Axillary lymph-node metabolic activity assessment on 18F-FDG-PET/CT in rheumatoid arthritis patients treated with biologic therapies.

Objective: Recent studies have provided new insights into the role of lymph nodes (LNs) in rheumatoid arthritis (RA). The aim of this study was to evaluate the metabolic activity of the axillary LNs in relation to that of the upper limb joints and the clinical assessment of disease activity in RA patients treated with biologic therapies.Method: 18F-fluorodeoxyglucose-positron emission tomography/computed tomography (18F-FDG-PET/CT) scans were acquired for 64 patients with RA at baseline and after 6 months of biologic therapy, and the patients' clinical status was evaluated. The maximum standardized uptake value (SUVmax), metabolic active volume, and total lesion glycolysis (TLG) were used to assess glucose metabolism in the LNs and 12 joints. Clinical evaluations included serum markers and the Disease Activity Score based on 28-joint count-erythrocyte sedimentation rate (DAS28-ESR).Results: Changes in the SUVmax and TLG for the axillary LNs correlated significantly with those of the ipsilateral wrist joints. There was a positive correlation between the changes in the three metabolic parameters of the axillary LNs and the changes in disease activity after treatment. After 6 months of biologic therapy, all metabolic parameters for the axillary LNs in patients with a DAS28-ESR < 3.2 were significantly lower than those of patients with a DAS28-ESR ≥ 3.2.Conclusion: A relationship between the glucose metabolism of the axillary LNs and the ipsilateral wrist joints was demonstrated by the 18F-FDG-PET/CT parameters. The metabolic activity and active volume of axillary LNs may reflect the therapeutic response to the biologic treatment of RA.

Biological significance of fluorine-18-α-methyltyrosine (FAMT) uptake on PET in patients with oesophageal cancer.

PURPOSE: (18)F-FAMT as an amino-acid tracer for positron emission tomography (PET) is useful for detecting human neoplasms. (18)F-FAMT is accumulated in tumour cells solely via L-type amino-acid transporter 1 (LAT1). This study was conducted to investigate the biological significance of (18)F-FAMT uptake in patients with oesophageal cancer. METHODS: From April 2008 to December 2011, 42 patients with oesophageal cancer underwent both (18)F-FAMT PET/CT and (18)F-FDG PET/CT before surgical treatment. The immunohistochemical analysis of LAT1, CD98, Ki-67, CD34, p53, p-Akt and p-mTOR was performed on the primary lesions. In vitro experiments were performed to examine the mechanism of (18)F-FAMT uptake. RESULTS: High uptake of (18)F-FAMT was significantly associated with advanced stage, lymph node metastasis and the expression of LAT1, CD98, Ki-67 and CD34. LAT1 expression yielded a statistically significant correlation with CD98 expression, cell proliferation, angiogenesis and glucose metabolism. In vitro experiments revealed that (18)F-FAMT was specifically transported by LAT1. CONCLUSIONS: The uptake of (18)F-FAMT within tumour cells is determined by the LAT1 expression and correlated with cell proliferation and angiogenesis in oesophageal cancer. The present experiments also confirmed the presence of LAT1 as an underlying mechanism of (18)F-FAMT accumulation.

Expansion of radiofrequency ablation volume by saturated NaCl saline injection in the area of vaporization.

BACKGROUND: Vaporization around the radiofrequency (RF) electrode after RF application (RFA) limits the RF ablation area. PURPOSE: To determine whether saturated saline injected into the area of vaporization after initial RFA extends ablation area after further RFA. MATERIAL AND METHODS: RFA was performed in 18 ex vivo porcine livers and four in vivo rabbit erector spinae muscles. An RF electrode was used to ablate an area with 40W of parallel current for 15 min. The ablation margin was determined using a thermocouple, and the radius of the ablated area was measured. After RF electrode removal, saturated saline was infused through a percutaneous ethanol injection needle into the site of the original RFA in 11 liver samples and two erector spinae muscles. Three minutes later, RFA was resumed for 15 min. The remaining seven control liver samples and two spinae muscles received RFA without saline injection. The radius of the final ablated area was then measured. RESULTS: In the ex vivo study, injection of saturated saline significantly decreased tissue impedance (87.7+/-9.4 to 51.1+/-9.7 Omega, P<0.0001), and increased the mean radius of the ablated area (15.9+/-3.0 to 25.0+/-3.6 mm, P<0.0001). These significant changes were not observed without injection of saturated saline. Similar trends were found in the in vivo study. CONCLUSION: Injection of saturated saline into the area of vaporization around the RF electrode, followed by additional RFA, caused concentric expansion of the final ablation area, facilitating more efficient tumor ablation.

