BACKGROUND

Raynaud's phenomenon (RP) associated with connective tissue disease (secondary RP) may be difficult to manage with conservative therapy. A combination of sympathetically mediated vasospasm and vaso-occlusion has been implicated as the etiology of digital ischemic phenomenon. Thoracic sympathetic outflow blocking has been performed with various techniques. However, there have been some limitations in all treatment options.

OBJECTIVE

We report on a patient with medically refractory digital ulceration and gangrene caused by scleroderma who was successfully treated with a continuous infusion of mepivacaine into the thoracic sympathetic ganglions as a means to improve finger circulation.

METHODS

We are reporting on a 32-year-old female patient suffering from a medically intractable gangrenous ulcer in the right third finger and the left second and third fingers, accompanied by aching pain (VAS, visual analogue scale, 5 - 6/10) and numbness in both forearms. She underwent continuous infusion of mepivacaine through the thoracic sympathetic catheter placed in T2 vertebral segment for 13 days on the right and for 11 days on the left and cervical epidural infusion of mepivcaine with fentanyl for 10 days after the medical treatment failed. Her finger temperature increased 2 degrees C - 5 degrees C during the thoracic sympathetic block with continuous infusion of mepivacine. Her finger wounds healed completely with 13 days of the continuous thoracic sympathetic block without any complications.

CONCLUSIONS

Continuous infusion of mepivacaine into the thoracic sympathetic ganglionic space led to the healing of the medically refractory gangrenous ulcer of the fingers in the patient with scleroderma.

OBJECTIVE

To assess the prevalence of Raynaud's phenomenon (RP) and of RP associated systemic sclerosis (SSc) in a large regional representative study.

METHODS

Ten thousand individuals aged between 14-65 years participated in face-to-face interviews. The stratified sample of the South-West Hungarian population was representative for age, sex and urban or rural residence. Individuals reporting complaints suggesting the presence of "clinically significant" RP were asked to undergo a clinical investigation. Patients showing complaints provoked by taking something out of the freezer (-20 degrees C) compartment of the refrigerator and/or whether they had experienced digital ulcers were sorted into this category.

RESULTS

The overall prevalence of RP was at least 578.9/10,000, and the prevalence of "clinically significant" RP could be calculated as at least 87.7/10,000 inhabitants. In this latter group 17.2% of the cases had either established SSc or anticentromere antibody or scleroderma capillary pattern on nailfold capillaroscopy. SSc with "clinically significant" RP and/or ulcers was identified in a prevalence of 9.1/10.000 individuals, whilst there was a prevalence of 14.7/10,000 of RP with either anticentromere antibody or scleroderma capillary pattern.

CONCLUSIONS

"Clinically significant" RP affects almost 1% of the population. We identified cases with early stages of scleroderma spectrum disorder showing either anticentromere autoantibody or scleroderma capillary pattern. The prevalence of SSc was found to be higher than expected. It is reasonable to screen "clinically significant" RP cases for scleroderma-related symptoms because this approach makes it possible to identify patients with both SSc and early scleroderma related symptoms.

Domoic acid, a potent excitotoxic analogue of glutamate and kainate, may cause seizures, amnesia, and sometimes death in humans consuming contaminated shellfish. Continuous behavioral observations and recordings of the electrocorticogram (ECoG, via bipolar, epidural electrodes) were obtained from nonanesthetized rats for 2 h after intraperitoneal injection with either saline, 2.2, or 4.4 mg/kg of domoic acid. Rats were then sacrificed for c-fos immunohistochemistry. Fast Fourier transformation (FFT) of the ECoG data to obtain the voltage as a function of frequency indicated that the lower frequency bands (theta, 4.75-6.75 Hz and delta, 1.25-4.50 Hz) were the first to respond, with a significant elevation by 30 min after the high dose of domoic acid. The lower dose of domoic acid also caused a significant elevation of ECoG voltage, but not until later in the session. Sixty minutes after dosing, the behavioral biomarkers of "ear scratching" and "rearing, praying" (RP) seizures became significantly elevated in the high-dose rats. The low-dose rats showed no significant alterations in behavior at any time during the session. In postmortem brains obtained immediately after the sessions, c-fos was activated in the anterior olfactory nucleus by both the low and high doses of domoic acid. However, only the high dose increased c-fos immunoreactivity in the hippocampus, affecting both the granule and pyramidal neurons. These data indicate that electroencephalographic and c-fos responses can be obtained at a dose of domoic acid that fails to activate the behavioral response most commonly used as a bioassay for this marine toxin: ear scratching with the ipsilateral foot.

