RESUMO
In our institution, a prospective observational trial testing micro-RNA (miRNA) and ARV7 mutational status in metastatic, castration resistant prostate cancer (mCRPC), is currently recruiting (PRIMERA trial, NCT04188275). A pre-planned interim analysis was performed when 50% of the planned accrual was reached. In this report, we explored the predictive value of Circulating Tumor Cell (CTC) detection in mCRPC patients undergoing 1st line therapy. Moreover, ARV7, ARFL, PSMA and PSA expression on CTC was reported to explore potential correlation with patient prognosis and response to therapy. PRIMERA is a prospective observational trial enrolling mCRPC patients undergoing standard treatment (ARTA + ADT) after I line ADT failure. Clinical and pathological features were collected. Outcomes selected for this preliminary analysis were time to castration resistance (TTCR), PSA at 8 weeks after ARTA therapy start, PSA drop at 8 weeks, Overall PSA drop, PSA nadir. Correlation between these outcomes and CTC detection was tested. Expression of ARV7, ARFL, PSA and PSMA was explored in CTC+ patients to assess their prevalence in this cohort and their impact on selected outcomes. Median TTCR was significantly shorter in CTC+ vs CTC- patients (32.3 vs 75 months, respectively, p = 0.03) and in ARFL+ vs ARFL- patients (30.2 vs 51.1 months, respectively, p = 0.02). ARV7, PSMA and PSA expression on CTC had no impact on median TTCR, nor on biochemical response to therapy. Patients in whom CTC and ARFL expression were detected had significant reduced TTCR. However, PSA response was not influenced by CTCs detection and specific biomarkers expression.
Assuntos
Antagonistas de Androgênios/uso terapêutico , Antígenos de Superfície/análise , Glutamato Carboxipeptidase II/análise , Células Neoplásicas Circulantes/química , Neoplasias de Próstata Resistentes à Castração/patologia , Receptores Androgênicos/genética , Humanos , Calicreínas/sangue , Masculino , Estudos Prospectivos , Antígeno Prostático Específico/sangue , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/genética , Neoplasias de Próstata Resistentes à Castração/mortalidadeRESUMO
CONTEXT: Cell-free nucleic acids circulating in plasma are considered a promising noninvasive tool for cancer monitoring. BRAF(V600E) mutation in cell-free DNA (cfDNA) could represent an appropriate marker for papillary thyroid carcinoma (PTC). OBJECTIVE: Our aim is to investigate the role of BRAF(V600E)-mutated allele in cfDNA as a marker for the diagnosis and follow-up of PTC. STUDY DESIGN: BRAF(V600E) allele was detected and quantified by an allele-specific real-time quantitative PCR assay in plasma from 103 patients affected by nodular goiter. As control populations, we enrolled 49 healthy subjects and 16 patients with non-nodular thyroid diseases. RESULTS: The percentage of circulating BRAF(V600E) was significantly different between patients and controls and throughout different cytological categories of ultrasound-assisted fine-needle aspiration. Patients with a histopathological diagnosis of PTC showed a higher percentage of circulating BRAF(V600E) (P = .035) compared to those with benign histology. In 19 patients, a second blood draw, taken 3-6 months after surgery, showed a lower percentage of BRAF(V600E) in cfDNA than the presurgical sample (P < .001). The diagnostic performance of circulating BRAF(V600E) was assessed by receiver operating characteristic curve analysis resulting in an area under the curve of 0.797. A cutoff value was chosen corresponding to maximum specificity (65%) and sensitivity (80%). On this basis, we evaluated the predictive value of BRAF(V600E) in Thy 3 patients with a resulting positive predictive value of 33% and a negative predictive value of 80%. CONCLUSIONS: The results of the present study provide encouraging data supporting the possibility to take advantage of circulating BRAF(V600E) in the management of PTC.
