RESUMO
OBJECTIVE: Affective disorders seem associated with aberrant intestinal microbiota but whether this pattern also occurs in individuals at increased heritable risk is unknown. We investigated associations between gut microbiota profiles and affective disorders by comparing monozygotic (MZ) twins concordant (affected twins with unipolar or bipolar disorder in remission) and discordant to affective disorders (high-risk) with MZ twins without affective disorders (low-risk). METHODS: Stool samples were collected from 128 MZ twins and the microbiome was profiled using 16S rDNA sequencing of the V3-V4 region. RESULTS: Affected twins had a lower diversity and an absence of a specific operational taxonomical unit (OTU) in comparison with low-risk twins. The high-risk twins exhibited the same pattern although the lower diversity was only at a trend level. The OTU belonged to the family Christensenellaceae. The findings were not explained by lifestyle factors (smoking, alcohol consumption, body mass index, or psychotropic medication). CONCLUSION: Affected twins in remission and high-risk twins presented aberrant gut microbiota with depletion of a specific OTU. If replicated, this reduced relative sequence absence may together with the globally altered microbiota composition act as a vulnerability marker by accentuating the effect of gene-environment interactions in individuals genetically disposed for an affective disorder.
Assuntos
Microbioma Gastrointestinal/genética , Transtornos do Humor/complicações , Transtornos do Humor/genética , Gêmeos Monozigóticos/genética , Adulto , Transtorno Bipolar/genética , Transtorno Bipolar/psicologia , Classificação/métodos , Dinamarca/epidemiologia , Doenças em Gêmeos/genética , Doenças em Gêmeos/psicologia , Feminino , Microbioma Gastrointestinal/fisiologia , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Transtornos do Humor/epidemiologia , Transtornos do Humor/psicologia , Indução de Remissão , Risco , Análise de Sequência de DNA/métodos , Gêmeos Monozigóticos/psicologiaRESUMO
A total of 38 patients (18 female/20 male) with childhood meningioma were recruited from the German registry HIT-Endo (1989-2009). In 5 cases meningioma occurred as second malignant neoplasm (SMN). Histologies were confirmed by reference assessment in all cases (SMN: 2 WHO I, 1 WHO II, 2 WHO III). The SMNs were diagnosed at a median age of 12.4 years with a median latency of 10.2 years after primary malignancy (PMN; 4 brain tumors, 1 lymphoblastic leukemia; median age at diagnosis 2.7 years). Meningioma occurred as SMN in the irradiated field of PMN (range 12-54 Gy). The outcome after treatment of SMN meningioma (surgery/irradiation) was favorable in terms of psychosocial status and functional capacity in 4 of 5 patients (1 death). We conclude that survivors of childhood cancer who were exposed to radiation therapy at young age harbor the risk of developing meningioma as a SMN at a particularly short latency period in case of high dose exposure.
Assuntos
Neoplasias Encefálicas/patologia , Neoplasias Meníngeas/secundário , Meningioma/secundário , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Adolescente , Antineoplásicos/uso terapêutico , Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/cirurgia , Criança , Pré-Escolar , Humanos , Lactente , Imageamento por Ressonância Magnética , Masculino , Neoplasias Meníngeas/diagnóstico , Neoplasias Meníngeas/tratamento farmacológico , Neoplasias Meníngeas/radioterapia , Neoplasias Meníngeas/cirurgia , Meningioma/diagnóstico , Meningioma/tratamento farmacológico , Meningioma/radioterapia , Meningioma/cirurgia , Leucemia-Linfoma Linfoblástico de Células Precursoras/radioterapia , Resultado do TratamentoRESUMO
AIMS: The aim of this study was to determine the survival of 15 different strains of Salmonella of selected serotypes during prolonged cold storage of beef. METHODS AND RESULTS: Fifteen strains of eight different serotypes of Salmonella were spiked onto fresh cuts beef portions, and the survival was followed during storage in a laboratory cooling system. Over a 14-day period, all strains were reduced significantly in numbers; however, strains of Salmonella Typhimurium DT104 and Salmonella Enteritidis PT4 and PT8 survived significantly longer than strains of the serovars Dublin, Derby, Infantis and Newport. For five selected strains, the observations were verified in a pilot plant cooling facility mimicking industrial cooling. No significant differences in reduction were found between the two cooling methods. CONCLUSIONS: A significant reduction in Salmonella can be obtained by dry aging of beef during cold storage but the survival is strain dependent. SIGNIFICANCE AND IMPACT OF THE STUDY: From a hygienic point of view, cold storage of unpacked beef, which is still performed in small slaughterhouses, is a good alternative to vacuum packaging.
