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1.
Artigo em Inglês | MEDLINE | ID: mdl-38637984

RESUMO

STUDY DESIGN: A single-center, prospective, comparative study. OBJECTIVE: This study aimed to investigate the efficacy and safety of the systemic transdermal diclofenac patch (DP) for immediate postoperative analgesia after lumbar spinal surgery. SUMMARY OF BACKGROUND DATA: Effective wound pain control after spinal surgery has been shown to lead to favorable outcomes. Using multimodal analgesia may decrease opioid use for postoperative pain. MATERIALS AND METHODS: Patients who underwent posterior lumbar spinal surgery between August 2022 and January 2023 were divided into two groups: patients who underwent surgery on even months and were treated with DP (DP [+] group) and those who underwent surgery on odd months and were not treated with DP (DP [-] group). The demographic data, morphine milligram equivalent (MMEs) within 24 hours, duration of hospitalization, number of rescue analgesics used, visual analog scale (VAS) scores of wound pain, deterioration in renal function, and other complications were compared. Subgroup analysis consisted of subgroups categorized based on surgical procedure (non-fusion or fusion surgery). RESULTS: In total, 111 and 113 patients were enrolled in the DP (+) and DP (-) groups, respectively. There was no significant difference in the deterioration of renal function in the DP (+) group. Overall comparisons demonstrated a significant difference between the DP (+) and DP (-) groups in the number of rescue analgesics used within 1 hour (P=0.046). In the non-fusion surgery subgroups, the MMEs within 24 hours, the number of rescue analgesics used within 1 and 3 hours, as well as the wound pain VAS at 1 and 3 hours postoperatively were significantly lower in the DP (+) group than in the DP (-) group (P=0.010, 0.015, 0.029, 0.005, and 0.048 respectively). CONCLUSION: Systemic transdermal DP may potentially offer safe and effective postoperative analgesia, especially in less invasive procedures such as non-fusion lumbar spinal surgery. LEVEL OF EVIDENCE: Level III.

2.
J Clin Med ; 12(17)2023 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-37685817

RESUMO

Adolescent idiopathic scoliosis (AIS), the most common pediatric musculoskeletal disorder, causes a three-dimensional spine deformity. Lenke type 5 AIS is defined as a structural thoracolumbar/lumbar curve with nonstructural thoracic curves. Although a rod curvature will affect clinical outcomes, intraoperative contouring of the straight rod depends on the surgeon's knowledge and experience. This study aimed to determine the optimum rod geometries to provide a pre-bent rod system for posterior spinal surgery in patients with Lenke type 5 AIS. These pre-bent rods will be beneficial for achieving proper postoperative outcomes without rod contouring based on surgeon experience. We investigated 20 rod geometries traced in posterior spinal reconstruction in patients with Lenke type 5 AIS. The differences between the center point clouds in each cluster were evaluated using the iterative closest point (ICP) method with modification. Before the evaluation using the ICP method, the point clouds were divided into four clusters based on the rod length using a hierarchical cluster analysis. Because the differences in the values derived from the ICP method were <5 mm for each length-based cluster, four representative rod shapes were generated from the length-based clusters. We identified four optimized rod shapes that will reduce operation time, leading to a decreased patient and surgeon burden.

3.
J Clin Med ; 12(2)2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36675427

RESUMO

Adolescent idiopathic scoliosis (AIS) is the most common pediatric spinal deformity. Early detection of deformity and timely intervention, such as brace treatment, can help inhibit progressive changes. A three-dimensional (3D) depth-sensor imaging system with a convolutional neural network was previously developed to predict the Cobb angle. The purpose of the present study was to (1) evaluate the performance of the deep learning algorithm (DLA) in predicting the Cobb angle and (2) assess the predictive ability depending on the presence or absence of clothing in a prospective analysis. We included 100 subjects with suspected AIS. The correlation coefficient between the actual and predicted Cobb angles was 0.87, and the mean absolute error and root mean square error were 4.7° and 6.0°, respectively, for Adam's forward bending without underwear. There were no significant differences in the correlation coefficients between the groups with and without underwear in the forward-bending posture. The performance of the DLA with a 3D depth sensor was validated using an independent external validation dataset. Because the psychological burden of children and adolescents on naked body imaging is an unignorable problem, scoliosis examination with underwear is a valuable alternative in clinics or schools.