Assuntos
Carcinoma/diagnóstico , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias da Glândula Tireoide/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma/sangue , Carcinoma/genética , Carcinoma/patologia , Carcinoma Papilar , DNA/análise , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas B-raf/sangue , Curva ROC , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide/sangue , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologiaRESUMO
The biochemistry of reactive oxygen species is an important field with wide implications. Both preventive and chain breaking antioxidants have a role in the limitation of oxidative stress that accompanies aging and diseases. The potent antioxidant activity of phenolic substances of red wine, in particular, have been proposed as an explanation for the "French Paradox" (the apparent incompatibility of a high fat diet with a low incidence of coronary heart diseases). A lot of researchers emphasize beneficial effects of red wine and insist on lower or no antioxidant effect of white wine. We have been studying the effect of both white and red wine on blood antioxidant capacity in humans. The white wine we have been testing was produced by an ancient Tuscany procedure (the same used for red winemaking) which includes fermentation with grapes juice together with peelings and seeds. A statistically significant increase in total antioxidant capacity (TAC) levels was observed after 2h from red or white wine ingestion. White wine effect appears to be faster than that of the red one, since a significant difference can also be reported after 1h. We can conclude that the big difference in the results of serum TAC due to white wine reported by us, in comparison to those reported by others relatively to white wine produced using the French method, can be explained by the difference in the winemaking procedure we adopted.
Assuntos
Antioxidantes/metabolismo , Vinho , Idoso , Idoso de 80 Anos ou mais , Humanos , Medições Luminescentes , Masculino , Pessoa de Meia-IdadeAssuntos
Carcinoma Basocelular/metabolismo , Imunossupressores/farmacologia , Tacrolimo/farmacologia , Fatores de Transcrição/antagonistas & inibidores , Fatores de Coagulação Sanguínea/efeitos dos fármacos , Fatores de Coagulação Sanguínea/metabolismo , Carcinoma Basocelular/patologia , Linhagem Celular Tumoral , Expressão Gênica/efeitos dos fármacos , Humanos , Receptores Patched , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas de Ligação a RNA , Receptores de Superfície Celular/efeitos dos fármacos , Receptores de Superfície Celular/metabolismo , Receptores Acoplados a Proteínas G/efeitos dos fármacos , Receptores Acoplados a Proteínas G/metabolismo , Proteínas Ribossômicas , Receptor Smoothened , Fatores de Transcrição/metabolismo , Proteína GLI1 em Dedos de ZincoRESUMO
Statistical data of Italy, issued on 1st January 2005, show that Tuscany has a high percentage of elderly subjects, and that to reach 100 years is a common possibility, mainly for women. Comparable values appear for longevity index (LI%), either calculated on the basis of the various resorts above sea level, or on the basis of the number of inhabitants. However, some differences are observed considering the various provinces: Grosseto Province has the second higher value of >65-year-old subjects, while its LI% is the lowest. Centenarity index (CI%) considerably differs from LI%: if ranking Tuscany according to the various heights of territorial positions, the first displays a negative, while the second one shows a positive correlation with the local height. If comparing for inhabitants number, CI% is maximal in provincial cities, while LI% is higher in smaller communities. All these data indicate that residents in small communities of the mountains reach easily 90 years, while 100 years are more frequently reached in larger cities in the plane areas. In conclusion, the highest province longevity (Lucca) coincides with the highest centenarian sex ratio, and it suggests that in this province the older women may enjoy particular forms of environmental protection, which helps them for reaching 100 years of age. Differences between the results of this study and others performed in Sardinia are discussed.
