Sp1 binding site. We looked for RFLP/HpaII polymorphism in the Bov-A2 element in bovine Zebu breeds by PCR-RFLP, and examined whether polymorphism in this element is associated with methylation. Among DNA samples from 135 Brazilian Zebu breed cattle, we identified an RFLP/HpaII polymorphism (T/C), which, based on a restriction methylation-sensitive assay employing HpaII and isoschizomer MspI enzymes (methylation-sensitive and -non-sensitive enzymes, respectively), appears to be a DNA methylation point. This is the first report of this polymorphism and on DNA methylation in the bovine CYP21 promoter region in Brazilian Zebu cattle.

The applicability of fingernail lead and cadmium levels as subchronic exposure biomarkers for preschool children.

Preschool children are exposed daily to metals in their homes and at daycare centers (DCC). Metal exposure and health effects are associated even at low levels, and children comprise a group of public health concern. Nail metals have been studied for exposure biomonitoring and compared to other biological media. The aim of this study was to explore the applicability of preschool fingernail lead and cadmium as subchronic exposure biomarkers. Nail lead and cadmium levels (NLL and NCL) of 602 preschool children (age: 1-4 years) who attended 21 DCC in São Paulo, Brazil, in 2013 were analyzed. Results were compared against blood lead and cadmium levels (BLL and BCL) found in a previous study. Inductively coupled plasma mass spectrometry (ICP-MS) analyses were performed for both samples. DCC and metal contaminated sites (MCS) were georeferencing. Logistic regression tests were applied to verify associations between nail metal levels and risk factors (sex, age, maternal education, secondary smoking, DCC geographic district, vehicle flow density, relative altitude and distance between DCC and nearest MCS) (p < 0.05). BLL was stratified by exposure level (low: <5 µg.dL-1; high: >13.9 µg.dL-1; medium: ≥5 µg.dL-1 and ≤13.9 µg.dL-1) and also tested the associations in order to verify if nail lead levels are affected by exposure intensity defined by blood metals concentrations. Radius distance and relative altitude of DCC to nearest MCS were associated with high NLL and NCL. Abnormal appearance of nails was associated with high NLL and low NCL. Lead and cadmium exposure magnitude had no significant impact on NLL. NLL should only be used for initial screening, and when financial resources are scarce, especially in areas located near contaminated sites. Preschool children were co-exposed to both lead and cadmium, reinforcing the need for broader studies evaluating exposure to environmental pollutants for more than one chemical element.

Association of coronary calcification with prognosis of Covid-19 patients without known heart disease.

Risk factors that determine the severity of Covid-19 have not been fully elucidated. The aim of this study was to evaluate the role of coronary artery calcification (CAC) as a risk factor for death or mechanical ventilation (MV) of patients without known heart disease infected with Covid-19. We analyzed 283 consecutive in-patients with acute respiratory symptoms with chest computed tomography (chest-CT), without previous heart disease, and criteria for Covid-19 (RT-PCR positive and/or typical clinical and chest-CT findings). CAC was classified by the number of coronary segments affected as absent (0), mild (1-3), and severe calcification (more than 3). The association between CAC, CAC severity, and death or MV due to severe respiratory failure was assessed by logistic regression. The mean age was 58.7±15.7 years and 54.1% were men. Patients with CAC were older, more likely to have hypertension, and less likely to be obese. CAC was present in 75 patients (26.5%), of which 42 had a mild calcification and 33 had severe calcification, and was associated with death (OR=2.35, 95%CI: 1.01-5.48) or MV (OR=2.72, 95%CI: 1.20-6.20) adjusted for multiple confounders, with significant and increased odds ratio for the severe form of CAC (death: OR=3.70, 95%CI: 1.20-11.42; MV: OR=3.30, 95%CI: 1.09-9.95). We concluded that CAC was an independent risk factor for death or MV in Covid-19 patients without previous heart disease, particularly for those with severe calcification. CAC can be easily visualized on common chest-CT, widely used in evaluation of moderate to severe Covid-19.

