Hepatocellular carcinoma after radiofrequency ablation therapy: dynamic CT evaluation of treatment.

The aim of this study is to analyze the dynamic CT findings after radiofrequency (RF) ablation for hypervascular hepatocellular carcinoma (HCC). RF ablation was used for 22 tumors in 20 patients. Peripheral enhancement was noted in 89% of regions within 1 month in the arterial phase. In 1-3 months, peripheral enhancement remained at 56% and was reduced to 22% by 3-6 months. It is difficult to determine the therapeutic result within 3 months due to continued peripheral enhancement in the arterial phase.

Synthesis of a terbium fluorescent chelate and its application to time-resolved fluoroimmunoassay.

A new nonadentate ligand, N, N, N1, N1-[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-phenylpyridine]tetrakis(acetic acid) (BPTA) for a Tb3+ fluorescent complex was synthesized. The Tb3+ complex is strongly fluorescent, having a large fluorescence quantum yield of 1.00 and very long fluorescence lifetime of 2.681 ms in 0.05 M borate buffer of pH 9.1. Streptavidin (SA) was labeled with BPTA by using its succinimidyl monoester, and the BPTA-Tb(3+)-labeled SA was used in sandwich-type time-resolved fluoroimmunoassay (TR-FIA) of alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) in human sera. The Tb(3+)-labeled SA was also used in competitive-type TR-FIA of bensulfuron-methyl (BSM) in water. The detection limits of these assays are 42 pg/mL for AFP, 70 pg/mL for CEA, and 0.4 ng/mL for BSM. In addition, a new simultaneous measurement method for AFP and CEA in a human serum sample was developed by using 4,4'-bis(1",1",2",2",3",3" -heptafluoro-4",6"-hexanedion-6"-ly)chlorosulfo- o-terphenyl (BHHCT)-Eu(3+)-labeled anti-AFP antibody, biotinylated anti-CEA antibody, and BPTA-Tb(3+)-labeled SA. The concentrations of AFP and CEA in 39 human serum samples were determined, and the results were compared with those of the independently determined AFP and CEA by TR-FIA with a single-label method. A good correlation was obtained with the correlation coefficients of 0.991 for AFP and 0.994 for CEA.

Shape of lens epithelial cells after intraocular lens implantation.

PURPOSE: To evaluate the morphological behavior of lens epithelial cells (LECs) after human cataract surgery with implantation of a poly(methyl methacrylate) (PMMA) or silicone intraocular lens (IOL). SETTING: Department of Ophthalmology, Fujita Health University, Banbuntane Hotokukai Hospital, Nagoya, Aichi, Japan. METHODS: Morphological observations of LECs in the patients with IOLs were made by light and transmission electron microscopy. The LECs were from 4 areas: (1) the region below the anterior capsule, touching the IOL; (2) the area between region 1 and the equatorial region; (3) the equatorial region; and (4) the central equatorial region and of the posterior capsule not touching the IOL. Case 1 had implantation of a single-piece IOL with a PMMA optic and haptics. Case 2 had a 3-piece IOL with a PMMA optic and polypropylene haptics. Case 3 had a 3-piece IOL with a silicone optic and polypropylene haptics. Areas 1 and 4 could not be observed in Case 2. RESULTS: The major difference between the patient with a PMMA IOL (Case 1) and the patient with a silicone IOL (Case 3) was that among the 4 areas observed, collagen fibers were present only in area 1 in Case 1 but in areas 2 or 3 as well in Case 3. CONCLUSIONS: Fibrous collagen fibers appeared in regions in which LECs adhered and there was capsule contact with the IOL optic. In addition fibrous collagen fibers appeared in more areas in the eye with the silicone IOL than in that with the PMMA IOL, perhaps because IOLs with silicone optics move slightly while in the capsular bag.

Total and pancreatic amylase measured with 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside.