Assuntos
Idoso de 80 Anos ou mais/estatística & dados numéricos , Altitude , Análise por Conglomerados , Emigração e Imigração/estatística & dados numéricos , Feminino , Humanos , Itália/epidemiologia , Masculino , Distribuição por SexoRESUMO
Usually, the effects of cognitive decline are not noted before the age of 70 years, which involve the intellectual capacities, the attention, the processes of elaboration and the memory. The studies on the cognitive disturbances of the elderly are numerous, and document the progressive increase of cerebral deterioration with advancing age. However, only a few studies refer to the significance of the cognitive disturbances in the clinical conditions and autonomy of the long living subjects. For this reason, we studied the cerebral deterioration of an adequate number of centenarians in correlation with their clinical conditions and autonomy. Our centenarian sample derived from the Italian multi-center study on centenarians (IMUSCE), which was an epidemiological study which identified 1173 centenarians (202 males, 971 females) in the age range of 100-109 years. From this sample, we analyzed 346 subjects as far as the cognitive functions and the degree of autonomy by using the psychometric tests of the mini-mental state examination (MMSE) and the instrumental activities of daily living (IADL) for the functional evaluations. In addition, we evaluated the clinical conditions of the subjects dividing them in three groups: Group A (those in good clinical conditions), Group B (those in discrete clinical conditions), and Group C (those in deteriorated clinical conditions). These analyses revealed that 187 (54.1%) of the 346 examined centenarians have shown an MMSE score in the normal range (score ratio from 1.0 to 0.63). The cognitive disorders are present in the centenarians in a clearly higher frequency (13.1%), than found in the common elderly (5.1%). The severe cognitive disorders do not allow a total autonomy or even a slight dependency. Only six subjects (1.7%) of the total sample were totally independent. These subjects had no cognitive disorders, and were in good clinical conditions. The results show that having an MMSE score in the normal range, and being in good clinical conditions are necessary but not sufficient prerequisites for a total autonomy in the IADL scores.
Assuntos
Atividades Cotidianas/psicologia , Cognição/fisiologia , Longevidade/fisiologia , Autonomia Pessoal , Idoso de 80 Anos ou mais , Transtornos Cognitivos/epidemiologia , Transtornos Cognitivos/psicologia , Feminino , Avaliação Geriátrica/métodos , Humanos , Itália/epidemiologia , Masculino , Memória/fisiologia , Entrevista Psiquiátrica Padronizada , Prevalência , Psicometria/métodos , Estudos RetrospectivosRESUMO
The human functional autonomy is reduced progressively with advancing age, although a considerable proportion of the centenarians maintain a good level of autosufficiency for the basic performance of the everyday life. Even if males have a lower probability to reach the centenarian age than females do, the male centenarians display generally better functional conditions than the females. Actually, there are no systemic studies on centenarian works or activities; nevertheless, the examples of several representative persons (artists, scientists, explorers, etc.) who remained active even after this age indicate that such a possibility is realistic, and that the maintenance of vital interest and passions, thus preserving competence and professional attitudes, is not in conflict with the reaching of extreme longevity.
Assuntos
Atividades Cotidianas , Criatividade , Longevidade/fisiologia , Atividade Motora/fisiologia , Autonomia Pessoal , Idoso de 80 Anos ou mais , Avaliação Geriátrica , HumanosRESUMO
Centenarians represent a group of population displaying the most rapid expansion. This progressive increase of the presence of centenarians is a multi-factorial phenomenon, which is due to the improvements of the environmental conditions and the life style and also to the progress of the medical science. In order to obtain a more reliable estimate of the longevity per gender and territorial entities, we propose two new indicators. (i) The ratio between the ultranonagenarians and the total population above 65 years old (called longevity index: LI%); (ii) the ratio between the centenarians and the total population above 90 years old (called centenarity index: CI%). An analysis of the data of the Italian National Office of Statistics (ISTAT, ) using these two indicators demonstrates that the subjects above the age of 90 are more frequent in the regions of Central and Northern Italy, which are more developed regarding the economic conditions and technological progress. Nevertheless, the Southern and Insular regions of Italy have a higher occurrence of centenarians among the ultranonagenarian population, and also a higher prevalence of male centenarians, as compared to the northern regions. This demonstrates that achievement of the threshold of 100 years old does not require only particular socio-economic conditions, but also an adequate climate and environment, as well as a favorable genetic composition.
