Case series of tracheal extubation in prone position after endoscopic retrograde cholangiopancreatography general anesthesia.

OBJECTIVE: The study aims to present a 15-patient case series of tracheal extubation in the prone position after endoscopic retrograde cholangiopancreatography (ERCP) general anesthesia. PATIENTS AND METHODS: Fifteen inpatients who underwent elective ERCP in our hospital were prospectively enrolled, and a series of case studies were conducted with the prone extubation technique after general anesthesia. All patients underwent routine operation of tracheal intubation under general anesthesia. After the surgery, when the train-of-four ratio (TOFr) ≥0.9, bispectral index (BIS) ≥80, tidal volume ≥6 ml/kg and the required actions could be performed, the endotracheal catheter was removed after sufficient negative pressure suction of oral secretions. After the endotracheal catheter was removed, the patient autonomously turned to the transport bed with the assistance of medical staff and was then admitted to the post-anesthesia care unit (PACU) for further observation. When the patient awoke, he had regained orientation, and presented stable vital signs, no nausea and vomiting, and no other discomfort symptoms, he/she was able to leave PACU and returned to the ward with a Steward score of ≥5. RESULTS: All 15 patients who underwent ERCP elective surgery were successfully extubated in the prone position after surgery. Transient hypoxemia with SpO2 below 90% occurred in 2 of the 15 patients and returned to normal with oxygen mask administration. 7 patients had coughs and were without special treatment. Another 1 patient showed transient abnormal hemodynamic fluctuations after extubation, mean airway pressure (MAP) was higher than 20% of the baseline value, and hemodynamics was stable after drug treatment. CONCLUSIONS: The prone extubation technique is feasible for ERCP general anesthesia patients. However, a larger sample size is needed to validate its safety and to verify whether there exist advantages of the extubation technique in a prone position over a supine position.

Flexible solar-cell from zinc oxide nanocrystalline sheets self-assembled by an in-situ electrodeposition process.

Zinc oxide nanocrystalline sheets were self-assembled on a flexible polymer substrate to act as the electrode of dye-sensitized solar cells by an in situ-construction electrodeposition process. It was discovered that the nanosheet-based solar cell exhibited better performance than a nanoparticle-based solar cell or a well-oriented nanowire-based solar cell. The nanosheet microstructure has advantages which include the depression of loss during photoelectron transport, the increase of dye compound adsorption, and the enhance of incident light capture. As a result, the performance of dye-sensitized solar cells can be obviously improved. This success provides a feasible bottom-up approach for integrating a solar cell together with nanodevices and microcircuits on a flexible substrate which can work with self-supplied solar energy.

White spot syndrome virus (WSSV) infects specific hemocytes of the shrimp Penaeus merguiensis.

White spot syndrome virus (WSSV) was specifically detected by PCR in Penaeus merguiensis hemocytes, hemolymph and plasma. This suggested a close association between the shrimp hemolymph and the virus. Three types of hemocyte from shrimp were isolated using flow cytometry. Dynamic changes of the hemocyte subpopulations in P. merguiensis at different times after infection were observed, indicating that the WSSV infection selectively affected specific subpopulations. Immunofluorescence assay (IFA) and a Wright-Giemsa double staining study of hemocyte types further confirmed the cellular localization of the virus in the infected hemocytes. Electron microscopy revealed virus particles in both vacuoles and the nucleus of the semigranular cells (SGC), as well as in the vacuoles of the granular cells (GC). However, no virus could be detected in the hyaline cells (HC). Our results suggest that the virus infects 2 types of shrimp hemocytes--GCs and SGCs. The SGC type contains higher virus loads and exhibits faster infection rates, and is apparently more susceptible to WSSV infection.

Analysis of three marine fish cell lines by rapd assay.

We tested the applicability of the random amplified polymorphic deoxyribonucleic acid (RAPD) analysis for identification of three marine fish cell lines FG, SPH, and RSBF, and as a possible tool to detect cross-contamination. Sixty commercial 10-mer RAPD primers were tested on the cell lines and on samples collected from individual fish. The results obtained showed that the cell lines could be identified to the correspondent species on the basis of identical patterns produced by 35-48% of the primers tested; the total mean similarity indices for cell lines versus correspondent species of individual fish ranged from 0.825 to 0.851, indicating the existence of genetic variation in these cell lines in relation to the species of their origin. Also, four primers, which gave a monomorphic band pattern within species/line, but different among the species/line, were obtained. These primers can be useful for identification of these cell lines and for characterization of the genetic variation of these cell lines in relation to the species of their origin. This supported the use of RAPD analysis as an effective tool in species identification and cross-contamination test among different cell lines.

Modification in framework region I results in a decreased affinity of chimeric anti-TAG72 antibody.