BACKGROUND: : Many different methods have been used to assay amylase activity, using nitrophenylated oligosaccharides as substrate; however, the hydrolysis steps in these methods are complex. METHODS: : We developed a new continuously monitoring assay for amylase activity in biological fluids, using 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside (GalG2CNP) as the substrate; this assay was used with anti-human salivary amylase monoclonal antibodies for specific determination of the pancreatic isoenzyme. Amylase converted GalG2CNP into beta-D-galactopyranosylmaltose and 2-chloro-4-nitrophenol, which was measured at 405 nm. RESULTS: : GalG2CNP was cleaved between 2-chloro-4-nitrophenol and beta-D-galactopyranosylmaltose and did not undergo transfer reactions. The within-assay CVs (n = 20) for total amylase (T-AMY) and pancreatic amylase (P-AMY) were 0.6-1.6% and 0.5-2.5%, respectively; and day-to-day CVs (n = 10) for T-AMY and P-AMY were 0.8-3.7% and 0.6-4.1%, respectively. T-AMY and P-AMY activities in serum or urine obtained by the proposed method correlated well with those determined by the 2-chloro-4-nitrophenyl 4-O-beta-D-galactopyranosyl-beta-maltotetraoside method or the modified IFCC method. CONCLUSIONS: : This novel assay for T-AMY and P-AMY measures both activities stoichiometrically, directly, and easily, and may be suitable for routine procedures.

A case of normotensive scleroderma renal crisis after high-dose methylprednisolone treatment.

A 68-year-old male was admitted for interstitial pneumonia associated with scleroderma. High-dose methylprednisolone was administered for treatment of the pneumonitis. Two weeks later, anemia, thrombocytopenia and progressive increase in BUN, creatinine and LDH were observed. Although the blood pressure remained normotensive, renal biopsy showed thrombosis of the polar arterioles and glomerular capillaries. The affected interlobular artery included concentric intimal thickening and thrombosis in the lumen. Our findings suggested that the antecedent use of high-dose corticosteroids is involved in precipitating normotensive renal crisis. Corticosteroids should be used in low doses and with great caution in scleroderma patients.

[New duct caliber measurement methods on magnetic resonance cholangiopancreatography].

PURPOSE: To report the feasibility of new duct caliber measurement methods using signal intensity on MR cholangiopancreatography (MRCP). MATERIALS AND METHODS: A phantom with 0.25-8 mm caliber ducts was imaged with a 1.5-Tesla MR system using the usual MRCP sequence. Each caliber was measured by conventional method (full width at half maximum, FWHM) and two new methods: the maximum intensity measurement method (MIM), in which caliber was calculated from maximum signal intensity, and the area intensity measurement method (AIM), in which caliber was calculated from the area under the signal intensity curve. Errors in calculated caliber were compared among the three methods of measurement. RESULTS: Mean measurement errors of each caliber for 1-8 mm ducts by MIM [14% (2.1-25%)] and by AIM [6.8% (0.4-16%)] were significantly lower than those by FWHM [30% (17-64%)]. AIM was significantly more accurate than MIM for caliber measurement of ducts over 1 mm. CONCLUSION: Two new caliber measurement methods for pancreatobiliary ducts in MRCP, MIM and AIM are accurate and should play an important role in clinical MRCP.

Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus.

The genome of the nucleopolyhedrovirus (NPV) (T3 strain) pathogenic for Bombyx mori (Bm) was sequenced and analysed. The BmNPV genome was 128,413 nucleotides long with a G+C content of 40% and contained 136 open reading frames (ORFs) encoding predicted proteins of over 60 amino acids. Although phenotypically different, the genome organizations of BmNPV and Autographa californica multinucleocapsid NPV (AcMNPV) were closely related. The BmNPV genome was over 90% identical to about three-quarters of the genome of AcMNPV. The relatedness of predicted amino acid sequences of corresponding ORFs between BmNPV and AcMNPV was about 90%. However, the BmNPV genome lacked homologues of the following AcMNPV ORFs: Ac3 (conotoxin), Ac7 (orf603), Ac48 (etm), Ac49 (pcna), Ac70 (hcf-1), Ac86 (pnk/pnl) and Ac134 (p94). In addition, BmNPV contained five ORFs related to Ac2. A high frequency of multiple 3 bp insertions was also found within BmNPV and AcMNPV coding sequences.