4.
Nanomaterials (Basel) ; 11(11)2021 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-34835865

RESUMO

Cyclic trinuclear complexes with group 11 metal(I) ions are fascinating and important to coordination chemistry. One of the ligands known to form these cyclic trinuclear complexes is pyrazolate, which is a bridging ligand that coordinates many transition metal ions in a Npz-M-Npz linear mode (Npz = pyrazolyl nitrogen atom). In these group 11 metal(I) ions, copper is the most abundant metal. Therefore, polynuclear copper(I) complexes are very important in this field. The cyclic trinuclear copper(I) complex [Cu(3,5-Ph2pz)]3 (3,5-Ph2pz- = 3,5-diphenyl-1-pyrazolate anion) was reported in 1988 as a landmark complex, but its photoluminescence properties have hitherto not been described. In this study, we report the photoluminescence and two different polymorphs of [Cu(3,5-Ph2pz)]3 and its derivative [Cu(3-Me-5-Phpz)]3 (3-Me-5-Phpz- = 3-metyl-5-phenyl-1-pyrazale anion). The substituents in [Cu(3-Me-5-Phpz)]3 cause smaller distortions in the solid-state structure and a red-shift in photoluminescence due to the presence of intermolecular cuprophilic interactions.

6.
Sci Rep ; 11(1): 13431, 2021 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-34183735

RESUMO

Coronavirus disease 2019 (COVID-19) is an emerging infectious disease that leads to severe respiratory failure (RF). It is known that host exposure to viral infection triggers an iron-lowering response to mitigate pathogenic load and tissue damage. However, the association between host iron-lowering response and COVID-19 severity is not clear. This two-center observational study of 136 adult hospitalized COVID-19 patients analyzed the association between disease severity and initial serum iron, total iron-binding capacity (TIBC), and transferrin saturation (TSAT) levels. Serum iron levels were significantly lower in patients with mild RF than in the non-RF group; however, there were no significant differences in iron levels between the non-RF and severe RF groups, depicting a U-shaped association between serum iron levels and disease severity. TIBC levels decreased significantly with increasing severity; consequently, TSAT was significantly higher in patients with severe RF than in other patients. Multivariate analysis including only patients with RF adjusted for age and sex demonstrated that higher serum iron and TSAT levels were independently associated with the development of severe RF, indicating that inadequate response to lower serum iron might be an exacerbating factor for COVID-19.


Assuntos
COVID-19/patologia , Ferro/sangue , Adulto , Idoso , COVID-19/complicações , COVID-19/virologia , Feminino , Ferritinas/sangue , Hospitalização , Humanos , Ferro/metabolismo , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Insuficiência Respiratória/etiologia , SARS-CoV-2/isolamento & purificação , Índice de Gravidade de Doença , Transferrina/análise
7.
FASEB J ; 35(5): e21420, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33774855

RESUMO

Although an aberrant reduction in pancreatic ß-cell mass contributes to the pathogenesis of diabetes, the mechanism underlying the regulation of ß-cell mass is poorly understood. Here, we show that diacylglycerol kinase δ (DGKδ) is a key enzyme in the regulation of ß-cell mass. DGKδ expression was detected in the nucleus of ß-cells. We developed ß-cell-specific DGKδ knockout (ßDGKδ KO) mice, which showed lower blood glucose, higher plasma insulin levels, and better glucose tolerance compared to control mice. Moreover, an increased number of small islets and Ki-67-positive islet cells, as well as elevated cyclin B1 expression in the islets, were detected in the pancreas of ßDGKδ KO mice. DGKδ knockdown in the ß-cell line MIN6 induced significant increases in bromodeoxyuridine (BrdU) incorporation and cyclin B1 expression. Finally, we confirmed that streptozotocin-induced hyperglycemia and ß-cell loss were alleviated in ßDGKδ KO mice. Thus, suppressing the expression or enzymatic activity of DGKδ that functions as a suppressor of ß-cell proliferation could be a novel therapeutic approach to increase ß-cell mass for the treatment of diabetes.


