A preliminary investigation into the comparison of dissolution/digestion techniques for the chemical characterization of polyurethane foam.

Due to their widespread use in domestic and commercial premises, polyurethane foams, as either fragmented or bulk foam, are types of evidence commonly found at crime scenes. The traditional approach to determining the evidential value of polyurethane foam (PF) involves comparing recovered and control fragments under low and high magnification, under various lighting conditions, as well as the comparison of their respective dye spectra. As with most forms of trace evidence, chemical comparison is also desirable. In this work, two approaches to chemically comparing foam fragments were investigated, i.e. inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis of the Tin (Sn) content in different foam types; and gas chromatography-flame ionization detection (GC-FID) analysis of soluble components in PFs mobilized by dichloromethane. Seven different foam types were studied and their Sn content was found to be different. They also produced characteristic GC-FID chromatographic profiles whose compounds were identified with gas chromatography-mass spectrometry (GC-MS) analysis. This study suggests that incorporating chemical data obtained from GC-FID/GC-MS and ICP-OES into a case involving PF could be advantageous, as this will enable the forensic scientist to broaden the comparison between control and recovered fragments, and further assess the strength of the evidence. However, ICP-OES analysis is a destructive technique with a relatively short sample turnaround time, whilst GC-FID analysis is more time-consuming and non-destructive, requiring corroboration with GC-MS data. The values of these two analytical techniques in the forensic chemical characterization of PFs are discussed.

Pitfalls of the application of microdialysis in clinical oncology: controversial findings with docetaxel.

Microdialysis is a novel and minimally invasive sampling technique, based on the diffusion of analytes from the interstitial compartment through a semi-permeable membrane, and enables direct assessment of tissue disposition and penetration of drugs. Variable antitumor responses may be associated with differences in tumor vascularity, capillary permeability or tumor interstitial pressure resulting in variable delivery of anticancer agents. In preparation of pharmacokinetic studies, aimed at measuring docetaxel concentrations in healthy and malignant tissues in vivo, in pre-clinical as well as clinical studies, in vitro recovery experiments were performed. In contrast to published data, the recovery experiments suggest that docetaxel has a very low recovery as a result of non-specific binding to currently available microdialysis catheters. Here we discuss our findings with docetaxel in a historical perspective and we report on our experience using polysorbate 80 to eliminate the non-specific binding and its effects on the recovery of docetaxel.

A preliminary risk assessment of organochlorines accumulated in fish to the Indo-Pacific humpback dolphin (Sousa chinensis) in the Northwestern waters of Hong Kong.

The Indo-Pacific humpback dolphin is considered threatened due to several factors including pollution in Hong Kong and the risks due to consumption of fish tainted with polychlorinated biphenyls (PCBs) and organochlorine pesticides were assessed. Six species of fish Collichthys lucida, Pseudosciaena crocea, Johnius sp., Thryssa sp., Mugil sp. and Trichiurus sp., which comprise the main prey species of humpback dolphins were collected for analyses. Risks due to total PCBs, total TEQs, PCB 118 and the pesticides were assessed with the use of toxicity reference values as the threshold reference benchmarks. The calculated risk quotients (RQs) showed that the risks associated with organochlorines were generally low. The highest RQ was associated with total TEQs suggesting that dioxin-like PCBs may pose the highest risk to the dolphins. The HCHs, total PCBs and heptachlor had comparatively high RQs and thus they should also be the priority organochlorines that would require further investigation.

Calcitonin receptor antibodies in the identification of osteoclasts.