The Drosophila Bruton's tyrosine kinase (Btk) homolog is required for adult survival and male genital formation.

We isolated a Drosophila fickleP (ficP) mutant with a shortened copulatory duration and reduced adult-stage life span. The reduced copulatory duration is ascribable to incomplete fusion of the left and right halves of the apodeme that holds the penis during copulation. ficP is an intronic mutation occurring in the Btk gene, a gene which encodes two forms (type 1 and type 2) of a Bruton's tyrosine kinase (Btk) family cytoplasmic tyrosine kinase as a result of alternative exon usage. The ficP mutation prevents the formation of the type 2 isoform but leaves expression of the type 1 transcript intact. Ubiquitous overexpression of the wild-type cDNA by using a heat shock 70 promoter during the late larval or pupal stages rescued the life span and genital defects in the mutant, respectively, establishing the causal relationship between the ficP phenotypes and the Btk gene mutation. The stage specificity of the rescuing ability suggests that the Btk gene is required for the development of male genitalia and substrates required for adult survival.

Cell biological analysis with respect to cause of fibrous opacification of the anterior capsule after cataract extraction.

It has been assumed that lens epithelial cells (LECs) existing at the capsulotomy edge have been traumatized through anterior capsulotomy in cataract extraction. In this study, the correlation between traumatized LECs remaining at the anterior capsulotomy edge and epidermal growth factor (EGF) found in the aqueous humor, a cell growth factor though to affect cell morphology, was determined. Anterior lens capsules with adhering LECs were obtained following anterior capsulotomy performed during cataract surgery to first confirm the presence of EGF receptors on LECs, which are needed for EGF to be biologically active. Besides, to identify any EGF receptors on traumatized LECs, I next intentionally traumatized the cells by pressing them with a forceps from the anterior capsular side. It has been found that the LECs containing EGF receptors were always those existing at the edge of the anterior capsular opening and LECs containing EGF receptors existed along the pressed region too. The present results indicate that traumatized LECs along the capsulotomy edge have undergone changes to manifest EGF receptors, thus allowing EGF from the aqueous humor to become more active. The physiological effect of EGF upon these LECs may therefore be one of the causative factors of fibrous opacification of the anterior capsulotomy edge after cataract extraction.

Cell-biological analysis of atopic cataractous lenses.

The number of people suffering from atopic dermatitis, a recent social problem believed to have arisen from environmental pollution and changes, continues to increase today, and these patients often encounter complications such as cataract and retinal detachment. In this study, I have conducted (1) a comparative study on the rate of cell proliferation between lens epithelial cells (LECs) obtained from 7 atopic cataractous lenses and from 1 normal lens, (2) a comparative study on cell density and alignment between LECs obtained from 5 nonatopic cataractous lenses and from 5 atopic cataractous lenses and (3) transmission electron microscopy of LECs obtained from 3 atopic cataractous lenses. My findings were as follows: (1) except for 1 case disclosing increased proliferative activity of the cells to become multilayered, LECs of atopic cataractous lenses showed diminished proliferative activity; (2) LECs of atopic cataractous lenses had decreased in cell density and revealed irregular cell alignment; (3) transmission electron microscopy of LECs of atopic cataractous lenses demonstrated multilayered cells, increased intercellular spaces, and degeneration and disappearance of some cells. A longer follow-up period and further studies using cells from additional atopic cataractous lenses are necessary before any conclusions can be drawn. However, obtaining human LECs especially of atopic cataract patients is not easy, and I do feel that my present study, although its number of patients may not be large enough, provides significant findings for further studies on the mechanism of atopic cataract formation.

Effect of epidermal growth factor upon morphological changes of human lens epithelial cells.