Intra-annual variation in rainfall and it's influence of the adult's Cyprideis spp (Ostracoda, Crustacea) on a eutrophic estuary (Guanabara Bay, Rio de Janeiro, Brazil).

Spatial and temporal distribution of two species of adult´s ostracods (Cyprideis sp. and Cyprideis salebrosa) were studied as a function of the rainfall patterns in the Guanabara Bay, Rio de Janeiro, Brazil. Samples were taken in ten stations, along six surveys representing three periods (Dry, Early and Late Rainy) for two years. Stations were nested in four areas (Outer, Central, EPA Guapimirim and Impacted). The bottom water (temperature, salinity, dissolved oxygen and oxygen saturation) were measured in each area to characterize the influence of seasonal variations by rainfall. Cyprideis sp. and Cyprideis salebrosa showed patterns distribution to seasonality/surveys (p = 0,002 and p <0,001, respectly). The spatial distribution of Cyprideis sp was significantly different areas studied (p <0.001) indicated well defined areas and distribution in along of the surveys. However, C. salebrosa showed homogeneous distribution in along of the areas within of each survey (p <0.001). Redundancy Analysis (RDA) for the two years evidenced environment preference of the Cyprideis sp. for areas with marine conditions (high influence for channel central) and C. salebrosa for brackish water (high influence of the rivers). This observation reinforces of the existence of areas created by the seasonality of pluviometric regime, a possible dispersion of the adult's ostracods and possibility the use with bioindicators.

How likely is Brazil to achieve its NDC commitments in the energy sector? A review on Brazilian low-carbon energy perspectives.

This paper offers perspectives on the development of low-carbon energy technology in Brazil, pinpointing changes that have occurred since our former publication in 2011. It takes a fresh approach in terms of how likely Brazil will achieve its Nationally Determined Contributions Commitments in the energy sector. Many countries have implemented national climate policies to accomplish their pledged NDC and contribute to the temperature objectives of the Paris Agreement on climate change. Based on official reports and databases of energy development projections in Brazil and the socioeconomic context, we discuss what can be expected for the future of the Brazilian energy sector, the probability of implementing selected technologies, and the prospects of reaching the NDC targets for 2025 and 2030. In addition, this paper provides an overview of the current stage of development of these technologies, main directions, and bottlenecks in Brazil. Analyses have shown that the Brazilian renewable matrix tends to remain significant, driven by the development of solar and mostly small hydroelectric power sources, as well as different types of biomass. In addition, the system will include the replacement of thermoelectric plants powered by diesel and fuel oil by natural gas plants. The prospects for Brazil's official energy plan for 2027 are aligned with the reference technology scenario, which represents the business as usual scenario. Despite this, low-carbon technologies could be implemented far beyond the NDC's goals, given the abundance of renewable natural resources in the country.

Neuronal plasticity of the enteric nervous system is correlated with chagasic megacolon development.

Chagas' disease is one of the few functional gastrointestinal disorders for which a causative agent has been identified. However, some pathological aspects of the chagasic megasyndromes are still incompletely understood. Chagasic megacolon is characterized by an inflammatory process, organ dilatation and neuronal reduction in both plexuses of the enteric nervous system (ENS). Although some studies on the ENS in Chagas' disease have been performed, the process of neuronal destruction and neuronal regeneration still remains unclear. Our hypothesis is that the regeneration process of the ENS may be involved with the mechanisms that prevent or retard organ dilatation and chagasic megacolon development. For that reason, we evaluated the neuronal regeneration with the marker GAP-43 in the colon's neuronal plexuses from chagasic patients with megacolon, and from non-infected individuals. Visual examination and quantitative analysis revealed an increased neuronal regeneration process in the dilated portion from chagasic patients when compared with the non-dilated portion and with non-infected individuals. We believe that this increased regeneration can be interpreted as an accentuated neuronal plasticity that may be a response of the ENS to avoid megacolon propagation to the entire organ and maintain the colon functional innervation.