[Aortic dissection after aortic valve replacement with an aorto-left atrial fistula; report of a case].

Aortocameral fistula is a rare complication of aortic dissection. We herein report a case of aortic dissection after aortic valve replacement (AVR) complicated with a fistula to the left atrium. A 76-year-old man who had undergone AVR 1 year previously, was admitted to our hospital because of facial edema and chest discomfort. On auscultation, a continuous murmur was heard at the left lower sternal border. Computed tomography revealed dissecting aneurysm of the ascending aorta and a fistula to the left atrium was suspected. Transesophageal echocardiography showed the fistula between the false lumen of the aneurysm and the left atrium. Ascending aorta replacement and closure of the fistula was performed. There was dense adhesion between the aortic root and the roof of the left atrium. It seems that postoperative adhesion plays an important role in formation of aortocameral fistula.

Thymic hyperplasia in patients with Graves' disease. Identification of thyrotropin receptors in human thymus.

Thymic size and density were studied in 23 untreated patients with Graves' disease and 38 control subjects using computed tomography. Both thymic size and density were higher in untreated patients with Graves' disease than in control subjects in the age-matched group. After treatment with antithyroid drugs, both thymic size and density were significantly reduced, with a concomitant decrease in thyrotropin receptor antibodies. PCR of human thymic cDNA using primers for human thyrotropin receptor amplified a fragment in a size expected for the receptor, and its nucleotide sequence was identical to human thyrotropin receptor cDNA in the thyroid. Northern blot analysis of human thymic poly(A)+ RNA demonstrated the presence of the full length form of thyrotropin receptor mRNA. Western blot analysis of human thymic membrane using anti-thyrotropin receptor peptide antibodies demonstrated a band of 100 kD that was also observed in the thyroid membrane. Immunohistochemistry of thymic tissue using mouse antihuman thyrotropin receptor monoclonal antibodies demonstrated the immunostaining of epithelial cells. These results indicate that thymic hyperplasia is apparently associated with Graves' disease and suggest that thymic thyrotropin receptor may act as an autoantigen that may be involved in the pathophysiology of development of Graves' disease.

[Giant cardiac metastasis of myxoid liposarcoma causing cardiac tamponade; report of a case].

We report a very rare case of cardiac metastasis of myxoid liposarcoma. A 55-year-old man presented with dyspnea. Two and a half years ago, he underwent resection of myxoid liposarcoma in the left thigh. Magnetic resonance imaging (MRI) revealed a giant tumor occupying the pericardiac cavity and pressing the heart and consequently causing cardiac tamponade. The patient underwent surgery through a left thoracotomy approach. The pericardiac cavity was filled with a giant tumor with a stalk from the right ventricle and 2 small nodules on the main pulmonary artery. He was relieved from the symptom: however, he had a recurrence of the tumor at the same site 5 months after the operation. He underwent surgery for the removal of the second tumor; however, he died 49 days after the operation. Although cardiac metastasis is a very rare condition, its awareness is essential for careful long-term follow-up for the early detection of a metastatic cardiac liposarcoma after the resection of the primary tumor.

[Surgical technique of aortic valve replacement for small aortic annulus in elderly patients].