A mouse/human chimeric B72.3P-1-6 antibody was produced by construction of a novel expression vector mpSV2neo-EP2-V-Crl containing the same gene fragments as the expression vector mpSV2neo-EP1-V-Crl (Xiang J., Roder J. and Hozunni N., submitted to Molec. Immun., 1991) except the promoter (P2) fragment in which the translation start codon ATG is retained. The expression vector was transfected into a heavy chain loss mutant cell line, B72.3M1. The translation of the chimeric heavy chain may start at the exogenous start codon ATG within the P2 fragment, which is 27 base pairs upstream of the endogenous start codon ATG in B72.3 heavy chain V region cDNA fragment, leading to an alteration in leader sequence cleavage sites and the formation of chimeric heavy chain with an elongation in the FR1 region. Chimeric B72.3P-1-6 antibody retained binding specificity to TAG72 antigen, but showed an eight-fold decrease in binding affinity to TAG72 compared with chimeric B72.3-1-3 antibody. This suggests that residues in FRI contribute to the correct folding of the antibody binding region of the B72.3 antibody.

Production of murine V-human Cr1 chimeric anti-TAG72 antibody using V region cDNA amplified by PCR.

A mouse/human chimeric B72.3-1-3 antibody was produced by construction of a novel expression vector mpSV2neo-EP1-V-Cr1. This vector contains the neo gene as a selection marker, the murine immunoglobulin heavy chain promoter and enhancer, the murine V region cDNA containing mRNA splicing joint sequences, amplified and cloned by the PCR technique directly from the B72.3 hybridoma RNA, and the human genomic Cr1 region. The expression vector containing the murine/human chimeric immunoglobulin heavy chain gene was transfected into heavy chain loss mutant cell line, B72.3Ml. Chimeric B72.3-1-3 antibody was produced at 2 micrograms/ml and retained full binding reactivity to TAG72 compared to the murine B72.3 parental antibody. Using this method, chimeric immunoglobulin molecules can be produced rapidly in comparison with the cDNA and genomic cloning techniques.

Synthesis and expression of metastasis-associated, Met-72/83 antigens.

In the present study, we report a more detailed biochemical analysis of the B16 melanoma, metastasis-associated, Met-72 antigen. Specifically, we have examined (1) the molecular forms of Met-72 isolated during synthesis, surface expression and 'shedding' and (2) the cell-surface expression of Met-72 during the cell cycle. These experiments show that the 72 kD species originally described has an isoelectric point of between 6.3 and 6.9, but is the desialylated derivative of an 83 kD native molecule whose isoelectric point ranges between pH 4.9 and 5.6. In addition, a 90 kD glycoprotein doublet was immunoprecipitated from biosynthetically labelled B16 melanoma cells, but does not appear to be a precursor of the 83 kD or 72 kD molecule. These findings have led us to interchangeably use the terminology Met-72 and Met 72/83. The latter terminology more accurately describes the physical forms which can be identified by different labelling procedures. When culture supernatants from 3H-leucine labelled cells were subjected to anti-Met-72 immunoprecipitation, a 35 kD species was identified as a possible 'shed' product of these cells. Met-72/83 expression during the cell cycle was analyzed by flow cytometry and found not to be restricted to any particular stage. In addition, experiments were performed to determine whether low levels of Met-72 expression on poorly metastatic B16 melanomal clones was a direct result of low levels of synthesis, or if other control mechanisms regulated intracellular pools of Met-72 prior to cell-surface expression.

Flow cytometric analysis of DNA in bone and soft-tissue tumors using nuclear suspensions.

Ninety-four bone and soft tissue tumors were analyzed for their DNA content using flow cytometry (FCM). A simple, rapid method for preparing isolated nuclear suspensions was used. Tissues, minced in a hypotonic solution containing detergent and propidium iodide as a fluorescent probe for DNA, provided in most instances high nuclear yields from only 0.02 to 0.03 g of solid tumor. Whereas all nonneoplastic samples had a diploid DNA content, various degrees of abnormal DNA distributions were detected in 90% of the neoplastic samples and were present in benign as well as malignant tumors. Our findings demonstrate that FCM DNA analysis is practical in most musculoskeletal tumors and support the observations of others that abnormal DNA content may serve as a general neoplastic marker in these tumors.

The lack of correlation between experimental metastatic potential and platelet aggregating activity of B16 melanoma clones viewed in relation to tumor cell heterogeneity.

Two widely used B16 melanoma cell lines of low and high lung colonizing potential (B16-F1 and B16-F10) were compared in their ability to induce platelet aggregation. The results of these experiments showed a reproducible difference in platelet aggregating activity of these two cell lines which directly correlated with their lung colonizing potentials. However, when clones were derived from these heterogeneous cell lines and tested for experimental metastatic potential, platelet aggregating ability and Met-72 expression, no correlation could be attached to the platelet aggregating activity of the clones. Results of these experiments provide direct evidence that platelet aggregation is not an accurate index of experimental metastatic potential of tumor cell clones, nor is it an essential trait of all metastatic cells. The ability of tumor cells to induce platelet aggregation is examined and discussed in the context of cellular heterogeneity.

Flow cytometric analysis and cytopathology of body cavity fluids.

