RESUMO
The gene coding for xylose isomerase from the thermophilic bacterium Fervidobacterium gondwanense was cloned and overexpressed in Escherichia coli. The produced xylose isomerase (XylA), which closely resembles counterparts from Thermotoga maritima and T. neapolitana, was purified and characterized. It is optimally active at 70 degrees C, pH 7.3, with a specific activity of 15.0 U/mg for the interconversion of glucose to fructose. When compared with T. maritima XylA at 85 degrees C, a higher catalytic efficiency was observed. Divalent metal ions Co2+ and Mg2+ were found to enhance the thermostability.
Assuntos
Aldose-Cetose Isomerases/metabolismo , Proteínas de Bactérias/metabolismo , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/enzimologia , Proteínas Recombinantes/metabolismo , Aldose-Cetose Isomerases/química , Aldose-Cetose Isomerases/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Southern Blotting , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Genes Bacterianos , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/genética , Meia-Vida , Cinética , Filogenia , Proteínas Recombinantes/química , Proteínas Recombinantes/genéticaRESUMO
Transformation of non-competent Escherichia coli JM109 was accomplished using pUC19 as donor plasmid and sepiolite as the acicular material to promote cell piercing via application of friction with a polystyrene stick or a magnetic bar on the surface of a hydrogel containing agar. An automatic spreading setup was built with a conventional stirring plate and compared to manual spreading. Several parameters were optimized, namely, the agar content of the hydrogel (2%), concentration of cells (OD=1.3 corresponding to 1.4×10(9) bacterial cells/mL), concentration of sepiolite (0.01%), manual versus mechanical spreading (automatic spreading more consistent) and spreading time (30s). Efficiency values up to 4.1×10(4) CFU/µg pUC19 were obtained. The method proved to be suitable for a rapid and low cost transformation of non-competent E. coli JM109, where higher values of efficiency do not need to be attained.
Assuntos
Escherichia coli/genética , Técnicas de Transferência de Genes , Plasmídeos/genética , Transformação Bacteriana , Escherichia coli/metabolismo , Plasmídeos/metabolismoRESUMO
SUMMARY: Curcuminoids, components of the rhizome of turmeric, show several beneficial biological activities, including anticarcinogenic, antioxidant, anti-inflammatory, and antitumor activities. Despite their numerous pharmaceutically important properties, the low natural abundance of curcuminoids represents a major drawback for their use as therapeutic agents. Therefore, they represent attractive targets for heterologous production and metabolic engineering. The understanding of biosynthesis of curcuminoids in turmeric made remarkable advances in the last decade, and as a result, several efforts to produce them in heterologous organisms have been reported. The artificial biosynthetic pathway (e.g., in Escherichia coli) can start with the supplementation of the amino acid tyrosine or phenylalanine or of carboxylic acids and lead to the production of several natural curcuminoids. Unnatural carboxylic acids can also be supplemented as precursors and lead to the production of unnatural compounds with possibly novel therapeutic properties. In this paper, we review the natural conversion of curcuminoids in turmeric and their production by E. coli using an artificial biosynthetic pathway. We also explore the potential of other enzymes discovered recently or already used in other similar biosynthetic pathways, such as flavonoids and stilbenoids, to increase curcuminoid yield and activity.