Assuntos
Envelhecimento/fisiologia , Indicadores Básicos de Saúde , Longevidade , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Itália , MasculinoRESUMO
Recent evidence suggests that interleukin-4 (IL-4) is related to mucosal tolerance by which an injurious immune response is prevented, suppressed or shifted to a non-injurious response. We investigated the expression of IL-4 and its splice variant isoform IL-4delta2 in gastric epithelial cells of healthy subjects and gastritis patients infected with Helicobacter pylori (H. pylori) with or without the cag pathogenicity island (cag-PAI). IL-4 and IL-4delta2 mRNAs were evaluated in microdissected gastric epithelium and in AGS cell lines co-cultured with H. pylori B128 or SS1 strains. IL-4 mRNA was consistently detected in microdissected gastric epithelial cells from healthy subjects. The IL-4 mRNA expression was low in H. pylori?infected patients, and markedly reduced in cag-PAI-positive ones. IL-4delta2 mRNA was expressed on gastric epithelium of H. pylori-infected patients, but not in healthy subjects. The IL-4delta2 expression was lower in cag-PAI-positive than in cag-PAI-negative H. pylori infected patients. AGS cells also produced IL-4 mRNA upon SS1 strain stimulation, whereas IL-4delta2 mRNA expression was detected in AGS co-cultured with either SS1 or B128 strains. An inverse correlation was documented between IL-4 and IL-4delta2 mRNA expression by microdissected gastric epithelial cells and the score of gastritis. IL-4, but not IL-4delta2, is expressed by gastric epithelium of healthy subjects, whereas IL-4delta2 and lesser IL-4 mRNA are detectable in the gastric epithelium of H. pylori-infected patients. Data suggest that gastric epithelial cells might regulate the balance between tolerance and immune response by the fine tuning of IL-4 and IL-4delta2 expression.
Assuntos
Mucosa Gástrica/metabolismo , Infecções por Helicobacter/metabolismo , Helicobacter pylori , Interleucina-4/biossíntese , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adulto , Linhagem Celular Tumoral , Separação Celular , Epitélio/metabolismo , Feminino , Mucosa Gástrica/citologia , Ilhas Genômicas/genética , Humanos , Interleucina-4/genética , Masculino , Microdissecção , Antro Pilórico , RNA/biossíntese , RNA/genética , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismoRESUMO
Mutations in genes encoding mitochondrial succinate dehydrogenase (SDH) are frequently involved in the development of neural crest-derived (NCD) tumors, such as pheochromocytomas (PHEOs) or paragangliomas (PGLs). In this study we report the results of sequencing analysis in leukocyte DNA of patients affected by PHEO/PGL who turned out to be SDH mutation carriers. A nonsense germline heterozygous mutation (Q109X) was found in the exon 4 of the SDHD gene in the index cases of six unrelated families affected by PHEO/PGL. Haplotype analysis showed the presence of a founder effect. Affected patients showed high clinical variability, ranging from monolateral to bilateral glomus tumors, variably associated or not with PGLs or PHEOs. A novel missense SDHD variant, T112I, was also found in one of our families. A new missense G106D mutation, involving a highly conserved amino acid, was found in two sisters affected by bilateral glomus tumors. A P81L mutation associated with abdominal and head and neck PGL was detected in three families. A G12S variant of the SDHD gene was found in one patient affected by a PHEO. The finding of this variant in 3 of 100 control subjects suggests that it is a polymorphism and not a mutation. A novel IVS2-1G>T variant was found at intron 2 of SDHD gene in one patient affected by a glomus tumor. All the tumors associated with SDHD mutations were benign. Conversely, the only mutation we found in SDHB gene (IVS3+1G>A) was associated with a malignant PHEO.
