Ring and unimodal angular-frequency distribution of THz emission from two-color femtosecond plasma spark.

We study angular and frequency-angular distributions of the terahertz (THz) emission of the low-frequency region (0.3-3 THz) from a two-color femtosecond plasma spark experimentally and in three-dimensional numerical simulations. We investigate the dependence of the angular shapes of the THz radiation on focusing conditions and pulse durations by using two laser facilities (pulse durations 35 and 150 fs) for different focusing geometries. Our experiments and simulations show that decrease in the numerical aperture from NA ≈0.2 to NA ≈0.02 results simultaneously in (I) squeezing of the THz angular distribution and (II) formation of the bright conical emission in the THz range. The moderate focusing NA ≈0.05, which forms the relatively narrow unimodal THz angular distribution, is identified as optimal in terms of angular divergence. Numerical simulations with carrier wave resolved show that bright THz ring structures appear at the frequencies ≥2 THz for longer focuses (NA ≈0.02), while for optimal focusing conditions NA ≈0.05 the conical emission develops at THz frequencies higher than 10 THz.

Distinct functional regions of the human polymeric immunoglobulin receptor.

The polymeric immunoglobulin receptor (pIgR) is a type I transmembrane protein that is expressed on the surfaces of glandular and intestinal epithelial cells. The extracellular portion of the pIgR is composed of six different domains. Domain 6 is involved in the enzymatic cleavage and release of the pIgR into the intestinal lumen as a free secretory component (fSC). A highly conserved 9-amino acid sequence is present in this region in various species. Although mutations in domain 6 are associated with particular diseases, such as IgA nephropathy and Epstein-Barr virus-related nasopharyngeal cancer, and the glutamic acid residues in the conserved 9-amino acid sequence are expected to be indispensable for the secretion of fSC, the importance of these residues has not been examined. In the present study, we attempted to examine the role of these residues in the enzymatic cleavage of the pIgR. The enzymatic cleavage of the pIgR was not affected by the presence of an alanine to valine substitution at position 580 or glutamine to alanine substitutions at positions 606 and/or 607, or the deletion of the whole 9-amino acid conserved sequence. Intriguingly, the 10 amino acid sequences flanking the N- and C-terminal ends of the conserved 9-amino acid sequence had opposite effects on pIgR cleavage. Namely, the N-terminal and C-terminal sequences enhanced and reduced pIgR cleavage efficiency, respectively. These results indicated that the pIgR can be divided into several functionally distinct regions.

Urinary excretion of the main metabolites of 3,4-methylenedioxymethamphetamine (MDMA), including the sulfate and glucuronide of 4-hydroxy-3-methoxymethamphetamine (HMMA), in humans and rats.

The urinary concentrations of the main metabolites of 3,4-methylenedioxymethamphetamine (MDMA; Ecstasy), specifically 4-hydroxy-3-methoxymethamphetamine sulfate (HMMA-Sul) and 4-hydroxy-3-methoxymethamphetamine glucuronide (HMMA-Glu), have been directly measured in both MDMA users and rats by an established liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) procedure. The concentrations of these conjugates in urine from MDMA users (n = 25) ranged from 6.5 to 202 microM (from 1.8 to 55.6 microg ml(-1)) for HMMA-Sul and from 1.3 to 87.0 microM (from 0.5 to 32.3 microg ml(-1)) for HMMA-Glu, and the ratio of HMMA-Sul to HMMA-Glu ranged from 1.6 to 9.9 (3.1 +/- 1.8). These results demonstrate that the sulfation is quantitatively more significant than the glucuronidation for HMMA in humans. In rats, in contrast, almost all the conjugated HMMA (>99%) was excreted as the glucuronide. These findings indicate that hydrolysis should be carefully made in urine analysis by gas chromatography (GC) or gas chromatography-mass spectrometry (GC-MS) by using either an acid or an enzyme possessing both sulfatase and beta-glucuronidase activities. It is concluded that a considerable interspecies variation exists in the conjugation of HMMA between humans and rats.

Coexpression of an unusual form of the EWS-WT1 fusion transcript and interleukin 2/15 receptor betamRNA in a desmoplastic small round cell tumour.

