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1.
Int J Oral Maxillofac Surg ; 44(12): 1480-5, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26149939

RESUMO

The association between XRCC6/Ku70, an upstream player in the DNA double-strand break repair system, and the risk of nasopharyngeal carcinoma (NPC) was examined. In this case-control study, 176 NPC patients and 352 cancer-free controls were genotyped, and the associations of XRCC6 promoter T-991C (rs5751129), promoter G-57C (rs2267437), promoter G-31A (rs132770), and intron 3 (rs132774) polymorphisms with NPC risk were evaluated. NPC tissue samples were also assessed for their XRCC6 mRNA and protein expression by real-time quantitative reverse transcription PCR and Western blotting, respectively. With regard to the XRCC6 promoter T-991C, the TC and CC genotypes were associated with a significantly increased risk of NPC compared with wild-type TT genotype (adjusted odds ratio 2.02 and 3.42, 95% confidence interval 1.21-3.32 and 1.28-8.94, P=0.0072 and 0.0165, respectively). The mRNA and protein expression levels for NPC tissues revealed significantly lower XRCC6 mRNA and protein expression in the NPC samples with TC/CC genotypes compared to those with the TT genotype (P=0.0210 and 0.0164, respectively). These findings suggest that XRCC6 may play an important role in the carcinogenesis of NPC and could serve as a chemotherapeutic target for personalized medicine and therapy.


Assuntos
Antígenos Nucleares/genética , Proteínas de Ligação a DNA/genética , Neoplasias Nasofaríngeas/genética , Adolescente , Idoso , Idoso de 80 Anos ou mais , Western Blotting , Carcinoma , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Genótipo , Humanos , Lactente , Autoantígeno Ku , Masculino , Carcinoma Nasofaríngeo , RNA Mensageiro/análise , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Inquéritos e Questionários , Taiwan
2.
Mol Psychiatry ; 20(10): 1205-11, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25421403

RESUMO

Despite optimal pharmacotherapy and cognitive-behavioral treatments, a proportion of patients with obsessive-compulsive disorder (OCD) remain refractory to treatment. Neurosurgical ablative or nondestructive stimulation procedures to treat these refractory patients have been investigated. However, despite the potential benefits of these surgical procedures, patients show significant surgery-related complications. This preliminary study investigated the use of bilateral thermal capsulotomy for patients with treatment-refractory OCD using magnetic resonance-guided focused ultrasound (MRgFUS) as a novel, minimally invasive, non-cranium-opening surgical technique. Between February and May 2013, four patients with medically refractory OCD were treated with MRgFUS to ablate the anterior limb of the internal capsule. Patients underwent comprehensive neuropsychological evaluations and imaging at baseline, 1 week, 1 month and 6 months following treatment. Outcomes were measured with the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS), the Hamilton Rating Scale for Depression (HAM-D) and the Hamilton Rating Scale for Anxiety (HAM-A), and treatment-related adverse events were evaluated. The results showed gradual improvements in Y-BOCS scores (a mean improvement of 33%) over the 6-month follow-up period, and all patients showed almost immediate and sustained improvements in depression (a mean reduction of 61.1%) and anxiety (a mean reduction of 69.4%). No patients demonstrated any side effects (physical or neuropsychological) in relation to the procedure. In addition, there were no significant differences found in the comprehensive neuropsychological test scores between the baseline and 6-month time points. This study demonstrates that bilateral thermal capsulotomy with MRgFUS can be used without inducing side effects to treat patients with medically refractory OCD. If larger trials validate the safety, effectiveness and long-term durability of this new approach, this procedure could considerably change the clinical management of treatment-refractory OCD.