Phase-contrast and transmission electron microscopic studies were performed to evaluate the effect of epidermal growth factor (EGF) upon the morphology of lens epithelial cells (LECs) obtained from human cataractous lenses, cultured on human anterior lens capsules, and divided into the following three groups: group 1 receiving no EGF supplement, group 2 supplemented with 1 ng/ml EGF, and group 3 supplemented with 10 ng/ml EGE Phase-contrast microscopy revealed that the cells in the group supplemented with 1 ng/ml EGF concentration had elongated, and those supplemented with 10 ng/ml EGF disclosed changes indicating fibroblast-like cells. Under transmission electron microscopy, the cells in the group supplemented with 1 ng/ml EGF concentration had become multilayered and a few cells had a ball-and-socket junction structure and nucleolar chromatin condensation, while those supplemented with 10 ng/ml EGF showed changes not only in the cells but also in the anterior lens capsules to present numerous microfibers. This is the first study to confirm the effect of EGF upon LECs cultured on anterior lens capsules, a condition closely resembling the intraocular environment, and these findings are significant.

Determination of alpha-amylase using 4-O-beta-D-galactopyranosylmaltotetraose (Gal-G4) as a substrate.

A new substrate, 4-O-beta-D-galactopyranosylmaltotetraose (Gal-G4) is applied for the determination of alpha-amylase in serum and urine in a coupled assay with alpha-glucosidase (EC 3.2.1.20), glucokinase (EC 2.7.1.2) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) as auxiliary enzymes. Gal-G4 having a 4-position of the non-reducing-end glucose residue modified by a beta-galactopyranose group is resistant for degradation by alpha-glucosidase as auxiliary enzyme. Moreover, this substrate is hydrolyzed at just one position by alpha-amylase in serum and urine. More than 99% of the products generated from Gal-G4 by alpha-amylase are identified 4-O-beta-D-galactopyranosylmaltose (Gal-G2), maltose, respectively. Glucose and maltose do not interfere the value of alpha-amylase activity at least up to 0.056 mmol/l (1 g/dl) glucose and 0.027 mmol/l (1 g/dl) maltose, respectively. We are now carrying out this work under the authority of The Enzyme committee of Japanese Society of Clinical Chemistry (JSCC) as a standard method for determination of alpha-amylase in clinical chemistry.

The relationship between morphological changes of lens epithelial cells and intraocular lens optic material.

To examine the morphological changes of lens epithelial cells (LECs) occurring directly beneath and at regions contacting various intraocular lens (IOL) optic materials, human LECs were cultured on human anterior lens capsules and were further incubated upon placing above the cells lens optics made of polymethylmethacrylate, silicone, and soft acrylic material. Observations as to the morphological changes of LECs under phase-contrast microscope and scanning electron microscope were performed on the 14th day of incubation. Gatherings of LECs were observed at regions contacting the soft acrylic material under phase-contrast microscope, and gatherings of LECs were observed accurately at the same regions mentioned above under scanning electron microscope. On the other hand, LECs in contact with two other optic materials did not show morphological changes. The results suggest that LECs attached to and proliferated on not only the anterior lens capsules but also the soft acrylic IOL optics. The model used in this study may be useful in studying the relationship between cellular movement of LECs and IOL optic material.

Versatility of the free anterolateral thigh flap for reconstruction of head and neck defects.

OBJECTIVE: The anterolateral thigh flap has many advantages in head and neck reconstruction. However, it has not yet come into widespread use because of the anatomic variations of its perforators. Herein, we describe a safe operative technique related to the patterns of the perforators and discuss its wide versatility. SETTING: A national cancer center hospital. PATIENTS: Thirty-eight anterolateral thigh flaps were transferred. Confirmation and dissection of the flap pedicle were simultaneously performed with tumor resection. The design and elevation of the flap were carried out immediately after the tumor resection was completed. RESULTS: From the study of the anatomic variations of the perforators, septocutaneous patterns were recognized in 10 cases (26.3%) and musculocutaneous patterns in 28 cases (73.7%). All flaps were easily and safely elevated with our techniques. Thirty-six flaps survived. Partial necrosis was noted owing to excessive thinning procedure in one patient and total necrosis was noted owing to venous thrombosis at the anastomosis part in another patient. CONCLUSIONS: We found that the anterolateral thigh flap has numerous advantages. It is possible to perform the flap elevation and the tumor resection simultaneously. The flap is generally thin and is suitable for reconstruction of intraoral defects. Combined flaps with neighboring tissues and other, distant flaps can be used. Furthermore, since our technique minimizes the problems of confirmation and dissection of the perforators, we conclude that this flap can be successfully used to repair a variety of large defects of the head and neck.

