[Application of a multi-material artifact reduction algorithm in a wide-detector CT in the evaluation of the portal venous angiography of postoperative TIPS and embolization].

Objective: To assess the effect of monochromatic images and metal artifact reduction (MAR) on the image quality of spectral CT portal venous angiography in patients with operation of after the performing transjugular intrahepatic portosystemic stent shunt(TIPS) and embolization. Methods: From December 2017 to April 2018, the examination data of 28 patients with portal hypertension due to cirrhosis who underwent portal vein angiography 1 month after TIPS and embolization were prospectively collected. After spectral CT scanning in revolution CT, the monochromatic energy levels(60 keV, 120 keV), 60 keV + 120 keV, 120kV-like + 120 keV fused images combined with MAR algorithm were reconstructed. Quantitative parameters such as image artifact index (AI) and qualitative visual evaluation scores were recorded and compared. Results: The 120 keV monochromatic images showed the lowest AI value(30.8±8.5, 18.2±4.3) and highest metal artifacts reduction effect. The 60 keV monochromatic images showed the highest AI value (57.3±15.7, 32.1±7.9) and the lowest metal artifacts reduction effect. The AI value of 60 keV + 120 keV fused images was lower than that of 60 keV images(26.2%, 24.7%). The difference of AI value between each group was statistically significant(all P<0.05). The interobserver agreement in the subjective image scores was moderate with kappa value of 0.824. The overall image quality score of 60 keV + 120 keV fused image and the noise score of 120 kV-like+120 keV were higher than the remaining groups. The differences of the subjective scores among each group were statistically significant(all P<0.05). Conclusion: The spectral CT with MAR algorithm can effectively improve the image quality of portal vein angiography after the TIPS and embolization therapy and the 60 keV + 120 keV fused images can eliminate artifacts and ensure a clear display of blood vessels.

[Application of adaptive statistical iterative reconstruction V (ASIR-V) in contrast-enhanced abdominal scanning with low-dose for liver cirrhosis].

Objective: To investigate the application of adaptive statistical iterative reconstruction-V (ASIR-V) in contrast-enhanced abdominal CT scanning with low-dose for liver cirrhosis. Methods: From June 2017 to May 2018, a total of 112 liver cirrhosis patients who underwent contrast-enhanced abdominal CT scanning were collected prospectively as the study group. According to Child-Pugh grading of liver function, the study group was divided into A, B and C groups. Thirty examiners with normal liver function who underwent contrast-enhanced abdominal CT scanning no abdominal diseases were collected as the control group. The control group applied 0 pre-ASIR-V. The study group applied 30%, 40% and 50% pre-ASIR-V in plain scanning, arterial and portal vein phase, respectively. The control and study group both combined with 60% post-ASIR-V. The difference of qualitative parameters (contrast to noise ratio of the liver, pancreas, spleen, abdominal aorta and portal vein), image noise and radiation dose were compared by One-way ANOVA. Subjective score of images were compared by Kruskal-Wallis H test. Results: The volume computed tomography dose index (CTDI(vol)), dose length product (DLP) and the effective dose (ED) of study group were lower than the control group in the same phase (F=13.354-28.192, P<0.01). And the ED were decreased by 1.12 (23.9%), 1.54 (33.5%), 2.14 mSv (46.7%). The CNR values of liver and portal vein in Child C group was 0.34-0.42 and 0.43-0.49 lower than that of Child A and control group, respectively (q=2.851-4.658, P<0.05). Image noise had no difference in study and control group. There were no statistical differences between each group of subjective score in arterial and portal vein phase. The mean score of Child C in portal phase was lower than 3, which affected the diagnosis. And there were significant difference among the control, Child A and Child C group(Z=26.734-29.218, P<0.05). Conclusions: According to the classification of liver function, liver cirrhosis combined with ASIR-V can ensure the image quality while reduce the radiation dose. When liver function is Child-Pugh A or B, preset 50%ASIR-V is recommended; 40%ASIR-V is recommended for Child-Pugh C.

[Application of preset adaptive statistical iterative reconstruction-V in dual-enhanced abdominal CT].

