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Adipocyte-cancer cell interactions promote tumor development and progression. Previously, we identified adipsin (CFD) and its downstream effector, hepatocyte growth factor (HGF), as adipokines that enhance adipocyte-breast cancer stem cell interactions. Here, we show that adipsin-dependent adipocyte maturation and the subsequent upregulation of HGF promote tumor invasion in breast cancers. Mature adipocytes, but not their precursors, significantly induced breast tumor cell migration and invasion in an adipsin expression-dependent manner. Promoters of tumor invasion, galectin 7 and matrix metalloproteinases, were significantly upregulated in cancer cells cocultured with mature adipocytes; meanwhile, their expression levels in cancer cells cocultured with adipocytes were reduced by adipsin knockout (Cfd KO) or a competitive inhibitor of CFD. Tumor growth and distant metastasis of mammary cancer cells were significantly suppressed when syngeneic mammary cancer cells were transplanted into Cfd KO mice. Histological analyses revealed reductions in capsular formation and tumor invasion at the cancer-adipocyte interface in the mammary tumors formed in Cfd KO mice. These findings indicate that adipsin-dependent adipocyte maturation may play an important role in adipocyte-cancer cell interaction and breast cancer progression.
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Adipocitos , Neoplasias de la Mama , Movimiento Celular , Factor D del Complemento , Invasividad Neoplásica , Animales , Femenino , Humanos , Ratones , Adipocitos/metabolismo , Adipocitos/patología , Neoplasias de la Mama/patología , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/genética , Línea Celular Tumoral , Técnicas de Cocultivo , Factor D del Complemento/metabolismo , Factor D del Complemento/genética , Factor de Crecimiento de Hepatocito/metabolismo , Factor de Crecimiento de Hepatocito/genética , Ratones NoqueadosRESUMEN
Trehalose is known to protect enzymes from denaturation. In the present study, we observed promotion of apple polyphenol oxidase (PPO) inactivation in a trehalose solution with thermal treatment. Crude PPO from Fuji apple was mixed with either sucrose or trehalose solutions, then the samples treated at 25 or 65 °C. In the presence of trehalose, PPO activities were markedly decreased upon treatment at 65 °C with increasing trehalose concentration. Furthermore, the reduction in PPO activity in the presence of trehalose was proportional to storage time after thermal treatment and thermal treatment time. Comparing PPO activities between treatment time 0 and 90 min at 65 °C, activities decreased 89 % for trehalose concentration of 0.2 M. These results indicates that trehalose acts not only as inhibitor but as promoter of inactivation of PPO. The Lineweaver-Burk plot indicated that trehalose acts on PPO as a non-competitive inhibitor during the 65 °C treatment. Two mechanisms of PPO inactivation in the presence of trehalose were suggested; one is the suppression of PPO activation cause by a thermal treatment, and another is the conformational change to inactivation form of PPO in conjunction with trehalose and a thermal treatment. Additionally, apple juice including 0.2 or 0.5 M trehalose with 65 °C treatment indicated slow browning than the juice with 0.2 or 0.5 M sucrose or without sugars. This result demonstrates that the preventing of browning with trehalose is a viable industrial food process.
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Objectives: To determine whether the tongue menthol test, which measures the cold sensation detection threshold (CDT) of the tongue, used before and after oxaliplatin administration is an objective evaluation method for oxaliplatin-induced peripheral neuropathy (OPN). Methods: The tongue menthol test was administered to patients both before and after undergoing chemotherapy containing oxaliplatin for colorectal cancer. The tongue menthol test was conducted by applying a menthol solution (a selective agonist of transient receptor potential cation channel subfamily M member 8 [TRPM8]) to the tongue and measuring the CDT. Results: The mean CDT before the first dose of oxaliplatin was 0.34% (0.005%-1%; n=38), and the mean CDT after the first dose was 0.32% (0.005%-1%; n=38). The CDT appeared to decrease after the first dose, but this difference was not significant. In patients who received five courses of oxaliplatin, changes in CDT values were compared before and after the five courses. In patients with Neurotoxicity Criteria of Debiopharm (DEB-NTC) grade 2 neuropathy, the pre-oxaliplatin administration CDT was compared between before grade exacerbation and when exacerbation occurred, and was found to decline when grade exacerbation occurred. Moreover, when the CDTs before and after administration were compared before grade exacerbation, there was a significant decrease in CDT after administration (P=0.04). Conclusions: By performing a menthol test in oxaliplatin-treated patients, it may be possible to objectively predict the exacerbation of peripheral neuropathy at an early stage.