BACKGROUND: The beta chain of the interleukin 2/15 receptor (IL-2/15Rbeta) is induced by the expression of the EWS-WT1. A case of desmoplastic small round cell tumour (DSRCT) expressing only an unusual EWS-WT1 treated by us is reported here. AIM: To characterise an unusual form of EWS-WT1. METHODS: Frozen tissue sections of the axillary tumour were examined using a laser-assisted microdissection technique and reverse transcriptase polymerase chain reaction. RESULTS: The novel fusion of exon 8 of EWS and the defective exon 10 of WT1 (-KTS) was detected. Although it was an unusual form, the coexpression of the present EWS-WT1, IL-2/15Rbeta and Janus kinase (JAK1) mRNA was detected in the tumour cells. IL-2 and signal transducers and activators of transcription (STAT5) mRNA were detected in both tumour and stromal cells. CONCLUSION: The induction of the IL-2/15 receptor signalling pathway may contribute to tumorigenesis in DSRCT through a paracrine or an autocrine system, even though the EWS-WT1 was an unusual form.

Overexpression of anti-apoptotic Mcl-1 in testicular germ cell tumours.

AIMS: To determine the expression of Mcl-1 in testicular germ cell tumours in order to clarify the role of this anti-apoptotic factor in these tumours. Various members of the Bcl-2 family have been implicated in the apoptotic mechanisms regulating germ cell apoptosis. Mcl-1 is an anti-apoptotic Bcl-2 family member and has recently been reported to be related to the progression of malignancy; however, the involvement of Mcl-1 in the development of germ cell tumours is still unknown. METHODS AND RESULTS: Mcl-1 expression in testicular germ cell tumours was investigated by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR). By immunohistochemistry, overexpression of Mcl-1 was present in all germ cell tumours that were studied, including embryonal carcinoma and yolk sac tumour, as well as choriocarcinoma and teratoma. In teratomas, Mcl-1 was widely distributed in the epithelial, myogenic, neural and mesenchymal components. RT-PCR analysis after microdissection revealed high levels of Mcl-1 mRNA in all tumour variants compared with non-neoplastic germ cells. CONCLUSION: Overexpression of anti-apoptotic Mcl-1 may function to enhance the viability of testicular germ cells, thereby leading to tumorigenesis.

Huge IgD plasmacytoma in the abdomen presenting coagulation necrosis.

A 65-year-old Japanese woman was diagnosed in 1996 with a pathological fracture of the left femur caused by immunoglobulin D-type myeloma (IgD myeloma). She responded well to combination chemotherapy followed by irradiation. The patient experienced renal failure and became dependent on haemodialysis. In 1999, large plasmacytomas developed in the abdomen and left humerus. The abdominal tumour appeared to induce gastroduodenal ulcers and jejunal obstruction. We initiated irradiation therapy without chemotherapy to prevent further growth of the plasmacytoma, although treatment-resistant gastroduodenal ulcers developed. Continued blood loss from the gastroduodenal ulcers resulted in a deterioration in the patient's health, which prevented successful haemodialysis. An autopsy showed that the plasmacytoma had undergone coagulation necrosis. We conclude that the use of combination chemotherapy with topical irradiation was an acceptable treatment measure against IgD plasmacytoma; irradiation without chemotherapy was the most likely cause of the coagulation necrosis seen in the plasmacytoma at autopsy.

Application of compact ultrasound imaging device to postmortem diagnosis.

In regions with low autopsy rates, forensic examiners often have to rely on external findings. Imaging techniques can assist the external examination and provide a more objective diagnosis. The SonoSite 180, a portable ultrasound device, was used for the examination of dead bodies. The influence of different degrees of decomposition was estimated. Even in cases with intestinal gas formation images of internal organs could be obtained with special techniques. Various pathological findings were detected by ultrasound and verified by autopsy (e.g. pericardial tamponade, cardiac hypertrophy, fatty liver, aortic aneurysm, metastatic liver, etc.). The experiences with the SonoSite 180 are promising. The device can be carried to the death scene or to the morgue and therefore serve as a valuable tool for medicolegal applications.