IGF2/Mboll polymorphism in Gir and Nelore cattle.

The imprinted gene insulin-like growth factor 2 (IGF2) carries out important functions in the development of placental mammals, during the embryonic and fetal stages and is located on bovine chromosome 29. The main aim of this study was to verify the occurrence of a polymorphism (C/T transversion) in exon 6 of the IGF2 gene in Gir and Nelore cattle, Zebu breeds of great economic importance in Brazil for dairy and beef production, respectively. A 193 bp fragment of bovine IGF2 exon 6 was amplified by PCR with specific primers for this region and digested with Mboll enzyme to analyze the polymorphic segment. In a total of 39 Gir animals (29 females and 10 males), the estimated frequencies of the C and T alleles were 0.42 and 0.58, respectively. The genotypic frequencies were 0.10 for CC, 0.26 for TT and 0.64 for CT. For 28 Nelore animals (17 females and 11 males), the estimated frequencies of the C and T alleles were 0.43 and 0.57, respectively. The genotypic frequencies were 0.18 for CC, 0.32 for TT, and 0.50 for CT. This is the first report of the occurrence of this polymorphism in these breeds. The polymorphic site can be a regulatory motif with functional significance to gene regulation of the IGF2 gene. This polymorphism could be used to investigate the allele-specific expression of the IGF2 gene, its epigenetic status, and its role in developmental, growth and reproductive traits.

Mirna biogenesis pathway is differentially regulated during adipose derived stromal/stem cell differentiation.

Stromal/stem cell differentiation is controlled by a vast array of regulatory mechanisms. Included within these are methods of mRNA gene regulation that occur at the level of epigenetic, transcriptional, and/or posttranscriptional modifications. Current studies that evaluate the posttranscriptional regulation of mRNA demonstrate microRNAs (miRNAs) as key mediators of stem cell differentiation through the inhibition of mRNA translation. miRNA expression is enhanced during both adipogenic and osteogenic differentiation; however, the mechanism by which miRNA expression is altered during stem cell differentiation is less understood. Here we demonstrate for the first time that adipose-derived stromal/stem cells (ASCs) induced to an adipogenic or osteogenic lineage have differences in strand preference (-3p and -5p) for miRNAs originating from the same primary transcript. Furthermore, evaluation of miRNA expression in ASCs demonstrates alterations in both miRNA strand preference and 5'seed site heterogeneity. Additionally, we show that during stem cell differentiation there are alterations in expression of genes associated with the miRNA biogenesis pathway. Quantitative RT-PCR demonstrated changes in the Argonautes (AGO1-4), Drosha, and Dicer at intervals of ASC adipogenic and osteogenic differentiation compared to untreated ASCs. Specifically, we demonstrated altered expression of the AGOs occurring during both adipogenesis and osteogenesis, with osteogenesis increasing AGO1-4 expression and adipogenesis decreasing AGO1 gene and protein expression. These data demonstrate changes to components of the miRNA biogenesis pathway during stromal/stem cell differentiation. Identifying regulatory mechanisms for miRNA processing during ASC differentiation may lead to novel mechanisms for the manipulation of lineage differentiation of the ASC through the global regulation of miRNA as opposed to singular regulatory mechanisms.

MicroRNA regulation of stem cell differentiation and diseases of the bone and adipose tissue: Perspectives on miRNA biogenesis and cellular transcriptome.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression through targeting and suppression of mRNAs. miRNAs have been under investigation for the past twenty years and there is a large breadth of information on miRNAs in diseases such as cancer and immunology. Only more recently have miRNAs shown promise as a mechanism for intervention with respect to diseases of the bone and adipose tissue. In mesenchymal stem cell (MSC) differentiation, alterations in miRNA expression patterns can differentially promote an osteogenic, adipogenic, or myogenic phenotype. This manuscript reviews the current literature with respect to miRNAs in the context of MSC function with a particular focus on novel avenues for the examination of miRNA associated with bone and adipose tissue biology and disease. Specifically we highlight the need for a greater depth of investigation on MSCs with respect to miRNA biogenesis, processing, strand selection, and heterogeneity. We discuss how these mechanisms facilitate both altered miRNA expression and function.