Recent reports have shown that aortic valve replacement in elderly patients over 65 years with atherosclerotic aortic stenosis and a small aortic annulus is possible by using a small sized bioprosthesis (Carpentier-Edwards pericardial valve). Here we present out surgical technique. Firstly, the native calcified aortic valve was removed completely to gain total exposure of the surrounding aortic root and sinus of Valsalva like Bentall procedure. Secondly, a small sized bioprosthesis was implanted with intermittent noneverting mattress 2-0 sutures with spaghetti and small polytetrafluoroethylene (PTFE) felt. Aortic annulus is the dilated by inserting Hegar dilator sizing from 25 to 27 mm. Therefore, aortic valve replacement for small aortic annulus in intra- or supra-annular position should be easily accomplished. Good surgical results and hemodynamic state were achieved in 25 consecutive cases using this technique.

Molecular cloning and expression of human ribonuclease 4 cDNA.

A cDNA coding for human ribonuclease 4 was isolated from a pancreas cDNA library and sequenced. This cDNA (996 bp) includes an entire open reading frame encoding mature protein (119 aa) following signal peptide (28 aa). Expression of mature protein in Escherichia coli showed an apparent molecular mass of about 16 kDa, which was slightly lower than the mature form of human RNase 1, in SDS-PAGE.

The three-dimensional structure of human RNase 4, unliganded and complexed with d(Up), reveals the basis for its uridine selectivity.

The RNase 4 family is unique among RNase enzymes, displaying the highest level of sequence similarity and encompassing the shortest polypeptide chain. It is the only one showing high specificity. The human representative is an intracellular and plasma enzyme, first isolated from colon adenocarcinoma cell line HT-29. The crystal structures of human recombinant RNase 4, unliganded and in complex with d(Up), have been determined, revealing in the unique active site an explanation for the uridine specificity. Arg101, at a position not involved in catalysis in the other RNase enzymes, penetrates the enzyme moiety shaping the recognition pocket, a flip that is mediated by the interaction with the (shorter chain) C-terminal carboxylate group, providing an anchoring point for the O4 atom of the substrate uridine. The bulky Phe42 side-chain forces Asp80 to be in the chi1=-72.49 degrees rotamer, accepting a hydrogen bond from Thr44, further converting the latter into a hydrogen bond acceptor. This favours an interaction with the -NH-donor group of uridine at position 3 over that with the =N-acceptor of cytidine. The two chemical groups that distinguish uracyl from cytosine are used by the enzyme to discriminate between these two bases.

Local massive venous invasion in colorectal cancer: CT-pathological correlation and its clinical implication.

OBJECTIVE: To identify CT findings of massive venous invasion (MVI) in colorectal cancer, compare them to pathological findings and evaluate its clinical implications. METHODS: Among 423 patients who received surgical resection of colorectal cancer, pre-operative CT of 26 patients (15 males, 11 females; mean age, 63.0 ± 12.1 years) with histopathologially proven MVI and 26 patients (14 males, 12 females; mean age, 71.1 ± 9.6 years) with histopathologically proven lymph node (LN) metastases were reviewed and compared with histopathological findings. We evaluated CT detectability of MVI and the morphologic differences between MVI and LN metastasis. All cases were followed up for at least 6 months after surgery. RESULTS: Pre-operative CT correctly diagnosed only one case as tumour thrombus. 9 lesions were not detected on CT, and others were misdiagnosed pre-operatively as regional LN metastasis (14 cases) and juxtatumoural abscess (2 cases). After reviewing these cases, MVIs were identifiable in 20 of 26 cases. MVI was depicted on CT as nodules (oval, lobulated), abscess-like or intravenous tumour thrombus. MVI was significantly larger than LN metastasis (p < 0.05), while contrast enhancement was significantly lower (p < 0.05), and MVI often had an enhanced rim. Ten patients had synchronous metastases, and six patients had metachronous distant metastases within 5 years. CONCLUSION: Many cases of MVI were distinguishable from LN metastases on pre-operative CT of colorectal cancer, but their appearances were varied, reflecting their histopathological behaviours. The distant metastatic rate was much higher in cases with MVI. ADVANCES IN KNOWLEDGE: Radiologists should be aware of CT findings of MVI in colorectal cancer as a sentinel sign of distant metastasis.