Osteoclasts are the cells responsible for bone resorption, and their number and rate of formation are critical in determining bone mass. To identify and quantify osteoclasts, as well as to study their formation in bone and in osteoclastogenic cultures, osteoclast-specific cell markers are required. Only the calcitonin receptor (CTR) expression unambiguously identifies osteoclasts and distinguishes them from macrophage polykaryons. However, present autoradiographic methods for CTR detection are cumbersome and time consuming. We have developed rabbit polyclonal antibodies specific for the C-terminal intracellular domain of the mouse and rat Cla CTR. These antibodies labeled HEK-293 cells stably transfected with CTR (but not untransfected HEK-293 cells). This labeling is abrogated by preabsorbing the antibodies with the recombinant antigen. The antibodies immunostained primary mouse and rat osteoclasts as well as osteoclasts in sections of mouse bone. Osteoclasts (both mononuclear and multinucleated) formed from mouse bone marrow or spleen cells cocultured with osteoblasts in the presence of 1,25 dihydroxyvitamin D3 and prostaglandin E2 were also specifically immunostained by the CTR antibodies. Cocultures incubated under conditions that did not allow osteoclastogenesis (i.e., omission of mediators or osteoblasts, or culture for less than 4 days) were not immunostained by CTR antibodies. Autoradiographic detection of 125I-labeled salmon calcitonin combined with CTR immunohistochemistry showed that both methods labeled the same cells. A CTR polyclonal antibody and monoclonal antibody F4/80 were used in combination to show immunofluorescence labeling of murine osteoclasts and macrophage populations, respectively, in marrow/osteoblast cocultures. These results indicate that simple and rapid CTR antibody-based methods can be used to identify osteoclasts, and can be used to characterize the antigenic profile of osteoclasts by using double immunofluorescence analysis.

The effects of endurance exercise on dystrophic mdx mice. II. Contractile properties of skinned muscle fibres.

Dystrophic (mdx) mice were subjected to a 15 week exercise programme consisting of endurance swimming. Single fibres from the extensor digitorum longus (EDL, fast-twitch) and soleus (SOL, mixed fast- and slow-twitch) muscles were attached to a sensitive force-recording apparatus, and activated in Ca(2+)- and Sr(2+)-buffered solutions. In addition to the normal well-defined fibre types in these muscles, a small number of fibres were also sampled from the soleus of both experimental groups, which were 'Intermediate' to the other two SOL fibre types. Type IIB fibres from the EDL and type IIA fibres from the soleus of the Swim group were significantly less sensitive to Ca2+ and Sr2+ compared with those fibres sampled from the sedentary (Sedent) group, suggesting that endurance exercise was able to modify Ca(2+)- and Sr(2+)-activated contractile characteristics. The swim-trained (Swim) group's increased incidence of SOL fibres with characteristics intermediate to those of the fast- and slow-twitch fibre types suggests a possible exercise-induced fibre type transformation as an adaptation to the functional demand.

Species difference in the specific receptors of platelet activating factor.

Relative potencies of platelet activating factor (PAF) and PAF analogs and several PAF receptor antagonists when inhibiting the [3H]PAF specific binding to human and rabbit platelet membranes and membrane fragments of human lung tissues were compared. In rabbit platelets, L-652,731 was found to be most potent in the list of PAF receptor antagonists with an equilibrium inhibition constant (Ki) of 9.83 (+/- 2.92) X 10(-9) M followed by L-653,150 greater than kadsurenone congruent to Ono-6240 greater than ginkgolide B greater than CV-3988 greater than L-651,142, whereas in human platelets the relative potencies of these PAF receptor antagonists were as follows: Ono-6240 greater than L-653,150 congruent to L-652,731 congruent to kadsurenone greater than ginkgolide B greater than CV-3988 greater than L-651,142. Ono-6240 was the most potent one with a Ki of 4.86 (+/- 1.44) X 10(-8) M which was roughly two times more potent than that in rabbit platelets, whereas the affinity of L-652,731 was about ten times less in human platelets (Ki = 1.03 (+/- 0.15) X 10(-7) M) compared to that in rabbit platelets (Ki = 9.83 (+/- 2.92) X 10(-9) M). These variations between species among PAF antagonists strongly suggest that there exists a species difference at or near the binding site of the receptor of platelet activating factor. The relative potency of these PAF receptor antagonists in human lung membranes differed very little from that in human platelets and was found to be Ono-6240 greater than L-653,150 congruent to kadsurenone congruent to L-652,731 greater than ginkgolide B greater than CV-3988 greater than L-651,142. Even though C16-PAF showed slightly higher potency in human lung, and CV-3988 and Ono-6240 showed slightly lower, the difference was too small to suggest that there is a difference in the PAF receptors between human platelets and human lung tissues.