Assuntos
Transtorno Obsessivo-Compulsivo/diagnóstico por imagem , Transtorno Obsessivo-Compulsivo/cirurgia , Adulto , Feminino , Humanos , Cápsula Interna/cirurgia , Imagem por Ressonância Magnética Intervencionista/métodos , Masculino , Testes Neuropsicológicos , Procedimentos Neurocirúrgicos/métodos , Procedimentos Cirúrgicos Ultrassônicos/métodos , Ultrassonografia de Intervenção/métodos
3.
Lett Appl Microbiol ; 59(1): 9-16, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24521100

RESUMO

UNLABELLED: We investigated the role of the Bradyrhizobium japonicum lpcC gene, encoding a mannosyl transferase, involved in the lipopolysaccharide (LPS) biosynthesis. The inactivation of the lpcC gene considerably altered the LPS structure and the cell surface properties. LPS analysis showed that the lpcC mutant JS715 had an abnormal LPS structure deficient in O-antigen. The cell surface hydrophobicity increased approximately threefold in JS715 compared to the wild type. The increased cell surface hydrophobicity is likely to be related with cell aggregation in the mutant culture. For the growth comparison, JS715 showed slower growth rate than the wild type. The motility of JS715 decreased in soft agar plates, but it showed enhanced biofilm-forming ability. Interestingly, JS715 was not able to nodulate the host legume soybean (Glycine max). This study shows not only that lpcC is involved in the biosynthesis of O-antigen in the B. japonicum LPS, but also that inactivation of the lpcC gene affects symbiotic capability of B. japonicum and surface-related properties such as cell hydrophobicity, biofilm formation and motility. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the role of the B. japonicum lpcC in nodulation with soybean and importance of cell surface hydrophobicity. The results also highlight that intact LPS is required for successful symbiosis between B. japonicum and soybeans. Our findings not only support previous studies emphasizing the necessity of LPS on the interaction between the two symbiotic partners, but also contribute to a better understanding of the symbiotic mechanisms.


Assuntos
Biofilmes , Bradyrhizobium/genética , Glycine max/microbiologia , Antígenos O/genética , Simbiose , Aderência Bacteriana , Bradyrhizobium/química , Bradyrhizobium/metabolismo , Técnicas de Inativação de Genes , Genes Bacterianos , Interações Hidrofóbicas e Hidrofílicas , Antígenos O/biossíntese , Nódulos Radiculares de Plantas/microbiologia , Propriedades de Superfície
4.
Poult Sci ; 92(6): 1664-9, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23687164

RESUMO

A total of 216 chicken offal samples (chicken liver = 72; chicken heart = 72; chicken gizzard = 72) from wet markets and hypermarkets in Selangor, Malaysia, were examined for the presence and density of Listeria monocytogenes by using a combination of the most probable number and PCR method. The prevalence of L. monocytogenes in 216 chicken offal samples examined was 26.39%, and among the positive samples, the chicken gizzard showed the highest percentage at 33.33% compared with chicken liver (25.00%) and chicken heart (20.83%). The microbial load of L. monocytogenes in chicken offal samples ranged from <3 to 93.0 most probable number per gram. The presence of L. monocytogenes in chicken offal samples may indicate that chicken offal can act as a possible vehicle for the occurrence of foodborne listeriosis. Hence, there is a need to investigate the biosafety level of chicken offal in Malaysia.


Assuntos
Listeria monocytogenes/isolamento & purificação , Carne/microbiologia , Animais , Galinhas , Moela das Aves/microbiologia , Coração/microbiologia , Fígado/microbiologia , Malásia , Reação em Cadeia da Polimerase/métodos
5.
Rev Sci Instrum ; 84(3): 034901, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23556837

RESUMO

In this paper, we describe the thermal conductivity measurement of single-walled carbon nanotubes thin film using a laser point source-based steady state heat conduction method. A high precision micropipette thermal sensor fabricated with a sensing tip size varying from 2 µm to 5 µm and capable of measuring thermal fluctuation with resolution of ±0.01 K was used to measure the temperature gradient across the suspended carbon nanotubes (CNT) film with a thickness of 100 nm. We used a steady heat conduction model to correlate the temperature gradient to the thermal conductivity of the film. We measured the average thermal conductivity of CNT film as 74.3 ± 7.9 W m(-1) K(-1) at room temperature.