Different lipid contents between aldosterone-producing and nonhyperfunctioning adrenocortical adenomas: in vivo measurement using chemical-shift magnetic resonance imaging.

Lipid contents in the 11 aldosterone-producing and 24 nonhyperfunctioning adrenocortical adenomas and 8 pheochromocytomas were assessed in vivo by using chemical-shift fast low-angle shot magnetic resonance imaging. T2 relaxation times were measured by using a spin echo sequence. The relative lipid contents of pheochromocytomas were significantly less than adrenocortical adenomas. The relative lipid contents of aldosterone-producing adenomas were significantly less than nonhyperfunctioning adenomas. The longer T2 relaxation times of aldosterone-producing adenomas may also represent less lipid contents compared with nonhyperfunctioning adenomas. These results strongly support the suggestion that there is a very close relationship between accumulated lipid droplets and functional aspects in the adrenocortical cells. Chemical-shift magnetic resonance imaging may be a useful method for the indirect analysis of the function of adrenocortical adenomas in vivo.

Gene arrangement and RNA transcription of the BamHI fragments K and M2 within the non-oncogenic Marek's disease virus serotype 2 unique long genome region.

We determined the nucleotide sequence of a 6593 bp fragment of the Marek's disease virus serotype 2 (MDV2) unique long region located in the right part of genomic BamHI-M2 and the adjacent part of BamHI-K fragments. Within this region five complete open reading frames (ORFs) were identified whose deduced amino acid sequences exhibited homology to the UL53 (glycoprotein K), UL54 (immediate early regulatory protein ICP27), and UL55 gene products of herpes simplex virus type 1 (HSV-1). Homologue to the HSV-1 UL56 was not detected. However, we identified a gene between the MDV2 UL54 and UL55 genes with homology to the first ORF (ORF-1) of equine herpesvirus type 1 and corresponding gene identified in pseudorabies virus. Two adjacent ORFs contained in the BamHI-K fragment, ORF 873s and ORF 873, were found by computer analysis to have the properties of an intron encoding a glycoprotein: ORF 873s encodes a 84 amino acid polypeptide with a stretch of a hydrophobic signal sequence in the C-terminus, and ORF 873 encodes a 873 amino acid polypeptide with a transmembrane domain and putative three N-linked glycosylation sites. All the identified genes were confirmed to be transcribed with 3'-coterminal transcripts and/or a unique transcript in the virus-infected cells. Especially, 3.5 kb mRNA of ORF 873s and ORF 873 are transcribed from a potential promoter region of ORF 873s, and splice donor and acceptor sites are used to splice the mRNA after cleavage of a 113 bp-nucleotide sequence.

Use of contrast-enhanced computed tomography to measure clearance per unit renal volume: a novel measurement of renal function and fractional vascular volume.

The iodinated contrast agents used for computed tomography (CT) have pharmacokinetics similar to inulin and can measure physiological indices, such as clearance per unit renal volume (alpha/V) and fractional vascular volume (fvv). Clinical experience with these techniques is, however, scanty, and the present study explored their potential in subjects with and without renal dysfunction. In a series of subjects, a single slice of kidney was scanned sequentially after the bolus injection of contrast material. Time-attenuation curves were constructed, and alpha/V and fvv were calculated using a Patlak graphic analysis. In the first part of the study, 50 normal kidneys in 35 subjects (aged 21 to 75 years) were studied. In the second stage, alpha/V was compared with glomerular filtration rate (GFR) measurements in 24 patients with diabetes (aged 28 to 84 years) with or without renal dysfunction. In normal kidneys, alpha/V averaged 0.49 +/- 0.11 mL/min/mL and fvv averaged 35% +/- 12%. These values agree with literature data obtained using other techniques. A negative correlation was seen between age and alpha/V (r = 0.66; P < 0.0001), but not fvv. In patients with diabetes, a strong correlation was observed between renal clearance values, calculated from CT and corrected for renal volume, and GFR (r = 0.87; P < 0.0001). Dynamic CT can provide quantitative renal physiological information on a regional basis noninvasively.