We propose a specific, reproducible and sensitive HPLC method for the determination of N(epsilon)-(carboxymethyl)lysine (CML) excreted in urine. Total CML was measured in acid hydrolysates of urine samples, while free CML was measured in acetonitrile-deproteinised urine samples using a RP-HPLC method with ortho-phtaldialdehyde (OPA)-derivatisation and fluorescence detection suited for automation. We compared the CML excretion of 51 non-proteinuric patients with diabetes mellitus (DM) (age 57+/-14 years, HbA1c 8.0+/-1.8%) to 42 non-diabetic controls (C) (age 45+/-17 years). The urinary excretion of total CML in diabetic patients was increased by approximately 30% (DM: 0.58+/-0.21; C: 0.45+/-0.14 microM/mmol creatinine; P<0.001). While urinary excretion of free CML was not significantly different, excretion of bound CML was increased (DM: 0.36+/-0.17; C: 0.27+/-0.14; P<0.05) in diabetic patients. CML excretion was correlated with protein and albumin excretion, but did not correlate with HbA1c, duration of DM or diabetic complications such as neuropathy or retinopathy. Furthermore, no age-dependent change of total CML excretion was found, while free CML excretion was lower in younger subjects. The specific and sensitive determination of CML by RP-HPLC of its OPA-derivative is well suited for automation and better than that of less defined glycoxidation products (AGEs).

OBJECTIVE

This study aimed to determine if temporomandibular disorders (TMD) correlate with alterations in body posture detectable through posturography.

METHODS

Thirty-five asymptomatic subjects and 35 TMD patients (34 males and 36 females; mean age, 27.7+/-8.6 years) constituted the matched control and TMD groups, respectively. Posturography was performed under four different experimental conditions: (a) eyes open with mandibular rest position (Eyes Open RP); (b) eyes open with dental occlusion (Eyes Open DO); (c) eyes closed with mandibular rest position (Eyes Closed RP); and (d) eyes closed with dental occlusion (Eyes Closed DO). The X, Y, and absolute centre of pressure displacements from the projection of a theoretical barycentre and the sway area, sway length, and sway velocity were recorded as static and dynamic posturographic parameters, respectively.

RESULTS

Generally, no differences were found in any of these parameters between the groups and between the RP and DO within either Eyes Open/Closed conditions. The only differences were found under Eyes Closed as compared to Eyes Open, irrespective of the RP/DO conditions for dynamic and not for static posturographic parameters.

CONCLUSIONS

This study failed to show detectable alterations in body posture in TMD patients.

Luminance contrast discrimination was measured in 14 patients with retinitis pigmentosa (RP) and 14 control observers with normal vision, using steady-pedestal and pulsed-pedestal paradigms [Pokorny, J., & Smith, V. C. (1997). Psychophysical signatures associated with magnocellular and parvocellular pathway contrast gain. Journal of the Optical Society of America A, 14, 2477-2486] to bias performance toward the magnocellular (MC) or parvocellular (PC) pathway, respectively. The aim was to determine the relative effects of retinal degeneration on MC- and PC-pathway function in RP. For five of the RP patients, contrast discrimination thresholds were within normal limits for both the steady-pedestal and pulsed-pedestal paradigms. The other nine RP patients showed threshold elevations for the steady-pedestal paradigm (presumed magnocellular mediation), whereas their thresholds for the pulsed-pedestal paradigm (presumed parvocellular mediation) were within normal limits for all but the two patients who had the most extreme threshold elevations using the steady-pedestal paradigm. A control experiment on four of the RP patients, using a greater number of pedestal contrasts, verified that the patients' thresholds for the pulsed-pedestal paradigm showed the pattern expected for contrast discrimination mediated by the PC pathway. The higher threshold elevations for the steady-pedestal paradigm than for the pulsed-pedestal paradigm indicate that the retinal degeneration that occurs in RP predominantly disrupts contrast discrimination under stimulus conditions that favor the MC pathway.