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The assembly of discrete active species to form periodical nanostructures is essential in realizing low-cost artificial enzymes that mimic natural enzymatic functions in extraordinary bio(chemo)selective reactions. In this study, we developed artificial bifunctional glucose/gluconic acid dehydrogenase from naturally abundant resources: l-aspartic acid (Asp) and montmorillonite (a subgroup of smectite natural clay minerals). ß-d-Glucose (Glc) was dehydrogenated to 2-keto-d-gluconate (2-KGA) at 25 and 30 °C in an aqueous acidic solution (pH = 3, 4, and 5). The reaction involved sequential steps that yielded d-gluconic acid (GA) as an intermediate. The second step of the dehydrogenation (GA to 2-KGA) occurred at a higher rate than the first (Glc to GA), which is comparable to the natural process. A negatively charged carboxylate in Asp was required for the dehydrogenation, which donates an electron pair (COO:-) to the hydroxyl group bonded to the C(1)-position of Glc. The acidic sites in clay served as coenzymatic sites (electron acceptor), promoting the Glc dehydrogenation as the Glc reduced by Asp approached the clay coenzymatic sites. The active coenzymatic structures were developed in 48 h (induction period) through the rearrangement of the adsorbed Asp and Glc molecules on montmorillonite in water (intermediate structure). The spontaneous assembling of the intermediate structures facilitated the one-pot dehydrogenation of Glc to 2-KGA via periodic "hydrated stacked layers" comprising clay nanosheets, Asp, and Glc. The facile synthetic route proposed here is inexpensive and would be beneficial without using both GDH and GADH enzymes bound to a cell membrane.
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Aminoácidos , Glucosa , Aceleración , Bentonita , Arcilla , Gluconatos , Glucosa/metabolismo , AguaRESUMEN
BACKGROUND: Eosinophilic enteritis is a chronic inflammatory disorder of the intestinal tract that is characterized by eosinophil infiltration. Cytomegalovirus (CMV), a common virus, has a broad infectivity range. CMV is retained in the host body after infection. Impairment of host immune defences may reactivate the latent CMV, leading to symptoms of overt disease. CASE PRESENTATION: A Japanese female in her 70 s was admitted to a hospital due to diarrhoea and then transferred to our hospital. Laboratory data showed hypoalbuminemia. Computed tomography (CT) revealed oedema of the small intestine. Lower gastrointestinal endoscopy revealed oedema of the submucosa, without any remarkable changes in the mucosa of the terminal ileum. Histological examination of the terminal ileum revealed infiltration of > 20 eosinophils per high-power field (HPF). These findings aided in diagnosing eosinophilic enteritis. We administered methylprednisolone (500 mg/day) for three days, followed by tapering prednisolone. However, the patient's general condition and hypoalbuminemia failed to improve. Immunoglobulin (Ig) G- CMV and IgM-CMV tests were positive. CMV antigenemia was extremely high. Therefore, we administered ganciclovir intravenously, which improved the patient's condition. Furthermore, azathioprine was administered to taper and discontinue prednisolone without relapse of eosinophilic enteritis. This treatment helped stabilize the patient's condition for approximately four years. CONCLUSION: We present a case of eosinophilic enteritis accompanied by CMV disease during prednisolone treatment. The patient's condition improved after administration of ganciclovir. Azathioprine aided in discontinuing prednisolone and stabilizing the patient's condition for approximately four years.