Assuntos
Encéfalo/enzimologia , Proliferação de Células , Diabetes Mellitus Experimental/complicações , Diacilglicerol Quinase/fisiologia , Hiperglicemia/prevenção & controle , Células Secretoras de Insulina/metabolismo , Animais , Hiperglicemia/etiologia , Hiperglicemia/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de Sinais
8.
J Vet Med Sci ; 81(8): 1129-1132, 2019 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-31270308

RESUMO

Veterinary vaccines are subjected to a safety testing using laboratory animals via intraperitoneal injection per batch. From April 2010 to March 2011, 7 guinea pigs in 4 batch tests exhibited unrecoverable weight loss and/or were found dead. Six guinea pigs had developed intussusception, whereas another one had developed an intestinal obstruction consequent to adhesion. A histopathology revealed that these lesions were associated with inflammatory foci. Other animals than the 7 guinea pig also developed similar inflammatory foci but did not develop bowel disorders. In the retesting of these batches, animals did not exhibited clinical signs, though inflammatory foci were detected. The clinical signs, detected in the primary test, might be due to bowel disorders secondary to an inflammatory response, rather than toxicity.


Assuntos
Doenças Inflamatórias Intestinais/veterinária , Testes de Toxicidade/veterinária , Vacinas/efeitos adversos , Animais , Cobaias , Doenças Inflamatórias Intestinais/etiologia , Injeções Intraperitoneais , Vacinas/administração & dosagem
9.
ACS Chem Biol ; 14(7): 1546-1555, 2019 07 19.
Artigo em Inglês | MEDLINE | ID: mdl-31246411

RESUMO

Plant pathogenic bacteria possess sophisticated mechanisms to detect the presence of host plants by sensing host-derived compounds. Ralstonia solanacearum, the causative agent of bacterial wilt on solanaceous plants, employs quorum sensing to control the production of the secondary metabolite ralfuranones/ralstonins, which have been suggested to be involved in virulence. Here, we report that d-galactose and d-glucose, plant sugars, activate the production of ralfuranones/ralstonins in R. solanacearum. As a result, two new derivatives, ralfuranone M (1) and ralstonin C (2), were found in the culture extracts, and their structures were elucidated by spectroscopic and chemical methods. Ralstonin C (2) is a cyclic lipopeptide containing a unique fatty acid, (2S,3S,Z)-3-amino-2-hydroxyicos-13-enoic acid, whereas ralfuranone M (1) has a common aryl-furanone structure with other ralfuranones. d-Galactose and d-glucose activated the expression of the biosynthetic ralfuranone/ralstonin genes and in part became the biosynthetic source of ralfuranones/ralstonins. Ralfuranones and ralstonins were detected from the xylem fluid of the infected tomato plants, and their production-deficient mutants exhibited reduced virulence on tomato and tobacco plants. Taken together, these results suggest that activation of ralfuranone/ralstonin production by host sugars functions in R. solanacearum virulence.


Assuntos
Galactose/metabolismo , Glucose/metabolismo , Lactonas/metabolismo , Doenças das Plantas/microbiologia , Ralstonia solanacearum/fisiologia , Solanaceae/microbiologia , Interações Hospedeiro-Patógeno , Solanum lycopersicum/metabolismo , Solanum lycopersicum/microbiologia , Percepção de Quorum , Ralstonia solanacearum/patogenicidade , Solanaceae/metabolismo , Nicotiana/metabolismo , Nicotiana/microbiologia
10.
J Obstet Gynaecol Res ; 44(6): 1174-1176, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29607582

RESUMO

Paraurethral leiomyomas are extremely rare, and few cases have been reported in the literature. Because of the rarity of the disease and its anatomical complexity, diagnosis and management is challenging for clinicians. Here, we herein report the case of a 50-year-old woman who complained of a protruding mass at the urethral opening. Magnetic resonance imaging revealed a sharply defined tumor measuring 43 × 36 × 26 mm in diameter, located in the anterior vaginal wall, compressing the urethra. Based on the perioperative diagnosis of leiomyoma, the paraurethral tumor was excised via the vaginal approach. The patient was discharged 4 days postoperatively without any complications. Pathological features confirmed the diagnosis of leiomyoma. This case implies that, although there is the possibility of complications due to the surgical procedure, tumor removal is a reasonable choice for management to exclude malignancy and relieve symptoms.