Assuntos
Mutação em Linhagem Germinativa , Paraganglioma/genética , Succinato Desidrogenase/genética , Sequência de Aminoácidos , Animais , Heterozigoto , Humanos , Dados de Sequência Molecular , Homologia de Sequência de Aminoácidos , Succinato Desidrogenase/químicaRESUMO
AIMS: To develop a quantitative real-time PCR (Rt PCR) assay for the early detection of Biscogniauxia nummularia, a xylariaceous fungus that causes strip-canker and wood decay on European beech (Fagus sylvatica L.). METHODS AND RESULTS: The molecular assay was based on TaqMan chemistry using species-specific primers and a fluorogenic probe designed on the ITS1 sequence of rRNA gene clusters. The specificity of the oligonucleotides and the probe were tested using the DNA of B. nummularia isolates from different geographic areas, of phylogenetically related species, and of some fungi commonly colonizing European beech bark and wood. A total of 31 symptomless and symptomatic shoots of European beech were collected from three forest sites in the Apennine Mountains of Italy. The percentage of positive detections of B. nummularia with the TaqMan assay was 78.6%, compared with only 14.3% of positive isolations on growth media for two sites. CONCLUSIONS: In shoots, the quantitative Rt PCR assay detected down to 8.0-fg fungal DNA per microgram of total DNA extracted. SIGNIFICANCE AND IMPACT OF THE STUDY: The assay developed in quantitative Rt PCR, by using TaqMan chemistry, revealed a rapid and sensitive method useful for the early detection of B. nummularia in symptomless European beech twigs.
Assuntos
Fagus/microbiologia , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase/métodos , Árvores/microbiologia , Xylariales/isolamento & purificação , Primers do DNA , DNA Fúngico/análise , DNA Fúngico/isolamento & purificação , Taq Polimerase , Xylariales/genéticaRESUMO
In somatostatinoma, a rare malignant somatostatin (SST)-secreting neoplasia, tumour regression is rarely observed, implying the need for novel antiproliferative strategies. Here, we characterized a long-term culture (SST-secreting cancer (SS-C cells)) established from a human somatostatinoma. High concentrations of SST and chromogranin A were released by SS-C cells and SST release was stimulated by depolarizing stimuli and inhibited by the SST analogue, octreotide. SS-C cells expressed mRNA for SST receptor (SSTR) subtypes 1, 2 and 4, being also able to bind native SST. Moreover, SS-C cells were positively stained with an antibody to SSTR2. SS-C cells also expressed interferon-gamma (IFN-gamma) receptor mRNA and measurable telomerase activity. Our findings indicate that in vitro exposure of SS-C cells to native SST-28, to octreotide, to IFN-gamma, or to 3'-azido-3'deoxythymidine (AZT), a telomerase inhibitor, results in inhibition of SS-C cell proliferation. Concomitant with growth inhibition, apoptosis was detected in SST-, octreotide-, IFN-gamma- or AZT-treated SS-C cell cultures. Taken together our results characterized native SST, SST analogues, IFN-gamma and a telomerase inhibitor as growth-inhibiting and proapoptotic stimuli in cultured human somatostatinoma cells. Based on these findings, the potential of SST analogues, IFN-gamma and AZT, alone or in combination, should be further explored in the medical treatment of somatostatinoma.