Assuntos
Vias Biossintéticas , Curcuma , Curcumina/análogos & derivados , Curcumina/metabolismo , Escherichia coli/metabolismo , Disponibilidade Biológica , Ácidos Carboxílicos/metabolismo , Curcuma/química , Curcuma/metabolismo , Escherichia coli/genética , Fenilalanina/metabolismo , Policetídeos/metabolismo , Tirosina/metabolismoRESUMO
Caffeic acid is a plant secondary metabolite and its biological synthesis has attracted increased attention due to its beneficial effects on human health. In this study, Escherichia coli was engineered for the production of caffeic acid using tyrosine as the initial precursor of the pathway. The pathway design included tyrosine ammonia lyase (TAL) from Rhodotorula glutinis to convert tyrosine to p-coumaric acid and 4-coumarate 3-hydroxylase (C3H) from Saccharothrix espanaensis or cytochrome P450 CYP199A2 from Rhodopseudomonas palustris to convert p-coumaric acid to caffeic acid. The genes were codon-optimized and different combinations of plasmids were used to improve the titer of caffeic acid. TAL was able to efficiently convert 3mM of tyrosine to p-coumaric acid with the highest production obtained being 2.62mM (472mg/L). CYP199A2 exhibited higher catalytic activity towards p-coumaric acid than C3H. The highest caffeic acid production obtained using TAL and CYP199A2 and TAL and C3H was 1.56mM (280mg/L) and 1mM (180mg/L), respectively. This is the first study that shows caffeic acid production using CYP199A2 and tyrosine as the initial precursor. This study suggests the possibility of further producing more complex plant secondary metabolites like flavonoids and curcuminoids.
Assuntos
Ácidos Cafeicos/metabolismo , Escherichia coli K12/metabolismo , Tirosina/metabolismo , Actinobacteria/enzimologia , Actinobacteria/genética , Amônia-Liases/genética , Amônia-Liases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Vias Biossintéticas , Ácidos Cumáricos/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , DNA Bacteriano/genética , Escherichia coli K12/genética , Genes Bacterianos , Engenharia Genética , Dados de Sequência Molecular , Propionatos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Rodopseudomonas/enzimologia , Rodopseudomonas/genética , Rhodotorula/enzimologia , Rhodotorula/genéticaRESUMO
Many cancers do not respond to chemotherapy on primary exposure to drugs, thus manifesting intrinsic drug resistance. Other cancers that do initially respond subsequently become resistant to the same drugs and simultaneously to other drugs to which the patient has had no previous exposure. This is a form of acquired drug resistance. There is a pressing need to better understand the mechanisms of drug resistance and to use this information to develop strategies for the chemosensitization of drug-resistant tumors. A goal of the pathology laboratory is to offer chemosensitivity tests that identify intrinsic or acquired resistance of tumors to specific drugs or classes of drugs to enable the clinician to tailor therapy to the biology of cancers in individual patients. Multidrug resistance is one type of drug resistance. It can be present in either an intrinsic or acquired form. The human gene that confers human multidrug resistance, the MDR1 gene, has been cloned and classified as a member of the MDR gene family. Its encoded protein, called Mdr1, is an energy-driven membrane efflux transporter that maintains intracellular concentrations of certain chemotherapeutic drugs at nontoxic levels. Useful model systems for studying multidrug resistance have been developed in several research laboratories. Applying selection pressure by exposing cultured cancer cells to escalating doses of natural product anti-cancer drugs allows cross-resistant cell lines to be produced which share patterns of drug resistance with human cancers. A common feature of these drug-resistant lines is the expression of Mdr1. Using techniques of genetic engineering, molecular probes have been developed that can be used to measure MDR1 mRNA and MDR1 gene amplification. Mdr can be measured by immunochemistry methods. Currently, such measurements are being used to stratify patients in clinical trials designed to determine if chemosensitization by inhibition of the pump function of Mdr is a clinically useful therapeutic strategy. If successful, Mdr/MDR1 mRNA laboratory testing might significantly increase the clinical laboratory's role in cancer patient management.