Objective: To analyze the effect of preset adaptive statistical iterative reconstruction-V (ASIR-V) on image quality and radiation dose in dual-enhanced abdominal CT and to investigate the optimal ASIR-V in clinic use. Methods: From February 13 to April 30 in 2017, one hundred and eighty patients who received up abdominal CT scan in the First Affiliated Hospital of Zhengzhou University were collected prospectively. All patients underwent arterial phase (AP) and portal venous phase (PVP) enhanced abdominal CT(120 kVp, noise index 10) and were randomly divided into 6 groups according to random number table (A-F, 30 cases in each group). In group A-F, 0-50% preset ASIR-V (an interval of 10%) was applied, respectively. Qualitative parameters (subjective image quality, diagnosis confidence and radiation dose) and quantitative parameters[image noise, CT number and contrast to noise ratio (CNR)]were measured and compared among the groups by using one-way analysis of variance or Kruskal-Wallis H test. Results: The CT dose index volume (CTDIvol) decreased with the increasing of preset ASIR-V. The effective radiation dose (ED) was significant different among groups (F=27.598, P<0.05), and the ED of group B-F dropped by 10.8%, 21.7%, 31.2%, 44.9% and 61.9% respectively when compared with that in group A. Group E showed the optimal image quality (Z=18.675, 27.548, 19.761, all P<0.05) and diagnosis confidence(Z=21.387, 17.693, 22.459, all P<0.05) in plain scan, AP and PVP phases. There was no significant differences in image noise and CT value of liver, pancreas and muscle among groups (F=1.468, 0.337, 0.592, 0.284, all P>0.05). There were significant differences in CNRs in liver and portal vein in PVP phase among the groups (F=3.980, 4.681, both P<0.05). Conclusion: In abdominal CT, 40% preset ASIR-V can provides the best image quality and it can reduce radiation dose for 44.9%.

[Application of half-dose spectral CT based on the automatic spectral imaging mode selection and adaptive statistical iterative reconstruction in the CT examination of upper abdomen in obese patients.]

Objective: To investigate the value of combined use of half-dose spectral CT based on the automatic spectral imaging mode selection (GSI Assist) and adaptive statistical iterative reconstruction (ASIR) in the CT examination of upper abdomen in obese patients as compared with conventional CT. Methods: Eight-two obese patients (body mass index≥29 kg/m(2)) were prospectively selected from October to December 2016, and contrast-enhanced CT during arterial phase (AP) and portal venous phase (PVP) were carried out in those patients.The patients were randomly assigned to the study group and control group with random number table (41 cases in each group). In the study group, half-dose spectral CT based on GSI Assist was applied and monochromatic images (40 to 70 keV, 10 keV as increment) were reconstructed using 50% ASIR (group A). In the control group, the fixed tube potential of 120 kVp was done with images reconstruction using 30% ASIR (group B). Quantitative parameters of radiation dose, CT value, contrast to noise ratio (CNR) and image noise were compared between the two groups by using two sample t test, while qualitative visual parameters (overall image quality as graded on a 5-point scale) were compared with Mann-Whitney U test. Results: There was significant difference in effective radiation dose between the two groups[(5.2±0.8) vs (10.4±1.7) mSv, t=-17.822, P<0.001], and it was decreased for 50% in group A. During the arterial phase (AP) and portal venous phase (PVP), at the energy level of 40 keV and 50 keV, higher CT values, higher or similar CNRs, higher image noise and lower overall image quality scores were found in group A when compared with group B. At the energy level of 60 keV, group A had higher CT values, higher or similar CNRs, similar overall image quality scores with higher or similar image noise as compared with group B. At the energy level of 70 keV, the two groups had similar CT values, CNRs and image noise, and higher overall image quality scores were found in group A. Conclusions: In obese patients, combined use of half-dose spectral CT based on GSI Assist and ASIR can reduce effective radiation dose up to 50% when compared with conventional upper abdominal CT.Monochromatic images at 70 keV can maintain CNR and improve overall image quality.

[Combined use of wide-detector and adaptive statistical iterative reconstruction-V technique in abdominal CT with low radiation dose].