There are data to suggest that both the humoral and cellular immune responses directed against Tat are beneficial in delaying HIV disease progression. We examined the association between the occurrence of Tat-specific binding antibodies (Abs) and different parameters of HIV-1 disease progression. We generated eight Tat proteins, derived from HIV-1 subtypes A, B, C, and D, and circulating recombinant form CRF01_AE. These proteins were used to screen for Tat-specific binding Abs by an ELISA. Using five Tat proteins, we investigated whether the occurrence of Tat-specific Abs within 2 years after seroconversion for the majority, affected disease progression over time among 126 participants using survival analysis and rate of CD4 decline. Of these, 52 participants with a sample at 1.5 and 4.5 years after seroconversion were further examined to study the effect of Tat-specific Ab loss or maintenance on disease progression. Finally, using all the eight Tat proteins, we also investigated whether specific Abs to these Tat proteins among 48 participants, grouped as rapid progressors (RP, n = 26) and long-term survivors (LTS, n = 22) according to their CD4 decline over time, affected disease progression. Survival analysis did not reveal any evidence of protection from progression by Tat-specific Abs. Comparison of rate of CD4 declines between individuals with and without Abs to any Tat protein showed only a small and borderline significant advantage of having Tat-specific Abs (p = 0.043). There was no correlation between either loss or maintenance of Tat-specific Abs and disease progression. Comparison of LTS with RP showed no evidence that Tat-specific Abs slows participants' disease progression. This study showed no evidence of a protective effect of having Tat-specific Abs among these Ugandan subjects.

BACKGROUND

Leaves and fruits of Passiflora species are widely used around the world in popular medicine, mainly as sedatives and tranquilisers. C-glycosyl flavonoids are the main components of these species.

OBJECTIVE

To investigate the constituent patterns and to develop a chromatographic method for the characterisation of the C-glycosyl flavonoids profile of the extracts of the leaves and the pericarp of South American Passiflora species.

METHODS

The chemical composition of extracts from the leaves and the fruits' pericarp of Passiflora edulis var. flavicarpa, P. edulis var. edulis, Passiflora alata, Passiflora tripartita var. mollissima, Passiflora quadrangularis, Passiflora manicata and Passiflora ligularis was evaluated for the presence of C-glycosyl flavonoids. Two separate HPLC methods were developed suitable for a diode array detector (DAD) and a MS detector. Separation by HPLC-DAD was achieved on a Luna C-18 column, using solvent A (tetrahydrofuran-isopropanol-acetonitrile) and solvent B (H₃PO₄ 0.5%) in an isocratic elution mode. In the HPLC-MS, the components were separated on a Luna RP-18A column by a gradient elution (water-acetonitrile-formic acid).

RESULTS

The presence of C-glycosyl flavonoids was identified in leaves and pericarp of P. edulis var. flavicarpa, P. alata, P. edulis var. edulis and P. tripartita var. molissima, but only in leaf extracts of P. quadrangularis and P. manicata and not at all in P. ligularis. The different species and varieties showed different major constituents. The C-glycosyl flavonoids identified more frequently were orientin, isoorientin, vitexin and isovitexin.

CONCLUSIONS

The methods established are simple and can be used as a tool for the characterisation and quality control of pharmaceutical preparations containing these Passiflora extracts.

OBJECTIVE

Retinitis pigmentosa (RP) is a genetically heterogeneous disease with over 60 causative genes known to date. Nevertheless, approximately 40% of RP cases remain genetically unsolved, suggesting that many novel disease-causing genes are yet to be identified. In this study, we aimed to identify the causative mutation for a large autosomal dominant RP (adRP) family with negative results from known retinal disease gene screening.

METHODS

Linkage analysis followed by whole-exome sequencing was performed. Stringent variant filtering and prioritization was carried out to identify the causative mutation.

RESULTS

Linkage analysis identified a minimal disease region of 8 Mb on chromosome 10 with a peak parametric logarithm (base 10) of odds (LOD) score of 3.500. Further whole-exome sequencing identified a heterozygous missense mutation (NM_000188.2:c.2539G>A, p.E847K) in hexokinase 1 (HK1) that segregated with the disease phenotype in the family. Biochemical assays showed that the E847K mutation does not affect hexokinase enzymatic activity or the protein stability, suggesting that the mutation may impact other uncharacterized function or result in a gain of function of HK1.

CONCLUSIONS

Here, we identified HK1 as a novel causative gene for adRP. This is the first report that associates the glucose metabolic pathway with human retinal degenerative disease, suggesting a potential new disease mechanism.

OBJECTIVE

The ARO 96-02 trial primarily compared wait-and-see (WS, arm A) with adjuvant radiation therapy (ART, arm B) in prostate cancer patients who achieved an undetectable prostate-specific antigen (PSA) after radical prostatectomy (RP). Here, we report the outcome with up to 12 years of follow-up of patients who retained a post-RP detectable PSA and received salvage radiation therapy (SRT, arm C).