Assuntos
Contaminação de Alimentos/análise , Microbiologia de Alimentos , Armazenamento de Alimentos/métodos , Carne/microbiologia , Salmonella enteritidis/isolamento & purificação , Salmonella typhimurium/isolamento & purificação , Temperatura Baixa , Indústria de Embalagem de Carne , Salmonella enteritidis/crescimento & desenvolvimento , Salmonella typhimurium/crescimento & desenvolvimento , VácuoRESUMO
BACKGROUND: In patients with active Crohn's disease (CD), treatment of intra-abdominal abscess usually comprises antibiotics and radiologically guided percutaneous drainage (PD) preceding surgery. The aim of this study was to investigate the risk of postoperative complications and identify the optimal time interval for surgical intervention after PD. METHODS: A multicentre, international, retrospective cohort study was carried out. Details of patients with diagnosis of CD who underwent ultrasonography- or CT-guided PD were retrieved from hospital records using international classification of disease (ICD-10) diagnosis code for CD combined with procedure code for PD. Clinical variables were retrieved and the following outcomes were measured: 30-day postoperative overall complications, intra-abdominal septic complications, unplanned intraoperative adverse events, surgical-site infections, sepsis and pathological postoperative ileus, in addition to abscess recurrence. Patients were categorized into three groups according to the length of the interval from PD to surgery (1-14 days, 15-30 days and more than 30 days) for comparison of outcomes. RESULTS: The cohort comprised 335 CD patients with PD followed by surgery. Median age was 33 (i.q.r. 24-44) years, 152 (45.4 per cent) were females, and median disease duration was 9 (i.q.r. 3.6-15) years. Overall, the 30-day postoperative complications rate was 32.2 per cent and the mortality rate was 1.5 per cent. After adjustment for co-variables, older age (odds ratio 1.03 (95 per cent c.i. 1.01 to 1.06), P < 0.012), residual abscess after PD (odds ratio 0.374 (95 per cent c.i. 0.19 to 0.74), P < 0.014), smoking (odds ratio 1.89 (95 per cent c.i. 1.01 to 3.53), P = 0.049) and low serum albumin concentration (odds ratio 0.921 (95 per cent c.i. 0.89 to 0.96), P < 0.001) were associated with higher rates of postoperative complications. A short waiting interval, less than 2 weeks after PD, was associated with a high incidence of abscess recurrence (odds ratio 0.59 (95 per cent c.i. 0.36 to 0.96), P = 0.042). CONCLUSION: Smoking, low serum albumin concentration and older age were significantly associated with postoperative complications. An interval of at least 2 weeks after successful PD correlated with reduced risk of abscess recurrence.