Objective: To investigate the image quality and radiation dose with wide-detector(80 mm) and adaptive statistical iterative reconstruction-V (ASIR-V) technique at abdominal contrast enhanced CT scan. Methods: In the first phantom experiment part, the percentage of ASIR-V for half dose of combined wide detector with ASIR-V technique as compared with standard-detector (40 mm) technique was determined. The human experiment was performed based on the phantom study, 160 patients underwent contrast-enhanced abdominal CT scan were prospectively collected and divided into the control group (n=40) with image reconstruction using 40% ASIR (group A) and the study group (n=120) with random number table. According to pre-ASIR-V percentage, the study group was assigned into three groups[40 cases in each group, group B: 0 pre-ASIR-V scan with image reconstruction of 0-100% post-ASIR-V (interval 10%, subgroups B0-B10); group C: 20% pre-ASIR-V with 20%, 40% and 60% post-ASIR-V (subgroups C1-C3); group D: 40%pre-ASIR-V with 40% and 60% post-ASIR-V (subgroups D1-D2)]. Image noise, CT attenuation values and CNR of the liver, pancreas, aorta and portal vein were compared by using two sample t test and One-way ANOVA. Qualitative visual parameters (overall image quality as graded on a 5-point scale) was compared by Mann-Whitney U test and Kruskal-Wallis H test. Results: The phantom experiment showed that the percentage of pre-ASIR-V for half dose was 40%. With the 40% pre-ASIR-V, radiation dose in the study group was reduced by 35.5% as compared with the control group. Image noise in the subgroups of B2-B10, C2-C3 and D1-D2 were lower (t=-14.681--3.046, all P<0.05) while CNR in the subgroups of B4-B10, C2-3 and D1-D2 were higher(t=2.048-9.248, all P<0.05)than those in group A, except the CNR of liver in the arterial phase (AP) in C2, D1 and D2 and the CNR of pancreas in AP in D1 (t=0.574-1.327, all P>0.05). The subjective image quality scores increased gradually in the range of 0-60% post-ASIR-V and decreased with post-ASIR-V larger than 70%. The overall image quality of subgroup B3-B8, C2-C3 and D1-D2 were higher than that in group A (Z=-2.229--6.533, all P<0.05). Conclusion: Compared with stand-detector together with ASIR technique, wide-detector combined with 40% pre-ASIR-V technique with 60% post-ASIR-V image reconstruction can reduce radiation dose while maintain good overall image quality.

Characterizing titanium dioxide and zinc oxide nanoparticles in sunscreen spray.

OBJECTIVE: Numerous commercial products contain titanium dioxide (TiO2 ) and zinc oxide (ZnO) nanoparticles (NPs); however, many of these are not labelled as containing NPs. This study sought to develop an effective means of characterizing TiO2 and ZnO NPs in sunscreen sprays, including the size, shape and composition of the particles as well as their aggregation/agglomeration characteristics. METHODS: Transmission electron microscopy (TEM) coupled with a window-type microchip K-kit/copper grid and X-ray diffraction (XRD) was used to characterize the oxide NPs. RESULTS: TME pre-treatment was performed using two approaches: using a conventional copper grid (requiring dilution) and using a K-kit (not requiring dilution). The use of K-kit in conjunction with XRD makes it possible to obtain direct measurements from samples that have not undergone pre-treatment, which could otherwise alter the nature of the samples, such as the degree of agglomeration. XRD was used to obtain information related to particle size and crystal structure. A strong correlation was observed between XRD and TEM measurements. CONCLUSION: The proposed measurement methods were shown to be highly effective in the characterization of oxide NPs in sunscreen sprays, providing consistent information related to NPs and their interactions in the formulations.

Microwave irradiation enhances gene and oligonucleotide delivery and induces effective exon skipping in myoblasts.

Microwave (MW) energy consists of electric and magnetic fields and is able to penetrate deep into biological materials. We investigated the effect of MW (2450 MHz) irradiation on gene delivery in cultured mouse myoblasts and observed enhanced transgene expression. This effect is, however, highly variable and critically dependent on the power levels, duration and cycle conditions of MW exposure. MW irradiation greatly enhances delivery of 2'O methyl-phosphorothioate antisense oligonucleotide (AON) targeting mouse dystrophin exon 23 and induces specific exon skipping in cultured myoblasts. Effective delivery of AON by MW irradiation is able to correct the dystrophin reading frame disrupted by a nonsense point mutation in the H2K mdx myoblasts, resulting in the restoration of dystrophin expression. MW-mediated nucleic acid delivery does not directly link to the increase in system temperature. The high variability in gene and oligonucleotide delivery is most likely the result of considerable irregularity in the distribution of the energy and magnetic field produced by MW with the current device. Therefore, achieving effective delivery of the therapeutic molecules would require new designs of MW devices capable of providing controllable and evenly distributed energy for homogenous exposure of the target cells.

