Tinnitus characteristics in patients with hyperacusis and vertigo (including Ménière's disease) vs. hyperacusis alone.

INTRODUCTION AND OBJECTIVES: To compare clinical and psychoacoustic tinnitus characteristics in patients with the comorbidity of hyperacusis, hyperacusis and vertigo, and with Ménière's disease (MD). MATERIALS AND METHODS: Three hundred and twenty-nine tinnitus patients underwent audiological and otoneurological evaluation. Records of 94 individuals younger than 65 years, 40 women and 54 men (mean age 41.8, range 24-64 years), who complained of tinnitus and hyperacusis, were analyzed. One hundred and thirty-one ears with tinnitus were identified: 67 in the group of patients with tinnitus and hyperacusis (group 1; 41 patients); 28 in the group fulfilling criteria of MD diagnosis (group 2; 28); and 36 in the group with tinnitus, hyperacusis and typical symptoms of vertigo (group 3; 25). RESULTS AND CONCLUSIONS: Mean value of interaural difference in canal paresis in group 1 was 6.3%; in group 2: 23.7%; and in group 3: 25.9%; p<.001. Mean tinnitus pitch value was significantly lower in group 3 (1679Hz; SD=1139) and group 2 (2250Hz; SD=1162) compared to group 1 (4538Hz; SD=3123; p=.012). Values of tinnitus intensity and other characteristics did not significantly differ between the groups. Tinnitus and hyperacusis were most frequently preceded by acoustic trauma. Tinnitus coinciding with hyperacusis and vertigo was observed in patients after head trauma. Mean tinnitus pitch was lower in the groups of patients with hyperacusis and peripheral labyrinthine lesion than in tinnitus sufferers with hyperacusis alone. Tinnitus sufferers with low tinnitus pitch should undergo vestibular system evaluation. Hyperacusis and vertigo are likely comorbidities in tinnitus patients after head trauma. Hyperacusis may coincide in tinnitus patients after head trauma.

Tinnitus characteristics in patients with hyperacusis and vertigo (including Ménière's disease) vs. hyperacusis alone

Introduction and objectives: To compare clinical and psychoacoustic tinnitus characteristics in patients with the comorbidity of hyperacusis, hyperacusis and vertigo, and with Ménière's disease (MD).Materials and methodsThree hundred and twenty-nine tinnitus patients underwent audiological and otoneurological evaluation. Records of 94 individuals younger than 65 years, 40 women and 54 men (mean age 41.8, range 24–64 years), who complained of tinnitus and hyperacusis, were analyzed. One hundred and thirty-one ears with tinnitus were identified: 67 in the group of patients with tinnitus and hyperacusis (group 1; 41 patients); 28 in the group fulfilling criteria of MD diagnosis (group 2; 28); and 36 in the group with tinnitus, hyperacusis and typical symptoms of vertigo (group 3; 25).Results and conclusionsMean value of interaural difference in canal paresis in group 1 was 6.3%; in group 2: 23.7%; and in group 3: 25.9%; p<.001. Mean tinnitus pitch value was significantly lower in group 3 (1679Hz; SD=1139) and group 2 (2250Hz; SD=1162) compared to group 1 (4538Hz; SD=3123; p=.012). Values of tinnitus intensity and other characteristics did not significantly differ between the groups. Tinnitus and hyperacusis were most frequently preceded by acoustic trauma. Tinnitus coinciding with hyperacusis and vertigo was observed in patients after head trauma.Mean tinnitus pitch was lower in the groups of patients with hyperacusis and peripheral labyrinthine lesion than in tinnitus sufferers with hyperacusis alone. Tinnitus sufferers with low tinnitus pitch should undergo vestibular system evaluation. Hyperacusis and vertigo are likely comorbidities in tinnitus patients after head trauma. Hyperacusis may coincide in tinnitus patients after head trauma. (AU)