Assuntos
Resistência a Medicamentos/genética , Genes , Glicoproteínas de Membrana/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Animais , Biomarcadores Tumorais/análise , Expressão Gênica , Marcadores Genéticos/análise , Humanos , Glicoproteínas de Membrana/análise , Neoplasias/análise , Neoplasias/tratamento farmacológico , Neoplasias/genéticaRESUMO
Classic multidrug resistance is mediated by a P-glycoprotein. Using monoclonal antibody C219 (MAb C219) in an immunohistochemical study, we found high levels of putative Golgi P-glycoprotein in normal columnar and transitional epithelium in subpopulations of patients with specific blood types. For example, Golgi staining was present in blood type A patients in 46% of normal colon samples (N = 21) and 88% of normal ureter samples (N = 17). In comparison, Golgi staining was present in blood group O patients in only 6% of normal colon samples (N = 34) and in 0% of normal ureter samples (N = 19). The association of MAb C219 Golgi staining with blood type A and lack of Golgi staining with blood type O was statistically significant in normal colon (P = .001) and normal ureter (P less than .0001). Inappropriate hyperexpression of P-glycoprotein was frequently found in colon carcinomas. Additional evidence that Golgi MAb C219 reactivity represents P-glycoprotein is presented. This includes (1) immunostaining of Golgi with two anti-P-glycoprotein MAbs, C219 and JSB-1, and (2) experiments in which Mab C219 Golgi reactivity was blocked by preincubation of MAb C219 with a specific P-glycoprotein epitope-containing peptide. The high degree of association of Golgi P-glycoprotein with blood type A may suggest a role for P-glycoprotein in processing or trafficking of specific blood group antigens.
Assuntos
Sistema ABO de Grupos Sanguíneos , Anticorpos Monoclonais , Reações Antígeno-Anticorpo , Colo/análise , Glicoproteínas de Membrana/análise , Ureter/análise , Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais/imunologia , Colo/patologia , Neoplasias do Colo/análise , Neoplasias do Colo/sangue , Neoplasias do Colo/patologia , Resistência a Medicamentos , Epitélio/análise , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Células Tumorais Cultivadas/análise , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
The relative increase in endometrial adenocarcinoma in women has increased the need for more objective criteria in the distinction of hyperplastic and neoplastic endometrium. The authors have used the ability of lectins to detect changes in surface glycoproteins to probe the differences among proliferative endometrium, endometrial adenomatous hyperplasia and adenocarcinomas. Paraffin-embedded sections of tissue obtained by Vakutage endometrial sampling technics were stained with each of 7 FITC lectin conjugates. Thirty-four specimens were examined (7 proliferative, 12 hyperplastic, and 15 adenocarcinomatous). Wheat germ agglutinin binding was detectable in all specimens with a distribution at the cell luminal border of glandular formations irrespective of diagnosis. However, adenocarcinoma cases showed distribution along the lumenal border and the cell periphery with loss of orientation of the lectin binding. Similar alterations and increased binding were noted for Concanavalin A. The WGA binding to sections was specifically inhibitable by oligosaccharides of N-acetyl-glucosamine. The results provide an objective criterion for detection of loss of cell orientation useful in the diagnosis of endometrial adenocarcinoma in tissue fragments.
Assuntos
Adenocarcinoma/diagnóstico , Hiperplasia Endometrial/diagnóstico , Lectinas/metabolismo , Neoplasias Uterinas/diagnóstico , Adenocarcinoma/imunologia , Especificidade de Anticorpos , Ligação Competitiva , Diagnóstico Diferencial , Feminino , Humanos , Lectinas/imunologia , Ligação Proteica , Neoplasias Uterinas/imunologiaRESUMO
BACKGROUND: Fine-needle aspiration breast biopsy has been used increasingly as an alternative to excisional biopsy. The purpose of this study is to evaluate the accuracy of fine-needle aspiration with histopathologic confirmation. METHODS: A retrospective study was performed using a computer database over a 5-year period. All women who had had fine-needle aspiration breast biopsy with histopathologic confirmation of the diagnosis were included. Fine-needle aspirations were interpreted as malignant, suspicious, or benign. Histopathologic diagnosis included core-needle biopsy, open excisional biopsy, or mastectomy specimen. RESULTS: A total of 697 patients fulfilled the criteria. Only 5 (0.7%) of the specimens were inadequate for study. There were 401 total malignant fine-needle aspiration diagnoses, with only 3 false-positive specimens. All three were ductal hyperplasia, one from a previously radiated breast. There were 125 suspicious readings; 84 of these were malignant and 41 were false-suspicious specimens. Most of the false-suspicious lesions were fibrocystic disease. Of the 166 lesions interpreted as benign, there were 13 false-negative specimens. The test had a 97% sensitivity, 78% specificity, 92% positive predictive value, and 92% negative predictive value. CONCLUSIONS: Fine-needle aspiration is a sensitive test that can be useful as an adjunct in the diagnosis of breast cancer. "Malignant" and "benign" interpretations are highly predictive but must be used only in the context of other diagnostic modalities. "Suspicious" lesions require further investigation.