Assuntos
Abscesso Abdominal , Doença de Crohn , Abscesso Abdominal/diagnóstico por imagem , Abscesso Abdominal/etiologia , Abscesso Abdominal/cirurgia , Adulto , Idoso , Doença de Crohn/complicações , Doença de Crohn/cirurgia , Drenagem , Feminino , Humanos , Estudos Retrospectivos , Listas de EsperaRESUMO
AIM: The risk of local recurrence following curative surgery for colorectal cancer (CRC) is up to 50%. A rigorous follow-up program may increase survival. Guidelines on suitable methods for scheduled follow up examinations are needed. This study evaluates a strict follow-up program including carcinogenic embryonic antigen (CEA), chest X-ray, abdominal ultrasound (US), computed tomography (CT) and (18)F-FDG positron emission tomography (FDG-PET). METHOD: A cohort of 132 patients, treated by surgery with curative intent for CRC, was included. Patients were followed prospectively with scheduled controls at 3, 6, 12 and 24 months after curative surgery. CEA, chest X-ray, US, CT and FDG-PET supplemented by clinical examination. The end-point was recurrence. Sensitivity and specificity was estimated 2 years after surgery. RESULTS: Of the 132 patients included in the study, 25 experienced recurrence, detected at scheduled controls (n = 18) and at intervals between them (n = 7). The results of CT and FDG-PET were correlated with recurrence. CT combined with FDG-PET had the highest specificity and sensitivity. CONCLUSION: A total of 72% of recurrences were detected at scheduled controls. The findings supported a strict follow-up program following curative surgery for colorectal cancer. FDG-PET combined with CT should be included in control programs.
Assuntos
Neoplasias Colorretais/patologia , Neoplasias Colorretais/cirurgia , Procedimentos Clínicos , Recidiva Local de Neoplasia/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno Carcinoembrionário/sangue , Feminino , Humanos , Fígado/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Exame Físico , Tomografia por Emissão de Pósitrons , Radiografia Abdominal , Radiografia Torácica , Sensibilidade e Especificidade , Sigmoidoscopia , Tomografia Computadorizada por Raios X , UltrassonografiaRESUMO
Intestinal dysbiosis in inflammatory bowel disease (IBD) patients depend on disease activity. We aimed to characterize the microbiota after 7 years of follow-up in an unselected cohort of IBD patients according to disease activity and disease severity. Fifty eight Crohn's disease (CD) and 82 ulcerative colitis (UC) patients were included. Disease activity was assessed by the Harvey-Bradshaw Index for CD and Simple Clinical Colitis Activity Index for UC. Microbiota diversity was assessed by 16S rDNA MiSeq sequencing. In UC patients with active disease and in CD patients with aggressive disease the richness (number of OTUs, p = 0.018 and p = 0.013, respectively) and diversity (Shannons index, p = 0.017 and p = 0.023, respectively) were significantly decreased. In the active UC group there was a significant decrease in abundance of the phylum Firmicutes (p = 0.018). The same was found in CD patients with aggressive disease (p = 0.05) while the abundance of Proteobacteria phylum showed a significant increase (p = 0.03) in CD patients. We found a change in the microbial abundance in UC patients with active disease and in CD patients with aggressive disease. These results suggest that dysbiosis of the gut in IBD patients is not only related to current activity but also to the course of the disease.
Assuntos
Doença de Crohn/etiologia , Doença de Crohn/patologia , Disbiose , Microbioma Gastrointestinal , Proteobactérias , Biodiversidade , Estudos de Casos e Controles , Doença de Crohn/diagnóstico , Progressão da Doença , Suscetibilidade a Doenças , Fezes/microbiologia , Humanos , Doenças Inflamatórias Intestinais/etiologia , Doenças Inflamatórias Intestinais/patologia , Metagenômica/métodos , RNA Ribossômico 16S/genética , Índice de Gravidade de DoençaRESUMO
In a genome-wide screen using differential methylation hybridization (DMH), we have identified a CpG island within the 5' region and untranslated first exon of the secretory granule neuroendocrine protein 1 gene (SGNE1/7B2) that showed hypermethylation in medulloblastomas compared to fetal cerebellum. Bisulfite sequencing and combined bisulfite restriction assay were performed to confirm the methylation status of this CpG island in primary medulloblastomas and medulloblastoma cell lines. Hypermethylation was detected in 16/23 (70%) biopsies and 7/8 (87%) medulloblastoma cell lines, but not in non-neoplastic fetal (n=8) cerebellum. Expression of SGNE1 was investigated by semi-quantitative competitive reverse transcription-polymerase chain reaction and found to be significantly downregulated or absent in all, but one primary medulloblastomas and all cell lines compared to fetal cerebellum. After treatment of medulloblastoma cell lines with 5-aza-2'-deoxycytidine, transcription of SGNE1 was restored. No mutation was found in the coding region of SGNE1 by single-strand conformation polymorphism analysis. Reintroduction of SGNE1 into the medulloblastoma cell line D283Med led to a significant growth suppression and reduced colony formation. In summary, we have identified SGNE1 as a novel epigenetically silenced gene in medulloblastomas. Its frequent inactivation, as well as its inhibitory effect on tumor cell proliferation and focus formation strongly argues for a significant role in medulloblastoma development.