Assuntos
Cromograninas/metabolismo , Neoplasias do Jejuno/patologia , Receptores de Somatostatina/metabolismo , Somatostatina/metabolismo , Somatostatinoma/patologia , Telomerase/metabolismo , Adulto , Fármacos Anti-HIV/farmacologia , Antineoplásicos Hormonais/farmacologia , Proliferação de Células/efeitos dos fármacos , Cromogranina A , Feminino , Humanos , Interferon gama/genética , Interferon gama/farmacologia , Neoplasias do Jejuno/metabolismo , Octreotida/farmacologia , RNA Mensageiro , Somatostatinoma/metabolismo , Telomerase/antagonistas & inibidores , Células Tumorais Cultivadas , Zidovudina/farmacologiaRESUMO
Laser-assisted microdissection (LMD) has been developed to procure precisely the cells of interest in a tissue specimen, in a rapid and practical manner. Together with real-time PCR and RT-PCR techniques, it is now feasible to study genetic alterations, gene expression features and proteins in defined cell populations from complex normal and diseased tissues. The process that brings from sample collection to the final quantitative results is articulated in several steps, each of which requires optimal choices in order to end up with high-quality nucleic acid or protein that allows successful application of the final quantitative assays. This review will describe shortly the development of LMD technologies and the principles they are based on. Trying to highlight the advantages and disadvantages of LMD, the main problems related to specimens collection and processing, section preparation and extraction of bio-molecules from microdissected tissue samples have been analysed.
Assuntos
Microdissecção/métodos , Microscopia Confocal/métodos , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/metabolismo , Linhagem Celular , Sistemas Computacionais , DNA/isolamento & purificação , Proteínas de Ligação a DNA/metabolismo , Perfilação da Expressão Gênica , Técnicas de Preparação Histocitológica , Técnicas Histológicas , Humanos , Neoplasias/metabolismo , Proteínas/isolamento & purificação , RNA/isolamento & purificação , Telomerase/metabolismoRESUMO
BACKGROUND: Mutations in genes coding for the mitochondrial complex II succinate dehydrogenase (SDH) subunits cause familial neural crest derived (NCD) tumours. METHODS: Index cases from six apparently unrelated families affected by NCD tumours were analysed for mutations in the SDHB, SDHC, and SDHD genes. RESULTS: The same nonsense germline heterozygous mutation (Q109X) in exon 4 of the SDHD gene was found in each of the six families. Overall, 43 heterozygotes were identified. These were evaluated for the presence of NCD tumours through radiological examination of the neck, thorax, and abdomen, and measurement of urinary metanephrines and plasma chromogranin A. A novel missense SDHD variant, T112I, which did not segregate with the Q109X mutation and was not associated with phenotypic manifestations, was observed in one of the families. Microsatellite analysis showed a common haplotype in all individuals heterozygous for the Q109X mutation, indicating a founder effect. Overall, 18 heterozygotes were clinically affected by at least one NCD tumour. Every affected patient inherited the germline mutation from the father, confirming SDHD maternal genomic imprinting. Penetrance of the paternally inherited mutation progressively increased from 33% to 83% at 30 and 60 years, respectively. Affected patients showed high clinical variability, ranging from monolateral to bilateral glomus tumours variably associated or not with paragangliomas or phaeochromocytomas. Loss of heterozygosity was observed in tumour cells isolated by laser capture microdissection. CONCLUSIONS: This study shows that a single founder SDHD mutation is present in an area of central Italy and that this mutation is associated with widely variable interfamilial and intrafamilial expressivity.