Assuntos
Biópsia por Agulha , Doenças Mamárias/patologia , Neoplasias da Mama/patologia , Mama/patologia , Doença da Mama Fibrocística/patologia , Mama/citologia , Neoplasias da Mama/cirurgia , Bases de Dados como Assunto , Reações Falso-Negativas , Reações Falso-Positivas , Feminino , Humanos , Mastectomia , Sistemas Computadorizados de Registros Médicos , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
The National Bladder Cancer Group undertook a study to determine the effectiveness and toxicity of mitomycin in patients who failed on thiotepa. A total of 117 patients with residual superficial transitional cell bladder cancer (Ta, T1, Tis) who had previously failed on intravesical thiotepa were treated with 40 mg of mitomycin instilled intravesically weekly for eight weeks. Four to six weeks after the last treatment, tumor response was evaluated by cystoscopy, biopsy of the site of the index lesion, and cytology. In 57 patients (48.7%), visible tumor had been ablated. Results of cystoscopy, biopsy, and cytology were negative in 32 (27.4%) patients. Eleven patients (9.4%) had no visible tumor and negative cytology unconfirmed by biopsy. In 14 patients (12%) who had complete destruction of the tumor at cystoscopy, and biopsy specimen was negative for tumor, cytology was positive, indicating a partial response. Six patients (5.1%) withdrew from the study before the first evaluation because of local toxicity (cystitis).
Assuntos
Mitomicinas/uso terapêutico , Neoplasias da Bexiga Urinária/cirurgia , Administração Tópica , Biópsia , Carcinoma in Situ/prevenção & controle , Ensaios Clínicos como Assunto , Cistite/induzido quimicamente , Cistoscopia , Avaliação de Medicamentos , Feminino , Humanos , Masculino , Mitomicinas/administração & dosagem , Mitomicinas/efeitos adversos , Mucosa/efeitos dos fármacos , Mucosa/patologia , Metástase Neoplásica/prevenção & controle , Recidiva Local de Neoplasia/prevenção & controle , Estadiamento de Neoplasias , Estudos Prospectivos , Tiotepa/efeitos adversos , Tiotepa/uso terapêutico , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/tratamento farmacológico , Urina/análiseRESUMO
Fine-needle aspiration (FNA) biopsy of a thyroid nodule was performed in 797 patients. Ninety-six patients had resection of the thyroid nodule performed subsequent to a one-time FNA biopsy. The surgical pathology of these 96 cases demonstrated a 5.8% false-negative rate and a 9.9% false-positive rate. As a consequence, we prospectively evaluated the routine practice of repeat FNA of cytologically benign thyroid nodules. Repeat FNA confirmed the original benign cytology in 183 (93%) of 196 patients. Seventeen of these 183 patients with benign FNA on both biopsies had resection of the nodule performed because of the development of suspicious clinical signs or in response to the patient's choice; 1 recurrent cyst was found to be carcinomatous. Of the 13 patients demonstrating a change in cytology on repeat FNA biopsy, 9 had a nodule that was classified as possibly malignant (suspicious); 6 of these patients underwent resection, and 1 patient was found to have a carcinomatous nodule. Four patients had nodules that were classified as probably malignant on repeat FNA biopsy; all of their nodules were resected, and three of them were found to be carcinomatous. This study demonstrates that, although one-time FNA biopsy of thyroid nodules is highly accurate, with a relatively low false-negative rate, repeat fine-needle biopsy improves on this diagnostic accuracy, thereby decreasing the risk of misdiagnosing a thyroid nodule that is malignant.