Assuntos
Leiomioma/cirurgia , Uretra , Miomectomia Uterina/métodos , Neoplasias Uterinas/cirurgia , Feminino , Humanos , Leiomioma/diagnóstico por imagem , Pessoa de Meia-Idade , Neoplasias Uterinas/diagnóstico por imagem
11.
Dent Mater J ; 36(6): 798-808, 2017 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-28652554

RESUMO

The suitability of various post and core materials for the 4-unit zirconia fixed partial denture abutment teeth was investigated to lower stress concentration. Artificial mandibular teeth (44 and 47) were used as the abutment teeth. There were two types of posts and cores: resin composite with glass fiber posts (RC), and cast platinum gold alloy (MC). Two experimental groups were analyzed. For Group RM, the premolars were restored with RC and the molars were restored with MC. For Group MR, the premolars were restored with MC and the molars were restored with RC. Static loading was applied to the occlusal surfaces, and the surface strain of the frameworks and roots was measured with a strain gauge. Group RM had a greater number of statistical differences between premolars and molars compared to Group MR. This result suggests that the Group MR post and core material combination reduces stress concentration.


Assuntos
Prótese Parcial Fixa , Técnica para Retentor Intrarradicular , Materiais Restauradores do Canal Radicular/química , Dente Pré-Molar , Projeto do Implante Dentário-Pivô , Análise do Estresse Dentário , Planejamento de Dentadura , Vidro , Ligas de Ouro , Humanos , Técnicas In Vitro , Modelos Dentários , Dente Molar , Platina , Zircônio/química
12.
J Vet Med Sci ; 77(8): 913-8, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25866403

RESUMO

Infectious bursal disease (IBD) is characterized by immunosuppression due to the depletion of lymphocytes in the atrophied bursa of Fabricius (BF). We have sometimes encountered contradictory findings: chickens infected with the vaccine IBD virus (IBDV) strain have sometimes exhibited a highly atrophied BF, but not immunosuppression. In this study, chickens administered vaccine or wild-type strains of IBDV were later vaccinated with the B1 strain of the Newcastle disease virus (NDV). Bursal changes were examined histologically with a focus on the bursal follicle. The immunoreactivity to NDV was also evaluated with the hemagglutination inhibition test. In gross examination, we observed a few chickens with a severely atrophied BF in vaccine strain-administered groups (vaccine groups), and the level of severity was the same as that in the wild-type strain-administered group (wild-type group). However, these chickens retained humoral antibody responses to NDV and were revealed to possess a higher number of bursal follicles than those of the wild-type group. These results indicated that macroscopic evaluation dose not accurately reflect the immunoreactivity and degree of bursal damage in IBDV-administered chickens. We also found non-immunosuppressed chickens in the wild-type group. These non-immunosuppressed chickens retained a significantly higher number of normal follicles and total follicles according to our statistical analysis. Furthermore, a high correlation coefficient between the NDV-HI titer and the number of normal follicles was found in the wild-type group. These results implied that the retained number of normal follicles is important for the immunoreactivity of chickens infected with IBDV.