Histologic criteria for the diagnosis of allied diseases of Hirschsprung's disease in adults.

A few reports in the literature have discussed the histologic criteria for the diagnosis of allied diseases of Hirschsprung's disease in adults, and studies report that intestinal neuronal dysplasia (IND) in adults may develop from IND in infants. The aim of this study was to examine the differences between the histological findings of IND in infants and those in adults, and to assess whether allied diseases of Hirschsprung's disease (HD) in adults should be considered as congenital or acquired diseases. For these purposes, we studied nine adult patients with severe constipation, and an adult patient with acute intestinal obstruction. We routinely examined the patients using barium enema, anorectal manometry and rectal mucosal biopsy. However, in patients suspected of allied diseases, we carried out full-thickness rectal biopsies. In seven operated cases, we also examined resected intestines. The tissue samples were examined using AChE-staining, NADPH-diaphorase staining, HE-staining, and silver impregnation. Histologically, we diagnosed two males as having HD, two males as having IND, five patients (two males and three females) as having hypoganglionosis, and one female as having a degeneration of the intramural plexus. The following conclusions were drawn: 1) Inflammations such as ulcerative colitis or ischemic colitis may cause IND TYPE B in adults whose histological findings are similar to those generally seen in infants; 2) It is suggested that IND is closely related to hypoganglionosis; 3) In hypoganglionosis, a patient with findings of elevated AChE-positive nerve fibers in the mucosa and AChE-positive nerve fibers in an arterial wall may belong to a subtype of IND; 4) Most of the allied diseases of HD in adults may occur as an acquired disease, not as a congenital disease.

Differential expression of secreted frizzled-related protein 4 in decidual cells during pregnancy.

During pregnancy, the uterus shows marked morphological and physiological changes under the regulation of ovarian steroid. To elucidate the molecular cues of these changes, we tried to identify the transcripts differentially expressed in the pregnant rat uterus by using the suppression subtractive hybridization method. Seven independent clones were isolated and one of the up-regulated genes was secreted frizzled-related protein 4 (sFRP4). sFRP4 contains a Wnt-binding domain and belongs to the secreted frizzled protein family whose members are assumed to function as modulators of the Wnt signal. The expression level of sFRP4 mRNA reached a peak in the pregnant uterus on day 12, when uterine decidualization was almost complete in the rat. In situ hybridization histochemistry revealed that sFRP4 transcripts were observed in the decidual cells. In addition, proliferating cell nuclear antigen (PCNA)-positive cells were shown to be overlapped in decidua, suggesting that sFRP4 mRNA expression was accompanied by the late phase of decidual cell proliferation. Moreover, sFRP4 and estrogen receptor-alpha transcripts were co-localized. Furthermore, we analyzed the regulation of sFRP4 by estrogen using 17 beta-estradiol-treated ovariectomized rats. sFRP4 mRNA was detected in the uterus at 48 h after estrogen treatment, especially in endometrial stroma where PCNA-positive cells were also observed. The results in this study led us to the notion that sFRP4 mRNA may be up-regulated after estrogen treatment in the late phase of uterine cell proliferation.

A highly potent 26,27-Hexafluoro-1a,25-dihydroxyvitamin D3 on calcification in SV40-transformed human fetal osteoblastic cells.