Assuntos
Biópsia por Agulha , Nódulo da Glândula Tireoide/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Reações Falso-Negativas , Reações Falso-Positivas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Neoplasias da Glândula Tireoide/patologia , Nódulo da Glândula Tireoide/cirurgiaRESUMO
Intrauterine aspiration (Vakutage), endocervical aspiration cytology and vaginal, ectocervical and endocervical (VCE) smears were used as outpatient procedures for diagnosis of endometrial cancer and hyperplasias in 840 patients. The results of the three technics were correlated with D&C or hysterectomy specimens. Ninety-three percent of the malignant lesions (82 adenocarcinomas, 5 mixed mesodermal tumors, 5 mixed adenosquamous carcinomas and 5 metastatic adenocarcinomas) were diagnosed by the Vakutage sample but only 67% by the VCE smears and 68% by endocervical aspiration cytology. The diagnostic accuracy of Vakutage in 50 cases of cystic hyperplasia was 88% (as compared to 14% for VCE smears and endocervical aspiration cytology) and 89% in 90 cases of adenomatous hyperplasias (as compared to 20% for the other two technics). Endometrial polyps were diagnosed exclusively from the Vakutage tissue sample in 83% of the 42 cases.
Assuntos
Adenocarcinoma/diagnóstico , Citodiagnóstico/métodos , Hiperplasia Endometrial/diagnóstico , Neoplasias Uterinas/diagnóstico , Biópsia por Agulha , Estudos de Avaliação como Assunto , Feminino , Humanos , Neoplasias do Colo do Útero/diagnóstico , Esfregaço VaginalRESUMO
Cutaneous and subcutaneous masses of the abdominal wall are uncommon. However, a variety of benign and neoplastic entities can be encountered in this region. We report a series of 22 fine-needle aspirations (FNA) of malignant cutaneous and subcutaneous lesions involving the abdominal wall. All of these lesions were metastatic neoplasms. There were 14 females and eight males, with an age range of 35-83 years (mean 65 years). Twenty-one had a previous history of malignancy. The mean interval between the primary diagnosis and FNA was 26 months (range 1.5-128 months). The sites of origin in order of decreasing frequency were colon (n = 4), ovary (n = 4), breast (n = 3), endometrium (n = 2), melanoma (n = 2), and one case each of cervix, urinary bladder, kidney, pancreas, gallbladder, and lymphoma. One case was a squamous-cell carcinoma of unknown origin. Fourteen of the 22 patients were dead at the end of this study, with a mean survival of 8.4 months (range 0.5-44 months) following FNA. One patient was alive with disease at 13 months, and seven patients were lost to follow-up. Based on this data and on review of the literature we conclude that the majority of malignant cutaneous and subcutaneous lesions of the abdominal wall subject to FNA biopsy are metastatic tumors which originate from intra-abdominal, pelvic, and retroperitoneal organs and that FNA is a highly useful technique in the assessment of these lesions of the abdominal wall.