Neuronal nitric oxide synthase activation is involved in insulin-mediated cardiovascular effects in the nucleus tractus solitarii of rats.

Neuronal nitric oxide synthases (nNOS) is distributed throughout the central nervous system (CNS) and has been proposed to modulate neuronal activity in the nucleus tractus solitarii (NTS). Here, we investigated whether the activation of nNOS is involved in insulin-induced cardiovascular responses in the NTS. Insulin (100 IU/ml) was unilaterally microinjected into the NTS, and the cardiovascular effects were evaluated before and after microinjection of the nNOS inhibitors 7-nitroindazole (7-NI) (5 pmol) and N(5)-(1-imino-3-butenyl)-l-ornithine (vinyl-L-NIO) (600 pmol). Western blot and immunohistochemical analyses were performed to determine nNOS phosphorylation levels after insulin or phosphoinositide 3-kinase (PI3K) inhibitor LY294002 microinjection into the NTS. Unilateral microinjection of insulin into the NTS produced prominent depressor and bradycardic effects in WKY rats. Pretreatment with the nNOS inhibitors 7-NI and Vinyl-L-NIO attenuated the cardiovascular response evoked by insulin in Wistar-Kyoto (WKY) rats. Moreover, Western blot analysis showed a significant increase in nNOS (16.5+/-0.4-fold; P<0.05; n=4) phosphorylation after insulin injection, whereas the PI3K inhibitor LY294002 abolished the insulin-induced effects. In situ nNOS phosphorylation was found to be increased in the NTS after insulin injection. Furthermore, co-immunoprecipitation assay showed Akt and nNOS can bind to each other as detected by phospho-Akt(S473) and phospho-nNOS(S1416) antibodies. In vitro kinase assay showed insulin activated Akt can directly phosphorylate nNOS(S1416). These results demonstrated that nNOS may couple with the activation of the insulin receptor, via the liberation of NO, in order to participate in central cardiovascular regulation of WKY rats.

bcl-2 overexpression inhibits cell death and promotes the morphogenesis, but not tumorigenesis of human mammary epithelial cells.

Overexpression of the B cell leukemia/lymphoma-2 (bcl-2) gene has been shown to confer a survival advantage on cells by inhibiting apoptosis. In epithelia, the bcl-2 gene is also related to development and differentiation, and the protein is strongly expressed in the embryo in the epithelial cells of the developing mammary gland. To investigate directly the effect of bcl-2 on human epithelial cells, we used an amphotropic recombinant retrovirus to introduce the gene into nontumorigenic cell lines developed from luminal epithelial cells cultured from milk. Here we demonstrate that while bcl-2 overexpression does not directly induce the tumorigenic phenotype, it provides a survival advantage to the mammary epithelial cells by inhibiting cell death at confluence or under conditions of serum starvation, bcl-2 can also affect the phenotype of the original epithelial cells, and promote epithelial-mesenchymal conversion, accompanied by loss of the cell adhesion molecules E-cadherin and alpha 2 beta 1 integrin. The extent of the epithelial-mesenchymal conversion varies with small differences in the phenotype of the parental line and with the level of expression of Bcl-2 and in some cases cell lines emerge with a mixed phenotype. The increased survival of Bcl-2-expressing cells at confluence results in multilayering, and the development of three- dimensional structures. Where a mixed phenotype is observed these structures consist of an outer layer of polarized epithelial cells separated by a basement membrane-like layer from an inner mass of fibroblastoid cells. Branching morphogenesis of bcl-2 transfectants is also observed in collagen gels (in the absence of fibroblast growth factors). The results strongly indicate that by increasing their survival under restrictive growth conditions, and by modifying the epithelial phenotype, bcl-2 can influence the specific morphogenetic behavior of mammary epithelial cells.

Function of WW domains as phosphoserine- or phosphothreonine-binding modules.

Protein-interacting modules help determine the specificity of signal transduction events, and protein phosphorylation can modulate the assembly of such modules into specific signaling complexes. Although phosphotyrosine-binding modules have been well-characterized, phosphoserine- or phosphothreonine-binding modules have not been described. WW domains are small protein modules found in various proteins that participate in cell signaling or regulation. WW domains of the essential mitotic prolyl isomerase Pin1 and the ubiquitin ligase Nedd4 bound to phosphoproteins, including physiological substrates of enzymes, in a phosphorylation-dependent manner. The Pin1 WW domain functioned as a phosphoserine- or phosphothreonine-binding module, with properties similar to those of SRC homology 2 domains. Phosphoserine- or phosphothreonine-binding activity was required for Pin1 to interact with its substrates in vitro and to perform its essential function in vivo.