Deletion analysis of four of eighteen late gene expression factor gene homologues of the baculovirus, BmNPV.

Nucleotide sequence analysis of the genome of the baculovirus Bombyx mori nuclear polyhedrosis virus (BmNPV) identified 18 homologues of the Autographa californica NPV (AcNPV) lefs (late expression factor genes). These BmNPV lefs showed high (73-98%) amino acid sequence identities to AcNPV lefs and were localized to similar positions in the genome. One lef, p35, was previously characterized in AcNPV and BmNPV deletion experiments. Functional deletion of each of the BmNPV lef homologues was attempted here by insertion of a beta-galactosidase gene cassette into the coding region of each lef. Four of 18 BmNPV lef (39K, ie-2, lef-7, and p35) deletion mutants were successfully isolated, indicating that the other 14 BmNPV lefs were likely essential for viral replication in cell culture. Further analysis showed that deletion of lef-7, p35, and ie-2 resulted in lower levels of viral DNA replication, indicating that the BmNPV lef-7, p35, and ie-2 products have stimulating effects on DNA replication. Deletion of 39K resulted in a significantly lower level of late gene transcription and extremely low (over 10(2)-fold less at 48-80 hr p.i.) production of progeny budded virus in BmN cells. In contrast, the deletion did not affect viral DNA replication, indicating that BmNPV 39K is involved in late gene transcription. Reduced late gene expression presumably affected production and/or release of progeny budded virus particles. This was corroborated by transmission electron microscopy, which showed that virus replication was abnormal in BmN cells infected with a BmNPV mutant lacking 39K and virion production was low. Even though 39K deletion resulted in a loss of oral infectivity, the 39K deletion mutant replicated in silkworm larvae when injected into the body cavity, as did the ie-2, lef7, and p35 deletion mutants. In addition, a BmNPV homologue of the baculovirus very late expression factor gene (vif-1) found in AcNPV was essential, implying an essential function of the BmNPV vif-1 homologue at a step before the onset of very late gene expression.

Presence of growth factor in human vitreous.

The aqueous humor and the vitreous not only maintain integrity of the eye but also play a vital role in providing nutrients to ocular tissues. While the presence of epidermal growth factor, basic fibroblast growth factor, and transforming growth factor-beta (TGF-beta) have been confirmed in human aqueous humor, only TGF-beta has been found to exist in human vitreous. In this study I obtained eyes for keratoplasty from 20 male and 20 female donors, or a total of 40 subjects; upon removing the corneal button. I extracted 500 microliters of the vitreous humor in order to confirm, using the ELISA method, the presence of transforming growth factor-alpha (TGF-alpha) and epidermal growth factor (EGF) in the vitreous. TGF-alpha ranging from 78 to 250 pg/ml, or an average of 130.65 +/- 40.89 pg/ml, was found in all eyes. On the other hand, EGF was not confirmed in all eyes. This is the first study to confirm the presence of TGF-alpha in the human vitreous.

Lens epithelial cells in postoperative aqueous humor.

The aqueous humor during or after cataract surgery often contains red blood cells (RBCs) from blood; inflammatory cells from white blood cells (WBCs); fibrin, pigment cells, and fibroblasts from either the iris or the ciliary body; lens epithelial cells (LECs); and corneal endothelial cells. Since LECs may be related to postoperative complications, including a fibrin reaction and a secondary cataract, confirmation of their presence postoperatively not only in the capsular bag, but also in the aqueous humor, is necessary for identifying these complications. Using aqueous humor obtained from pig eyes, I attempted to identify LECs immunohistochemically in the aqueous humor, following phacoemulsification (PEA) and aspiration of the cortex, and found the presence of cells that react to keratin antibody, the marker for epithelial cells. Although these cells could be lens, conjunctival, or corneal epithelial cells, they were found following PEA and aspiration of the cortex, and therefore are believed to be LECs.