Assuntos
Temperatura Alta , Nanotecnologia/métodos , Nanotubos de Carbono/química , Condutividade Térmica , Eletrônica , Elétrons , Desenho de Equipamento , Filtração , Análise de Fourier , Teste de Materiais/métodos , Óptica e Fotônica , Cimento de Policarboxilato/química , Reprodutibilidade dos Testes , Temperatura , Sensação Térmica
6.
Poult Sci ; 91(10): 2686-90, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22991558

RESUMO

This study aimed to determine the prevalence Listeria monocytogenes in raw chicken meat samples at hypermarkets and wet markets. Chicken drumsticks, breasts, and thighs were randomly selected. The most probable number (MPN) PCR method was used to quantify the L. monocytogenes in the samples. Listeria monocytogenes was detected in 20% of the samples. Occurrence of L. monocytogenes was highest in breast (42.03%) followed by drumstick (11.27%) and thigh (7.14%). Samples from hypermarkets showed higher occurrence (25.71%) of L. monocytogenes compared with wet markets (14.29%). The density of L. monocytogenes found in samples ranged from <3.0 to 16 MPN•g(-1). The presence of L. monocytogenes in raw chicken meat is unwanted but unpreventable. Thus, further research on the processing method to reduce and eliminate this kind of bacteria in chicken meat before consumption is necessary. The presence of L. monocytogenes in chicken samples suggests the importance of this pathogen in chicken. Thus, more study is needed to find ways to eliminate this pathogen from poultry.


Assuntos
Microbiologia de Alimentos , Listeria monocytogenes/isolamento & purificação , Carne/microbiologia , Animais , Galinhas , Comércio , Malásia , Reação em Cadeia da Polimerase/métodos
7.
Clin Neuropathol ; 30(6): 328-32, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22011740

RESUMO

AIMS: The stereotactic brain biopsy is an essential diagnostic procedure in modern neurologic patient management. A side-cutting biopsy needle is one of the most widely used needle types. Recently we found a characteristic tissue artifact named "peripheral compressing artifact" in the brain tissues biopsied using a side-cutting needle of Leksell's system. We investigate prevalence, possible cause and its clinical implication of this type of artifact. MATERIALS AND METHODS: We examined the biopsies from 80 patients (44 cases of gliomas, 13 lymphomas, 7 germ cell tumors, 2 other tumors, 1 metastatic carcinoma, 4 non-tumorous conditions such as demyelinating disease and 8 non-diagnostic) in the stereotactic biopsy group with a suspected brain tumor, who underwent a stereotactic brain biopsy using side-cutting needle of Leksell's system. We also evaluated 16 cases of open brain biopsies without Leksell's system as a control group. RESULTS: The artifact is a semi-circular or band-like tissue compression in the periphery of the biopsied tissue. This artifact was found in 30 (37.5%) out of 80 cases and 57 (11.9%) out of 477 biopsied pieces. It might be produced during rotating of the inner cannula of the biopsy needle. Histologically, it might be misinterpreted as "hypercellular", "spindle", "well circumscribed", or rarely as "pseudopalisading" especially in glioma. CONCLUSIONS: Awareness of this artifact would help making the appropriate pathological diagnosis for glioma.


Assuntos
Artefatos , Neoplasias Encefálicas , Biópsia , Encéfalo , Glioma , Humanos , Agulhas , Técnicas Estereotáxicas
8.
Lett Appl Microbiol ; 50(5): 452-6, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20158607

RESUMO

AIMS: To reveal the effects of the O-polysaccharide antigen of Bradyrhizobium japonicum LPS on biofilm formation and motility. METHODS AND RESULTS: Wild type and O-antigen-deficient mutant strains of B. japonicum were tested for biofilm formation on polyvinyl chloride (PVC) surfaces and motility on semi-solid (0.3%) agar media. After 7 days of incubation, the amount of biofilms formed by the mutant was c. 3.5-fold greater than that of the wild type. Unlike biofilm formation, the motility assay revealed that the mutant strain was less motile than the wild type. CONCLUSIONS: This study shows enhanced biofilm formation and decreased motility by the O-antigen-deficient mutant, suggesting that the lack of the O-polysaccharide of the rhizobial LPS is associated with biofilm-forming ability and movement. SIGNIFICANCE AND IMPACT OF THE STUDY: LPS plays an important role in both pathogenic and beneficial bacteria. It has also been reported that LPS deficiency negatively affects biofilm formation. However, our results demonstrate that the O-antigen-deficient mutant enhances biofilm formation, presumably through a significant increase in hydrophobicity. It is notable that the hydrophobicity of cell walls might be a key regulator in controlling biofilm development in B. japonicum.