26,27-hexafluoro-1a,25-dihydroxyvitamin D3 (F6-D3) has been reported to be 5-10 times more potent than 1a,25-dihydroxyvitamin D3[1,25(OH)2D3] in biological systems in vivo and in vitro. However, the effect of F6-D3 on bone formation has yet to be clarified. In the present study, we investigated the effect of F6-D3 on SV40-transfected human fetal osteoblastic cells (SV-HFO) and found it to be about 100 times greater than that of 1,25(OH)2D3 in stimulating calcification. F6-D3 was also about 100 times more effective than 1,25(OH)2D3 in enhancing the expression of mRNA for alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). In the presence of 10?8 M F6-D3 and 10?6 M 1,25(OH)2D3, the calcification began on day 9 and increased up to day 19. Expression of mRNA for ALP and OCN reached a maximum on day 4 and thereafter declined. On the other hand, when osteoblastic cells were incubated with a low level of [1b-3H]-F6-D3- or [1b-3H]-1,25(OH)2D3, each radioactive peak could not be detected. However, on the incubation of osteoblastic cells and radioactive substrate in the presence of ketoconazole, a selective inhibitor of CYP24, a clear peak for each substrate was detected. This suggested that F6-D3 as well as 1,25(OH)2D3 is metabolized by CYP24. Osteoblastic cells were incubated with 10?8 M[1b-3H]-F6-D3 or 10?8 M[1b-3H]-1,25(OH)2D3 for 4, 9, and 14 days. A small peak of 1,25(OH)2D3 was observed and thereafter its level decreased. In addition, two unknown peaks increased when the culture period was extended. In the case of F6-D3, peaks of F6-D3 and 26,27-hexafluoro-23-oxo-1a,25(OH)2D3(23-oxo-F6) were clearly detected, the latter being about 4 times higher than the former. Both peaks was retained up to day 14. The amount of unlabeled F6-D3 and 23-oxo-F6 calculated from the specific radioactivity in the cells may be similar to the amount of 1,25(OH)2D3 and its metabolites. The strong activity of F6-D3 in stimulating calcification may be due to the fact that F6-D3 is much more potent than 1,25(OH)2D3 in enhancing the expression of mRNA for ALP, OCN, and OPN and that the amount of F6-D3 and 23-oxo-F6 accumulated in the cells is much greater than that of 1,25(OH)2D3 and its metabolite.

Sol-Gel transition behavior of pure iota-carrageenan in both salt-free and added salt states.

This paper describes how strongly the gelation process of iota-carrageenan is affected by addition of metallic ions from the creep and creep recovery, dynamic viscoelasticity (DVE) and DSC measurements. Creep results at T = 25 degrees C indicate that below a polymer concentration C of 3.0 wt % the salt-free system behaves as a viscous solution, and it starts to exhibit viscoelasticity as C exceeds 3.0 wt %. In the range C = 5.0-7.0 wt %, the salt-free system shows gellike behavior whereas the added salt system, measured in the low C range 1.0-2.5 wt %, showed gellike behavior at the same temperature. The sol-gel transition temperature T(c) was determined using Winter's criterion as the temperature at which both G'(omega) and G' '(omega) follow power law behavior with the same exponent n. DSC measurements reveal that salt-free and added salt systems take different types of thermal behavior within the same temperature range. The temperature T(c) is quite close to the gelation temperature T(m) determined from DSC measurement. The Eldrige-Ferry plot was performed to estimate activaton enthalpy, which shows that physical cross-links in the salt-free iota-carrageenan is not strong in comparison with those of samples which contains metal ions. We conclude from the data analysis of C dependence of the plateau modulus using the theory developed by Jones and Marques for rigid networks based on the fractal theories that addition of metallic ions gives rise to a rigid fiber like structure even at low C of iota-carrageenan in contrast to the salt-free system for which a flexible structure has been maintained at higher C.

An in vitro DNA virus for in vitro protein evolution.

In vitro virus is a molecular construct for in vitro protein evolution, which requires some mechanism to link phenotype to genotype. The first in vitro virus was realized by bonding a nascent protein with its coding mRNA via puromycin in in vitro translation. We report a new construct of in vitro DNA virus. The virion was a covalent cDNA-protein fusion, and virion formation did not require any modification of mRNA. Due to intactness of mRNA, this type of in vitro DNA virus will take the next step toward in vitro autonomous evolution, just like in vivo viral evolution in a cellstat.

Rapid functional analysis of protein-protein interactions by fluorescent C-terminal labeling and single-molecule imaging.

Detection of protein-protein interactions is a fundamental step to understanding gene function. Here we report a sensitive and rapid method for assaying protein-protein interactions at the single-molecule level. Protein molecules were synthesized in a cell-free translation system in the presence of Cy5-puro, a fluorescent puromycin, using mRNA without a stop codon. The interaction of proteins thus prepared was visualized using a single-molecule imaging technique. As a demonstration of this method, a motor protein, kinesin, was labeled with Cy5-puro at an efficiency of about 90%, and the processive movement of kinesin along microtubules was observed by using total internal reflection microscopy. It took only 2 h from the synthesis of proteins to the functional analysis. This method is applicable to the functional analysis of various kinds of proteins.