Gravitational compression of colloidal gels.

We study the compression of depletion gels under the influence of a gravitational stress by monitoring the time evolution of the gel interface and the local volume fraction, φ, inside the gel. We find φ is not constant throughout the gel. Instead, there is a volume fraction gradient that develops and grows along the gel height as the compression process proceeds. Our results are correctly described by a non-linear poroelastic model that explicitly incorporates the φ-dependence of the gravitational, elastic and viscous stresses acting on the gel.

Traveling surface acoustic wave (TSAW) microfluidic fluorescence activated cell sorter (µFACS).

We report a microfluidic fluorescence activated cell-sorting (µFACS) device that employs traveling surface acoustic waves (TSAW) to sort cells at rates comparable to conventional jet-in-air FACS machines, with high purity and viability. The device combines inertial flow focusing and sheath flow to align and evenly space cells, improving the sorting accuracy and screening rate. We sort with an interdigital transducer (IDT) whose tapered geometry allows precise positioning of the TSAW for optimal cell sorting. We sort three different cell lines at several kHz, at cell velocities exceeding one meter per second, while maintaining both sorting purity and cell viability at around 90% simultaneously.

FLJ10540-elicited cell transformation is through the activation of PI3-kinase/AKT pathway.

A significant challenge in the post-genomic era is how to prioritize differentially expressed and uncharacterized novel genes found in hepatocellular carcinoma (HCC) microarray profiling. One such category is cell cycle regulated genes that have only evolved in higher organisms but not in lower eukaryotic cells. Characterization of these genes may reveal some novel human cancer-specific abnormalities. A novel transcript, FLJ10540 was identified. FLJ10540 is overexpressed in HCC as examined by quantitative reverse transcription-polymerase chain reaction and immunohistochemistry. The patients with higher FLJ10540 expression had a poor survival than those with lower FLJ10540 expression. Functional characterization indicates that FLJ10540 displays a number of characteristics associated with an oncogene, including anchorage-independent growth, enhanced cell growth at low serum levels and induction of tumorigenesis in nude mice. FLJ10540-elicited cell transformation is mediated by activation of the phosphatidylinositol 3'-kinase (PI3K)/AKT pathway. Moreover, FLJ10540 forms a complex with PI3K and can activate PI3K activity, which provides a mechanistic basis for FLJ10540-mediated oncogenesis. Together, using a combination of bioinformatics searches and empirical data, we have identified a novel oncogene, FLJ10540, which is conserved only in higher organisms. The finding raises the possibility that FLJ10540 is a potential new therapeutic target for HCC treatment. These findings may contribute to the development of new therapeutic strategies that are able to block the PI3K/AKT pathway in cancer cells.

Characterization of titanium dioxide and zinc oxide nanoparticles in sunscreen powder by comparing different measurement methods.

Numerous consumer products, such as cosmetics, contain nanoparticles (NPs) of titanium dioxide (TiO2) or zinc oxide (ZnO); however, this raises questions concerning the safety of such additives. Most of these products do not indicate whether the product includes NPs. In this study, we characterized metal oxide NPs according to size, shape, and composition as well as their aggregation/agglomeration characteristics. In order to comprehend quickly the characterization of metal oxide NPs, we employed single particle inductively coupled plasma (SP-ICPMS) to help quantify the size of metal oxide NPs; then, we use transmission electron microscopy (TEM) to corroborate the results. The crystal size and structure was measured by X-ray diffraction (XRD), there are two crystal phase of TiO2 NPs in sunscreen powder showed in XRD. However, SP-ICPMS proved highly effective in determining the size of NPs, the results of which remarkably good agreement with the TEM measurements. Pre-treatment included a conventional copper grid (requiring sample dilution) to evaluate the size, shape and composition of primary particles or plastic embedding (without the need for sample dilution) to evaluate the aggregate/aggregation of native NOAAs. The proposed method is an effective and fast approach to the characterization of oxide NPs in cosmetic sunscreen powder. These findings outline an alternative approach to the analysis of NPs in powder-form matrix.