Assuntos
Neoplasias Cerebelares/patologia , Metilação de DNA , Meduloblastoma/patologia , Proteína Secretora Neuroendócrina 7B2/genética , Azacitidina/análogos & derivados , Azacitidina/farmacologia , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células , Neoplasias Cerebelares/genética , Ilhas de CpG/genética , Metilases de Modificação do DNA/antagonistas & inibidores , Decitabina , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Inibidores Enzimáticos/farmacologia , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Meduloblastoma/genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Transcrição Gênica/efeitos dos fármacosRESUMO
Supratentorial primitive neuroectodermal tumors (sPNET) and atypical teratoid/rhabdoid tumors (AT/RT) of the CNS represent a biological and clinical enigma, despite advances in both molecular techniques and clinical management for these two rare embryonal brain tumors of childhood. Epigenetic changes hold great potential as possible disease mechanisms and may be manipulated therapeutically. We thus studied aberrant methylation of the genes RASSF1A and CASP8 and its consequence on expression in cell lines and primary tumors using a combination of semiquantitative methylation specific PCR (MSP), bisulfite sequencing and RT-PCR. In all, 17 samples of autopsy-derived normal appearing brain served as controls. Opposed to control tissues 19/24 sPNET and 4/6 AT/RT demonstrated aberrant methylation for the RASSF1A promoter region. Treatment of cell lines using 5-Aza-2'-deoxycytidine (5AZA) alone or in combination with trichostatin A (TSA) succeeded in re-establishing expression of RASSF1A in cell lines derived from a renal rhabdoid, an AT/RT and a medulloblastoma. A 5' CpG-rich region of CASP8 was methylated in normal tissues and in tumors. However, CASP8 showed inconsistent expression patterns in normal and tumor tissues. Our results indicate that aberrant methylation of the RASSF1A promoter region may be of importance in the origin and progression of sPNET and AT/RT while the analysed 5'-CpG rich region of the CASP8 gene does not seem to play an important role in these tumors. Further studies of epigenetic changes in these rare tumors are warranted as their biology remains obscure and treatment efforts have been rather unsuccessfull.
Assuntos
Neoplasias Encefálicas/genética , Metilação de DNA , Inativação Gênica , Tumores Neuroectodérmicos Primitivos/genética , Tumor Rabdoide/genética , Teratoma/genética , Proteínas Supressoras de Tumor/genética , Adolescente , Adulto , Azacitidina/análogos & derivados , Azacitidina/farmacologia , Caspase 8 , Caspases/genética , Criança , Pré-Escolar , Ilhas de CpG , Metilação de DNA/efeitos dos fármacos , Decitabina , Epigênese Genética , Feminino , Inativação Gênica/efeitos dos fármacos , Humanos , Ácidos Hidroxâmicos/farmacologia , Lactente , Masculino , Regiões Promotoras GenéticasRESUMO
Previous studies have shown cellular insulin resistance in conventionally treated insulin-dependent diabetics. To determine whether insulin resistance is also present in insulin-dependent diabetics before the commencement of insulin therapy, we studied nine newly diagnosed untreated insulin-dependent diabetics and nine control subjects. Insulin binding to adipocytes, monocytes, and erythrocytes was normal in the diabetic individuals. Basal (noninsulin stimulated) glucose transport rate was normal, whereas the maximal insulin responsiveness of glucose transport was severely impaired (P less than 0.02). Insulin sensitivity as judged by left or rightward shifts in the insulin dose-response curves was unchanged. Moreover, the basal lipogenesis rate measured at a glucose concentration of 0.5 mmol/liter was decreased in the diabetics (P less than 0.05), and the maximal insulin responsiveness of lipogenesis was also reduced (P less than 0.05). We conclude that fat cells from untreated insulin-deficient diabetics are insulin resistant. The major defects are (1) reduced maximal insulin responsiveness of glucose transport and conversion to lipids that are postbinding abnormalities, and (2) reduced basal glucose conversion to lipids.