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Infecciones por Citomegalovirus , Hipoalbuminemia , Azatioprina/uso terapéutico , Citomegalovirus , Infecciones por Citomegalovirus/complicaciones , Infecciones por Citomegalovirus/diagnóstico , Infecciones por Citomegalovirus/tratamiento farmacológico , Enteritis , Eosinofilia , Femenino , Ganciclovir/uso terapéutico , Gastritis , Humanos , Hipoalbuminemia/tratamiento farmacológico , Hipoalbuminemia/etiología , Prednisolona/uso terapéuticoRESUMEN
Adipose tissue is a component of the tumor microenvironment and is involved in tumor progression. We have previously shown that adipokine adipsin (CFD) functions as an enhancer of tumor proliferation and cancer stem cell (CSC) properties in breast cancers. We established the Cfd-knockout (KO) mice and the mammary adipose tissue-derived stem cells (mADSCs) from them. Cfd-KO in mADSCs significantly reduced their ability to enhance tumorsphere formation of breast cancer patient-derived xenograft (PDX) cells, which was restored by the addition of Cfd in the culture medium. Hepatocyte growth factor (HGF) was expressed and secreted from mADSCs in a Cfd-dependent manner. HGF rescued the reduced ability of Cfd-KO mADSCs to promote tumorsphere formation in vitro and tumor formation in vivo by breast cancer PDX cells. These results suggest that HGF is a downstream effector of Cfd in mADSCs that enhances the CSC properties in breast cancers.
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The genus Pestalotiopsis are endophytic fungi that have recently been identified as cellulolytic system producers. We herein cloned a gene coding for a xylanase belonging to glycoside hydrolase (GH) family 10 (PesXyn10A) from Pestalotiopsis sp. AN-7, which was isolated from the soil of a mangrove forest. This protein was heterologously expressed by Pichia pastoris as a host, and its enzymatic properties were characterized. PesXyn10A was produced as a glycosylated protein and coincident to theoretical molecular weight (35.3 kDa) after deglycosylation by peptide-NfF-glycosidase F. Purified recombinant PesXyn10A exhibited maximal activity at pH 6.0 and 50 °C, and activity was maintained at 90 % at pH 5.0 and temperatures lower than 30 °C for 24 h. The substrate specificity of PesXyn10A was limited and it hydrolyzed glucuronoxylan and arabinoxylan, but not ß-glucan. The final hydrolysis products from birchwood xylan were xylose, xylobiose, and 1,23-α-D-(4-O-methyl-glucuronyl)-1,4-ß-D-xylotriose. The addition of metallic salts (NaCl, KCl, MgCl2, and CaCl2) activated PesXyn10A for xylan degradation, and maximal activation by these divalent cations was approximately 160 % at a concentration of 5 mM. The thermostability of PesXyn10A significantly increased in the presence of 50 mM NaCl or 5 mM MgCl2. The present results suggest that the presence of metallic salts at a low concentration, similar to brackish water, exerts positive effects on the enzyme activity and thermal stability of PesXyn10A.
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The number of patients undergoing robotic surgery for rectal cancer has rapidly increased in Japan, since the government approved the procedure for national insurance coverage in April 2018. Robotic surgery has the potential to overcome some limitations of laparoscopic surgery, especially in the narrow pelvis, providing a three-dimensional view, articulated instruments, and a stable camera platform. Although meta-analyses and randomized controlled trials have failed to demonstrate the superiority of robotic surgery over laparoscopic surgery with respect to the short-term clinical outcomes, the published findings suggest that robotic surgery may be potentially beneficial for patients who are obese, male, or patients undergoing sphincter-preserving surgery for rectal cancer. The safety and feasibility of robotic surgery for lateral lymph node dissection, the standard procedure for locally advanced lower rectal cancer in Japan, have been demonstrated in some retrospective studies. However, additional prospective, randomized trials are required to determine the actual benefits of robotic surgery to ameliorate the urogenital and oncological outcomes. The cost of this approach is a long-standing principal concern. A literature search showed that the cost of robotic surgery for rectal cancer was 1.3-2.5 times higher per patient than that for the laparoscopic approach. We herein describe our surgical technique using a da Vinci Surgical System (S/Si/Xi) with 10 years of experience in performing robotic surgery. We also review current evidence regarding short-term clinical and long-term oncological outcomes, lateral lymph node dissection, and the cost of the procedure.
