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With the continuous progress in ultralarge virtual libraries which are readily accessible, it is of great interest to explore this large chemical space for hit identification and lead optimization using reliable structure-based approaches. In this work, a novel growth-based screening protocol has been designed and implemented in the structure-based design platform CONTOUR. The protocol was used to screen the ZINC database in silicoand optimize hits to discover 11β-HSD1 inhibitors. In contrast to molecular docking, the virtual screening process makes significant improvements in computational efficiency without losing chemical equities through partitioning 1.8 million ZINC compounds into fragments, docking fragments to form key hydrogen bonds with anchor residues, reorganizing molecules into molecular fragment trees using matched fragments and common substructures, and then regrowing molecules with the help of developed intelligent growth features inside the protein binding site to find hits. The growth-base screening approach is validated by the high hit rate. A total of 50 compounds have been selected for testing; of these, 15 hits having diverse scaffolds are found to inhibit 11β-HSD1 with IC50values of less than 1 μM in a biochemical enzyme assay. The best hit which exhibits an enzyme IC50of 33 nM is further developed to a novel series of bicyclic 11β-HSD1 inhibitors with the best inhibition of enzyme IC50of 3.1 nM. The final lead candidate exhibits IC50values of 7.2 and 21 nM in enzyme and adipocyte assays, respectively, displayed greater than 1000-fold of selectivity over 11β-HSD2 and two other related hydroxysteroid dehydrogenases, and can serve as good starting points for further optimization to develop clinical candidates.

The neurotrophins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), and NT4/5 are all found in the developing cerebellum. Granule cells, the major target neurons of mossy fibers, express BDNF during mossy fiber synaptogenesis. To determine whether neurotrophins contribute to the development of cerebellar afferent axons, we characterized the effects of neurotrophins on the growth of mossy fiber neurons from mice and rats in vitro. For a mossy fiber source, we used the basilar pontine nuclei (BPN), the major source of cerebellar mossy fibers in mammals. BDNF and NT4/5 increased BPN neuron survival, neurite outgrowth, growth cone size, and elongation rate, while neither NT3 nor NGF increased survival or outgrowth. In addition, BDNF and NT4/5 reduced the size of neurite bundles. Consistent with these effects, in situ hybridization on cultured basilar pontine neurons revealed the presence of mRNA encoding the TrkB receptor which binds both BDNF and NT4/5 with high affinity. We detected little or no message encoding the TrkC receptor which preferentially binds NT3. BDNF and NT4/5 also increased TrkB mRNA levels in BPN neurons. In addition to previously established functions as an autocrine/paracrine trophic factor for granule cells, the present results indicate that cerebellar BDNF may also act as a target-derived trophic factor for basilar pontine mossy fibers. © 1999 John Wiley & Sons, Inc. J Neurobiol 40: 254–269, 1999

Body temperature and metabolic responses to 2 h treadmill exercise in dogs given glucose intravenously (25–30 mg·kg-1· min-1 throughout the run) were compared with those measured in the same animals with elevated plasma FFA concentrations (soya bean oil ingestion+intravenous heparin) and in control experiments (24 h fasting). In comparison with control conditions enhanced glucose availability for the working muscles caused a reduction in the exercise-induced increases in both rectal (by 0.9± 0.11° C) and muscle (by 0.9±0.16° C) temperatures, a lower rate of oxygen uptake (by 16%) and an elevated respiratory exchange ratio. A tendency towards enhanced body temperature responses to exercise, accompanied by increases inVO2 and cardiac frequency was noted in dogs with elevated plasma FFA concentrations as compared with the control animals. The estimated amount of heat effectively dissipated from the body, expressed as a fraction of heat load (thermoregulatory efficiency) was significantly higher in dogs infused with glucose (0.962±0.0035), than in the controls (0.947± 0.0043) and those with elevated plasma FFA concentrations (0.931±0.0029).

Thermoregulatory reactions evoked by selective preoptic-anterior hypothalamic (PO/AH) heating in conscious rabbits were associated with significant changes in renal function. Urine flow rate decreased from a control value of 0.92±(S.E.) 0.08 to 0.47±0.07 ml/min after 10–20 min of heating, urine osmolality increased from 273±34 to 417±46 Osm/kg H2O, and free water clearance per 100 ml GFR decreased from 1.11±0.46 to -0.50±0.23 ml/min. These changes were followed by a gradual recovery despite continued heating. Clearances of exogenous creatinine and p-aminohippurate fell transiently during the first 10 min of heating and then returned to normal.

To compare clinical and psychoacoustic tinnitus characteristics in patients with the comorbidity of hyperacusis, hyperacusis and vertigo, and with Ménière's disease (MD).