METHODS

For the study, 388 patients with pT3-4pN0 prostate cancer with positive or negative surgical margins were recruited. After RP, 307 men achieved an undetectable PSA (arms A + B). In 78 patients the PSA remained above thresholds (median 0.6, range 0.05-5.6 ng/mL). Of the latter, 74 consented to receive 66 Gy to the prostate bed, and SRT was applied at a median of 86 days after RP. Clinical relapse-free survival, metastasis-free survival, and overall survival were determined by the Kaplan-Meier method.

RESULTS

Patients with persisting PSA after RP had higher preoperative PSA values, higher tumor stages, higher Gleason scores, and more positive surgical margins than did patients in arms A + B. For the 74 patients, the 10-year clinical relapse-free survival rate was 63%. Forty-three men had hormone therapy; 12 experienced distant metastases; 23 patients died. Compared with men who did achieve an undetectable PSA, the arm-C patients fared significantly worse, with a 10-year metastasis-free survival of 67% versus 83% and overall survival of 68% versus 84%, respectively. In Cox regression analysis, Gleason score ≥8 (hazard ratio [HR] 2.8), pT ≥ 3c (HR 2.4), and extraprostatic extension ≥2 mm (HR 3.6) were unfavorable risk factors of progression.

CONCLUSIONS

A persisting PSA after prostatectomy seems to be an important prognosticator of clinical progression for pT3 tumors. It correlates with a higher rate of distant metastases and with worse overall survival. A larger prospective study is required to determine which patient subgroups will benefit most from which treatment option.

Cordyceps militaris has been artificially cultivated in China, and the great amounts of produced medium residue were discarded after the harvest. The aims of this work were to analyze the structure of the residue polysaccharide (RPS) of C. militaris SU-12, and to investigate the pharmacological effects of RPS on lipid metabolism and oxidative stress. RPS was composed of glucose, arabinose and mannose with a ratio of 62:1.6:1 by gas chromatography analysis, and the Mw (weight-average molecular weight), Mn (number-average molecular weight) and Mz (z-average molecular weight) of RPS were 2.86×10(3), 6.85×10(2), and 1.97×10(4)Da, respectively. The mice experiments demonstrated that RPS could reduce the levels of blood and liver lipid, and improve the glutamate pyruvate transaminase and antioxidant activity. The histopathological observations of mice livers indicated that RPS could attenuate liver cell injury. Results suggest that the RPS might be used as a potential antihyperlipidemic, hepatoprotective and antioxidant product.

Sequential application of solvent extraction, gel permeation chromatography, and RP-HPLC in combination with taste dilution analyses, followed by LC-MS and 1D/2D-NMR experiments and thiolytic degradation, revealed that, besides theobromine and caffeine, the flavan-3-ols epicatechin, catechin, procyanidin B-2, procyanidin B-5, procyanidin C-1, [epicatechin-(4beta-->8)](3)-epicatechin, and [epicatechin-(4beta-->8)](4)-epicatechin were among the key compounds contributing to the bitter taste as well as the astringent mouthfeel imparted upon consumption of roasted cocoa. In addition, a series of quercetin, naringenin, luteolin, and apigenin glycopyranosides as well as a family of not previously identified amino acid amides, namely, (+)-N-[4'-hydroxy-(E)-cinnamoyl]-L-aspartic acid, (+)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-aspartic acid, (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-glutamic acid, (-)-N-[4'-hydroxy-(E)-cinnamoyl]-L-glutamic acid, (-)-N-[4'-hydroxy-(E)-cinnamoyl]-3-hydroxy-L-tyrosine, (+)-N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-aspartic acid, and (+)-N-(E)-cinnamoyl-L-aspartic acid, have been identified as key astringent compounds of roasted cocoa. Furthermore, (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-3-hydroxy-l-tyrosine (clovamide), (-)-N-[4'-hydroxy-(E)-cinnamoyl]-L-tyrosine (deoxyclovamide), and (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-tyrosine, reported previously as antioxidants, have been found as contributors of cocoa's astringent taste. By means of the half-tongue test, the taste thresholds of flavan-3-ols and glycosides have been determined.