Assuntos
Músculos Abdominais/patologia , Biópsia por Agulha , Metástase Neoplásica/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica/diagnósticoRESUMO
Only 15% of primary malignant tumors are confined to the gland at the time of presentation. Fine-needle aspiration (FNA) is valuable in confirming the malignant nature of these unresectable lesions. Benign pancreatic lesions and metastatic neoplasms can also be evaluated by fine-needle aspiration. We undertook a retrospective study to evaluate the efficacy of FNA in assessing pancreatic masses. Three hundred and sixty-four radiologically guided FNAs of the pancreas performed between 1986-1996 were reviewed. Surgical material was also evaluated when available and compared to the FNA material. There were 223 men and 141 women. Their ages ranged from 23-90, with a mean of 64 yr. Two hundred and twelve patients (58%) had primary malignant tumors, 183 had adenocarcinomas, 15 had mucinous cystadenocarcinomas, 12 had neuroendocrine tumors, and 2 had pleomorphic giant-cell carcinomas. Ninety-one (43%) had available surgical material which showed adenocarcinoma. Ninety-one patients (25%) had benign aspirates, including 53 showing benign ductal epithelium, 23 showing pancreatitis/inflammation. 10 showing pseudocysts, and 7 showing serous cystadenomas. Surgical material was available in 24 (26%) of these patients. Two of these showed adenocarcinoma. Sixteen aspirates (4%) were suspicious for malignancy, 13 (81%) of which showed adenocarcinoma on follow-up biopsies. Twenty-two aspirates (6%) showed metastatic neoplasms. Twenty-three (6%) had unsatisfactory specimens. Ten (43%) of these had follow-up biopsies, 3 of which were malignant. FNA of primary benign and malignant pancreatic masses is highly sensitive (98%) and specific (100%). Eighty-one percent of the suspicious lesions showed adenocarcinoma on follow-up biopsy. FNA of metastatic neoplasms to the pancreas is also very accurate. This technique can be useful in avoiding unnecessary surgery.
Assuntos
Pâncreas/patologia , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/secundário , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia por Agulha , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
A variety of inflammatory and neoplastic scalp lesions are encountered in surgical pathology. However, the literature on fine-needle aspirations (FNAs) of the scalp is exceedingly rare. We report on a series of 70 FNAs involving cutaneous and subcutaneous lesions on the scalp. A total of 70 fine-needle aspirations of cutaneous and subcutaneous scalp lesions was reviewed to identify patterns of metastasis to the scalp and to demonstrate the effectiveness of FNA in diagnosing these lesions. There were 42 male and 28 female patients, ranging in age from 29-91 yr (mean, approximately 61 yr). Sixty-one patients had a previous history of malignancy. Of these aspirates, 59 were neoplastic, consistent with the patient's known primary. One case was an abscess, and the remaining case was unsatisfactorvy for cytologic evaluation. Follow-up biopsy revealed granulomatous inflammation. The most common primary tumor to metastasize to the scalp was lung carcinoma, which was seen in 18 cases, followed by hematopoietic malignancies in 14 cases. Melanoma was identified in 6 cases, head and neck tumors in 5 cases, renal malignancies in 4 cases, gastrointestinal tumors in 3 cases, sarcoma in 3 cases, breast and prostate malignancy in 2 cases each, and an olfactory neuroblastoma and meningioma in 1 case each. The remaining 9 aspirates were from patients who did not have a previous history of malignancy. These included 6 benign aspirates consisting of 3 epidermal inclusion cysts, 2 lipomas, and 1 neurofibroma. Two aspirates were malignant and included 1 primary squamous-cell carcinoma and 1 metastatic adenocarcinoma of unknown origin. The remaining case was unsatisfactory for cytologic evaluation. Follow-up biopsy of this lesion showed noncaseating granulomas. Of the aspirates from patients with a previous history of malignancy, 97% were neoplastic. Lung carcinoma and hematopoietic malignancies were the most common neoplasms that metastasized to the scalp. Since the scalp is a common site for metastasis, awareness of this fact is useful to both oncologists and dermatologists. It must be understood that FNA can provide a rapid and accurate diagnosis in the evaluation of scalp masses.
Assuntos
Couro Cabeludo/patologia , Neoplasias Cutâneas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia por Agulha/métodos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Neoplasias Cutâneas/diagnóstico , Neoplasias Cutâneas/secundárioRESUMO
With improved radiologic techniques fine-needle aspiration (FNA) is becoming a rapid, effective diagnostic method in evaluating a wide range of liver masses. Review of six hundred two radiologically guided liver aspirates performed over a ten-year period forms the basis of this report.