Assuntos
Infecções por Birnaviridae/veterinária , Bolsa de Fabricius/patologia , Vírus da Doença Infecciosa da Bursa/imunologia , Doenças das Aves Domésticas/virologia , Animais , Infecções por Birnaviridae/imunologia , Infecções por Birnaviridae/patologia , Infecções por Birnaviridae/virologia , Bolsa de Fabricius/imunologia , Galinhas/imunologia , Galinhas/virologia , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/patologia , Vacinas Virais/imunologia
13.
Eur J Pharmacol ; 718(1-3): 376-82, 2013 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-23973646

RESUMO

Treatment with atypical antipsychotic drugs is known to increase the risk of glucose intolerance and diabetes. However, the mechanism of this effect is unclear. Since central adenosine 5'-monophosphate-activated protein kinase (AMPK) plays an important role in regulating nutrient homeostasis, the present study was performed to examine the involvement of central AMPK in the glucose intolerance induced by olanzapine, an atypical antipsychotic drug, in mice. Acute intraperitoneal treatment with olanzapine dose-dependently increased blood glucose levels in the glucose tolerance test. Intracerebroventricular administration of olanzapine also increased blood glucose levels in the glucose tolerance test. The glucose intolerance induced by both intraperitoneal and intracerebroventricular treatment with olanzapine was significantly attenuated by intracerebroventricular pretreatment with the AMPK inhibitor compound C. Intracerebroventricular treatment with the AMPK activator AICAR increased blood glucose levels in the glucose tolerance test, and this increase was inhibited by compound C. Moreover, the hypothalamic level of phosphorylated AMPK after glucose injection was significantly increased by intracerebroventricular pretreatment with olanzapine. Olanzapine did not affect plasma glucagon and insulin levels. Our results indicate that acute treatment with olanzapine causes glucose intolerance through the activation of hypothalamic AMPK. The present study suggests that the inhibition of central AMPK activity may have a therapeutic effect on the metabolic disturbance induced by atypical antipsychotic drugs.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Benzodiazepinas/farmacologia , Glucose/metabolismo , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Animais , Ativação Enzimática/efeitos dos fármacos , Glucagon/sangue , Teste de Tolerância a Glucose , Homeostase/efeitos dos fármacos , Insulina/sangue , Masculino , Camundongos , Camundongos Endogâmicos ICR , Olanzapina , Fosforilação/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia
14.
Avian Dis ; 57(1): 140-2, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23678743

RESUMO

This report describes a case of total anomalous pulmonary venous connection (TAPVC) in a 5-wk-old male white leghorn chicken that presented with growth retardation. This chicken was a specific-pathogen-free chicken bred in an isolator. At 5 wk of age, the chicken was euthanatized and autopsied. Macroscopically, the right ventricle and right atrium were significantly enlarged whereas the left atrium was small and blind-ending with no connection to the pulmonary veins. The pulmonary veins were connected directly to the right atrium. The above abnormality was accompanied by an ostium secundum-type atrial septal defect. No other malformations were observed. TAPVC is a very rare congenital cardiac abnormality that has not been reported in avian species to date.


Assuntos
Galinhas/anormalidades , Comunicação Interatrial/veterinária , Veias Pulmonares/anormalidades , Animais , Átrios do Coração/anormalidades , Átrios do Coração/patologia , Comunicação Interatrial/patologia , Ventrículos do Coração/anormalidades , Ventrículos do Coração/patologia , Masculino , Veias Pulmonares/patologia
15.
J Biosci Bioeng ; 115(1): 15-9, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22929984

RESUMO

A small heat shock protein, AgsA, possesses chaperone activity that can reduce the amount of heat-aggregated protein in vivo, and suppress the aggregation of chemical- and heat-denatured proteins in vitro. Therefore, we examined the ability of AgsA to stabilize the activity of several enzymes by using this chaperone activity. We observed that AgsA can stabilize the enzymatic activities of Renilla (Renilla reniformis) luciferase, firefly (Photinus pyralis) luciferase, and ß-galactosidase, and showed comparable or greater stabilization of these enzymes than bovine serum albumin (BSA), a well-known stabilizer of enzyme activities. In particular, AgsA revealed better stabilization of Renilla luciferase and ß-galactosidase than BSA under disulfide bond-reducing conditions with dithiothreitol. In addition, AgsA also increased the enzymatic performance of ß-galactosidase and various restriction enzymes to a comparable or greater extent than BSA. These data indicate that AgsA may be useful as a general stabilizer of enzyme activities.