Cuminum cyminum L. roots, stems and leaves, and flowers were investigated for their essential oils, total phenolics, flavonoids, and tannins contents, individual phenolic compounds, and antioxidant activities. The essential oil was investigated by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS), whereas identification and quantification of individual target polyphenolic compounds was performed by reversed-phase high-performance liquid chromatography (RP-HPLC). Essential oil yields were 0.03% in roots, 0.1% in stem and leaves, and 1.7% in flowers. Major components of the oils were bornyl acetate (23%), α-terpinene (34%), and γ-terpinene (51%) in roots, stems and leaves, and flowers, respectively. In all C. cyminum organs, total phenolics content ranged from 11.8 to 19.2 mg of gallic acid equivalents per gram of dry weight (mg of GAE/g of DW). Among the polyphenols studied, 13 were identified in roots, 17 in stem and leaves, and 15 in flowers. The major phenolic compound in the roots was quercetin (26%), whereas in the stems and leaves, p-coumaric, rosmarinic, trans-2-dihydrocinnamic acids and resorcinol were predominant. In the flowers, vanillic acid was the main compound (51%). The antioxidant activities of C. cyminum essential oils and acetone extracts obtained from the three organs were assessed using four tests [1,1-diphenyl-2-picrylhydrazyl (DPPH), β-carotene/linoleic acid, reducing power, and chelating power assays]. The acetone extract of flowers was strongly effective as a DPPH radical scavenger, lipid peroxidation inhibitor, and reducing agent, with IC(50) values of 4, 32, and 8 μg/mL, respectively. Moreover, the acetone extract of stems and leaves showed the highest chelating power. However, the essential oils exhibited moderate activities in the different tests.

Fructosamine 3-kinase (FN3K), an enzyme initially identified in erythrocytes, catalyses the phosphorylation of fructosamines on their third carbon, leading to their destabilization and their removal from protein. We show that human erythrocytes also contain FN3K-related protein (FN3K-RP), an enzyme that phosphorylates psicosamines and ribulosamines, but not fructosamines, on the third carbon of their sugar moiety. Protein-bound psicosamine 3-phosphates and ribulosamine 3-phosphates are unstable, decomposing at pH 7.1 and 37 degrees C with half-lives of 8.8 h and 25 min respectively, as compared with 7 h for fructosamine 3-phosphates. NMR analysis indicated that 1-deoxy-1-morpholinopsicose (DMP, a substrate for FN3K and FN3K-RP), like 1-deoxy-1-morpholinofructose (DMF, a substrate of FN3K), penetrated erythrocytes and was converted into the corresponding 3-phospho-derivative. Incubation of erythrocytes with 50 mM allose, 200 mM glucose or 10 mM ribose for 24 h resulted in the accumulation of glycated haemoglobin, and this accumulation was approx. 1.9-2.6-fold higher if DMP, a competitive inhibitor of both FN3K and FN3K-RP, was present in the incubation medium. Incubation with 50 mM allose or 200 mM glucose also caused the accumulation of ketoamine 3-phosphates, which was inhibited by DMP. By contrast, DMF, a specific inhibitor of FN3K, only affected the glucose-dependent accumulation of glycated haemoglobin and ketoamine 3-phosphates. These data indicate that FN3K-RP can phosphorylate intracellular, protein-bound psicosamines and ribulosamines, thus leading to deglycation.

A sensitive HPLC assay for all-trans-retinol, alpha-tocopherol, and gamma-tocopherols in human serum and plasma is reported. Sample preparation is performed in one step and involves precipitation of proteins and extraction of lipids with two volumes of an ethanol-chloroform mixture (3:1, v/v) without I.S. addition. After removal of the precipitated protein, 20 microl aliquots of the supernatant (equivalent to 6.7 microl of serum or plasma) were injected into the HPLC system and analyzed using fluorometric detection. RP-HPLC was performed using a C(18) S3 ODS2 column with a methanol-water step gradient (97:3 to 100) at 1.0 ml/min. The quantification limit expressed as nanograms of analyte per milliliter of serum or plasma was approximately 30 ng for all-trans-retinol, 300 ng for alpha-tocopherol and 250 ng for gamma- and delta-tocopherol. The method was validated and applied to human serum and plasma from a total of 120 subjects. This procedure requires a small volume of serum or plasma and can therefore be a valuable tool for measuring low concentrations of these vitamins in preterm infants with sensitivity, precision and accuracy.

BACKGROUND

There exists a current lack of information about the composition of the different types of plasma. No direct comparisons between apheresis plasma (AP) and recovered plasma (RP) derived from in-line-filtered whole blood (WB) have been published to date.

METHODS

Sixty AP units, 100 RP units from in-line-filtered WB held for 3 hours at 20 degrees C between donation and freezing, and an additional 100 RP units held for 15 hours at 20 degrees C before freezing were analyzed for coagulation factors and inhibitors, total protein, immunoglobulin G (IgG), and hemostasis and proteolysis activation markers. The influence of twice freezing and thawing on clotting factors V, VIII, and XI was also examined.