Assuntos
Biópsia por Agulha , Neoplasias Hepáticas/diagnóstico , Fígado/patologia , Carcinoma Hepatocelular/diagnóstico , Carcinoma Neuroendócrino/diagnóstico , Carcinoma de Células Escamosas/diagnóstico , Colangiocarcinoma/diagnóstico , Feminino , Humanos , Fígado/diagnóstico por imagem , Linfoma/diagnóstico , Masculino , Melanoma/diagnóstico , Sarcoma/diagnóstico , Tomografia Computadorizada por Raios X , UltrassonografiaRESUMO
Renal masses secondary to metastases are not common. Few comprehensive reviews exist, which consist primarily of autopsy and radiologic reports. The purpose of this study was to review the types and incidences of various neoplasms which metastasize to the kidney and to determine the usefulness of fine-needle aspiration (FNA) in diagnosing them. Two hundred and sixty-one radiologically guided FNAs of renal lesions over a 9-yr period were reviewed. The diagnoses of the 261 renal FNAs were as follows: 136 (52%) were malignant, 111 (43%) were benign, and 14 (5%) were unsatisfactory. Of the 136 positive FNAs, 28 (21%) revealed metastatic tumors. The overall incidence of renal FNAs displaying metastatic tumors was 11%. Among the 28 patients with metastases to the kidney, 23 patients were men and 5 were women, with the mean age being 58 yr. Twenty-five patients (89%) had prior history of a primary malignancy, including lung carcinoma (11 cases, 39%), lymphoma (8 cases, 29%), hepatocellular carcinoma (3 cases, 11%), and one case each of breast, pancreatic, and cervical cancer. In the remaining 3 patients (11%), with metastatic adenocarcinoma (2 cases) and squamous-cell carcinoma (1 case), the primary tumor site remained unknown despite an extensive clinical workup. Overall survival after FNA was poor, with a mean of 9.8 mo. FNA is useful in the diagnosis of masses in the kidney secondary to metastatic disease. This information is of clinical importance, principally in the exclusion of a primary malignancy, but also to avoid unnecessary surgery and to plan for subsequent patient care.
Assuntos
Biópsia por Agulha , Neoplasias Renais/diagnóstico , Neoplasias Renais/secundário , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/patologia , Criança , Estudos de Avaliação como Assunto , Feminino , Humanos , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/patologia , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/patologia , Linfoma/diagnóstico , Linfoma/patologia , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia , Valor Preditivo dos Testes , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologiaRESUMO
Salmonellosis, one of the most common food and water-borne diseases, has a major global health and economic impact. Salmonella cells present high infection rates, persistence over inauspicious conditions and the potential to preserve virulence in dormant states when cells are viable but non-culturable (VBNC). These facts are challenging for current detection methods. Culture methods lack the capacity to detect VBNC cells, while biomolecular methods (e.g. DNA- or protein-based) hardly distinguish between dead innocuous cells and their viable lethal counterparts. This work presents and validates a novel bacteriophage (phage)-based microbial detection tool to detect and assess Salmonella viability. Salmonella Enteritidis cells in a VBNC physiological state were evaluated by cell culture, flow-cytometry and epifluorescence microscopy, and further assayed with a biosensor platform. Free PVP-SE1 phages in solution showed the ability to recognize VBNC cells, with no lysis induction, in contrast to the minor recognition of heat-killed cells. This ability was confirmed for immobilized phages on gold surfaces, where the phage detection signal follows the same trend of the concentration of viable plus VBNC cells in the sample. The phage probe was then tested in a magnetoresistive biosensor platform allowing the quantitative detection and discrimination of viable and VBNC cells from dead cells, with high sensitivity. Signals arising from 3 to 4 cells per sensor were recorded. In comparison to a polyclonal antibody that does not distinguish viable from dead cells, the phage selectivity in cell recognition minimizes false-negative and false-positive results often associated with most detection methods.