Assuntos
Diabetes Mellitus Tipo 1/metabolismo , Glucose/metabolismo , Tecido Adiposo/metabolismo , Adulto , Transporte Biológico , Eritrócitos/metabolismo , Feminino , Humanos , Resistência à Insulina , Corpos Cetônicos/sangue , Lipídeos/biossíntese , Masculino , Monócitos/metabolismo , Receptor de Insulina/metabolismoRESUMO
Early detection of colorectal cancer (CRC) improves patient survival. Plasma tissue inhibitor of metalloproteinases 1 (TIMP-1) measurements by enzyme-linked immunosorbent assay (ELISA) have been suggested as a new method for the early detection of CRC. To further investigate the nature of TIMP-1 in plasma, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI TOF MS) was used. TIMP-1 measurements of plasma from 16 healthy donors and 14 CRC patients were performed using TIMP-1 monoclonal antibody in SELDI TOF MS and ELISA. SELDI TOF MS applying an antibody to TIMP-1 revealed that human plasma TIMP-1 has a mass of 25.1 kDa and exhibits several isoforms. Both methods showed increased plasma TIMP-1 values for cancer patients as compared to healthy individuals. The p values for the separation of the groups were 0.0019 for ELISA and <0.0001 for SELDI TOF MS. CRC did not fundamentally affect the appearance of TIMP-1 as evaluated by SELDI TOF MS.
Assuntos
Doadores de Sangue , Neoplasias Colorretais/sangue , Inibidor Tecidual de Metaloproteinase-1/sangue , Ensaio de Imunoadsorção Enzimática , Humanos , Isoformas de Proteínas/sangue , Valores de Referência , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Cysticercosis of the central nervous system is the main cause of late-onset epilepsy in tropical countries. The case of a 13-year-old German girl with a generalized seizure following long-term sojourns in the Tropics is reported. Cranial imaging showed two cerebral lesions with central calcifications. Serological, molecular and cultural examination of cerebrospinal fluid and blood was negative for various parasites, fungi and bacteria including mycobacteria. Histopathological examination after neurosurgical resection revealed calcareous bodies pathognomonic for platyhelminths, in particular tapeworms. Taken together, the radiological and histopathological findings indicate infection with cysticerci, the larvae of Taenia solium.
Assuntos
Encefalopatias/diagnóstico por imagem , Encéfalo/patologia , Neurocisticercose/diagnóstico por imagem , Convulsões/etiologia , Adolescente , Animais , Encéfalo/parasitologia , Encefalopatias/complicações , Encefalopatias/parasitologia , Feminino , Alemanha , Humanos , Imageamento por Ressonância Magnética , Neurocisticercose/complicações , Neurocisticercose/parasitologia , Taenia solium , Tomografia Computadorizada por Raios X , Viagem , Medicina TropicalRESUMO
AIM: To assess the potential use of plasma and urine levels of tissue inhibitor of metalloproteinases 1 (TIMP-1) in urothelial cancer. METHODS: TIMP-1 levels were determined in urine and plasma from healthy donors (n=26), patients with bacterial bladder infection (n=24), urothelial bladder adenoma (n=3) or adenocarcinoma (n=7). RESULTS: Free and total TIMP-1 in plasma were weakly but significantly correlated with age; urinary TIMP-1 was not. A strong correlation between free and total TIMP-1 in plasma was observed, with an average ratio of 0.85. No correlation between total TIMP-1 in urine and plasma was found (p=0.55). No significant differences in free or total TIMP-1 in plasma were found between healthy individuals, patients with cystitis or bladder cancer (p=0.4). Urinary TIMP-1 levels were significantly increased in patients with cystitis (p=0.001). No apparent differences in TIMP-1 levels were found in patients with bladder cancer at different stages. CONCLUSION: Our previous observation of a weak but significant correlation between plasma TIMP-1 and age was confirmed. Likewise, an association between free and total TIMP-1 in plasma with a ratio of 0.85 was established. No correlation between plasma and urine TIMP-1 was found. Measurement of TIMP-1 in plasma and/or urine is apparently not useful for the identification of bladder cancer.