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PURPOSE: This study aimed to clarify the short- and long-term outcomes of robotic surgery with or without lateral lymph node dissection (LLND) for rectal cancer at Fujita Health University Hospital, Aichi, Japan, during a self-pay period. METHODS: We retrospectively evaluated 115 consecutive patients who underwent robotic surgery for rectal cancer between September 2009 and March 2018, with a median follow-up period of 48 months. Total mesorectal excision was completed by two certified surgeons using a da Vinci S, Si, or Xi Surgical System with an entirely robotic single-docking technique. The surgical and pathological outcomes, morbidity, and oncological results were examined. RESULTS: Lateral lymph node dissection was performed in 26 patients (22.6%). Neither conversion to open surgery nor perioperative blood transfusion occurred. Ten patients (8.7%) experienced Clavien-Dindo grade III postoperative complications. Pathologically, both the distal and radial margins were negative in all cases. The 5-year relapse-free survival rates for stages I, II, III, and IV were 93.5%, 100%, 83.8%, and not reached, respectively. CONCLUSIONS: Even in the period before coverage by the health insurance system, robotic surgery for rectal cancer performed by experienced surgeons was safe and technically feasible, with favorable perioperative results and long-term oncological outcomes, including rates of the relapse-free survival.
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Neoplasias del Recto/cirugía , Procedimientos Quirúrgicos Robotizados/métodos , Estudios de Cohortes , Supervivencia sin Enfermedad , Femenino , Humanos , Escisión del Ganglio Linfático , Masculino , Estadificación de Neoplasias , Neoplasias del Recto/mortalidad , Neoplasias del Recto/patología , Estudios Retrospectivos , Factores de Tiempo , Resultado del TratamientoRESUMEN
The polyethyleneimine-modified polymers, polystyrene-divinylbenzene-based (TAs) and polymethacrylate-based polymers (TAm), were used as palladium scavengers to eliminate residual palladium species after palladium on carbon-catalyzed Sonogashira-type coupling reaction. Since both TAs and TAm indicated relatively favorable elimination abilities toward residual palladium species in the reaction mixture, the affinities of TAs and TAm for palladium species were used as supports for palladium catalysts. The TAm-supported palladium catalyst (Pd/TAm) indicated better catalyst properties for the chemoselective hydrogenation compared to those of the corresponding TAs-supported palladium catalyst (Pd/TAs). Aromatic benzyl ethers; aromatic and aliphatic N-Cbzs; and aromatic carbonyl groups were smoothly hydrogenated in the presence of 1-5 mol % of Pd/TAm in MeOH or 2-PrOH. In contrast, the hydrogenation of aromatic ketones was selectively suppressed in morpholine which act as appropriate catalyst poison and solvent. Furthermore, Pd/TAm-catalyzed chemoselective hydrogenation was applicable to continuous-flow reaction.
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Hydrothermal reaction is known to be one of the most efficient procedures to extract hemicelluloses from lignocellulosic biomass. We investigated the molecular structure of xylooligosaccharides released from corn cob in a continuous flow type hydrothermal reactor designed in our group. The fraction precipitable from the extract with four volumes of ethanol was examined by 1H-NMR spectroscopy and MALDI-TOF MS before and after enzymatic treatment with different purified enzymes. The released water-soluble hemicellulose was found to correspond to a mixture of wide degree of polymerization range of acetylarabinoglucuronoxylan fragments (further as corn cob xylan abbreviated CX). Analysis of enzymatic hydrolyzates of CX with an acetylxylan esterase, GH3 ß-xylosidase, GH10 and GH11 xylanases revealed that the main chain contains unsubstituted regions mixed with regions of xylopyranosyl residues partially acetylated and occasionally substituted by 4-O-methyl-d-glucuronic acid and arabinofuranose esterified with ferulic or coumaric acid. Single 2- and 3-O-acetylation was accompanied by 2,3-di-O-acetylation and 3-O-acetylation of Xylp residues substituted with MeGlcA. Most of the non-esterified arabinofuranose side residues were lost during the hydrodynamic process. Despite reduced branching, the acetylation and ferulic acid modification of pentose residues contribute to high yields and high solubility of the extracted CX. It is also shown that different enzyme treatments of CX may lead to various types of xylooligosaccharides of different biomedical potential.