Assuntos
Segregação de Cromossomos , Códon sem Sentido , Proteínas de Membrana/genética , Paraganglioma/genética , Adulto , Idoso , Análise Mutacional de DNA , Feminino , Efeito Fundador , Predisposição Genética para Doença , Impressão Genômica , Haplótipos , Humanos , Itália , Perda de Heterozigosidade , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade , Paraganglioma/diagnóstico , Linhagem , Fenótipo , Succinato DesidrogenaseRESUMO
The traditional mediterranean diet is associated with a hope for longer survival. It has also been shown that the red wine possesses a protective effect against the oxidative stress. We studied TAC, the DHEAS and the IGF-1 in a group of 26 healthy centenarians, 17 women and 9 men, of the age range of 100--105 years. Furthermore, we analyzed also serum urate and bilirubin levels between drinkers and abstainers. Most of centenarian subjects have been moderate wine consumers (<500 ml/day of red wine). These subjects were subdivided as follows: (i) Group A: those who had maintained the style of their dietary habits as compared to the previous years (n=3 males, 10 females); (ii) Group B: those who actually consumed a diet being deficient compared to that of the previous years, but remained moderate drinkers of red wine (n=3 males, 4 females); and (iii) Group C: those who actually consumed a diet being deficient compared to that of the previous years, and at the same time, were abstainers in wine consumption (n=3 males, 3 females). The results show that in men three of the studied parameters decreased from Group A to C to considerable extents, as follows (mean+/-S.D.). TAC: 302.4+/-32.3; 142.0+/-24.1 and 96.4+/-20.1 micromol/l; DHEAS: 3.35+/-0.81; 2.52+/-0.18 and 1.34+/-0.14 micromol/l; IGF-1: 85.7+/-6.7; 76.6+/-6.7 and 65.6+/-2.6 ng/ml, respectively. For the same parameters, the results in the women were: TAC: 258.4+/-12.2; 182.1+/-14.0 and 107.6+/-10.0 micromol/l; DHEAS: 3.85+/-0.16; 2.34+/-0.19 and 2.05+/-0.04 micromol/l; IGF-1: 89.7+/-6.7; 76.6+/-4.7 and 64.2+/-2.7 ng/ml, respectively. We did not find any significant difference in the other serum parameters between drinkers (n=14) and abstainers (n=3) (urate: 267.6+/-52.9, and 289.5+/-80.1; bilirubin: 9.81+/-4.29 and 7.18+/-2.89 micromol/l, respectively). Our data suggest that the deteriorated diet caused a reduction of TAC, DHEAS and IGF-1 in the centenarians. However, red vine consumption exerted a protective effect against this trend, even if this protection is not reaching statistical significance in some cases (in men), which is due most probably to the lower number of male subjects in the study.
Assuntos
Antioxidantes/metabolismo , Sulfato de Desidroepiandrosterona/sangue , Dieta , Fator de Crescimento Insulin-Like I/metabolismo , Vinho , Idoso de 80 Anos ou mais , Feminino , Humanos , Itália , MasculinoRESUMO
AIMS: Real-time PCR, based on TaqMan chemistry, was used to detect Biscogniauxia mediterranea, a fungal pathogen that after a long endophytic phase may cause charcoal disease in oak trees. METHODS AND RESULTS: Specific primers and probe were designed and tested on axenic cultures of B. mediterranea and other fungi commonly colonizing oaks. Twig samples were collected in Tuscany from apparently healthy oaks (Quercus cerris, Quercus ilex and Quercus pubescens) growing near trees infected with the fungus. Twigs were divided into two groups: one for isolation in agar plates, and one for real-time PCR after DNA extraction. The detection limit of the assay was 0.01 pg/DNA, whereas the amounts of fungal DNA detected in asymptomatic tissue were >0.5 pg microg(-1) total DNA extracted. In the apparently healthy twigs the frequency of isolation found on agar was 25.0%, much lower than that with real-time PCR (96.4%). CONCLUSIONS: Real-time PCR is a sensitive and fast technique able to specifically detect and quantify the DNA of B. mediterranea in oak tissue. SIGNIFICANCE AND IMPACT OF THE STUDY: This diagnostic method is a precise tool to localize fungi in symptomless plant tissues and promises to advance our understanding of fungal infection during their latent phase.