A total of 75 samples of body cavity fluids from 71 patients were analyzed by both flow cytometry (FCM), to detect cells with an abnormal DNA content (aneuploidy), and by conventional cytopathology. Samples included 27 pleural fluids, 35 peritoneal fluids, 11 peritoneal washings and 2 pericardial fluids. For cytologic examination, the samples were prepared using standard techniques. Samples for FCM analysis were centrifuged and exposed to a hypotonic solution containing detergent and propidium iodide, a DNA intercalating fluorescent stain. Aneuploidy as well as cytologic malignancy were found in 17 samples. Forty-seven samples had normal DNA histograms by FCM and were also cytologically negative. Four samples suspicious by cytology but normal by FCM were from patients with renal-cell carcinoma (two samples from the same patient), endometrial adenocarcinoma without metastasis and chronic lymphocytic leukemia. Three samples abnormal by FCM but negative by cytology were from patients with ovarian cystadenoma, cirrhosis and uterine leiomyoma. FCM showed aneuploidy in four cytologically negative samples from patients with histologically proven malignancy (lymphoma, colonic adenocarcinoma, cervical squamous cell carcinoma, and endometrial adenosquamous carcinoma). Based on these results, FCM analysis combined with conventional cytopathology yielded 100% sensitivity, 100% predictive value of a negative result and 94% specificity. This rapid and quantitative FCM analysis of body cavity fluids can be a very useful adjunct to conventional diagnostic cytopathology.

In vitro modulation of the metastatic phenotype. I. Analysis of differentiation forms of the B16 melanoma expressing Met-72 determinants and metastatic activity.

In vitro cultures of a highly metastatic B16 melanoma clone (BL6-10) were found to undergo dramatic changes in morphology and differentiation upon transfer to another culture medium. Specifically, BL6-10 melanoma cells which had been originally selected and adapted for growth in Eagles' Hanks' amino acid supplemented media with 10 per cent newborn calf serum were amelanotic and epitheliod in shape. When these cells were shifted into Dulbecco's modified Eagles medium with 10 per cent fetal calf serum, they became highly melanotic and of spindle/dendritic morphology within 4 days of culture. These morphological changes as well as other parameters were all characteristic of established criteria of melanoma differentiation. Alterations in the differentiation state of our highly metastatic variant, BL6-10, did not result in any change in tumorigenicity but did have profound effects on metastatic potential. All of the morphological and functional characteristics of the differentiated melanoma were found to be reversible by re-plating the cells in their original growth medium and 4 days of in vitro growth. These studies have allowed us to follow and more firmly establish Met-72 antigen expression as a surface marker for metastatic cells of the B16 melanoma, and have provided direct experimental evidence that the less differentiated, Met-72 positive melanoma form is the dominant cell type capable of metastatic potential.

High levels of Met-72 antigen expression: correlation with metastatic activity of B16 melanoma tumor cell variants.

In the present study, both poorly and highly metastatic clones derived from the C57BL/6 mouse B16 melanoma were used with cyclophosphamide in an attempt to elicit host antibody responses against cell surface markers expressed on highly metastatic tumor variants. The immunizations, performed in both syngeneic and xenogeneic combinations in Lewis rats, resulted in the production of 3 mouse and 2 rat monoclonal antibodies (MoAb) that preferentially react with highly metastatic clones derived from the B16 melanoma. These MoAb all immunoprecipitated a 72,000-dalton, cell surface-expressed glycoprotein, referred to as Met-72. In this study, 2 of the mouse anti-Met-72 MoAb were examined in detail for a) tumor specificity, b) reactivity against normal mouse tissue by in vivo absorption, and c) their ability to discriminate highly metastatic clones derived from the B16 melanoma.

Melanoma cells growing in aggregates on a non-adhesive poly(HEMA) substrate exhibit polykaryocytosis but do not develop an increased metastatic capability.

B16-F1 melanoma cells were plated onto plastic tissue-culture dishes rendered non-adhesive for cells by coating with 0.12 per cent poly(2-hydroxyethyl methylacrylate), poly(HEMA). These growth conditions caused the normally flat, adherent B16-F1 cells to grow as single cells in suspension. Within 24 hours, the rounded cells formed aggregates and grew at a slower rate than control cells grown at the same density on untreated plastic dishes. Microscopic observations provided evidence that polykaryocytosis was occurring among the aggregates. Following replating onto standard adhesive tissue-culture plastic, 20-30 per cent of the aggregates were observed to contain varying numbers of multinucleated giant cells (polykaryocytes). The study has revealed a previously undescribed propensity of certain B16-F1 cells cultivated as aggregates in suspension to develop into polykaryocytes, most probably as a result of spontaneous tumor cell-tumor cell fusion. The possible relevance of this behavior in vitro to events in tumor progression is discussed. This study, however, does not support the findings of others that the metastatic capability of B16-F1 cells is increased by such non-adherent culture conditions. No increase in metastatic potential was observed for B16-F1 cells, or for a low metastatic clone (F1-7) derived from it, grown for 72 or 96 hours in a spherical configuration compared to control cells grown in a flat, adherent monolayer.