Survival of Gardnerella vaginalis in human urine.

The authors studied the survival of Gardnerella vaginalis in human urine and determined conditions for optimum recovery on agar media. Gardnerella counts declined by greater than 99.9% in urine held at 37 degrees C for 24 hours, whereas the falloff was negligible at 4 degrees C. Viability was lost after 6 hours in urine with pH of 5, and only 0.01% cells survived in urine with pH of 7. In contrast, greater than 90% cells survived exposure at pH of 6. Dialysis to remove small molecular weight (less than 14,000) inhibitors did not enhance survival. Co-cultivation with Ureaplasma urealyticum and the addition of glycogen improved survival. Maximum recovery from urine required anaerobic incubation on enriched agar medium (pH 6.5-7.5) for at least 48 hours. Gardnerella vaginalis survives poorly in human urine at 37 degrees C. Culture for these bacteria requires prolonged anaerobic incubation.

Release of platelet activating factor and its involvement in the first phase of carrageenin-induced rat foot edema.

Platelet activating factor (PAF), a potent lipid-like vasoactive agent, induced rat foot edema when it was injected subplantarly. The edema reached its maximum 1 h after PAF challenge. Indomethacin did not inhibit the peak edematous response whereas both PAF antagonists, kadsurenone and L-652,731, inhibit the PAF-induced rat foot edema (PFE). Both PAF antagonists also partially block the first phase of the carrageenin-induced rat foot edema (CFE). Using the inhibition of [3H]PAF receptor binding to prepared rabbit platelet membranes, release of PAF or PAF-like materials in carrageenin-injected rat hindpaw was observed. These results suggest that the released PAF or PAF-like materials together with the released histamine and kinin evoke the first phase hindpaw edema in the rats. Indomethacin or PAF antagonist, administered alone, does not block the first phase or the second phase of CFE, respectively. However, PAF antagonist potentiated the inhibitory effects of indomethacin suggesting that the released PAF may also be involved in the biosynthesis of prostaglandins to initiate the second phase of rat CFE.

L-659,989: a useful probe in the detection of multiple conformational states of PAF receptors.

L-659,989 is a potent, specific and competitive platelet-activating factor (PAF) receptor antagonist. The 2,5-tritium labeled L-659,989, similar to [3H]PAF, specifically binds to rabbit platelet membranes with an equilibrium dissociation constant (KD) of 1.60 (+/- 0.20) nM in 10 mM MgCl2. However, guanosine 5'-triphosphate (GTP) and several cations affect the specific binding of [3H]PAF and of [3H]L-659,989 to rabbit platelet membranes in different ways. K+, Mg2+, Ca2+ and Mn2+ potentiate the specific binding of both ligands. Na+ and Li+ inhibit the specific [3H]PAF binding, but enhance the binding of [3H]L-659,989; GTP reduces the [3H]PAF binding but has no effect on the binding of [3H]-L-659,989. Ni2+ inhibits the [3H]L-659-989 binding, but has no effect on the binding of [3H]PAF. In the presence of 150 mM NaCl, [3H]L-659,989 exhibits identical KD and detectable binding sites (Bmax) values as those in the presence of 10 mM MgCl2, while KD And Bmax values of [3H]PAF are dramatically reduced in the presence of 150 mM NaCl compared to those in 10 mM MgCl2. These results suggest the existence of multiple conformational states of the PAF specific receptor and that PAF and L-659,989 bind differently to those states. In the presence of 150 mM NaCl and 1 mM GTP, receptors appear to exist in a single conformational state with an equilibrium dissociation constant (KB) of 0.93 microM for PAF as derived from the Schild plot. In isolated rabbit platelets pretreated with 10 microM ETH 227, a Na(+)-specific ionophore, the detectable [3H]PAF binding sites drop from 260 to 100 binding sites per platelet, but the binding sites for [3H]L-659,989 remain roughly the same. The Na+ binding sites which modulate the conformation of PAF receptors are therefore protected from extracellular Na+ until ionophore is added, and are probably located on the cytoplasmic side of the plasma membrane.