Assuntos
Inibidor Tecidual de Metaloproteinase-1/sangue , Inibidor Tecidual de Metaloproteinase-1/urina , Neoplasias da Bexiga Urinária/sangue , Neoplasias da Bexiga Urinária/urina , Adenocarcinoma/diagnóstico , Adenoma/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Infecções Bacterianas/diagnóstico , Creatinina/urina , Cistite/sangue , Cistite/urina , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doenças da Bexiga Urinária/diagnósticoRESUMO
The pig acute phase protein (APP) response to experimental Streptococcus suis (S. suis) infection was mapped by the measurement of the positive APPs C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp) and major acute phase protein (pig-MAP) and the negative APPs albumin and apolipoprotein (Apo) A-I. The aim was to elucidate the differences in the acute phase behaviour of the individual APPs during a typical bacterial septicaemic infection. Pigs were inoculated subcutaneously with live S. suis serotype 2 and blood was sampled before and on various days post inoculation (p.i.), until the pigs were killed and autopsied on day 14 p.i. Clinical signs (fever and lameness) were observed in four of the five inoculated pigs from day 2 p.i., and these pigs also had arthritic lesions at autopsy. CRP and SAA showed fast increases in serum concentrations, CRP being elevated from days 1 to 12 p.i. and peaking at 10 times the day 0-levels on day 1 p.i. SAA rose quickly to peak levels of 30-40 times the day 0-level on days 1-2 and returned to pre-inoculation level on day 5 p.i. Hp and pig-MAP showed slightly slower responses, both peaking around 5 days p.i. Hp was increased throughout the experiment with maximum levels around 10 times the day 0-levels, and pig-MAP was elevated on days 1-12 p.i. with peak levels of around seven times the day 0-levels. Apo A-I was decreased from days 1 to 8 and showed minimum levels of about 40% of day 0-levels around 1-2 days p.i. No clear pattern of changes in albumin levels could be identified. One pig, showing clinical signs on day 2 only, also showed an APP response, although of a relatively short duration, whereas three pigs presenting clinical signs for several days had a more protracted acute phase response. Remarkably, the one pig showing no clinical signs and no arthritic lesions showed an APP response comparable to that of the other, clinically affected pigs. Thus, both acute clinical and subclinical S. suis infection could be revealed by the measurement of one or more of the APPs CRP, SAA, Hp, pig-MAP and Apo A-I. The combined measurement of two or three APPs, including proteins with slow and fast kinetics, should be used to achieve the highest sensitivity for the detection of ongoing S. suis infection during a prolonged time period. A diagnostic tool based on such APP-measurements could considerably improve strategic control procedures for this important infection.
