Application of indocyanine green in pediatric surgery.

The use of near-infrared (NIR) fluorescence imaging with indocyanine green (ICG) has gained popularity in many fields in adult surgery, such as sentinel lymph node mapping, intra-operative solid tumor identification, and organ perfusion assessment. However, the clinical application of ICG in pediatric surgery is just at the beginning. This review paper presents the advantages, current applications and potential developments of NIR fluorescence imaging with ICG in our field.

Specific epitopes of the structural and hypothetical proteins elicit variable humoral responses in SARS patients.

BACKGROUND: Severe acute respiratory syndrome (SARS) is an infectious disease which was caused by a novel coronavirus (SARS-CoV). SARS has caused an outbreak in the world during 2003 and 2004, with 8098 individuals being infected and a death toll of 774 in 28 regions around the world. Specific humoral responses to viral infection remain unclear. OBJECTIVE: To analyse the antigenicity of the SARS-CoV genome and identify potential antigenic epitopes in the structural proteins. METHODS: Potential antigenic epitopes were identified in the structural proteins (nucleocapsid, membrane, spike, and small envelope proteins) and hypothetical proteins (SARS3a, 3b, 6, 7a, and 9b) that are specific for SARS-CoV. A peptide chip platform was created and the profiles of antibodies to these epitopes were investigated in 59 different SARS patients' sera obtained 6-103 days after the onset of the illness. Serial sera from five additional patients were also studied. RESULTS: Epitopes at the N-terminus of the membrane protein and the C-terminus of nucleocapsid protein elicited strong antibody responses. Epitopes on the spike protein were only moderately immunogenic but the effects were persistent. Antibodies were also detected for some putative proteins, noticeably the C-termini of SARS3a and SARS6. CONCLUSIONS: Important epitopes of the SARS-CoV genome that may serve as potential markers for the viral infection are identified. These specific antigenic sites may also be important for vaccine development against this new fatal infectious disease.

Reflection on SARS precautions in a severe intellectual disabilities hospital in Hong Kong.

BACKGROUND: Hong Kong went through a battle with a new respiratory disease, severe acute respiratory syndrome (SARS), from March to June 2003. All clinical settings, including rehabilitative and infirmary setting, have actively involved in fighting against the infection. The intent of this paper was to reflect on the SARS precautionary measures that had been taken in a severe intellectual disabilities hospital in Hong Kong. METHODS: A review on six SARS precautionary measures were conducted. They were assessment of risk, formulation of operational guidelines, implementation of infection control measures, education and training of staff, conducting audits and carrying out environmental improvement work. RESULTS: Patients were at risk of getting infected from carers, visitors, volunteers, and staff and patients of general hospitals. A SARS Quarantine Unit, isolation ward, was opened to isolate patients who might have had close contact with SARS patients during a stay in a general hospital or when they returned from home leave. Undoubtedly, both staff and relatives participated in preventing the patients from being infected. No day leave and home leave was reported and the number of hospitalization in general hospital was decreased during the critical period. Three infection control audits were conducted and improvement work was carried out subsequently. CONCLUSION: The practice of grouping within a standard isolation room is recommended to continue in the future. Moreover, intensive infection control training for all staff is of highest importance to safeguard the health of both staff and patient.

Genomic organization and characterization of a three-gene rat adult beta-globin haplotype.

The isolation and detailed characterization of a three-beta-globin gene (GloB) haplotype in the Sprague-Dawley (S-D) rat is described. An enriched library, lambda SDHelib, was screened with a human GloB probe, humbg44, and from which a beta minor gene, Rathbbz, was isolated, sequenced and characterized. A S-D rat GloB-specific probe, Ratbgze12, derived from the Rathbbz gene, was then used to screen a S-D rat genomic library, lambda SDglib. The clone T1510 was isolated and identified to include the entire Rathbbz gene and part of another GloB gene, Rathbby, which was 5' upstream from Rathbbz. Chromosomal walking upstream using the riboprobe, rnaT71, led to the isolation of an overlapping clone, Ta49, which was shown to include two full-length GloB genes; the most 5' was Rathbbx followed by Rathbby. Sequence data suggests that Rathbbx is a beta major gene, whereas Rathbby is a hybrid gene of Rathbbx and Rathbbz. Genomic hybridization confirmed this particular three-gene haplotype in the S-D rat. This haplotype, a1, may be the prototype of the GloB cluster in rat.

Two new rat alpha-globin sequences as identified by the conserved region PCR.