RESULTS

AP contains substantially greater activities of factor (F) V, FVIII, F IX, and FXI than RP frozen within 3 hours after WB donation. Prolonged holding of RP at 20 degrees C for more than 15 hours caused an additional reduction in FVIII, FXI, and protein S activities. Significantly greater levels of prothrombin fragments 1 and 2, platelet factor 4, and neutrophil elastase were found in RP compared with AP. IgG was lower in AP compared with RP. Twice freezing and thawing caused a marked drop in FV, FVIII, and FXI activity.

CONCLUSIONS

Higher FVIII and F IX potencies in AP compared with RP can be expected to result in greater yields when used for purification of these clotting factors. AP is presumably more efficient than RP for treating coagulopathies. RP, however, may contain higher IgG levels than AP.

A sensitive and simple method for high-performance liquid chromatography (HPLC) determination of traces of chromium species in lake sediments after preconcentration by cloud point extraction (CPE) has been developed. Simultaneous preconcentration of Cr(III) and Cr(VI) in sediment samples was achieved by CPE with 1-(2-thiazolylazo)-2-naphthol (TAN) as the chelating agent and non-ionic surfactant octylphenoxypolyethoxyethanol (Triton X-114) as the extractant. Baseline separation of the TAN chelates of Cr(III) and Cr(VI) was realized on a RP-C(18) column by using a mixture of methanol-water (69:31, v/v) solution and 4.5 mmol L(-1) CTMAB buffered with 0.03 mol L(-1) NaAc-HAc solution (pH 5.5) as the mobile phase at a flow rate of 0.8 mL min(-1). The variables affecting the complexation and extraction steps were examined. The precision (R.S.D.) for seven replicate injections of a mixture of 100 microg L(-1) of Cr(III) and Cr(VI) was 1.2 and 0.9% for the retention time, 4.7 and 2.7% for the peak area, respectively. The concentration factor was 45 for Cr(III) and 40 for Cr(VI). The detection limit (LOD) of this method, calculated as three times the standard deviation of the blank signals was 7.5 microg L(-1) for Cr(III) and 3.5 microg L(-1) for Cr(VI), respectively. The proposed procedure was applied to the speciation of chromium in sediment samples with satisfactory results.

BACKGROUND

Since the demand for (18)F-fluorinated peptides for quantitative in vivo receptor imaging using PET has increased, a new chemoselective two-step (18)F-labeling strategy based on hydrazone formation between an unprotected hydrazine-functionalized peptide and an (18)F-labeled aldehyde was developed.

METHODS

First, 4-[(18)F]fluorobenzaldehyde ([(18)F]FB-CHO) was prepared from 4-formyl-N,N,N-trimethylanilinium triflate via direct no-carrier-added (18)F-fluorination (dimethyl sulfoxide, 90 degrees C, 5 min) and purified by RP-HPLC. Hydrazone formation between [(18)F]FB-CHO and 6-hydrazinonicotinic acid (HYNIC) and the unprotected HYNIC-functionalized peptides (HYNIC-d-Phe(1))-Tyr(3)-Thr(8)-octreotide and (HYNIC-Arg(1))-substance P was evaluated with respect to the dependence of radiochemical yield on pH, precursor concentration and temperature. The stability of [(18)F]FB-CH=N-HYNIC-Tyr(3)-Thr(8)(NH(2))-octreotide in aqueous solution at various pH (4.0, 5.5 and 7.5) as well as the in vivo stability of [(18)F]FB-CH=N-HYNIC-Tyr(3)-Thr(8)-octreotide in mouse blood (30 min p.i.) was investigated.

RESULTS

Yields of the hydrazone formation were independent of pH between pH 0.5 and 5.5. Optimal labeling yields of 85% were obtained with a precursor concentration of 2.1 mM at 70 degrees C for 10 min. The labeling products were stable at pH 7.5 at 37 degrees C, while in more acidic media (pH 4.0) the product slowly decomposed to form up to 31+/-2% [(18)F]FB-CHO within 5 h. Metabolite studies showed no detectable degradation of [(18)F]FB-CH=N-HYNIC-Tyr(3)-Thr(8)-octreotide in mouse blood (30 min p.i.).

CONCLUSIONS

In conclusion, chemoselective hydrazone formation between unprotected HYNIC-functionalized peptides and [(18)F]FB-CHO is a fast and straightforward radiolabeling method leading to high yields under mild acidic conditions. In addition, it represents a powerful and versatile radiolabeling strategy that is applicable to a variety of radionuclides and peptide precursors already available for (99m)Tc labeling.

