Gordana Prijic

Pharmacological characterization of rat amylin receptors: implications for the identification of amylin receptor subtypes

BACKGROUND AND PURPOSE Amylin (Amy) is an important glucoregulatory peptide and AMY receptors are clinical targets for diabetes and obesity. Human (h) AMY receptor subtypes are complexes of the calcitonin (CT) receptor with receptor activity‐modifying proteins (RAMPs); their rodent counterparts have not been characterized. To allow identification of the most clinically relevant receptor subtype, the elucidation of rat (r) AMY receptor pharmacology is necessary.

Tetracycline Treatment Retards the Onset and Slows the Progression of Diabetes in Human Amylin/Islet Amyloid Polypeptide Transgenic Mice

OBJECTIVE Aggregation of human amylin/islet amyloid polypeptide (hA/hIAPP) into small soluble β-sheet–containing oligomers is linked to islet β-cell degeneration and the pathogenesis of type 2 diabetes. Here, we used tetracycline, which modifies hA/hIAPP oligomerization, to probe mechanisms whereby hA/hIAPP causes diabetes in hemizygous hA/hIAPP-transgenic mice. RESEARCH DESIGN AND METHODS We chronically treated hemizygous hA/hIAPP transgenic mice with oral tetracycline to determine its effects on rates of diabetes initiation, progression, and survival. RESULTS Homozygous mice developed severe spontaneous diabetes due to islet β-cell loss. Hemizygous transgenic animals also developed spontaneous diabetes, although severity was less and progression rates slower. Pathogenesis was characterized by initial islet β-cell dysfunction followed by progressive β-cell loss. Islet amyloid was absent from hemizygous animals with early-onset diabetes and correlated positively with longevity. Some long-lived nondiabetic hemizygous animals also had large islet-amyloid areas, showing that amyloid itself was not intrinsically cytotoxic. Administration of tetracycline dose-dependently ameliorated hyperglycemia and polydipsia, delayed rates of diabetes initiation and progression, and increased longevity compared with water-treated controls. CONCLUSIONS This is the first report to show that treating hA/hIAPP transgenic mice with a modifier of hA/hIAPP misfolding can ameliorate their diabetic phenotype. Fibrillar amyloid was neither necessary nor sufficient to cause diabetes and indeed was positively correlated with longevity therein, whereas early- to mid-stage diabetes was associated with islet β-cell dysfunction followed by β-cell loss. Interventions capable of suppressing misfolding in soluble hA/hIAPP oligomers rather than mature fibrils may have potential for treating or preventing type 2 diabetes.

Isolation, Structural Elucidation, and Synthesis of Lepteridine From Ma̅nuka (Leptospermum scoparium) Honey

Ma̅nuka honey, made from the nectar of Leptospermum scoparium, has garnered scientific and economical interest due to its nonperoxide antibacterial activity. Biomarkers for genuine ma̅nuka honey are increasingly in demand due to the presence of counterfeit ma̅nuka honey. This work reports the identification of a compound previously unreported in ma̅nuka honey by HPLC, and determination of the structure of the as 3,6,7-trimethyllumazine using NMR, MS, IR, and UV/vis spectroscopy. This assignment was confirmed by total synthesis. The natural product, renamed lepteridine, was only observed in ma̅nuka honeys and could potentially serve as a biomarker for genuine ma̅nuka honey.

Leptosperin is a distinct and detectable fluorophore in Leptospermum honeys.

New Zealand manuka (Leptospermum scoparium) honey exhibits two unique fluorescence signatures that distinguish it from other honey types. One of these is the MM1 fluorescence marker (270-365nm excitation-emission) which we show is due to a Leptospermum nectar-derived compound, leptosperin. Synthetic or honey-purified leptosperin not only displayed an identical fluorescence spectrum, but supplementation of leptosperin into clover or artificial honeys generated the MM1 fluorescence signature. There was a quenching effect of the honey matrix on leptosperin fluorescence but otherwise leptosperin was chemically stable over prolonged storage at 37°C. Leptosperin was also present in the woody-fruited Australian Leptospermum species at elevated concentrations but virtually absent in Leptospermum subtenue suggesting its elevated expression developed following the mid-Miocene separation of the genus. These findings suggest that fluorescence spectroscopy could offer a rapid and high-throughput screening method for identification of Leptospermum honeys using the MM1 fluorescence marker.