Introducción y objetivos: Comparar las características clínicas y psicoacústicas del tinnitus en pacientes con comorbilidad de hiperacusia, hiperacusia y vértigo, y con enfermedad de Ménière (EM).Materiales y métodosTrescientos veintinueve pacientes con tinnitus se sometieron a evaluación audiológica y otoneurológica. Se analizaron los registros de 94 pacientes menores de 65 años, 40 mujeres y 54 hombres (edad media 41,8, rango 24-64 años), que se quejaron de tinnitus e hiperacusia. Se identificaron 131 oídos con acúfenos: 67 en el grupo de pacientes con acúfenos e hiperacusia (grupo 1; 41 pacientes); 28 en el grupo que cumplía criterios de diagnóstico de enfermedad de Ménière (grupo 2; 28); y 36 en el grupo con acúfenos, hiperacusia y síntomas típicos de vértigo (grupo 3; 25).Resultados y conclusionesEl valor medio de la diferencia interaural en paresia del canal en el grupo 1 fue del 6,3%; en el grupo 2 del 23,7%; y en el grupo 3 del 25,9%; p<0,001. El valor medio del tono del tinnitus fue significativamente menor en el grupo 3 (1679Hz; DE=1139) y el grupo 2 (2250Hz; DE=1162), en comparación con el grupo 1 (4538Hz; DE=3123; p=0,012). Los valores de la intensidad del tinnitus y otras características no difirieron significativamente entre los grupos. El tinnitus y la hiperacusia fueron precedidos con mayor frecuencia por traumatismos acústicos. Se observó tinnitus coincidente con hiperacusia y vértigo en pacientes después de traumatismo craneoencefálico.El tono medio del tinnitus es menor en los grupos de pacientes con hiperacusia y lesión laberíntica periférica que en los que padecen tinnitus con hiperacusia. (AU)

The impact of transcription factors Znf1, Sip4, Adr1, Tup1, and Hap4 on xylose alcoholic fermentation in the engineered yeast Saccharomyces cerevisiae.

Lignocellulosic biomass is an attractive sustainable platform for fuel ethanol production. Xylose is a second after glucose most abounded sugar in lignocellulosic hydrolysates. Effective conversion of xylose to ethanol is one of key prerequisite for the development of an efficient conversion of biomass to ethanol. Engineered Saccharomyces cerevisiae strains are able to xylose fermentation. However, the yield and productivities of xylose fermentation remains lower in comparison with glucose fermentation. In this work, we studied impact of transcription factors Znf1, Sip4, Adr1, Tup1, and Hap4 on xylose catabolism. We have isolated znf1Δ, adr1Δ, tup1Δ and hap4Δ mutants, and strains overexpressing SIP4, ADR1 and HAP4 genes on the background of xylose-fermenting strain of S. cerevisiae aiming to explore involvement of these transcription factors in regulation of xylose growth and fermentation. It was shown that hap4Δ reveal 1.8-fold increase of ethanol production from xylose as compared to that of parental strain. The hap4Δ mutant accumulates 10.38 g l-1 of ethanol with an overall ethanol yield reaching 0.41 g g-1 of consumed xylose. While the other constructed strains revealed a decrease in ethanol production from this pentose.

The role of Mig1, Mig2, Tup1 and Hap4 transcription factors in regulation of xylose and glucose fermentation in the thermotolerant yeast Ogataea polymorpha.

Glucose is a preferred carbon source for most living organisms. The metabolism and regulation of glucose utilization are well studied mostly for Saccharomyces cerevisiae. Xylose is the main pentose sugar released from the lignocellulosic biomass, which has a high potential as a renewable feedstock for bioethanol production. The thermotolerant yeast Ogataea (Hansenula) polymorpha, in contrast to S. cerevisiae, is able to metabolize and ferment not only glucose but also xylose. However, in non-conventional yeasts, the regulation of glucose and xylose metabolism remains poorly understood. In this study, we characterize the role of transcriptional factors Mig1, Mig2, Tup1 and Hap4 in the natural xylose-fermenting yeast O. polymorpha. The deletion of MIG1 had no significant influence on ethanol production either from xylose or glucose, however the deletion of both MIG1 and MIG2 reduced the amount of ethanol produced from these sugars. The deletion of HAP4-A and TUP1 genes resulted in increased ethanol production from xylose. Inversely, the overexpression of HAP4-A and TUP1 genes reduced ethanol production during xylose alcoholic fermentation. Thus, HAP4-A and TUP1 are involved in repression of xylose metabolism and fermentation in yeast O. polymorpha and their deletion could be a viable strategy to improve ethanol production from this pentose.

The role of peroxisomes in xylose alcoholic fermentation in the engineered Saccharomyces cerevisiae.