BACKGROUND

Delirium is a common neuropsychiatric syndrome associated with poor outcomes. Evidence supports a neuroinflammatory etiology, but the role of the inflammatory marker C-reactive protein (C-RP) remains unclear. We investigated the relationship between C-RP and delirium and its severity as well as interaction with medical diagnosis.

METHODS

From an existing database (710 patients over 70 years old admitted to a Medical Acute Admissions Unit) we analyzed data which included C-RP levels, delirium (using the Confusion Assessment Method), and other clinical and demographic factors. Primary diagnoses were grouped (cardiovascular, musculoskeletal, infection, metabolic, and other).

RESULTS

There was a strong association between elevated C-RP and delirium (t = 5.09; p < 0.001), independent of other potential risk factors for delirium (odds ratio (OR) = 1.32 (95% CI: 1.10-1.58) p = 0.003). There was no significant association between C-RP and delirium severity, and between C-RP and delirium in the populations with cardiovascular disease, infection upon admission, or from the metabolic group despite an OR of 2.24 (95% CI: 0.92-5.45). There was an association in the musculoskeletal group (OR 2.19 (95% CI: 1.19-4.02)).

CONCLUSIONS

There is an association between elevated C-RP and delirium. This is strongest in patients admitted with musculoskeletal disease but not in others, implying that C-RP is involved in the genesis of delirium in musculoskeletal disease, but that other factors or processes may be more important in those with cardiovascular disease or infection.

Peripheral airways resistance (Rp) has been shown to be increased in asymptomatic asthmatic patients with normal spirometric values, and to be correlated with airways hyperresponsiveness to methacholine. We investigated whether Rp in asthmatic subjects with exercise-induced bronchospasm (EIB) would rise in response to cool, dry air. Using a wedged bronchoscope technique, we challenged an isolated lung segment with high flows (500 to 1,000 ml/min) of cool (22 degrees C) dry 5% CO2 in air for 5 min in eight asthmatic subjects with EIB and eight normal subjects. Baseline Rp and Rp following challenge were measured with saturated air at 37 degrees C at a flow rate of 100 ml/min. Baseline Rp was significantly greater in the asthmatic (0.09; [0.05 to 0.23] cm H2O/ml/min; median [interquartile range]) than in the normal subjects (0.05; [0.03 to 0.07] cm H2O/ml/min) (p = 0.04). The asthmatic, but not the normal subjects, had a significant absolute maximal increase in Rp following cool, dry air (0.10 [0.03 to 0.15] cm H2O/ml/min) (p < 0.01). In the asthmatic subjects, baseline Rp correlated with airways hyperresponsiveness to exercise (r = -0.76, p = 0.03). We conclude that the peripheral airways of asthmatic individuals with EIB are responsive to cool, dry air, and may play an important role in EIB.

OBJECTIVE

To compare the anxiolytic activities and flavonoid compositions of the two populations of the species Passiflora edulis, Passiflora edulis 'edulis' with purple fruit and Passiflora edulis 'flavicarpa' with yellow fruit.

METHODS

Four samples for each population of Passiflora edulis were collected from different districts of China. Swiss albino mice were used as experimental animals in elevated plus-maze (EPM) test to assay the anxiolytic effects of ethanol extracts of the samples. The conventional parameters and ethological items of the behavior of the mice were recorded and analyzed. Flavonoid compositions of the samples were analyzed by RP-HPLC monitored with diode array detection and the chromatograms were compared.

RESULTS

The ethanol extracts of the samples of Passiflora edulis 'flavicarpa' displayed anxiolytic activity at 400 mg/kg, while those of Passiflora edulis 'edulis' exhibited sedative effect at 400 mg/kg. The chromatograms of the samples belonging to similar population of Passiflora edulis were identical, but those belonging to different population were distinct from each other. The series of peaks between 16 and 24 min in the chromatograms of Passiflora edulis 'flavicarpa' did not appear in those of Passiflora edulis 'edulis', either did the peaks between 54 and 90 min in chromatograms of Passiflora edulis 'edulis' not appear in those of Passiflora edulis 'flavicarpa'. The six major flavonoid compounds isolated from the leaves of Passiflora edulis 'flavicarpa', lucenin-2, vicenin-2, isoorientin, isovitexin, luteolin-6-C-chinovoside, and luteolin-6-C-fucoside, had not been detected in Passiflora edulis 'edulis'.

CONCLUSIONS

Passiflora edulis 'flavicarpa' is extremely different from Passiflora edulis 'edulis' and they should be distinguished when pharmacological studies are performed on them. The aerial part of Passiflora edulis 'flavicarpa' is possible to be utilized as the resource of Passionflower Extract.