We report three cases of light chain escape (LCE) at a single institution in Poland, including an interesting case of biclonal monoclonal gammopathy of undetermined significance (MGUS) that satisfied the criteria for progression to light chain multiple myeloma (LCMM) with a rapid rise in serum free light chain (FLC) levels, following steroidal treatment for simultaneous temporal artery inflammation and polymyalgia rheumatica (PMR). In the three cases discussed, progression of the disease by light chain escape was associated with rapid and severe renal impairment, highlighting the necessity for prompt detection of such free light chain-only producing clones in order to prevent the possible development of irreversible end-organ damage. Interestingly, monitoring of these three patients by serum free light chain assay (sFLC) and retrospective heavy/light chain analysis (HLC) detected this clonal evolution prior to clinical relapse and suggests that these assays represent important additional tools for more accurate monitoring of multiple myeloma patients.

Multiple myeloma (MM) and its benign precursor monoclonal gammopathy of undetermined significance (MGUS) are diseases characterised by the production of monoclonal immunoglobulins. Clonal heterogeneity in MM has become a well-accepted phenomenon; however dogma would suggest the proteins produced by these clones remain consistent. Free light chain (FLC) escape is one exception to this rule, but is comparatively poorly documented and to our knowledge has not been identified in MGUS patients. Here we report 2 cases of MM patients with intact immunoglobulin and FLC producing clones that have different sensitivities to treatment leading to escape. In addition we report an MGUS patient whose routine corticosteroid treatment for polymyalgia rheumatica (PMR) impacted on the intact immunoglobulin but not the FLC producing clones and led to an escaping FLC clone which was subsequently diagnosed as myelomic.

Multiple myeloma (MM) and its benign precursor monoclonal gammopathy of undetermined significance (MGUS) are diseases characterised by the production of monoclonal immunoglobulins. Clonal heterogeneity in MM has become a well-accepted phenomenon; however dogma would suggest the proteins produced by these clones remain consistent. Free light chain (FLC) escape is one exception to this rule, but is comparatively poorly documented and to our knowledge has not been identified in MGUS patients. Here we report 2 cases of MM patients with intact immunoglobulin and FLC producing clones that have different sensitivities to treatment leading to escape. In addition we report an MGUS patient whose routine corticosteroid treatment for polymyalgia rheumatica (PMR) impacted on the intact immunoglobulin but not the FLC producing clones and led to an escaping FLC clone which was subsequently diagnosed as myelomic.Serum protein electrophoresis (SPE) and immunofixation electrophoresis (IFE) were performed using Hydrasys 2 apparatus (SEBIA).  Serum free light chain (FLC) levels were measured nephelometrically on a Siemens BNTM  II using polyclonal antisera assays, Freelite®(The Binding Site Group Limited, Birmingham, UK).Two intact immunoglobulin MM patients (Patient 1 IgAκ: 14.4g/L, κ FLC: 1320 mg/L, age: 65, sex: male; Patient 2 IgAλ: 8g/L, λ FLC: 9510 mg/L, age: 48, sex: male) were monitored through the course of their disease for 762 and 1330 days respectively. Following cyclophosphamide, thalidomide and dexamethasone (CTD) treatment Patient 1 achieved a VGPR (∼90% reduction in IgAk and 65% reduction in FLC) which was stable for 270 days. Subsequently whilst only a trace of IgAκ was present, dFLC levels increased from 460mg/L to 15194mg/L. Patient 2 achieved a CR following treatment with vincristine, doxorubicin and dexamethasone (VAD) and autologous stem cell transplant (ASCT) which was stable for 330 days. As with Patient 1 relapse was characterised by a substantial increase in dFLC from 8.61mg/L to 3168mg/L. In both patients the velocity of change and sensitivity to treatment of the intact immunoglobulin and FLC suggested the presence of heterogeneous clones.A rare biclonal MGUS patient, IgGκ (3.9g/L) and λ FLC (316mg/L), was identified during routine laboratory investigations (age: 71, sex: female) and followed annually in accordance with local guidelines (low/moderate risk MGUS). 16 months following MGUS diagnosis the patient was started on oral methylprednisolone for PMR. The treatment resolved the PMR and coincidently caused a reduction in the IgGκ serum concentration (trace quantities) and a normalisation of the FLC κ/λ ratio. A year post steroidal treatment whilst the IgGκ monoclonal protein concentration remained stable, dFLC levels increased from 9.6 mg/L to 1052 mg/L (κ/λ ratio: 0.008), indicating the emergence of a λ FLC clone although the patient remained asymptomatic. 4 months later and almost 4 years following diagnosis, the patient progressed to symptomatic disease with severe renal impairment (creatinine 6.19 mg/dL; eGFR 7.03 ml/min/1.73m2) anemia (Hb 9.0g/dL) and 70% clonal plasma cells present in the bone marrow. The dFLC concentration had further increased to 9726 mg/L however the IgGκ monoclonal protein was no longer detectable by IFE indicating the biclonal MGUS had progressed to a λ light chain multiple myeloma.Routine monitoring of MM patients to detect FLC escape is recommended by international guidelines, in light of the MGUS patient FLC escape leading to MM, we suggest routine evaluation of FLC levels in MGUS may also be beneficial.Endean: The Binding Site Group Ltd: Employment. Harding:The Binding Site: Employment.

Endean: The Binding Site Group Ltd: Employment. Harding:Binding Site: Employment.

Abstract 5008