Xylose is a second-most abounded sugar after glucose in lignocellulosic hydrolysates and should be efficiently fermented for economically viable second-generation ethanol production. Despite significant progress in metabolic and evolutionary engineering, xylose fermentation rate of recombinant Saccharomyces cerevisiae remains lower than that for glucose. Our recent study demonstrated that peroxisome-deficient cells of yeast Ogataea polymorpha showed a decrease in ethanol production from xylose. In this work, we have studied the role of peroxisomes in xylose alcoholic fermentation in the engineered xylose-utilizing strain of S. cerevisiae. It was shown that peroxisome-less pex3Δ mutant possessed 1.5-fold decrease of ethanol production from xylose. We hypothesized that peroxisomal catalase Cta1 may have importance for hydrogen peroxide, the important component of reactive oxygen species, detoxification during xylose alcoholic fermentation. It was clearly shown that CTA1 deletion impaired ethanol production from xylose. It was found that enhancing the peroxisome population by modulation the peroxisomal biogenesis by overexpression of PEX34 activates xylose alcoholic fermentation.

Discovery of Potent Inhibitors of 11ß-Hydroxysteroid Dehydrogenase Type 1 Using a Novel Growth-Based Protocol of in Silico Screening and Optimization in CONTOUR.

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 silico and optimize hits to discover 11ß-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 11ß-HSD1 with IC50 values of less than 1 µM in a biochemical enzyme assay. The best hit which exhibits an enzyme IC50 of 33 nM is further developed to a novel series of bicyclic 11ß-HSD1 inhibitors with the best inhibition of enzyme IC50 of 3.1 nM. The final lead candidate exhibits IC50 values of 7.2 and 21 nM in enzyme and adipocyte assays, respectively, displayed greater than 1000-fold of selectivity over 11ß-HSD2 and two other related hydroxysteroid dehydrogenases, and can serve as good starting points for further optimization to develop clinical candidates.

Discovery of BI 135585, an in vivo efficacious oxazinanone-based 11ß hydroxysteroid dehydrogenase type 1 inhibitor.

A potent, in vivo efficacious 11ß hydroxysteroid dehydrogenase type 1 (11ß HSD1) inhibitor (11j) has been identified. Compound 11j inhibited 11ß HSD1 activity in human adipocytes with an IC50 of 4.3nM and in primary human adipose tissue with an IC80 of 53nM. Oral administration of 11j to cynomolgus monkey inhibited 11ß HSD1 activity in adipose tissue. Compound 11j exhibited >1000× selectivity over other hydroxysteroid dehydrogenases, displays desirable pharmacodynamic properties and entered human clinical trials in 2011.

High incidence of intact or fragmented immunoglobulin in urine of patients with multiple myeloma.

In this prospective study we determined the incidence of intact/fragmented immunoglobulin and Bence Jones protein in urine immunofixation using Sebia reagents and HydrasysTM 2 apparatus and compared the results to concentrations of serum free light chains (FLC) assessed using Siemens BNTM II nephelometer and the immunoassay Freelite (Binding Site) in 289 patients with multiple myeloma at diagnosis. It was found that in one third of IgG, IgA and IgD myeloma patients, intact/fragmented immunoglobulin can be detected in urine and is connected with impaired renal function and reduced survival. Urine immunofixation detects monoclonal protein (FLC and intact/fragmented immunoglobulin) in 66-79% of IgG and IgA myeloma patients while serum FLC immunoassay detect it in 82-94% of IgG and IgA myeloma patients. However, the latter method is inadequate for detection of intact/fragmented immunoglobulin in urine. Serum FLC immunoassay and urine immunofixation are complementary methods in diagnosing and monitoring monoclonal protein in patients with myeloma.

Light Chain Escape in 3 Cases: Evidence of Intraclonal Heterogeneity in Multiple Myeloma from a Single Institution in Poland.

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.

Structure-based design and synthesis of 1,3-oxazinan-2-one inhibitors of 11ß-hydroxysteroid dehydrogenase type 1.

Structure based design led directly to 1,3-oxazinan-2-one 9a with an IC(50) of 42 nM against 11ß-HSD1 in vitro. Optimization of 9a for improved in vitro enzymatic and cellular potency afforded 25f with IC(50) values of 0.8 nM for the enzyme and 2.5 nM in adipocytes. In addition, 25f has 94% oral bioavailability in rat and >1000× selectivity over 11ß-HSD2. In mice, 25f was distributed to the target tissues, liver, and adipose, and in cynomolgus monkeys a 10 mg/kg oral dose reduced cortisol production by 85% following a cortisone challenge.

Flow cytometric immunophenotypic characteristics of plasma cell leukemia.