Phagocytized apoptotic cells in subcutaneous panniculitis-like T-cell lymphoma.

We reported on a case of subcutaneous panniculitis-like T-cell lymphoma (SPTCL) with multiple erythematous nodular lesions on the extremities, trunk and face. Histological examination of an excised lesion revealed a dense infiltrate of markedly atypical T-lymphoid cells expressing the CD8+ phenotype located in the subcutaneous tissue with histiocyte-phagocytizing apoptotic cells. The 'bean-bag' histiocytic cells, the characteristic finding of SPTCL, are considered to be products of haemophagocytosis. In our case the 'bean-bag' cells were produced by phagocytosis of apoptotic bodies, as confirmed by electron microscopy. It is suspected that 'bean-bag' cells are related not to haemophagocytosis but to phagocytosis of apoptotic cells in the CD8+ T-cell type of SPTCL.

[Immunohistochemical analysis with anti-cytokeratin antibody in the prostatic epithelium].

PURPOSE: We performed immunohistochemical studies of the prostatic epithelium using three different anti-cytokeratin monoclonal antibodies (35 beta-H11, RCK108, and 34 beta-E12), and also investigated the immunoreactivity of various prostatic lesions with basal cell specific anti-cytokeratin antibody (34 beta-E12). MATERIALS AND METHODS: One hundred and thirty one prostatic specimens were obtained at surgery or biopsy. H-E stained sections were available for review in all cases. They were classified according to histopathology; benign prostatic hyperplasia (BPH), prostatic cancer (PCA), atrophic acini, atypical adenomatous hyperplasia (AAH), and prostatic intraepithelial neoplasia (PIN). ABC or LSAB method was utilized for immunohistochemical staining with 3 anti-cytokeratin monoclonal antibodies. RESULTS: 35 beta-H11 was mainly stained in the luminal cells and RCK108 was stained both in the luminal and the basal cells in BPH. 35 beta-H11 showed highly positive staining in the prostatic cancer regardless of degree of differentiation. RCK108 tended to be less stained in the prostatic cancer cells with lower grades of tumor differentiation compared to those with higher grades. 34 beta-E12 was stained only in the basal cells, but neither in the normal luminal cells nor the cancer cells. Using 34 beta-E 12, basal cells were positively stained in most of the cases with BPH, while not in PCA. Atrophic acini and AAH was stained with 34 beta-E12 as positively as BPH. Basal cells were discontinuously or negatively stained in many cases with high-grade PIN. CONCLUSIONS: The luminal cells in BPH were highly positively stained using 35 beta-H11 or RCK108. RCK108 tended to be less stained in the prostatic cancer cells with lower grades of tumor differentiation. Positive staining of 34 beta-E12 strongly suggested a benign lesion, therefore immunohistochemistry using this antibody would be useful as an aid for pathological diagnosis.

Different expression of hepatic and renal cytochrome P450s between the streptozotocin-induced diabetic mouse and rat.

1. Since limited information is available about alterations of cytochrome P450 levels in diabetic animals other than rat, expression of P450s in the liver and kidney of the streptozotocin (STZ)-induced diabetic mouse was investigated. 2. The mRNA levels of CYP2B10, 3A11, 4A10 and 4A14 in the liver were increased in the STZ-induced diabetic mouse of both sexes. The CYP2B9 mRNA level was increased in the liver of the male diabetic mouse. These alterations were observed even at 2 weeks after administration. Insulin treatment restored these changes. The findings were consistent with changes reported in rat. 3. The levels of hepatic CYP1A2 and 2E1 and renal 2E1 and 4A did not change in the diabetic mouse at any time-point examined. No changes were seen in CYP2A- or 2C-related proteins in the diabetic mouse. These findings were in contrast to those in rat. 4. The results indicate that mouse P450s respond to insulin-dependent diabetes mellitus differently from those of the rat, and suggest that the expression of P450s in diabetes is not generally the same across animal species.