Assuntos
Infecções Estreptocócicas/veterinária , Streptococcus suis/imunologia , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Proteínas de Fase Aguda/imunologia , Animais , Apolipoproteína A-I/imunologia , Temperatura Corporal/imunologia , Proteína C-Reativa/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Haptoglobinas/imunologia , Imunodifusão/veterinária , Coxeadura Animal/imunologia , Proteína Amiloide A Sérica/imunologia , Organismos Livres de Patógenos Específicos , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/microbiologia , SuínosRESUMO
Part-skim Mozzarella cheese was manufactured from milk hydrolyzed with fungal phospholipase A1 prior to renneting. The phospholipase treatment reduced fat losses in whey and cooking water and increased cheese yield as a result of improved fat and moisture retention in the cheese curd. The amount of phospholipids in the whey was reduced because of improved retention of lysophospholipids in the cheese curd. Water binding in the fresh curds and young cheeses up to 3 wk of storage was investigated by a 1H nuclear magnetic resonance spin-spin relaxation technique. In the fresh curds, 2 dominant water fractions were present, characterized by average spin-spin relaxation times (T2) of 14 and 86 to 89 ms, respectively. These 2 fractions of low- and high-molecular-mobility water were similar in all cheeses and presumed to represent water associated with the casein matrix and water present in the pores. A few hours after manufacture, cheeses made with phospholipase showed decreased T2 of the high-mobility fraction, indicating improved water-holding capacity. It is suggested that lysophospholipids released from the fat globule membranes act as surface-active agents in the cheese curd, helping emulsification of water and fat during processing and reducing syneresis. During 3 wk of storage after manufacture, the mobility of both water fractions increased in all cheeses, but was highest in the cheeses made with phospholipase. The increase in mobility during the first weeks of storage has earlier been ascribed to structural changes in the protein matrix, which in principle could be accelerated because of the higher moisture content. However, the microstructure of phospholipase-treated cheese was investigated by confocal laser scanning microscopy and found to be very similar to the control cheese during processing and up to 28 d of storage. In addition, flowability, stretchability, and browning were acceptable and similar in all the manufactured cheeses. Thus, phospholipase hydrolysis of cheese milk improved the cheese yield without changing the cheese microstructure, and resulted in cheese with functional properties that were identical to traditional Mozzarella cheese.
Assuntos
Queijo/análise , Indústria de Laticínios/métodos , Tecnologia de Alimentos/métodos , Fosfolipases A1/metabolismo , Animais , Queijo/normas , Manipulação de Alimentos/métodos , Fusarium/enzimologia , Hidrólise , Espectroscopia de Ressonância Magnética/métodos , Leite/química , Leite/metabolismo , Proteínas do Leite/análise , Fosfolipídeos/análise , Fatores de Tempo , Água/análise , Água/metabolismo , Proteínas do Soro do LeiteRESUMO
Medulloblastoma (MB) represents the most frequent malignant brain tumor in children. Most MBs appear sporadically; however, their incidence is highly elevated in two inherited tumor predisposition syndromes, Gorlin's and Turcot's syndrome. The genetic defects responsible for these diseases have been identified. Whereas Gorlin's syndrome patients carry germ-line mutations in the patched (PTCH) gene, Turcot's syndrome patients with MBs carry germ-line mutations of the adenomatous polyposis coli (APC) gene. The APC gene product is a component of a multiprotein complex controlling beta-catenin degradation. In this complex, Axin plays a major role as scaffold protein. Whereas APC mutations are rare in sporadic MBs, a hot-spot region of beta-catenin (CTNNB1) mutations was identified in a subset of MBs. To find out if Axin is also involved in the pathogenesis of sporadic MBs, we analyzed 86 MBs and 11 MB cell lines for mutations in the AXIN1 gene. Using single-strand conformation polymorphism analysis, screening for large deletions by reverse transcription-PCR, and sequencing analysis, a single somatic point mutation in exon 1 (Pro255Ser) and seven large deletions (12%) of AXIN1 were detected. This indicates that AXIN1 may function as a tumor suppressor gene in MBs.