Assuntos
Fungos/isolamento & purificação , Quercus/microbiologia , Primers do DNA , DNA Fúngico/análise , Fungos/genética , Itália , Reação em Cadeia da Polimerase/métodos , Quercus/genéticaRESUMO
Liver function is well maintained with increasing age. The aim of our study was to investigate if long-term moderate (Assuntos
Consumo de Bebidas Alcoólicas/epidemiologia
, Fígado/fisiologia
, Vinho/efeitos adversos
, Adulto
, Idoso de 80 Anos ou mais
, Alanina Transaminase/sangue
, Fosfatase Alcalina/sangue
, Aspartato Aminotransferases/sangue
, Bilirrubina/sangue
, Butirilcolinesterase/sangue
, Feminino
, Humanos
, Estilo de Vida
, Masculino
, Pessoa de Meia-Idade
, Tempo
, gama-Glutamiltransferase/sangue
RESUMO
Somatostatin analogs are effective in inhibiting growth of human breast cancer cell lines. These antiproliferative effects are mediated by specific receptors located on cell membranes. The somatostatin receptor subtype 2 (sst2) is the principal mediator of somatostatin effects in normal and cancer cells, and its presence has already been demonstrated in breast cancer. The purpose of our study was to evaluate the clinical relevance of the expression of sst2 by quantifying its mRNA in a large group of infiltrating breast cancers and their corresponding normal tissues. The expression of sst2 mRNA was measured with quantitative real time RT-PCR in 169 breast cancers and in their corresponding unaffected tissues. We evaluated the association of sst2 expression with the commonest clinical-pathologic features of breast cancer. The correlation with a marker of cell proliferation (Ki-67) and with receptor concentration was also evaluated. In cancer tissues, we found that the absolute concentrations of sst2 mRNA were significantly higher in estrogen receptor (ER)-positive samples (P=0.002) as well as in lymph-node-negative cancers (P=0.04) (Student's t-test or one-way ANOVA). In addition, sst2 mRNA was significantly higher in breast cancers than in corresponding unaffected tissues (P=0.0002). However, when the clinical-pathologic parameters were considered, this gradient maintained its statistical significance only in tumors expressing positive prognostic markers, such as the presence of ER (P=0.0005) and progesterone receptors (PgR) (P=0005), and the lack of lymph-node involvement (P=0.0003). The same difference was also significant in postmenopausal women (P=0.001) and in T1 patients (P=0.001). In addition, sst2 mRNA expression was significantly higher (P=0.008) in low-proliferating breast cancers. Finally, we found that the quantitative expression of sst2 mRNA was directly related to the PgR concentration in breast cancer tissues (P<0.001). Our data seem to indicate that an upregulation of sst2 gene expression is a common feature of breast cancers which, on the basis of conventional predictive parameters, are expected to have a better prognosis. Featuring a possible role of somatostatin analogs in combined endocrine therapies for breast cancer, our results seem to confirm that the sst2 status of the tumor should be previously investigated.
Assuntos
Neoplasias da Mama/genética , Carcinoma Ductal/genética , Carcinoma Lobular/genética , RNA Mensageiro/genética , Receptores de Somatostatina/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/metabolismo , Carcinoma Ductal/metabolismo , Carcinoma Lobular/metabolismo , Proliferação de Células , Feminino , Humanos , Hibridização In Situ , Linfonodos/patologia , Invasividade Neoplásica/patologia , Estadiamento de Neoplasias , Prognóstico , RNA Mensageiro/metabolismo , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/metabolismo , Receptores de Somatostatina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
AIMS: To develop a real-time polymerase chain reaction (PCR) method for rapid detection and quantification of Oenococcus oeni in wine samples for monitoring malolactic fermentation. METHODS AND RESULTS: Specific primers and fluorogenic probe targeted to the gene encoding the malolactic enzyme of O. oeni were developed and used in real-time PCR assays in order to quantify genomic DNA either from bacterial pure cultures or wine samples. Conventional CFU countings were also performed. The PCR assay confirmed to be specific for O. oeni species and significantly correlated to the conventional plating method both in pure cultures and wine samples (r = 0.902 and 0.96, respectively). CONCLUSIONS: The DNA extraction from wine and the real-time PCR quantification assay, being performed in ca 6 h and allowing several samples to be concurrently processed, provide useful tools for the rapid and direct detection of O. oeni in wine without the necessity for sample plating. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid quantification of O. oeni by a real-time PCR assay can improve the control of malolactic fermentation in wines allowing prompt corrective measures to regulate the bacterial growth.