Convenient and efficient in vitro folding of disulfide-containing globular protein from crude bacterial inclusion bodies.

We investigated how the folding yield of disulfide-containing globular proteins having positive net charges from crude bacterial inclusion bodies was affected by additives in the folding buffer. In screening folding conditions for human ribonucleases and its derivative, we found that addition of salt (about 0.4 M) to a folding buffer increased the folding yield. This suggested that electrostatic interaction between polyanionic impurities such as nucleic acids and cationic unfolded protein led to the formation of aggregates under the low-salt conditions. Since inclusion bodies were found to contain nucleic acids regardless of the electrostatic nature of the expressed protein, the electrostatic interaction between phosphate moieties of nucleic acids and basic amino acid residues of a denatured protein may be large enough to cause aggregation, and therefore the addition of salt in a folding buffer may generally be useful for promotion of protein folding from crude inclusion bodies. We further systematically investigated additives such as glycerol, guanidium chloride, and urea that are known to act as chemical chaperons, and found that these additives, together with salt, synergistically improved folding yield. This study, suggesting that the addition of salt into the folding buffer is one of the crucial points to be considered, may pave the way for a systematic investigation of the folding conditions of disulfide-containing foreign proteins from crude bacterial inclusion bodies.

Tissue-specific expression of pancreatic-type RNases and RNase inhibitor in humans.

The tissue-specific expression of five human pancreatic-type RNases and RNase inhibitor was analyzed by Northern hybridization against poly(A)+ RNA prepared from 16 normal tissues. The widespread expression of RNase 1 was observed in almost all of the tissues. RNase 4 and angiogenin showed a similar distribution of expression abundantly present in the liver. This suggested the identity of the cell types producing these two molecules. However, no relativity appeared to be present between the vascularization of the tissues and the angiogenin expression. A narrow range of expression of the eosinophil-derived neurotoxin gene was observed. This localization seems related to the phagocytic cells in the tissues. The undetectable level of the eosinophil cationic protein mRNA in normal tissues suggests that the differentiation of eosinophils, triggered by inflammation and/or atopy, is required. The expression of RNase inhibitor was found to be ubiquitous. The regulatory function of inhibitor against RNases in the cell should be considered in studying the physiological significance of the pancreatic-type RNase family.

Age-dependent decline in parenchymal perfusion in the normal human pancreas: measurement by dynamic computed tomography.

Parenchymal perfusion of the normal human pancreas using dynamic computed tomography (CT) was evaluated and correlated with patient demographic characteristics. The results of 23 patients (10 men and 13 women; age range, 25-71 years) who underwent enhanced CT of the upper abdomen and perfusion measurement were retrospectively reviewed. They had no evidence of pancreatic disease or diffuse liver disease, clinically or radiographically. Regions of interest were drawn in the pancreatic body and within the aorta. Pancreatic parenchymal perfusion per volume was then calculated by dividing the peak gradient of the pancreatic time-attenuation curve by the peak aortic CT number in increase. Perfusion in these patients ranged from 0.554 to 1.698 ml min(-1) ml(-1) (mean +/- SD, 0.963 +/- 0.064) and showed a negative correlation with the patient's age (r = 0.699, p < 0.0005). Pancreatic parenchymal density before contrast material injection (mean +/- SD, 48.86 +/- 0.978) was not correlated with perfusion measured by dynamic CT or age. No differences were observed in perfusion or density between men and women. In conclusion, parenchymal perfusion of the normal human pancreas measured by dynamic CT appears to decline with age.

In vitro and in vivo release of poly(DL-lactic acid) microspheres containing neurotensin analogue prepared by novel oil-in-water solvent evaporation method.