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Técnicas de Cultivo Celular por Lotes/métodos , Reactores Biológicos , Polisacáridos/química , Polisacáridos/metabolismo , Zea mays/metabolismo , Acetilación , Acetilesterasa/metabolismo , Técnicas de Cultivo Celular por Lotes/instrumentación , Biomasa , Reactores Biológicos/microbiología , Glucuronatos/análisis , Glucuronatos/metabolismo , Ácido Glucurónico/metabolismo , Estructura Molecular , Oligosacáridos/análisis , Oligosacáridos/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Xilanos/análisis , Xilanos/metabolismo , Xilosidasas/metabolismoRESUMEN
The carbohydrate esterase family 1 (CE1) in CAZy contains acetylxylan esterases (AXEs) and feruloyl esterases (FAEs). Here we cloned a gene coding for an AXE belonging to CE1 from Irpex lacteus (IlAXE1). IlAXE1 was heterologously expressed in Pichia pastoris, and the recombinant enzyme was purified and characterized. IlAXE1 hydrolyzed p-nitrophenyl acetate, α-naphthyl acetate and 4-methylumbelliferyl acetate, however, it did not show any activity on ethyl ferulate and methyl p-coumarate. We also examined the activity on partially acetylated and feruloylated xylan extracted from corncob by hydrothermal reaction. Similarly, ferulic and p-coumaric acids were not liberated, and acetic acid was only detected in the reaction mixture. The results indicated that IlAXE1 is an acetylxylan esterase actually reacted to acetyl xylan. However, since IlAXE1 was unable to completely release acetic acid esterifying xylopyranosyl residues, it is assumed that acetyl groups exhibiting resistance to deacetylation by IlAXE1 are present in corn cob xylan.
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A 63-year-old woman was admitted to our hospital with a right lower abdominal mass and general fatigue. Preoperative examination suggested a large ovarian tumor or cecal carcinoma. However, her intraoperative diagnosis was colon cancer; we therefore performed an ileocecal resection with oophorectomy. The tumor was pathologically diagnosed as adenosquamous carcinoma T4bN1M-stage IIIa. We administrated CapeOX adjuvant chemotherapy for 6 months. Adenosquamous carcinoma is extremely rare, at around 0.1% of all colorectal cancers, and usually has a poor prognosis. The patient is still alive without recurrence after 84 post-operative months, even with later developments of metachronous early colorectal cancer and breast cancer. We herein report a rare case of cecal ASC with good prognosis.
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We reviewed the clinical records of 302 patients with low rectal carcinoma to evaluate the effectiveness of lateral lymph node(LLN)dissection in cases of skip metastases to the LLN. Patients were divided into 4 groups according to nodal status: group N(133 cases), no metastasis in both the mesorectum lymph node(MLN)and LLN; groupM(100 cases), metastasis in the MLN and no LLN metastasis; group L(15 cases), which was defined as skip metastasis, with no metastasis in the MLN and LLN metastasis; and group ML(54 cases), node metastases in both the MLN and LLN. LLN metastasis was found in 22.8% of all patients, and skip metastasis was found in 10.1%. The recurrence rate in group L(40.0%, 6 cases)was significantly lower than that in group ML(75.9%, 41 cases)and not different from that in group M(43.0%, 43 cases). The initial recurrence sites in group L were the lung(3 cases)and local recurrence(3 cases). In group L, 20.0% had hematogenous recurrence, which was lower than in group ML. There was no significant difference between group L(5-year survival rate: 57.1%)and group M(71.7%)in terms of prognosis. Multivariate analysis revealed that histology was a risk factor for LLN metastasis in lowrectal cancer without MLN metastasis. The positive predictive value of LLN metastasis based on computed tomography was 43.9%. It was believed that LLN dissection was important, especially for non-differentiated cancer, in consideration of skip metastases to the LLN.