Risk to breeding success of fish-eating Ardeids due to persistent organic contaminants in Hong Kong: evidence from organochlorine compounds in eggs.

Eggs of two Ardeid species, the Little Egret (Egretta garzetta) and the Black-crowned Night Heron (Nycticorax nycticorax), were collected from two egretries located in the New Territories of Hong Kong with one located near the internationally acclaimed wetland reserve, the Mai Po Marshes, and the other in a remote site (A Chau). The eggs were analysed for organochlorine (OC) compounds including the DDTs, PCBs, hexachlorocyclohexanes (HCHs) and the chlordanes (CHLs). All of the OCs under investigation were detected in the eggs of both species with significantly higher levels in the Little Egret (DDTs, 560-2200; PCBs, 270-1700; CHLs, 81-470 ng g(-1) wet weight) than the Night Heron (DDTs, 210-1200; PCBs, 85-600; CHLs 59-75 ng g(-1) wet weight). The DDTs consisted mainly of DDE with levels ranging from 85% to 95% of the total. The HCHs were at about the same levels in both species (8.4-30 ng g(-1) wet weight). All of the OCs had linear concentration probability distributions on a log-normal basis which were used to evaluate exposure associated with these compounds as part of a probabilistic risk analysis. A linear dose/response relationship for the percentage reduction in the survival of young associated with DDE in eggs was developed. This probabilistic relationship was used to establish the threshold level (1000 ng g(-1) wet weight) at which there was a significant level of reduction in the survival of young above zero and the variability in DDE concentrations at this effect level. Using a threshold level of 1000 ng g(-1), the calculated Risk Quotient (RQ) had a 12.4% probability of RQ exceeding unity with the Night Heron, and 40.9% with the Little Egret. These results indicate that the DDTs in eggs would be expected to be associated with adverse effects on the survival of young of both species, particularly the Little Egret.

False 'hematuria' due to bacteriuria.

OBJECTIVE: To determine the effects of bacteriuria on dipstick urinalysis for microhematuria. DESIGN: Selection of urine samples with dipstick readings of at least one plus (+) for blood, followed by microscopic examination and bacterial culture, from patients with signs and symptoms of urinary tract diseases. SETTING: A large teaching hospital in Melbourne, Australia. PATIENTS: Two hundred forty-six inpatients and outpatients being investigated for signs and symptoms of urinary tract diseases. RESULTS: Seventeen (7%) of 246 patients having within-reference erythrocyte counts had polymicrobic cultures. One hundred nine patients had slight hematuria (1 x 10(7) to 6 x 10(7) cells/L). Of these, 21 bacteriuric patients had significantly higher dipstick findings than 88 abacteriuric patients (P < .005, Fisher's Exact Probability Test, two-tailed). Hydroperoxidase was detected in 80% of isolates (in 27 of 27 gram-negative bacilli cultures, in 6 of 6 staphylococci cultures, and in 3 of 12 streptococci cultures) on agar medium and in 69% of isolates in urine. CONCLUSION: These results show that patients without hematuria but with bacteriuria may give a false-positive dipstick reading for blood.

Determination of polychlorinated biphenyls in human blood serum by SPME.

We report a direct immersion SPME-GC/mu ECD method for the determination of non-volatile pollutants (Aroclor 1254) in human blood plasma using an enzymatic proteolytic approach to overcome the protein fouling problem. A nonspecific serine protease, Proteinase K, is used to remove the fouling interference. Significant improvement in relative repeatability from 28.9% to 3.4% (n = 10) was obtained. Analyte recovery up to 93% and detection limit of 1.0 ppb (total PCB) were achieved.

A preliminary risk assessment of trace elements accumulated in fish to the Indo-Pacific Humpback dolphin (Sousa chinensis) in the northwestern waters of Hong Kong.