Assuntos
Biofilmes , Bradyrhizobium/fisiologia , Antígenos O/metabolismo , Bradyrhizobium/genética , Mutação , Antígenos O/genética
9.
Eur J Anaesthesiol ; 25(8): 620-6, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18339215

RESUMO

BACKGROUND AND OBJECTIVE: Numerous in vitro studies have shown that volatile anaesthetics react with desiccated carbon dioxide (CO2) absorbents to produce carbon monoxide (CO). The effects of anaesthetic concentration, fresh gas flow rate, and the hydration of absorbent or the excretion of CO2 by patients on CO production have also been investigated. This work aims to identify the most significant one of these factors on CO concentration in a low-flow anaesthesia system, without control of the hydration of the absorbents. METHODS: A simulated clinical circle anaesthetic breathing system was used to study the CO concentration under various conditions. Desflurane was used at three different concentrations. Two CO2 flow rates and three fresh gas flow rates were used. The absorbent temperatures and hydration were measured simultaneously. RESULTS: Desflurane degraded to produce CO in the breathing tube, when the CO2 absorbents were not dried beforehand. In this imitation clinical low-flow setting, fresh gas flow affected the CO production more than the CO2 did (31.7% vs. 9.5%). The actual desflurane partial pressure was not a significant factor. The CO2 flow rate explained 18.2% and 54.0% of the variation of the absorbent hydration changes (%) and temperature, respectively. CONCLUSIONS: In clinical practice, the CO2 production varies among patients and is uncontrollable, but markedly affects CO production. The only controllable factor is the fresh gas flow rate if the ultimate goal is to reduce the undesirable exposure of patients to CO from the breathing tube according to this bench model without counting the oxygen consumption.


Assuntos
Anestesia com Circuito Fechado , Anestésicos Inalatórios/química , Dióxido de Carbono/química , Monóxido de Carbono/análise , Isoflurano/análogos & derivados , Absorção , Desflurano , Umidade , Isoflurano/química , Oxigênio/metabolismo , Pressão Parcial , Análise de Regressão , Temperatura
10.
Annu Rev Phys Chem ; 58: 565-84, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17291185

RESUMO

A combination of single-molecule spectroscopy and analysis with simulations is used to provide detailed information about the structural and dynamic properties of a fluorescent polymer MEH-PPV (poly[2-methoxy-5-(2'-ethylhexyloxy)-1,4-phenylenevinylene]) immersed in a nematic and smectic solvent. In nematic solvents, single-polymer molecules are oriented strongly along the solvent director, much more so than the solvent molecules, confirming Onsager's old prediction. The diffusion anisotropy parallel and perpendicular to the solvent director, however, is less than two, which is similar to that of a spherical colloid in a nematic solvent. In smectic solvents, there is a second orientation of the dissolved polymer perpendicular to the solvent director, which we hypothesize is caused by the polymer occupying the interlayer volume. The research discussed here emphasizes the importance of organization in complex fluids and suggests that the interplay of order on different length scales could be exploited to fabricate novel nanostructured materials.

11.
Opt Express ; 15(14): 8713-8, 2007 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-19547206

RESUMO

We have analyzed the linearity performance of analog fiber-optic links based on electroabsorption modulators (EAM) operating at high optical power. The negative feedback caused by photocurrent generation improves the modulator linearity in the gain saturation regime. In the absence of laser relative intensity noise (RIN), the link spur-free dynamic range (SFDR) increases with the power of four-thirds of the input optical power after gain saturation occurs. A multi-octave SFDR of more than 135 dB/Hz2/3 has been found to be achievable with sufficiently high power.

12.
Izv Akad Nauk Ser Biol ; (4): 453-9, 2005.
Artigo em Russo | MEDLINE | ID: mdl-16212267

RESUMO

Preliminary selective blockade of micro, delta1, delta2, kappa1, and kappa2 opioid receptors proved to have no effect on the incidence of ventricular arrhythmias during a 10-min coronary occlusion and subsequent reperfusion in ketamine-anesthetized rats. We propose that the endogenous opioid system has no considerable role in regulation of heart resistance to the arrhythmogenic effect of short-term local ischemia and subsequent reperfusion.