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Neoplasias del Recto , Humanos , Escisión del Ganglio Linfático , Ganglios Linfáticos , Metástasis Linfática , Recurrencia Local de Neoplasia , Pronóstico , Estudios RetrospectivosRESUMEN
We evaluated the effectiveness of chemoradiotherapy(CRT)by reviewing 11 clinicalcases of analsquamous cellcarcinoma( SCC). Radiotherapy(RT)consisted of 40 Gy delivered to pelvic and bilateral inguinal lesions, and a perianal booster dose of 20 Gy in fractions of 2.0 Gy per day, 5 days per week. 5-fluorouracil(5-FU)and mitomycin C were administered twice every 4weeks as standard chemotherapy. On the first day of RT, patients received a single bolus dose of 10mg/m2 mitomycin C, and a continuous 24-hour infusion of 750mg/m2 5-FU for 5 days. One patient with a T3 tumor was orally administered S- 1 during RT because of his poor generalcondition, and 1 patient with a T2 tumor did not receive 1 course of 5-FU and MMC owing to an adverse event. Grade 3 adverse effects occurred in 3 patients, but all 11 patients completed CRT. The anal lesions of 10 patients had complete response after CRT. Recurrence of anal lesions occurred in 4 patients, including 2 patients who were not treated with standard CRT. Of 8 patients who received CR via standard CRT, 2 patients had recurrence of anal lesions more than 60 months after completion of CRT. CRT is believed to be safe and effective for improving the prognosis of anal squamous cell carcinoma; however, sufficient and appropriate follow-up is necessary after complete response.
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Protocolos de Quimioterapia Combinada Antineoplásica , Neoplasias del Ano , Carcinoma de Células Escamosas , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias del Ano/tratamiento farmacológico , Neoplasias del Ano/radioterapia , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/radioterapia , Quimioradioterapia , Fluorouracilo/administración & dosificación , Humanos , Mitomicina/administración & dosificación , Estudios RetrospectivosRESUMEN
Rectovaginal fistula caused by a tension-free vaginal mesh (TVM) is a rare condition. Moreover, a rectovaginal fistula is a challenging issue to address for surgeons regardless of causes. Due to a low rate of occurrence, treatment modality for a rectovaginal fistula caused by a TVM has previously received little attention. A successful surgery using several key techniques to address a rectovaginal fistula caused by a TVM is herein reported. A 78-year-old woman who underwent a TVM for a rectocele three months ago was referred to our hospital with a two-month history of anal bleeding. Mesh protruding into both the vagina and the rectum was confirmed. The patient was operated on under diagnosis of a rectovaginal fistula caused by TVM. TVM was removed by transvaginal dissection of the rectovaginal septum with division of both anterior and posterior arms of the TVM. Layer-to-layer sutures of rectal and vaginal walls were crossly performed with a drain placed in the rectovaginal septum after saline irrigation followed by a covering sigmoid colostomy. The wound healed without infection after surgery, and a water-soluble contrast enema demonstrated the healing of the rectovaginal fistula two months after surgery. No recurrent fistula was confirmed 15 months after stoma closure.
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In the presence of palladium on carbon (Pd/C) as a catalyst, hydrogenation of aliphatic nitriles in cyclohexane efficiently proceeded at 25-60 °C under ordinary hydrogen gas pressure to afford the corresponding tertiary amines. However, the use of rhodium on carbon (Rh/C) led to the highly selective generation of secondary amines. Hydrogenation of aromatic nitriles and cyclohexanecarbonitrile selectively produced secondary amines in the presence of either Pd/C or Rh/C.