Association of coronary calcification with prognosis of Covid-19 patients without known heart disease

Risk factors that determine the severity of Covid-19 have not been fully elucidated. The aim of this study was to evaluate the role of coronary artery calcification (CAC) as a risk factor for death or mechanical ventilation (MV) of patients without known heart disease infected with Covid-19. We analyzed 283 consecutive in-patients with acute respiratory symptoms with chest computed tomography (chest-CT), without previous heart disease, and criteria for Covid-19 (RT-PCR positive and/or typical clinical and chest-CT findings). CAC was classified by the number of coronary segments affected as absent (0), mild (1-3), and severe calcification (more than 3). The association between CAC, CAC severity, and death or MV due to severe respiratory failure was assessed by logistic regression. The mean age was 58.7±15.7 years and 54.1% were men. Patients with CAC were older, more likely to have hypertension, and less likely to be obese. CAC was present in 75 patients (26.5%), of which 42 had a mild calcification and 33 had severe calcification, and was associated with death (OR=2.35, 95%CI: 1.01-5.48) or MV (OR=2.72, 95%CI: 1.20-6.20) adjusted for multiple confounders, with significant and increased odds ratio for the severe form of CAC (death: OR=3.70, 95%CI: 1.20-11.42; MV: OR=3.30, 95%CI: 1.09-9.95). We concluded that CAC was an independent risk factor for death or MV in Covid-19 patients without previous heart disease, particularly for those with severe calcification. CAC can be easily visualized on common chest-CT, widely used in evaluation of moderate to severe Covid-19.

Intra-annual variation in rainfall and its influence of the adults Cyprideis spp (Ostracoda, Crustacea) on a eutrophic estuary (Guanabara Bay, Rio de Janeiro, Brazil) / Variação intra-annual na chuva e a sua influência nos adultos de Cyprideis spp (Ostracoda, Crustacea) em um estuário eutrofizado (Guanabara Bay, Rio de Janeiro, Brasil)

Abstract Spatial and temporal distribution of two species of adult´s ostracods (Cyprideis sp. and Cyprideis salebrosa) were studied as a function of the rainfall patterns in the Guanabara Bay, Rio de Janeiro, Brazil. Samples were taken in ten stations, along six surveys representing three periods (Dry, Early and Late Rainy) for two years. Stations were nested in four areas (Outer, Central, EPA Guapimirim and Impacted). The bottom water (temperature, salinity, dissolved oxygen and oxygen saturation) were measured in each area to characterize the influence of seasonal variations by rainfall. Cyprideis sp. and Cyprideis salebrosa showed patterns distribution to seasonality/surveys (p = 0,002 and p <0,001, respectly). The spatial distribution of Cyprideis sp was significantly different areas studied (p <0.001) indicated well defined areas and distribution in along of the surveys. However, C. salebrosa showed homogeneous distribution in along of the areas within of each survey (p <0.001). Redundancy Analysis (RDA) for the two years evidenced environment preference of the Cyprideis sp. for areas with marine conditions (high influence for channel central) and C. salebrosa for brackish water (high influence of the rivers). This observation reinforces of the existence of areas created by the seasonality of pluviometric regime, a possible dispersion of the adult's ostracods and possibility the use with bioindicators.