Assuntos
Cardiopatias/metabolismo , Miocárdio/metabolismo , Peptídeos Opioides/metabolismo , Receptores Opioides/metabolismo , Animais , Antagonistas de Entorpecentes/administração & dosagem , Ratos , Ratos Wistar
13.
Nano Lett ; 5(9): 1757-60, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16159219

RESUMO

We examine the effect of polymer chain segmentation on the recently discovered ability of nematic solvents to elongate and align polymer chain solutes. Coordinated single molecule spectroscopy and beads-on-a-chain simulations are used to study the orientational and conformational order of a series of segmented conjugated polymers, dissolved in the nematic liquid crystal 5CB. The order parameters for alignment and elongation are both observed to decrease with increasing segmentation, reflecting an interplay among conformational entropy, solvation anisotropy, and bending energy of the chain.

14.
Chromosoma ; 111(3): 176-83, 2002 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12355207

RESUMO

In an attempt to amplify cervid satellite II DNA from the genomes of Indian muntjac and Chinese muntjac, a pair of primers derived from the white tailed deer satellite II DNA clone (OvDII) yielded a prominent approximately 1 kb polymerase chain reaction (PCR) product (in addition to the expected 0.7 kb satellite II DNA fragments) in both species. The approximately 1 kb products were cloned, sequenced, and analyzed by Southern blotting and fluorescence in situ hybridization (FISH). This revealed that the approximately 1 kb cloned sequences indeed represent a previously unknown cervid satellite DNA family, which is now designated as cervid satellite IV DNA. Approximately 1 kb PCR clones were also obtained from the genomes of the black tailed deer and Canadian woodland caribou with similar primer pairs. Extremely high sequence conservation (over 90% homology) was observed among the clones generated from all four deer species and PCR-Southern hybridization experiments further verified the co-amplification of two kinds of satellite DNA sequences with the same pair of primers. This satellite DNA was found to co-localize with centromeric proteins at the kinetochore by a simultaneous FISH and immunofluorescence study. Due to its high sequence conservation and close association with kinetochores, the newly identified satellite DNA may have a functional centromeric role.


Assuntos
Sequência Conservada/genética , Cervo Muntjac/genética , Animais , Sequência de Bases , Southern Blotting , Mapeamento Cromossômico , Hibridização in Situ Fluorescente , Técnicas In Vitro , Cinetocoros/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Rena/genética , Alinhamento de Sequência , Análise de Sequência de DNA
15.
Cancer J ; 7(6): 498-502, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11769862

RESUMO

PURPOSE: Local-regional recurrence rates of 30%-50% have been reported after resection of high-risk malignant melanomas (multiple node involvement, extracapsular spread, deep invasion, recurrent disease, and/or microscopically involved margins). Recently, we have been offering elective radiation therapy, after definitive surgery, to selected patients who have high-risk malignant melanomas. We herein report our initial results. PATIENTS AND METHODS: From 1993 to 1999, 40 patients who underwent surgery for high-risk malignant melanomas (multiple involved lymph nodes [21 patients]; close or microscopically involved surgical margins [nine patients]; extracapsular extension [six patients]; previously resected, recurrent disease [three patients]; and/or primary tumors more than 4 mm thick [four patients]) received elective radiation therapy. Thirty-six patients received 3000 cGy in five fractions (600 cGy per fraction given twice weekly), and four patients received 3600 cGy in six fractions. RESULTS: At a median follow-up of 18.4 months (range, 3.8-74.1 months), the actuarial 5-year local-regional control rate was 84%. Systemic recurrence rates in these patients were similar to those reported for this subset of patients, and the actuarial overall survival rate at 5 years was 39%. Acute toxicity was limited to erythema of the skin and, in one instance, probable cellulitis, with no late sequelae. DISCUSSION: Elective radiation therapy (600 cGy per fraction for five or six fractions) effectively controlled residual subclinical disease after surgery; however, better adjuvant systemic therapies need to be designed to eliminate distant metastases and to alter survival rates.