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We reviewed the clinical records of 14 cases who received neoadjuvant chemoradiotherapy(CRT)to evaluate the clinical effectiveness of the CRT for T4b rectal cancer. The preoperative radiotherapy consisted of 40-50 Gy delivered in fractions of 1.8-2.0 Gy per day, 5 days per week. A treatment with intravenous 5-fluorouracil, or oral tegafur-uracil(UFT)with l-leucovorin, or oral S-1, or capecitabine or intravenous irinotecan with oral S-1, was administered during radiotherapy. One patient died due to pelvic abscess at 69 days after CRT. Complete response(CR)or partial response(PR)was observed in 8 cases, 1 month after CRT. Curative surgery was performed in 10 patients. Among 10 patients who underwent curative surgery, both urinary and anal function were preserved in 5 patients. Although no lymph node metastasis was found in 9 patients of 10 patients who underwent curative surgery, recurrence was found in 5 patients, and local recurrence was found in 4 of these patients. Recurrence occurred in all patients who had recurrence within 1 year. Preoperative CRT was expected to be an effective treatment to improve the resection rate and prognosis for T4b rectal cancer. However, it was thought that it was necessary to be careful about local recurrence, especially within 1 year after surgery.
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Quimioradioterapia , Terapia Neoadyuvante , Neoplasias del Recto/terapia , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Humanos , Persona de Mediana Edad , Invasividad Neoplásica , Neoplasias del Recto/patologíaRESUMEN
Our laboratory has four separate methods for the analysis of trenbolone, stilbenes, resorcyclic acid lactones, and estradiol in bovine urine. The method described in this paper was in response to a client request to consolidate the methods preferably into one method. A multiresidue semi-quantitative method was developed and any suspect positive samples detected by the method were subjected to the method of standard addition to accurately quantify the concentration of the positive analyte. Samples were enzymatically hydrolyzed using ß-glucuronidase after which, supported liquid extraction on HM-N cartridges was performed, followed by solvent exchange into methyl tert-butyl ether (MTBE). The samples were evaporated and reconstituted into 10% methanol in water and loaded onto a SampliQ OPT SPE. The cleaned-up extract was further cleaned up on a SampliQ NH2 cartridge. The SPE eluate was split into two for analysis by gas chromatography-mass spectrometry (GC-MS) using electron ionization (EI) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) using both positive and negative electrospray. It was found that with the exception of estradiol and trenbolone all compounds could be analyzed by both GC-MS and LC-MS/MS, providing a semi-quantitative method. It is recommended that quantification is achieved using standard addition. Of the 13 compounds successfully monitored, the limits of detection (LODs), and the limits of quantification (LOQs) obtained were within the Codex limits for the target concentrations. As far as the authors are aware, the use of supported liquid extraction has not been reported for bovine urine analysis. © 2016 Her Majesty the Queen in Right of Canada. Drug Testing and Analysis © 2016 John Wiley & Sons, Ltd.
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Cromatografía Liquida/métodos , Cromatografía de Gases y Espectrometría de Masas/métodos , Lactonas/orina , Esteroides/orina , Estilbenos/orina , Espectrometría de Masas en Tándem/métodos , Anabolizantes/orina , Animales , Bovinos , Estradiol/orina , Límite de Detección , Extracción en Fase Sólida/métodos , Detección de Abuso de Sustancias/métodos , Acetato de Trembolona/orinaRESUMEN
Laparoscopic cholecystectomy (LC) does not require advanced techniques, and its performance has therefore rapidly spread worldwide. However, the rate of biliary injuries has not decreased. The concept of the critical view of safety (CVS) was first documented two decades ago. Unexpected injuries are principally due to misidentification of human factors. The surgeon's assumption is a major cause of misidentification, and a high level of experience alone is not sufficient for successful LC. We herein describe tips and pitfalls of LC in detail and discuss various technical considerations. Finally, based on a review of important papers and our own experience, we summarize the following mandatory protocol for safe LC: (1) consideration that a high level of experience alone is not enough; (2) recognition of the plateau involving the common hepatic duct and hepatic hilum; (3) blunt dissection until CVS exposure; (4) Calot's triangle clearance in the overhead view; (5) Calot's triangle clearance in the view from underneath; (6) dissection of the posterior right side of Calot's triangle; (7) removal of the gallbladder body; and (8) positive CVS exposure. We believe that adherence to this protocol will ensure successful and beneficial LC worldwide, even in patients with inflammatory changes and rare anatomies.