The aim of this prospective study was to define the flow cytometric characteristics of simultaneously investigated bone marrow and peripheral blood plasma cells antigens expression in 36 plasma cell leukemia (PCL) patients. The immunophenotypic profile of plasma cells was determined with a panel of monoclonal antibodies. The antigen expression intensity was calculated as relative fluorescence intensity (RFI). Bone marrow plasma cells showed expression of particular antigens in the following proportion of cases: CD49d 100%, CD29 94%, CD54 93%, CD44 83%, CD56 60%, CD18 26%, CD11b 29%, CD11a 19%, CD117 27%, CD71 30%, CD126 100% and CD19 0%, while the expression of those antigens on peripheral blood plasma cells was present in the following percentage of patients: CD49d 100%, CD29 96%, CD54 93%, CD44 95%, CD56 56%, CD18 50%, CD11b 53%, CD11a 29%, CD117 26%, CD71 28%, CD126 100% and CD19 0%. The expression of CD54 was significantly higher than that of adhesion molecules belonging to the integrin b2 family: CD11a, CD18 and CD11b, on both bone marrow and peripheral blood cells (p < 0.01). Expression of CD18, CD11a and CD11b was differential between two cell compartments: lower on bone marrow and higher on peripheral blood cells. We found that plasma cells in the bone marrow of patients with plasma cell leukaemia showed significantly greater granularity and size than those in the peripheral blood (p = 0.0001 and p = 0.04, respectively). However, no differences in cell size or granularity were revealed between bone marrow plasma cells from patients with PCL and multiple myeloma. In conclusion, impaired expression of adhesion molecules such as CD11a/CD18 (LFA-1) or CD56 may explain hematogenic dissemination characterizing PCL. The following pattern of adhesion molecule expression according to the proportion of plasma cells expressing a given antigen in peripheral blood and bone marrow and arranged in diminishing order may be established: CD49d > CD44 > CD54 > CD29 > CD56 > CD18 > CD11b > CD11a. Immuno-phenotyping of plasma cells in PCL, as in multiple myeloma, might be useful in detecting minimal residual disease in cases with aberrant antigen expression and for selecting therapeutic agents towards specific membrane targets.

Flow cytometric immunophenotypic characteristics of 36 cases of plasma cell leukemia.

Prospective flow cytometric analysis of antigens expression on bone marrow and peripheral blood plasma cells of 36 plasma cell leukemia (PCL) patients enabled to establish the following immunophenotype of leukemic plasma cell: CD38(++), CD138(+), CD54(+), CD49d(+), CD29(+), CD44(+), CD126(+), CD19(-), CD45(-). In one-third of patients PCL cells express CD56, CD71 and CD117. Expression of CD54 on plasma cells was higher as compared to expression of adhesion molecules CD11a, CD18 and CD11b (p<0.01). Expression of CD18, CD11a, CD11b was lower on bone marrow and higher on peripheral blood cells. In conclusion, impaired expression of adhesion molecules such as CD11a/CD18 or CD56 may explain hematogenic dissemination characterizing PCL.

Discovery and optimization of adamantyl carbamate inhibitors of 11ß-HSD1.

Synthesis of 2-adamantyl carbamate derivatives of piperidines and pyrrolidines led to the discovery of 9a with an IC(50) of 15.2 nM against human 11ß-HSD1 in adipocytes. Optimization for increased adipocyte potency, metabolic stability and selectivity afforded 11k and 11l, both of which were >25% orally bioavailable in rat.

Spirocyclic ureas: orally bioavailable 11 beta-HSD1 inhibitors identified by computer-aided drug design.

Structure-guided drug design led to the identification of a class of spirocyclic ureas which potently inhibit human 11beta-HSD1 in vitro. Lead compound 10j was shown to be orally bioavailable in three species, distributed into adipose tissue in the mouse, and its (R) isomer 10j2 was efficacious in a primate pharmacodynamic model.

Clinicopathological correlates of plasma cell CD56 (NCAM) expression in multiple myeloma.