TGFß promotes mesenchymal phenotype of pancreatic cancer cells, in part, through epigenetic activation of VAV1.

The highly homeostasis-resistant nature of cancer cells leads to their escape from treatment and to liver metastasis, which in turn makes pancreatic ductal adenocarcinoma (PDAC) difficult to treat, especially the squamous/epithelial-to-mesenchymal transition (EMT)-like subtype. As the molecular mechanisms underlying tumour heterogeneity remain elusive, we investigated whether epigenetic regulation might explain inter-individual differences in the progression of specific subtypes. DNA methylation profiling performed on cancer tissues prior to chemo/radiotherapy identified one hypermethylated CpG site (CpG6882469) in the VAV1 gene body that was correlated with demethylation of two promoter CpGs (CpG6772370/CpG6772811) in both PDAC and peripheral blood. Transforming growth factor ß treatment induced gene-body hypermethylation, dissociation of DNMT1 from the promoter, and VAV1 expression via SMAD4 and mutant KrasG12D. Pharmacological inhibition of TGFß-VAV1 signalling decreased the squamous/EMT-like cancer cells, promoted nuclear VAV1 localization, and enhanced the efficacy of gemcitabine in prolonging the survival of KPfl/flC mice. Together, the three VAV1 CpGs serve as biomarkers for prognosis and early detection, and the TGFß-VAV1 axis represents a therapeutic target.

Activation of the alpha2beta1 integrin prevents c-erbB2-induced scattering and apoptosis of human mammary epithelial cells in collagen.

Constitutive overexpression of c-erbB2 in the mammary epithelial cell line MTSV1-7 has been shown to result in epithelial-mesenchymal conversion, anchorage-independent growth and loss of organized morphogenesis in collagen. To elucidate the events leading to this drastic change, MTSV1-7 cells and its subclone HB2 (which shows a more strictly epithelial phenotype) were transfected with the hybrid trk-neu receptor consisting of the extracellular domain of the trkA nerve growth factor (NGF) receptor and the transmembrane and cytoplasmic domains of c-erbB2 (neu). In cells expressing this construct, c-erbB2 homodimerization can be mimicked by addition of NGF. In trk-neu transfectants of HB2 cells, modest expression led to increased cell proliferation upon NGF treatment. When clones with higher expression levels were grown in collagen, NGF instead induced cell scattering, diminished viability and dramatically increased apoptosis. Interestingly, both the dissociation of colonies and loss of cell viability could be completely reversed by treatment of the cells with antibodies that activate the adhesive capacity of the alpha2beta1 integrin. Long-term NGF treatment of high-expressing transfectants generated fibroblastic clones displaying a reduced expression of integrin alpha2 and E-cadherin, and extensive apoptosis in collagen. These results, which indicate that strong c-erbB2 signalling may lead to downregulation and/or inactivation of the alpha2beta1 integrin, promoting apoptosis in collagen, provide one possible explanation to the increased apoptosis frequently seen in early tumour development.

The midgut chymotrypsins of shrimps (Penaeus monodon, Penaeus japonicus and Penaeus penicillatus).

The midgut chymotrypsins (EC 3.4.4.5) of three species of shrimps, Penaeus monodon, Penaeus japonicus and Penaeus penicillatus were purified and studied in detail to clarify previous ambiguity in their identification. In each of the species there are two major forms of chymotrypsin, both single-chained with three disulfide bonds. One has a pI of 3.2 and Mr 27,000 or 28,000, while the other has a pI of 3.0 and Mr 25,000 or 26,000. The N-terminal amino acid sequences of the P. monodon enzymes are homologous to those of the crab (Uca pugilator) collagenase and to the other chymotrypsins. However, the active sites of the shrimp chymotrypsins are different from that of the well studied bovine alpha-chymotrypsin in some respects: (1) in spite of showing the typical specificity of chymotrypsin, the shrimp enzymes are more stringently selective for substrates with extended polypeptide chain; (2) some titration agents of alpha-chymotrypsin, including t-cinnamoylimidazole, 4-nitrophenyl guanidinobenzoate and its fluorescent derivative, do not react with the shrimp enzymes, neither do some of the alpha-chymotrypsin inhibitors: Tosyl-PheCH2Cl, methyl-4-nitrobenzenesulfonate and benzeneboronic acid; (3) the shrimp chymotrypsins are more reactive than the bovine enzyme toward native protein substrates including collagen; (4) the kinetic-salt-effects of the shrimp enzyme toward N-succinyl- and acetyl-Ala-Ala-Pro-Phe-4-nitroanilide mainly reflect electrostatic rather than hydrophobic interactions between the substrates and the enzyme. The shrimp enzymes are acid-labile but resistent to autolysis. Our results suggest that most Crustacea decapods contain chymotrypsins as one of the major digestive endopeptidases.