Assuntos
Neoplasias Encefálicas/genética , Deleção de Genes , Meduloblastoma/genética , Proteínas/genética , Proteínas Proto-Oncogênicas/fisiologia , Proteínas Repressoras , Transdução de Sinais/genética , Proteínas de Peixe-Zebra , Adolescente , Adulto , Proteína Axina , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Perda de Heterozigosidade , Masculino , Pessoa de Meia-Idade , Polimorfismo Conformacional de Fita Simples , Proteínas Proto-Oncogênicas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/fisiologia , Proteínas WntRESUMO
Inactivating mutations in the PTCH gene, a human homologue of the Drosophila segment polarity gene patched, have been identified recently in patients with nevoid basal cell carcinoma syndrome. These patients are predisposed to various neoplasias including basal cell carcinomas and medulloblastomas (MBs). To determine the involvement of PTCH in sporadic MBs, which represent the most frequent malignant brain tumors in children, we screened for PTCH alterations in an unselected panel of 64 biopsy samples from 62 patients and four continuous MB cell lines, all derived from patients with sporadic MBs. Using single-strand conformational polymorphism analysis, we screened exons 2-22 and detected nonconservative PTCH mutations in 3 of 11 samples from sporadic cases of the desmoplastic variant of MB but none in 57 MBs with classical (nondesmoplastic) histology. In two of the tumors with mutations and in two additional desmoplastic cases, loss of heterozygosity was found at 9q22. These findings suggest that PTCH represents a tumor suppressor gene involved in the development of the desmoplastic variant of MB.
Assuntos
Neoplasias Cerebelares/genética , Meduloblastoma/genética , Proteínas de Membrana/genética , Adolescente , Animais , Criança , Pré-Escolar , Feminino , Regulação Neoplásica da Expressão Gênica , Variação Genética , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Mutação , Receptores Patched , Receptor Patched-1 , Polimorfismo Conformacional de Fita Simples , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Receptores de Superfície Celular , Análise de Sequência de DNA , Células Tumorais CultivadasRESUMO
Whether patients diagnosed with primary breast cancer are offered adjuvant systemic therapy following surgical removal of the tumor is based on prognosis. Prognosis is estimated in every patient using established prognostic variables. Unfortunately, when using the currently available prognostic parameters a significant proportion of patients are over-treated. Thus, in order to improve stratification of breast cancer patients, additional prognostic factors need to be identified. Tissue inhibitor of metalloproteinases-1 (TIMP-1) is one of the promising candidates for new prognostic markers in breast cancer, as a number of studies have demonstrated an association between high tumor-tissue levels of TIMP-1 mRNA as well as TIMP-1 protein and a poor prognosis of breast cancer patients. TIMP-1 is a member of the TIMP family, currently comprising four members (TIMP-1-4), and its main function is inhibition of the activity of various matrix metalloproteinases (MMPs). The association between high levels of protease inhibitor and poor prognosis may be somewhat surprising, as proteolytic activity plays a pivotal role in cancer cell invasion and metastasis. However, the recent discovery of other biological functions of TIMP-1 such as growth-stimulating functions, as well as anti-apoptotic and pro-angiogenetic effects, may in part explain this paradox. The purpose of this review is to give an update on the current status of TIMP-1 in breast cancer, emphasizing the prognostic utility of the inhibitor. In addition, the suggested tumor-stimulatory roles of TIMP-1 will be outlined.
Assuntos
Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/diagnóstico , Inibidor Tecidual de Metaloproteinase-1/análise , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Biomarcadores Tumorais/genética , Feminino , Humanos , Prognóstico , RNA Mensageiro/análise , Inibidor Tecidual de Metaloproteinase-1/genéticaRESUMO
Phosphorylase kinase from human polymorphonuclear leukocytes was investigated in a gel filtered crude preparation (17,000 x g supernatant). It was found to exist in two forms, one (the phosphorylated form) more active than the other (the dephosphorylated form). Interconversion between the two forms was carried out by a cyclic AMP dependent protein kinase and phosphoprotein phosphatase, respectively. The ratio of activity measured at pH 8.0 and 6.0 was 0.36 for the non-activated and 0.83 for the activated form, which is in contrast to the behaviour of phosphorylase kinase from muscle. Km app for the substrate phosphorylase b was 650 U/ml and 85 U/ml for the non-activated and activated form, respectively, whereas Km app for ATP was 0.03 mM and identical for the two forms. The non-activated form of phosphorylase kinase was activated by Ca2+ in the range 10(-7)--5 . 10(-6) M, which may have physiological importance, whereas the activated form was insensitive to variations in Ca2+ concentration between 10(-9) and 10(-3) M.