Assuntos
Estabilidade Enzimática , Proteínas de Choque Térmico Pequenas/metabolismo , Chaperonas Moleculares/metabolismo , Animais , Dissulfetos/metabolismo , Ditiotreitol/farmacologia , Estabilidade Enzimática/efeitos dos fármacos , Proteínas de Choque Térmico Pequenas/farmacologia , Temperatura Alta , Luciferases de Vaga-Lume/metabolismo , Luciferases de Renilla/metabolismo , Chaperonas Moleculares/farmacologia , Soroalbumina Bovina/metabolismo , Soroalbumina Bovina/farmacologia , beta-Galactosidase/metabolismo
16.
Eur J Pharmacol ; 652(1-3): 117-20, 2011 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-21130084

RESUMO

We examined the involvement of tetrodotoxin (TTX)-resistant sodium channels in the peripheral mechanisms of the cough reflex in mice. We also examined the possibility of using ambroxol as an effective antitussive agent, and found that it produced antitussive effects through the inhibition of TTX-resistant sodium channels. The inhalation of fenvalerate, at concentrations of 0.3, 1 and 3µg/ml, for 5min produced coughs in a concentration-dependent manner. Pretreatment with tetrodotoxin, at a dose of 1µg/kg, s.c., slightly but significantly reduced the number of fenvalerate (3µg/ml)-induced coughs. However, the number of fenvalerate-induced coughs in tetorodotoxin-treated mice was still significantly greater than those in vehicle (0.4% DMSO) alone inhaled mice. On the other hand, pretreatment with tetrodotoxin, at a dose of 1µg/kg, s.c., almost completely reduced the number of citric acid (0.25M)-induced coughs to the level in vehicle (saline) alone inhaled mice. Pretreatment with ambroxol, at doses of 10, 30, 100 and 300mg/kg, p.o., dose-dependently and significantly reduced the number of fenvalerate (3µg/ml)-induced coughs. The present findings indicate that TTX-resistant sodium channels may play an important role in the enhancement of C-fiber-mediated cough pathways. Furthermore, ambroxol may prove to be a useful cough suppressant.


Assuntos
Antitussígenos/uso terapêutico , Tosse/tratamento farmacológico , Fibras Nervosas Amielínicas/efeitos dos fármacos , Canais de Sódio/metabolismo , Tetrodotoxina/uso terapêutico , Ambroxol/farmacologia , Ambroxol/uso terapêutico , Animais , Antitussígenos/farmacologia , Tosse/induzido quimicamente , Relação Dose-Resposta a Droga , Masculino , Camundongos , Camundongos Endogâmicos ICR , Fibras Nervosas Amielínicas/metabolismo , Nitrilas , Piretrinas , Tetrodotoxina/farmacologia
18.
Nihon Hoshasen Gijutsu Gakkai Zasshi ; 65(12): 1611-7, 2009 Dec 20.
Artigo em Japonês | MEDLINE | ID: mdl-20124738

RESUMO

We measured the physical characteristics of 2-million(2M)and 3-million(3M)color liquid crystal displays(LCD)whose aperture ratio was increased and compared them with conventional models. The results showed the influence of the increased aperture ratio on the physical characteristics of the LCDs. We evaluated resolution by means of modulation transfer function(MTF)and evaluated granularity by means of noise power spectrum(NPS). Each of the measurements was done with a high-resolution single-lens reflex-type digital camera. A decrease of MTF depending on sub-pixel structures was recognized. A decrease in the cross sub-pixel direction was recognized in the 2M model, and a decrease in the sub-pixel direction was recognized in the 3M model. As for NPS, a reduction was recognized in the sub-pixel and the cross sub-pixel direction in both models. As a result, an improvement in granularity was recognized. The improvement in granularity was large with the color LCDs whose aperture ratio was increased. The increase of an aperture ratio influenced both MTF and NPS, and the results depended on the shape and size of the sub-pixel cells.