Lepteridine as a unique fluorescent marker for the authentication of manuka honey

The recent discovery of two unique manuka marker fluorescence wavelengths (MM1 and MM2) potentially offers a rapid and cost-effective approach for manuka honey authentication using spectroscopy. The fluorophore responsible for the MM1 marker has been identified as leptosperin. We investigated whether lepteridine may be responsible for the MM2 fluorescence. We quantified the lepteridine in manuka honey and manuka nectar, which ranged between 5-52mg/kg and 80-205mg/kg, respectively. Notably, the fluorescent spectrum of synthetic lepteridine matched the MM2 fluorescence signature. Fluorescence quenching was observed in the honey matrix but otherwise, lepteridine was stable over prolonged storage at 37°C. Lepteridine was also found in Australian Leptospermum honeys and nectars. Lepteridine concentration was positively correlated with concentrations of the MM1 fluorescence marker leptosperin in honeys. These findings identify lepteridine as the principle compound responsible for MM2 fluorescence, and support the utility as a marker compound for manuka honey authentication.

A unique case of neural amyloidoma diagnosed by mass spectrometry of formalin-fixed tissue using a novel preparative technique

We report here a unique amyloidoma of the radial nerve which could not be subtyped by available techniques, including immunohistochemistry and standard clinical and laboratory evaluation. In order to identify the amyloid monomer, we developed a novel preparative procedure designed to optimize conditions for liquid chromatography tandem mass spectrometry analysis of formalin-fixed/paraffin-embedded (FFPE) tissue. Subsequent mass spectrometric analysis clearly identified kappa light chain as the monomer, with no evidence of lambda light chain. Manual interpretation of the matched spectra revealed no evidence of polyclonality. This study also enabled detailed characterisation of twelve likely amyloid matrix components. Finally, our analysis revealed extensive hydroxylation of collagen type I but, unexpectedly, an almost complete lack of hydroxylated residues in the normally heavily-hydroxylated collagen type VI chains, pointing to structural/functional alterations of collagen VI in this matrix that could have contributed to the pathogenesis of this very unusual tumour. Given the high quality of the data here acquired using a standard quadrupole-time of flight tandem mass spectrometer of modest performance, the robust and straightforward preparative method described constitutes a competitive alternative to more involved approaches using state-of-the-art equipment.

Fluorescence: A Novel Method for Determining Manuka Honey Floral Purity

Manuka honey, harvested from Leptospermum scoparium, is New Zealands most recog‐ nised honey type and commands a premium due to health‐related benefits. However, the plants distribution, relative to other species flowering simultaneously, allows hon‐ eybees to incorporate alternative nectars into the honey. Melissopalynological analysis in New Zealand is often unrepresentative due to the presence of many pollen‐bearing sources; consequently, alternative means of categorising manuka honey were examined. RP‐HPLC revealed that manuka honey contains distinct compounds, of which were rela‐ tively enriched and not present in the other New Zealand monofloral honeys. These main candidate compounds were isolated and have been described by mass spectrometry and nuclear magnetic resonance, synthesised to confirm structure, and as standards. These compounds, Leptosperin and Lepteridine, are a methyl syringate glycoside and pteridine derivative, respectively. Examination of these compounds revealed unique fluorescence signatures, this fluorescence could be detected in manuka honey samples the signal used to confirm that a honey was solely or predominantly consisted of L. scoparium nectar. Commercial manuka honeys were assessed by traditional analytical techniques, and comparisons were made with fluorescence signature; the fluorescence technique deter‐ mined the authenticity of the honeys accurately.

Human Amylin Transgenic Mice Show Elevated Core Body Temperature

Aberberga-Augskalne L PP087 Abou-Raya A PP059 Abou-Raya S PP059 AbuElKheir H PP059 Ackermans M OP21 Adamu V PP068 Aechundia-Cabanez H PP021 Afrashteh B PP001 Agapakis D PP043 Agrawal R OP25, PP010, PP091 Ahmed M PP084 Ahn C PP022, OP30 Aitken J OP02 Aitken J OP15 Aivars J PP087 Akbaba G PP035 Akbarov PP085, PP086 Akbarov Z PP062, PP063 Akbarzadeh M PP029 Akubude D PP068 Al Bannai F PP084 Al Daghri N PP016 Alashek W PP060 Albassas A PP012 Alebouyeh M PP029 Alexe O PP030 Alexia K PP019 Alexiadou K PP029, OP61, PP099 Alfadda A PP012, PP016 Alfieri A OP06 Alhammad A PP016 Alikhanova N PP062, PP063, PP085, PP086 Alnaamy M PP012 Alomaim W PP012 Amashukeli M OP23 Andreucci A OP33 Angelakas I PP079 Angelopoulos P PP031 Arambasic J OP01 Aranza-Doniz C PP020 Archundia-Cabanez H PP070 Argyrakopoulou G OP29 Aronne L OP31 Arribas C OP18 Asatiani K OP23, PP076 Aslanoglou D PP080, PP079, PP078 Avgeri A OP03, OP05