Assuntos
Melanoma/radioterapia , Neoplasias Cutâneas/radioterapia , Adulto , Idoso , Feminino , Humanos , Metástase Linfática , Masculino , Melanoma/mortalidade , Melanoma/patologia , Melanoma/secundário , Pessoa de Meia-Idade , Dosagem Radioterapêutica , Fatores de Risco , Neoplasias Cutâneas/mortalidade , Neoplasias Cutâneas/patologia , Taxa de Sobrevida
16.
Hepatogastroenterology ; 47(35): 1260-3, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11100327

RESUMO

BACKGROUND/AIMS: Laparoscopic liver resection is feasible for both benign and malignant disease with today's laparoscopic techniques and technology. Location of the tumor at the edge of segment 3, 4, 5, or 6 of our patients makes them an ideal candidate for laparoscopic resection. METHODOLOGY: There were 9 patients who underwent laparoscopic subsegmentectomy for hepatocellular carcinoma with cirrhosis. They were classified as Child A in 6 and B in 3 patients. Hepatitis B was found in 5 and Hepatitis C in 4 cases. Preoperative diagnosis of hepatocellular carcinoma was completed in 7 and definitive histologic diagnosis from frozen section in 2 cases. All 9 patients underwent subsegmentectomy and removal of the tumor with non-tumor cirrhotic liver with a distance of 10 mm at the least margin. Laparoscopic ultrasound allows exact localization of lesions and achievement of adequate resection margin. RESULTS: Those patients resumed a full diet on the 2nd-3rd day after the operation and were discharged home on day 4-7 with no complications but one had prolonging discharge due to ascitis from a drainage tube. Finally, the ascitis was controlled by medications for 1 week. All patients had high postoperative satisfaction. CONCLUSIONS: Laparoscopic liver resection is a procedure of significant risk and technically demanding. Therefore, it should be performed only by experienced liver surgeons with a high level of laparoscopic skill and in the carefully selected patient.


Assuntos
Carcinoma Hepatocelular/cirurgia , Hepatectomia/métodos , Laparoscopia , Cirrose Hepática/complicações , Neoplasias Hepáticas/cirurgia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
17.
J Microbiol Methods ; 43(2): 67-72, 2000 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-11121604

RESUMO

The green fluorescent protein encoded by gfp gene and the luminescent protein encoded by luxAB genes were used as markers to detect p-nitrophenol (PNP)-degrading Moraxella sp. G21r and polychlorinated biphenyl (PCB)-degrading Ralstonia eutrophas H850Lr cells, respectively, in mixed liquid cultures and in soil samples using a most-probable-number (MPN) assay. Population estimates for both gfp-marked G21r and lux-marked H850Lr by using MPN assays were similar to viable colony counts. The MPN assay with microtiter plates permitted the simultaneous detection of fluorescent and luminescent bacteria in soil samples faster than conventional plate counting.


Assuntos
Técnicas Bacteriológicas , Cupriavidus necator/isolamento & purificação , Proteínas Luminescentes/isolamento & purificação , Moraxella/isolamento & purificação , Microbiologia do Solo , Proteínas de Fluorescência Verde
18.
Genes Chromosomes Cancer ; 29(3): 240-55, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-10992299

RESUMO

Approximately one quarter of the identified human serpin genes are cancer-related and clustered mainly at two distinct loci: 6p25 and 18q21. We have studied a novel serpin gene cluster at 3q26 containing at least two recently identified members: the pancreas-specific protease inhibitor, pancpin (PI14), and the brain-associated protease inhibitor, neuroserpin (PI12). In this, unlike a previous study, both PI14 and PI12 at 3q26 were found to consist of 9 exons and 8 introns and to share a perfectly conserved gene organization whose pattern is very different from that of the ov-serpin family. This distinct pattern appears identical in the genomic structures of human plasminogen activator inhibitor-1 (PAI1) at 7q21 and protease nexin 1 (PI7) at 2q33-35, confirming that these four genes in three different chromosomes form a discrete subset within the serpin superfamily. As in the other three members whose gene expression is altered during tumorigenesis, PI12 expression was found to be down-regulated in tumor brain tissues and in two brain cancer cell lines: U-87 MG and H4. By screening genomic libraries, we isolated two overlapping clones showing that the marker SGC32223 (centromere) is located within intron F of PI12 and the marker WI-10077 (telomere) is located downstream of the 3'-flanking region of PI14. This finding indicates that the distance between human PI14 and PI12 is approximately 100 kb, and hence we speculate that other tissue-specific cancer-related serpin genes are likely to reside within this 3q26.1 cluster region.