[Visual evoked potentials elicited by pseudorandom stimulation in macular degeneration].

PURPOSE: To investigate the effect of a central scotoma on the amplitude, latency, and temporal frequency characteristics(TFCs) of the visual evoked potentials(VEPs) elicited by a pseudorandom binary stimulus(PRBS). METHOD: Patients with age-related macular degeneration(AMD) were selected, and VEPs were recorded from 26 eyes with AMD(17 eyes with visual acuity of less than 0.2, and 9 eyes with visual acuity between 0.3 and 0.9). Nine eyes of age-matched normal volunteers served as controls. To acquire the PRBS-VEPs, one eye was stimulated with a PRBS stimulus. The first order kernel was calculated from a cross correlation between PRBS and VEPs. The Fourier transformed first-order kernel was used as the TFC of the VEPs. RESULTS: The P2 latency of the first order kernels was delayed(p < 0.05), and the P2-N2 amplitude was reduced(p < 0.01) in AMD. A depression of the TFC values in the 6-18 Hz band was prominent in the patients with AMD(p < 0.01). CONCLUSION: The TFC, were strongly correlated with the visual acuity of patients with macular degeneration.

Sex-associated expression of mouse hepatic and renal CYP2B enzymes by glucocorticoid hormones.

The expression of Cyp2b9 and Cyp2b10 genes was investigated in kidney, liver, and cultured hepatocytes of adult C57BL/6NCrj mice. The constitutive expression level of CYP2B mRNA in kidney was higher in female than in male mice, as it was in the liver where more CYP2B9 than CYP2B10 was expressed in the females, and more CYP2B10 was expressed in the males. After treatment with dexamethasone (Dex), induction of CYP2B10 mRNA and protein in the kidneys was far greater in male than in female mice. In contrast to Dex, phenobarbital (PB), pregnenolone-16 alpha-carbonitrile (PCN), and 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) did not induce the expression of the Cyp2b gene in the kidneys of either sex. In the liver, PB, PCN, and DDT induced both CYP2B9 and CYP2B10 in both sexes to the same extent, whereas Dex induced only CYP2B10 and simultaneously suppressed CYP2B9. Dex-inducible expression of CYP2B mRNA was decreased by 11 beta-[4-dimethylamino]phenyl-17 beta-hydroxy-17-[1-propynyl]estra-4,9-dien-3-one (RU-486), in both the kidneys and liver from male mice, and in cultured hepatocytes. However, RU-486 itself induced the expression of CYP2B mRNA in female liver and cultured hepatocytes. Interestingly, RU-486 increased PB-inducible expression of these species in cultured hepatocytes. Gonadectomy increased the expression of CYP2B mRNA in untreated male liver, but suppressed Dex-induced expression in the kidneys of both sexes. These observations suggest that (a) there are multiple regulatory pathways in the expression of Cyp2b genes, one of which used by Dex is mediated via the glucocorticoid receptor, which is different from that used by PB, and (b) sex hormones play a role in the regulation of the sex-dependent expression of Cyp2b genes in the mouse.

Relationship between skeinoid fibers and stromal matrix in gastrointestinal stromal tumors: morphometric analysis with quick-freezing and deep-etching method.

Our previous study of a gastrointestinal autonomic nerve tumor with skeinoid fibers (SF) using the quick-freezing and deep-etching method, suggested that the distance between one radix and a neighboring radix (DRNR) in pre-existing meshwork structures around the tumor cells is consistent with the periodicity of the SF. Therefore, measurement of the DRNR in the meshwork could clarify the significance of the pericellular matrix for SF development. In the present study, we analyzed the meshwork in three cases of gastrointestinal stromal tumor (GIST), which showed different immunohistochemical stainings, but confirmed to have smooth muscle differentiation (SMD) by immunohistochemistry and/or electron microscopy. The DRNR from the three cases of GIST showed similar histogram patterns (a peak of 20-30 nm, mean values of 28.02, 25.74 and 26.45 nm), which were significantly shorter than the periodicity of SF (a peak of 40-45 nm, mean value of 42.14). Although we need further studies with additional GIST cases, we speculate that the pericellular matrix of GIST with SMD is not suitable for SF development.