Resumo A distribuição espacial e temporal de duas espécies de ostracodes adultos (Cyprideis sp. e Cyprideis salebrosa) foi estudada em função do padrão pluviométrico na Baía de Guanabara, Rio de Janeiro, Brasil. As amostras foram coletadas em dez estações ao longo de seis campanhas, representando três períodos (Seco, Pré e Pós Chuvoso) por dois anos. As estações foram agrupadas em quatro áreas (Externo, Central, EPA Guapimirim e Impactado). A água do fundo (temperatura, salinidade, oxigênio dissolvido e saturação de oxigênio) foi medida em cada área para caracterizar a influência das variações sazonais pela chuva. Cyprideis sp. e Cyprideis salebrosa mostrou distribuição de padrões para sazonalidade/campanhas (p = 0,002 e p <0,001, respctivamente). A distribuição espacial de Cyprideis sp. foi significativamente diferente das áreas estudadas (p <0,001) indicou áreas bem definidas e distribuição ao longo das campanhas. No entanto, C. salebrosa mostrou distribuição homogênea ao longo das áreas dentro de cada campanha (p <0,001). A Análise de Redundância (RDA) para os dois anos evidenciou a preferência ambiental dos Cyprideis sp. para áreas com condições marinhas (alta influência para o canal central) e C. salebrosa para águas salobras (alta influência dos rios). Essa observação reforça a existência de áreas criadas pela sazonalidade do regime pluviométrico, uma possível dispersão dos adultos de ostracodes e a possibilidade do uso como bioindicadores.

Gas phase RNA and DNA ions 2. Conformational dependence of the gas-phase H/D exchange of nucleotide-5'-monophosphates.

The conformational dependence of the gas-phase hydrogen/deuterium (H/D) exchange of nucleotide-5-monophosphate anions with the H/D exchange reagent D2S is reported here. The electrospray-generated [M-H]- anions of adenosine-5'-monophosphate, adenosine-5'-carboxylic acid, ribitol-5-phosphate, and 2-deoxy-ribitol-5-phosphate were reacted with D2S in the gas phase. Their reactivity (adenosine-5'-monophosphate exchanged 2 of 5 labile hydrogens, adenosine-5'-carboxylic acid exchanged 1 of 4, ribitol-5-phosphate exchanged 2 of 3, and 2-deoxy-ribitol-5-phosphate exchanged 1 of 2) suggests that the hydroxyl group in the 2 position of the ribose sugar and the amino hydrogen on the nucleobase do not exchange readily with D2S. Semiempirical molecular orbital calculations suggest that the labile hydrogens in these positions are thermodynamically facile to exchange but as a conformation inaccessible to the presumed phosphate anion, consistent with a mechanism in which the phosphate anion complexes with the exchange reagent and assists H/D exchange at a neighboring site.

Competitive binding to the oligopeptide binding protein, OppA: in-trap cleanup in an Fourier transform ion cyclotron resonance mass spectrometer.

This communication demonstrates that gentle infrared laser heating can remove unwanted buffer adducts from a gas-phase protein complex without dissociating the complex itself. Specifically, noncovalent complexes of the oligopeptide-binding protein, OppA, bound to either (Ala)3 or LysTrpLys were electrosprayed from aqueous buffer solution into a 9.4 tesla Fourier transform ion cyclotron resonance mass spectrometer. In addition to the intact complexes, several additional buffer adduct species were produced under the conditions of the experiment. Irradiation of the trapped ion population with a continuous-wave infrared CO2 laser at relatively low power (2.5 W) for 1 s dissociated the buffer adducts but retained the intact protein:peptide complexes. Adduct-free complex(es) were then readily identified, and signal-to-noise ratio also increased by an order of magnitude because the same number of protein ions are distributed over fewer species. Higher IR power (5 W for 1 s) dissociated the adduct-free complex(es) without internal fragmentation. The present in-trap clean-up technique may prove especially useful for identifying and screening the combinatorial library ligands most strongly bound to a receptor in the gas phase.

Gas-phase hydrogen/deuterium exchange of positively charged mononucleotides by use of Fourier-transform ion cyclotron resonance mass spectrometry.