In order to assess the potential risks associated with consumption of contaminated prey items to the Indo-Pacific Humpback dolphin (Sousa chinensis), fish species (Collichthys lucida, Pseudosciaena crocea, Johnius sp., Thryssa sp., Mugil sp. and Trichiurus sp.) representing the main food items of the dolphin were collected from the northwestern waters of Hong Kong, including the Sha Chau and Lung Kwu Chau Marine Park, which form the main habitat of the dolphin in Hong Kong. Within these waters, there are several potential sources of pollution including significant inputs from the Pearl River catchment, several major sewage outfalls and a series of mud pits that receive contaminated dredged sediments. Concentrations of thirteen trace elements (Ag, As, Cd, Co, Cr, Cs, Cu, Hg, Mn, Ni, Se, V, and Zn) in the fish tissue were analyzed by inductively coupled plasma mass spectrometer (ICP-MS). An assessment of the risks of adverse effects on the dolphin due to consumption of tainted fish was undertaken using two toxic reference benchmarks, namely the reference dose (RfD) and toxicity reference value (TRV). The risk quotient (RQ) calculated for each element showed that the risks from consumption of fish were generally low and within safe limits. The risks associated with arsenic, cadmium and mercury were, however, elevated. The highest calculated RQ was associated with total arsenic; however, the majority of arsenic in marine organisms tends to be in the non-toxic organic form, and the actual risk to the dolphin due to this metalloid is likely to be lower.

Determination of microcystins in cyanobacterial blooms by solid-phase microextraction-high-performance liquid chromatography.

A solid-phase microextraction (SPME) method was applied to study microcystin (MC) profiles in a natural Microcystis sp. bloom in a freshwater pond in Guangzhou, China. Three dominant MC variants, namely MC-LR, MC-YR, and MC-RR, were quantified. Simultaneous study of their total, extracellular, and intracellular profiles was made possible using SPME coupled to high-performance liquid chromatography. The total and intracellular concentrations of MC-LR in the bloom were 8.67 x 10(-2) microg/ ml and 1.93 mg/g, respectively. The corresponding concentrations of MC-YR were 1.20 x 10(-3) microg/ml and 0.06 mg/g, respectively, and those of MC-RR were 5.57 x 10(-2) microg/ml and 1.49 mg/g, respectively. Only MC-LR was detectable in the extracellular phase (1.49 x 10(-2) microg/ml) of the bloom, and its concentration was 14% of the intracellular content. Mass balance consideration revealed that only 71.1% of total MC-LR, 36.0% of total MC-YR, and 67.4% of total MC-RR within the cyanobacterial cells were released into the aqueous phase immediately after cell lysis.

Polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in sediments from Hong Kong.

Concentrations of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were determined in 14 sediment samples collected from four sites in the Mai Po Marshes Nature Reserve (within a RAMSAR Site) and from another six sites in Victoria Harbour and along the Hong Kong coastline. Elevated levels of PCDDs, and particularly OCDD, were detectable in all samples collected from the Mai Po Marshes and five of the six sites. In contrast to PCDDs, PCDFs were mainly found in sediment samples collected from industrial areas (Kwun Tong and To Kwa Wan) in Victoria Harbour. PCDD/ F levels and congener profiles in the samples from the Mai Po Marshes Nature Reserve in particular show strong similarities to those reported in studies which have attributed similar elevated PCDD concentrations to nonanthropogenic PCDD sources.

The isolation and characterization of kadsurenone from haifenteng (Piper futokadsura) as an orally active specific receptor antagonist of platelet-activating factor.

A natural product, kadsurenone, was isolated from the Chinese herbal preparation haifenteng (Caulis piperis futokadsurae) and characterized as an orally-active specific antagonist of the platelet-activating factor (PAF). Kadsurenone inhibits the specific binding of 3H-PAF to a receptor preparation from rabbit platelet membrane in a competitive and reversible manner, its Ki being 3.88 X 10(-8) M. It inhibits the aggregation of rabbit platelets in plasma induced by PAF with a pA2 of 6.28, but not those induced by arachidonic acid, epinephrine, ADP or A-23187. It inhibits the aggregation of isolated human neutrophils with a pA2 of 6.32. It also inhibits PAF-induced degranulation and release of beta-D-glucuronidase at 2-24 microM in vitro. In the rat, kadsurenone at 8-40 mg/kg i.p. inhibits the increases of plasma lysosomal enzymes and haematocrit induced by intravenous PAF. In the guinea pig, kadsurenone at 25-50 mg/kg p.o. reduces the increase of cutaneous vascular permeability induced by PAF. These results indicate that kadsurenone is a specific and effective receptor antagonist of PAF in several in vitro and in vivo systems.