Two types of Russell's viper revealed by variation in phospholipases A2 from venom of the subspecies.

This study compared the phospholipases A2 (PLA2S) present in four commercially available venoms of Russell's viper subspecies by HPLC fractionation and partial sequence analysis. A potent heterodimeric PLA2 neurotoxin (designated a Russtoxin) was found in the venoms of all Russell's vipers except Daboia russelli (Sri Lanka and South India). The venom PLA2S of D. r. russelli (southern India) used in a previous study appear to be the same as those of D. r. pulchella (Sri Lanka), while the venom PLA2S of D. r. russelli (Pakistan) and D. r. siamensis (Burma and Thailand) resemble those of D. r. formosensis (Taiwan). This study provides evidence for the presence of two types of Russell's viper. Daboia russelli formosensis (Taiwan). D. r. siamensis (Thailand and Burma) and D. r. russelli (Pakistan) represent one type whose venom contains PLA2S having an Asn residue at the N-terminus, while D. r. pulchella (South India and Sri Lanka) represents the other type, whose venom contains PLA2S with a N-terminal residue Ser.

Induction of sister chromatid exchanges and micronuclei by titanium dioxide in Chinese hamster ovary-K1 cells.

Titanium dioxide (TiO2) has color properties of extreme whiteness and brightness, is relatively inexpensive, and is extensively used as a white pigment in a variety of materials. TiO2, an effective blocker of ultraviolet light, is frequently added to sunscreens and cosmetic creams. However, the genotoxicity of TiO2 remains to be controversial. In this report, we have demonstrated that TiO2 can be transported into Chinese hamster ovary-K1 (CHO-K1) cells. The effects of TiO2 on induction of sister chromatid exchanges (SCE) and micronuclei (MN) were then studied in these cells. The SCE frequency in CHO-K1 cells treated with TiO2 at a nonlethal dose range (0 to 5 microM) for 24 h was significantly and dose-dependently increased. By the conventional MN assay, TiO2 at the dose ranged from 0 to 20 microM slightly increased the MN frequency in CHO-K1 cells. However, in the cytokinesis-block MN assay, the number of MN per 1000 binucleated cells was significantly and dose-dependently enhanced in CHO-K1 cells treated TiO2 at the same dose range for 24 h. These results suggest that TiO2 is a potential genotoxic agent.

Relationship between the characteristics of immunopathological expression of hepatitis C virus antigen and hepatocytic injury.

Biopsied liver tissues from 352 cases were tested for hepatitis C virus (HCVAg) with improved PAP immunohistologic chemical method. Furthermore, corresponding seroantibody to hepatitis C virus was also tested. The total HCVAg positive rate was 9.1%. The HCVAg positive rate in chronic persistent hepatitis (CPH) was 5%. The HCVAg positive rate in chronic active hepatitis (CAH) was 11.2%. The HCVAg positive rate raised gradually along with the severity of hepatocytic injury. HCVAg may be seen in necrotic liver cells exfoliating into the liver sinus, indicating a close relationship between HCVAg and hepatocytic injury. Expression of HCVAg was mostly of the nucleus type in CPH cases and was mostly of the plasma type in CAH cases. The periphery of nucleus type-expressed positive cells generally had no marked inflammatory cell infiltration. The periphery of plasma type-expressed positive cells had a certain amount of inflammatory cell infiltration. Along with the severity of hepatocytic injury, HCVAg expressed itself in a positive correlation according to the nucleus and plasma types. The HCVAg positive cells were located mostly in the lobular peripheral band and rarely located in the venoperipheral band. It was possible that this had some relation with the lobular microcirculation of blood and blood supply. In this study, there was no obvious correlation between the HCVAg positive rate in hepatic tissues and the anti-HCV positive rate in sera. Neither the patients with HCVAg positive liver tissues nor the patients with seropositive anti-HCV had any history of blood transfusion and the use of blood products.(ABSTRACT TRUNCATED AT 250 WORDS)