Assuntos
Cromossomos Humanos Par 3/genética , Família Multigênica/genética , Proteínas de Neoplasias , Inibidores de Serina Proteinase/genética , Sequência de Aminoácidos , Animais , Bacteriófago P1/genética , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Linhagem Celular , Cromossomos Artificiais Bacterianos/genética , Clonagem Molecular , DNA Complementar/isolamento & purificação , Éxons , Humanos , Íntrons , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Complexo de Endopeptidases do Proteassoma , Estrutura Secundária de Proteína , Ratos , Inibidores de Serina Proteinase/metabolismo , Serpinas/genética , Serpinas/metabolismo , Células Tumorais Cultivadas
19.
J Biomed Mater Res ; 52(1): 142-7, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10906685

RESUMO

Previous studies have shown that osteoclasts are derived from mononuclear cells of hemopoietic bone marrow and peripheral blood. The purpose of this study was to demonstrate the presence of multinucleated osteoclasts after adding alveolar mononuclear cells to new-born rat calvaria osteoblasts in vitro. To utilize osteoclast-free bone, fetal calvariae were obtained from newborn Wistar-rats and cultured in DMEM medium for 14 days. On the day of osteoblast culture, alveolar mononuclear cells were isolated from newborn Wistar rats with a serial washing method and then co-cultured with the calvarial osteoblasts. Bone resorption characteristics were observed both with light and scanning electron microscopy. When alveolar mononuclear cells were cultured for 14 days on the calvarial osteoblasts in response to 1 alpha, 25-dihydroxyvitamin D3, they formed tartrate-resistant acid phosphatase (TRAP)-positive mononuclear and multinucleated cells. Resorption pits were seen in the 7-14 days long-term cultures. These results indicate that osteoclasts can be derived from alveolar mononuclear cells in vitro when a suitable microenvironment is provided by calvarial osteoblasts and vitamin D(3).


Assuntos
Leucócitos Mononucleares/citologia , Pulmão/citologia , Osteoclastos/citologia , Animais , Diferenciação Celular , Núcleo Celular , Células Cultivadas , Ratos , Ratos Wistar
20.
Proc Natl Acad Sci U S A ; 97(1): 67-72, 2000 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-10618372

RESUMO

The serpins are a family of proteinase inhibitors that play a central role in the control of proteolytic cascades. Their inhibitory mechanism depends on the intramolecular insertion of the reactive loop into beta-sheet A after cleavage by the target proteinase. Point mutations within the protein can allow aberrant conformational transitions characterized by beta-strand exchange between the reactive loop of one molecule and beta-sheet A of another. These loop-sheet polymers result in diseases as varied as cirrhosis, emphysema, angio-oedema, and thrombosis, and we recently have shown that they underlie an early-onset dementia. We report here the biochemical characteristics and crystal structure of a naturally occurring variant (Leu-55-Pro) of the plasma serpin alpha(1)-antichymotrypsin trapped as an inactive intermediate. The structure demonstrates a serpin configuration with partial insertion of the reactive loop into beta-sheet A. The lower part of the sheet is filled by the last turn of F-helix and the loop that links it to s3A. This conformation matches that of proposed intermediates on the pathway to complex and polymer formation in the serpins. In particular, this intermediate, along with the latent and polymerized conformations, explains the loss of activity of plasma alpha(1)-antichymotrypsin associated with chronic obstructive pulmonary disease in patients with the Leu-55-Pro mutation.


Assuntos
alfa 1-Antiquimotripsina/química , Cromatografia de Afinidade , Humanos , Pneumopatias Obstrutivas/sangue , Pneumopatias Obstrutivas/genética , Modelos Moleculares , Dados de Sequência Molecular , Mutação , Plasmídeos , Conformação Proteica , Estrutura Secundária de Proteína , Difração de Raios X , alfa 1-Antiquimotripsina/sangue , alfa 1-Antiquimotripsina/genética
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