A strong two-photon induced phosphorescent Golgi-specific in vitro marker based on a heteroleptic iridium complex.

A new heteroleptic iridium complex demonstrated low cytotoxicity and near-infrared excitation (via two-photon absorption) for target-specific in vitro Golgi imaging in various cell lines (HeLa and A549 cells) with two-photon absorption cross section (~350 GM) in DMSO.

A community study of sleep bruxism in Hong Kong children: association with comorbid sleep disorders and neurobehavioral consequences.

BACKGROUND: The prevalence of childhood sleep bruxism (SB) varied from 5% to 46% among various studies. In addition to local facial and dental adverse consequences, accumulating evidence suggests that childhood SB could be associated with comorbid sleep and systemic neurobehavioral disturbances. This study attempted to investigate the prevalence and clinical correlates of SB in a large community sample. METHODS: This study was part of an ongoing epidemiologic study about sleep problems among Hong Kong Chinese children. A total of 9172 questionnaires were distributed to children of grades 1-6 from 13 randomly selected primary schools. Parents of the children were asked to complete and return the Hong Kong children sleep questionnaire, which aimed to explore the sleep problems and patterns of their children. RESULTS: Six thousand three hundred and eighty-nine questionnaires with valid answers to SB were received and the response rate was 69.7%. The mean age of the recruited children was 9.2±1.8years (50.6%, boys). The prevalence of SB with teeth grinding frequency more than thrice weekly over the past year was 5.9%. SB was more prevalent among boys with decreasing prevalence across age. SB was associated with chronic medical diseases, sleep-related breathing problem, upper respiratory infection, and other parasomnia features, especially sleep talking (OR (95%CI)=4.07 (2.33-7.11)). Children with SB were more likely noticed by their parents to be hyperactive (OR (95%CI)=1.61 (1.25-2.07)) and bad-tempered (OR (95%CI)=1.69 (1.35-2.12)) and had deterioration in their academic performance (OR (95%CI)=1.22(1.03-1.43)). CONCLUSIONS: Almost 6% of Hong Kong primary schoolchildren suffered from frequent SB. The condition was most prevalent among young boys. SB was found to be associated with a variety of medical conditions, neuropsychiatric sequelae, and comorbid sleep conditions, especially sleep talking and sleep related breathing problems. Further prospective studies will need to clarify the longitudinal course of childhood SB and its response to treatment.

A simple method allowing DIC imaging in conjunction with confocal microscopy.

Current optical methods to collect Nomarski differential interference contrast (DIC) or phase images with a transmitted light detector (TLD) in conjunction with confocal laser scanning microscopy (CLSM) can be technically challenging and inefficient. We describe for the first time a simple method that combines the use of the commercial product QPm (Iatia, Melbourne Australia) with brightfield images collected with the TLD of a CLSM, generating DIC, phase, Zernike phase, dark-field or Hoffman modulation contrast images. The brightfield images may be collected at the same time as the confocal images. This method also allows the calculation of contrast-enhanced images from archival data. The technique described here allows for the creation of contrast-enhanced images such as DIC or phase, without compromising the intensity or quality of confocal images collected simultaneously. Provided the confocal microscope is equipped with a motorized z-drive and a TLD, no hardware or optical modifications are required. The contrast-enhanced images are calculated with software using the quantitative phase-amplitude microscopy technique (Barone-Nugent et al., 2002). This technique, being far simpler during image collection, allows the microscopist to concentrate on their confocal imaging and experimental procedures. Unlike conventional DIC, this technique may be used to calculate DIC images when cells are imaged through plastic, and without the use of expensive strain-free objective lenses.