Based on the notion that regions of structural genes which encode critical domains of the corresponding proteins are highly conserved among closely related species, oligonucleotide primers were designed and used to amplify the alpha-globin sequence(s) of the Sprague-Dawley (SD) rat. Data of these amplified sequence constructs showed that two new rat alpha-globin specific sequences have been identified. Southern hybridization confirmed the presence of these sequences in the rat genome.

A class of amphipathic proteins associated with lipid storage bodies in plants. Possible similarities with animal serum apolipoproteins.

The lipid-storing tissues of plants contain many small (0.2-1 microns) lipid (normally triacylglycerol) droplets which are surrounded and stabilized by a mixed phospholipid and protein annulus. The proteinaceous components of the lipid storage bodies are termed oleosins and are not associated with any other cellular structures. The major oleosins of rapeseed and radish have been isolated by preparative SDS-PAGE and are respectively classes of 19 kDa and 20 kDa proteins. Both protein classes were N-terminally blocked for direct sequencing, but were partially sequenced following limited proteolytic digestion. The major rapeseed oleosin was made up of at least two 19 kDa polypeptides, termed nap-I and nap-II, which have closely related but different amino acid sequences. A single 20 kDa oleosin, termed rad-I, was found in radish. A near full length cDNA clone for a major rapeseed oleosin was sequenced and found to correspond almost exactly to the sequence of nap-II. The sequences of nap-I and rad-I show very close similarity to one another, as do the sequences of nap-II and the previously determined sequence for the major oleosin from maize. All four oleosins have a large central hydrophobic domain flanked by polar N- and C-terminal domains. Secondary structure predictions for the four oleosins are similar and a novel model is proposed based on a central hydrophobic beta-strand region flanked by an N-terminal polar alpha-helix and a C-terminal amphipathic alpha-helix. The possibility that oleosins exhibit structural and functional similarities with some animal apolipoproteins is discussed.

Low-resolution epitope characterisation in a family of seed apolipoproteins using polyclonal antibodies.

Polyclonal antibodies raised against a range of seed apolipoproteins from the family Cruciferae have been used for the first time for low resolution epitope characterisation. Antibodies were raised against the major seed apolipoproteins of Brassica napus, Sinapis alba and Raphanus sativum. In each case, the antibodies recognized, in addition to the 19-20 kDa apolipoprotein to which they were raised, similar 19-20 kDa apolipoproteins from a wide range of species in the family Cruciferae, but not in other plant families. Homologous or heterologous two-sites (sandwich) assays were performed with the format [antibody A - test apolipoprotein - antibody B - 2 degrees antibody]. The results showed a drastically reduced antibody B binding by apolipoproteins preincubated with an antibody A. This indicated the presence of a single major epitope on many of the apolipoproteins. The antigenicity of native and denatured apolipoproteins was similar, although the antigenicity of the former was much more readily destroyed by proteinase attack. It is concluded that there are relatively few major epitopes present on the Cruciferae apolipoproteins and it is suggested that these epitopes are localized on the small hydrophilic surface-exposed C- and N-terminal domains of the apolipoproteins.

An immunologically related family of apolipoproteins associated with triacylglycerol storage in the Cruciferae.

The major apolipoproteins associated with oil-storage bodies have been isolated from the mature seeds of six different species of the family Cruciferae. The apolipoproteins were all of molecular mass 19-20 kDa. They were highly abundant in mature seed tissue, accounting for up to 20% total seed proteins, and were localized exclusively on the membranes of oil-storage bodies. Antibodies were raised in rabbits and mice against the six purified apolipoproteins. In each case, the antibodies specifically recognized 19-20 kDa polypeptides on immunoblots of total seed proteins from 15 different species of the Cruciferae. The extent of the immunological cross-reactivity among the six purified seed apolipoproteins of the Cruciferae was investigated quantitatively using enzyme-linked immunosorbent assay. Very high levels of cross-reactivity were obtained, in contrast to a complete lack of cross-reactivity observed when the major seed apolipoprotein of a non-crucifer, Glycine max, was assayed. Peptide mapping studies showed that the different crucifer seed apolipoproteins gave rise to similar proteolytic cleavage products following treatment with Staphylococcus aureus V8 protease, Lysobacter enzymogenes Lys-C endoprotease, and trypsin. The patterns of immunogenic proteolytic cleavage products of the different apolipoproteins were also similar. We propose that there is a family of abundant 19-20 kDa apolipoproteins in mature seeds of oil-bearing Cruciferae. These apolipoproteins are all major components of the membranes of oil-storage bodies. The apolipoproteins are therefore very closely related with respect to their structure, function, and immunological properties.