The aim of this prospective, long-term study was to define the flow cytometric characteristics of plasma cell CD56 expression as well as determine the clinical characteristics of 204 multiple myeloma (MM) patients and 26 plasma cell leukemia (PCL) patients with regard to CD56 expression. CD56 expression intensity was determined by measurement of antigen molecules on the cell defined as Antibodies Bound per Cell (ABC) and calculation of Relative Fluorescence Intensity (RFI). CD56 expression was found in 66% of MM and 54% of PCL cases. The RFI values for individual MM patients ranged from 7.6 to 27.4 while ABC values on MM plasma cells from 2255 to 58469. There was a correlation between the proportion of all bone marrow CD38(++)/CD138(+) cells with CD56 expression and ABC and RFI indices. With regard to CD56 expression positive patients, the CD56(-) MM patients presented lower frequency of osteolysis (p = 0.01). The median survival was 48 months in CD56(+) patients and 43 months (p = 0.84) in CD56(-) cases. In conclusion, CD56 expression carries no distinct adverse prognosis and the lack of CD56 expression does not define a unique clinicopathological or prognostic entity in MM. A remarkable heterogeneity of CD56 expression intensity in CD56(+) patients imposes the necessity of determining CD56 expression intensity in candidates to antibody-based therapy.

Correlation of osteoprotegerin and sRANKL concentrations in serum and bone marrow of multiple myeloma patients.

INTRODUCTION: Recent studies suggest that multiple myeloma (MM) triggers osteoclastogenesis by disrupting the balance between the receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG), its natural antagonist. MATERIAL/METHODS: Determinations of bone marrow (BM) and serum OPG and sRANKL concentrations were performed in 133 MM patients and 42 healthy subjects by the ELISA method using Osteoprotegerin ELISA and sRANKL ELISA kits. RESULTS: MM patients had elevated serum levels of OPG compared with controls (p<0.0001) and OPG levels were higher in patients with renal failure and patients with hipercalcemia (p<0.001 and p=0.04, respectively). Serum OPG levels correlated with age, serum beta 2-microglobulin, and BM OPG concentrations and did not correlate with the presence of osteolysis or with stage of disease. sRANKL serum levels in MM patients and in controls were not statistically different (p=0.42). In MM patients, serum OPG and sRANKL levels were similar at diagnosis and in the plateau phase of disease. There was a correlation between BM and serum sRANKL concentrations (p<0.001). Median values of the sRANKL/OPG ratio for BM and serum of MM patients were 0.14 and 0.11, respectively. The median value of the sRANKL/OPG ratio for the serum of controls was 0.11. CONCLUSIONS: In 20% of MM patients, serum OPG levels are elevated, and this may be a compensative reaction related to increased bone destruction. There is not statistically significant relationship between sRANKL serum and BM levels and the main clinical and laboratory parameters of the disease. Determination of BM and the serum sRANKL/OPG ratio seems to have no clinical value.

C-kit receptor (CD117) expression on plasma cells in monoclonal gammopathies.

The surface expression of CD117 antigen (c-kit) on plasma cells from 158 multiple myeloma (MM), 12 plasma cell leukemia (PCL), 7 MGUS, 7 IgM lymphoplasmacytic lymphoma patients and 10 healthy subjects has been analyzed by flow cytometry using triple staining with the monoclonal antibodies CD138, CD117 and CD38. The antigen expression intensity was calculated as relative fluorescence intensity (RFI) and for direct quantitative analysis the QuantiBRITE test (Becton Dickinson) was applied. Antibody bounding capacity (ABC) was calculated using QuantiCALC software. CD117 antigen was present in 49/158 MM, 5/12 PCL and 5/7 MGUS patients. The RFI values ranged from 0.2 to 20.2 in particular MM patients (mean: 11.0+/-5.3; median 11.5) while the number of CD117 binding sites (ABC) on MM plasma cells ranged from 637 to 6217 (mean: 3029+/-1568; median 2946) (r=0.8328). In responsive to chemotherapy c-kit positive MM patients the percentage of CD117+ plasma cells in the bone marrow decreased significantly while in c-kit negative MM patients the percentage of CD117+ cells in bone marrow did not change and remained in the normal limits. When comparing the clinical and biological disease characteristics (monoclonal protein isotype, albumin, beta2-microglobulin, lactate dehydrogenase, stage of disease, response to chemotherapy, survival time) of c-kit positive and c-kit negative cases, no significant differences were found. In CD117 positive PCL cases expression of CD117 was detected in bone marrow plasma cells as well as in peripheral blood plasma cells. Normal plasma cells and those in IgM lymphoplasmacytic lymphoma did not show reactivity for the CD117 antigen. We conclude that it may be rationale to consider usefulness of therapy with tyrosine kinase inhibitors in the management of c-kit positive plasma cell proliferations. In one third of MM and PCL patients c-kit antigen could be considered as a "tumor associated marker" and together with CD38 and CD138 it may be of value for the identification of the malignant clone in minimal residual disease as it was first suggested by Spanish authors.