Assuntos
Apresentação de Dados , Cristais Líquidos
19.
In Vitro Cell Dev Biol Anim ; 42(3-4): 63-9, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16759150

RESUMO

We have developed a simple method for rapid detection of mycoplasma contamination in cell cultures using SYBR Green-based real-time polymerase chain reaction (PCR). To detect eight common contaminant mollicutes, including Mycoplasma (M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. salivarium, M. pirum) and Acholeplasma laidlawii, four primers were prepared based on the 23S rRNA regions. Using these primers and a minimum of 100 fg of mycoplasma genomic DNA, the 23S rRNA regions of these eight mycoplasma species were consistently amplified by real-time PCR. In contrast, no specific amplification product was observed using DNA templates prepared from various mammalian cell lines. Frozen and cultured samples of several cell lines were tested for mycoplasma contamination to evaluate the utility of this method. Of 25 samples that tested positive for mycoplasma by Hoechst staining, which requires two passages of cell cultures started from frozen samples, mycoplasma was detected by real-time PCR in 24 samples of cell extracts prepared directly from frozen samples. When cultured samples were used for this assay, the accuracy of the diagnoses was further improved. Thus, this technique, which is simple, rapid, and sensitive enough for practical application, is suitable for handling many samples and for routine screening for mycoplasma contamination of cell cultures.


Assuntos
Corantes Fluorescentes/metabolismo , Infecções por Mycoplasma , Mycoplasma/genética , Compostos Orgânicos/metabolismo , Reação em Cadeia da Polimerase/métodos , Animais , Benzotiazóis , Técnicas de Cultura de Células , DNA Bacteriano/análise , Diaminas , Humanos , Camundongos , Quinolinas , Ratos , Sensibilidade e Especificidade , Suínos
20.
Inorg Chem ; 45(4): 1698-713, 2006 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-16471983

RESUMO

Copper(I) complexes with tripodal nitrogen-containing neutral ligands such as tris(3,5-diisopropyl-1-pyrazolyl)methane (L1') and tris(3-tertiary-butyl-5-isopropyl-1-pyrazolyl)methane (L3'), and with corresponding anionic ligands such as hydrotris(3,5-diisopropyl-1-pyrazolyl)borate (L1-) and hydrotris(3-tertiary-butyl-5-isopropyl-1-pyrazolyl)borate (L3-) were synthesized and structurally characterized. Copper(I) complexes [Cu(L1')Cl] (1), [Cu(L1')(OClO3)] (2), [Cu(L1')(NCMe)](PF6) (3a), [Cu(L1')(NCMe)](ClO4) (3b), [Cu(L1')(CO)](PF6) (4a), and [Cu(L1')(CO)](ClO4) (4b) were prepared using the ligand L1'. Copper(I) complexes [Cu(L3')Cl] (5) and [Cu(L3')(NCMe)](PF6) (6) with the ligand L3' were also synthesized. Copper(I) complexes [Cu(L1)(NCMe)] (7) and [Cu(L1)(CO)] (8) were prepared using the anionic ligand L1-. Finally, copper(I) complexes with anionic ligand L3- and acetonitrile (9) and carbon monoxide (10) were synthesized. The complexes obtained were fully characterized by IR, far-IR, 1H NMR, and 13C NMR spectroscopy. The structures of both ligands, L1' and L3', and of complexes 1, 2, 3a, 3b, 4a, 4b, 5, 6, 7, and 10 were determined by X-ray crystallography. The effects of the differences in (a) the fourth ligand and the counteranion, (b) the steric hindrance at the third position of the pyrazolyl rings, and most importantly, (c) the charge of the N3 type ligands, on the structures, spectroscopic properties, and reactivities of the copper(I) complexes are discussed. The observed differences in the reactivities toward O2 of the copper(I) acetonitrile complexes are traced back to differences in the oxidation potentials determined by cyclic voltammetry. A special focus is set on the carbonyl complexes, where the 13C NMR and vibrational data are presented. Density functional theory (DFT) calculations are used to shed light on the differences in CO bonding in the compounds with neutral and anionic N3 ligands. In correlation with the vibrational and electrochemical data of these complexes, it is demonstrated that the C-O stretching vibration is a sensitive probe for the "electron richness" of copper(I) in these compounds.


Assuntos
Boratos/química , Cobre/química , Compostos Organometálicos/química , Pirazóis/química , Ligantes , Estrutura Molecular
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