Poly(DL-lactic acid) (PLA) microspheres containing a neurotensin analogue [NA; H(CH3)-Arg-Lys-Pro-Trp-tert-Leu-Leu-OEt.3HCl] were prepared by a novel oil-in-water (o/w) solvent evaporation method, and the release behaviors were evaluated in vitro. About 20% of the loaded NA was released initially, and the subsequent release lasted for a month from microspheres prepared with PLA of molecular weight 2000 (PLA 2000). A smaller initial release from PLA 4000 and PLA 6000 microspheres was found, but a lag time of 2-3 weeks during which the drug was not released was observed with PLA 4000 and PLA 6000 microspheres. The addition of relatively hydrophilic monoglycerides decreased the lag time, and a fairly constant release of NA was achieved. The pharmacokinetic behavior of NA from PLA 2000 microspheres was studied in rats. The release of the drug after a subcutaneous injection exhibited pseudo-zero-order kinetics for 1 month. The initial release of the drug from the microspheres was reflected in a sharp increase of the plasma levels of the de-ester form of NA [H(CH3)-Arg-Lys-Pro-Trp-tert-Leu-Leu-OH], and the subsequent steady-state levels agreed well with the predicted levels obtained from analysis of constant-infusion kinetics.

Preparation of neurotensin analogue-containing poly(dl-lactic acid) microspheres formed by oil-in-water solvent evaporation.

This report describes the preparation of injectable microspheres containing a neurotensin analogue (NA), which is a hexapeptide with neurotensin activity. NA, a hydrophilic drug, was successfully entrapped into poly(dl-lactic acid) microspheres prepared by a novel oil-in-water solvent evaporation method. The preparation method was investigated with regard to the partition of NA into the oily phase and the rapid phase separation of the polymer. Successful entrapment was achieved with the following conditions: (1) an alkaline water phase, (2) addition of fatty acid salt in the oily phase, and (3) addition of a water-miscible solvent in the oily phase. Under these conditions, NA was completely entrapped into the microspheres at poly(dl-lactic acid):NA molar ratios of greater than 3.

Enhancement of the oral bioavailability of cinnarizine in oleic acid in beagle dogs.

The present study was an attempt to develop a new dosage form of cinnarizine, which is slightly soluble in water, using lipid as a vehicle. The solubility of cinnarizine in several organic solvents was determined. It was found that cinnarizine dissolved well in oleic and linoleic acids. The bioavailability of cinnarizine from the oral administration of an oleic acid solution in a hard capsule was investigated and compared with that of a cinnarizine tablet, using beagle dogs. When cinnarizine was administered in a capsule, the bioavailability was greatly enhanced [i.e., the maximum concentration (Cmax) and AUC values were 2.9 and 4.0 times larger than those of a cinnarizine tablet, respectively]. Meanwhile, the tmax value (the time to reach Cmax) was unchanged. The absorption of cinnarizine from an oleic acid solution was considered to depend on the action of bile salts. This was supported by the results of a dissolution test using a bile salts solution as the dissolution test medium.

Enhancement of bioavailability of cinnarizine from its beta-cyclodextrin complex on oral administration with DL-phenylalanine as a competing agent.

The present investigation is concerned with an improvement of the bioavailability of cinnarizine by administering its beta-cyclodextrin complex together with another compound which competes with the beta-cyclodextrin molecule in complex formation in aqueous solution (competing agent). The bioavailability of cinnarizine on oral administration of the cinnarizine-beta-cyclodextrin inclusion complex was enhanced by the simultaneous administration of DL-phenylalanine as a competing agent, e.g., the AUC was 1.9 and 2.7 times as large as those of the cinnarizine-beta-cyclodextrin complex alone and cinnarizine alone, respectively. The enhancement of AUC and Cmax completely depended on the dose of DL-phenylalanine. It was found from these results that DL-phenylalanine acted as a competing agent in the GI tract and the minimum effective dose required of DL-phenylalanine might be 1 g for 50 mg of cinnarizine in the cinnarizine-beta-cyclodextrin complex. Evaluating the competing effect of DL-phenylalanine in vitro using an absorption simulator, it was found that the decreased penetration rate of cinnarizine through the artificial lipid barrier with addition of beta-cyclodextrin was restored with the addition of DL-phenylalanine.