The gas-phase structures of protonated (deoxy)nucleoside-5'- and 3'-monophosphates (mononucleotides) have been examined by the use of gas-phase hydrogen/deuterium (H/D) exchange and high-field Fourier-transform ion cyclotron resonance mass spectrometry. These nucleotides were reacted with three different deuterating reagents: ND3, D2O, and D2S, of which ND3 was the most effective. All mononucleotides fully exchanged their labile hydrogen for deuterium with ND3 with the exception of deoxycytidine-3'-monophosphate, deoxyadenosine-5'-monophosphate, adenosine-5'-monophosphate, and adenosine-3'-monophosphate. Semiempirical calculations demonstrate the presence of hydrogen bonding upon protonation of the purine mononucleotides which may lead to incomplete H/D exchange. H/D exchange rates differed between the deoxymononucleotides and the ribomononucleotides, suggesting that the 2'-OH group plays an important role in the exchange process. Reactions of nucleosides and mononucleotides with D2O demonstrate that a structure-specific long-lived ion-molecule complex between D2O and the mononucleotide involving the phosphate group is necessary for exchange to overcome the high-energy activation barrier. In contrast, a structure-specific long-lived ion-molecule complex between the mononucleotides and ND3 is not required for exchange to occur.

Quantifying the frequency of radiogenic thyroid cancer per clonogenic cell in vivo.

We used quantitative cell transplantation to evaluate the frequencies of the formation of radiogenic thyroid cancer per clonogenic rat thyroid epithelial cell in vivo. Irradiation of thyroid cells with 5 Gy 137Cs gamma rays before transplantation significantly increased the incidence of thyroid carcinoma formation in such grafts compared to similar grafts of unirradiated thyroid cells. We calculated the frequencies of radiogenic cancer by subtracting cancer incidences in unirradiated groups from incidences in irradiated groups and dividing by the number of clonogens grafted. The highest observed frequencies of radiogenic thyroid cancer so calculated were 0.141 and 0.046 cancers per surviving irradiated clonogenic cell. These cancer frequencies occurred in grafts containing averages of three and ten clonogens per site, respectively, and represent one cancer per approximately 7 and approximately 22 irradiated clonogens. We conclude that the highest observed frequencies of radiogenic cancer are likely to be the best estimates of the "real" frequency per irradiated clonogen in that virtually all the methodological sources of inaccuracy tend to decrease the observed frequency compared to the "real" frequency. Radiogenic initiation of cancer is thus a highly common cellular event among surviving irradiated clonogenic thyroid cells. To examine the role of endocrine-mediated tumor promotion on the expression of radiogenic cancer, we attenuated the intensity of thyrotropin (TSH)-mediated tumor promotion in some groups of recipient animals. We found that the incidence rates for radiation-associated cancer were significantly higher in rats with higher serum TSH levels compared to rats with lower TSH levels. We conclude from these data that (1) radiogenic thyroid cancer occurs with a high frequency and (2) chronic TSH stimulation accelerates progression of radiogenic neoplasms to overt carcinomas and promotes development of later-arising carcinomas in grafts of unirradiated thyroid clonogens.

Gas-phase reactions of glycine, alanine, valine and their N-methyl derivatives with the nitrosonium ion, NO+.

The gas-phase reactions of the nitrosonium ion, NO+ with the amino acids glycine, alanine and valine and their N-methyl derivatives were investigated under chemical ionization mass spectrometric (CIMS) conditions. Two products were observed in all cases: the formation of the iminium ion and the formation of an [M-H]+ ion. The latter product is consistent with a reaction channel involving hydride abstraction by NO+, and was confirmed by (i) examining the Ar+CI mass spectra of the same amino acids under similar source conditions and (ii) examining the unimolecular fragmentation reactions of the [M + H]+ ions of the N-nitroso-N-methyl derivatives of each of the amino acids in a tandem mass spectrometer. Further insights into the reaction of glycine with NO+ were obtained by performing ab initio calculations (at the MP2/6-31G* parallel HF/6-31G* level). These results indicate that four reactions are thermodynamically viable for glycine: (i) hydride abstraction; (ii) iminium ion formation (with concomitant loss of HONO and CO); (iii) diazonium ion formation; and (iv) diazonium ion formation followed by loss of N2. Possible reasons why reactions (iii) and (iv) are not observed are discussed, and comparisons with solution reactivity and the gas-phase reactivity of NO+ are also made.