In the first of this three paper series, an in vitro latex coagulation was shown to arise from aggregation of rubber particles (RP) and lutoid membranes. RP aggregation was shown to be induced by a specific Hevea latex lectin-like protein (HLL) present on the lutoid membrane. In this second paper, a binding protein (BP) ligand counterpart for HLL was identified. This RP-HLLBP, having a specific interaction, with HLL was isolated from RP and characterized. The protein was extracted from the small RP in the presence of a surfactant (0.2% Triton-X-100) and further purified to homogeneity. Purification steps included acetone precipitation, heat-treatment, and column chromatography. The presence of RP-HLLBP was monitored by its ability to compete with erythrocytes in the hemagglutination inhibition (HI) assay. The purified RP-HLLBP had an HI titre of 1.37 microgml(-1), a pI value of 5.4, optimum activity at pH 5-8 and was thermostable up to 60 degrees C. On SDS-PAGE a single glycoprotein with M(r) of 24 kDa was detected while on native PAGE the major Mr was about 120 kDa. The purified RP-HLLBP was shown to inhibit latex coagulation. Chitinase, but no other glycosidase tested, abolished its HI action and inhibited HLL-induced RP aggregation in a competitive dose dependent manner. This indicated the presence of, and role for, N-acetylglucosamine residues in the binding recognition. The Hevea latex lectin-like protein can thus be referred to as a Hevea latex lectin. Based on protein identification by peptide mass fingerprinting, the RP-HLLBP was confirmed to be the small rubber particle protein (SRPP). This work has unambiguously determined the role of an intrinsic RP glycoprotein (RP-HLLBP or SRPP) as a key component in formation of the rubber latex coagulum.

Lamotrigine (LTG) is a novel triazine anticonvulsant currently undergoing clinical trials. LTG N-glucuronide, the major human metabolite of LTG, was isolated from human urine by means of XAD-2 column chromatography and semi-preparative HPLC. The structure of the suspected lamotrigine 2-N-glucuronide was proven by mass spectroscopy and NMR spectroscopy, along with chemical and enzymatic hydrolysis studies. High resolution fast atom bombardment mass spectrometry and Electrospray tandem mass spectrometry of the glucuronide gave an M+ ion at 432.0 amu and a fragment ion at 256.0 (M - 176)+ amu. The proton NMR of the glucuronide indicated the presence of a glucuronic acid moiety. A downfield anomeric proton (5.35-5.60 ppm) implied direct attachment to the aromatic triazine ring. Carbon-13 NMR of the glucuronide revealed an upfield shift (delta = -7.0 ppm) of the C-3 carbon of the triazine ring compared to LTG, indicating attachment of the glucuronide to the N-2 position. Chemical degradation or rearrangement of the glucuronide occurs at neutral pH to produce an unknown product (RP-1), while at basic pH a different unknown product (RP-2) is formed. The glucuronide is unusually stable at acidic pH. Treatment of the glucuronide with beta-glucuronidase resulted in hydrolysis to LTG, and enzymatic hydrolysis was inhibited by saccharo-1,4-lactone.

OBJECTIVE

To compare the predictive accuracy (PA) of existing models in estimating risk of biochemical recurrence (BCR) vs aggressive recurrence (BCR with a prostate-specific antigen, PSA, doubling time, DT, of <9 months).

METHODS

The study included 1550 men treated with radical prostatectomy (RP) between 1988 and 2007 within the Shared Equal Access Regional Cancer Hospital database. The PA of nine different risk stratification models for estimating risk of BCR and risk of aggressive recurrence after RP was assessed using the concordance index, c.

RESULTS

The 10-year risks of BCR and aggressive recurrence were 47% and 9%, respectively. Across all nine models tested, the PA was a mean (range) of 0.054 (0.024-0.074) points higher for predicting aggressive recurrence than for predicting BCR alone (c = 0.756 vs 0.702). Similar results were obtained in four sensitivity analyses: (i) defining patients with BCR but unavailable PSADT (220) as having aggressive recurrence; (ii) defining these patients as not having aggressive recurrence; (iii) defining aggressive recurrence as a PSADT of <6 months; or (iv) defining aggressive recurrence as a PSADT of <12 months. The improvement in PA was greater for preoperative than for postoperative models (0.053 vs 0.036, P = 0.03).

CONCLUSIONS

Across nine different models the prediction of aggressive recurrence after RP was more accurate than the prediction of BCR alone. This is probably because current models mainly assess cancer biology, which correlates better with aggressive recurrence than with BCR alone. Overall, all models had relatively similar accuracy for predicting aggressive recurrence.