Open-label, phase II, multicenter study of lasofoxifene plus abemaciclib for treating women with metastatic ER+/HER2- breast cancer and an ESR1 mutation after disease progression on prior therapies: ELAINE 2.

BACKGROUND: Acquired ESR1 mutations in estrogen receptor-positive (ER+) metastatic breast cancer (mBC) drive treatment resistance and tumor progression; new treatment strategies are needed. Lasofoxifene, a next-generation, oral, endocrine therapy and tissue-specific ER antagonist, provided preclinical antitumor activity, alone or combined with a cyclin-dependent kinase 4/6 inhibitor (CDK4/6i) in ESR1-mutated mBC. PATIENTS AND METHODS: In the open-label, phase II, ELAINE 2 trial (NCT04432454), women with ESR1-mutated, ER+/human epidermal growth factor receptor 2-negative (HER2-) mBC who progressed on prior therapies (including CDK4/6i) received lasofoxifene 5 mg/day and abemaciclib 150 mg b.i.d until disease progression/toxicity. The primary endpoint was safety/tolerability. Secondary endpoints included progression-free survival (PFS), clinical benefit rate (CBR), and objective response rate (ORR). RESULTS: Twenty-nine women (median age 60 years) participated; all but one were previously treated with a CDK4/6i (median duration 2 years). The lasofoxifene-abemaciclib combination was well tolerated with primarily grade 1/2 treatment-emergent adverse events (TEAEs), most commonly diarrhea, nausea, fatigue, and vomiting. One patient (with no prior CDK4/6i) discontinued treatment due to grade 2 diarrhea. No deaths occurred during the study. Median PFS was 56.0 weeks [95% confidence interval (CI) 31.9 weeks-not estimable; ∼13 months]; PFS rates at 6, 12, and 18 months were 76.1%, 56.1%, and 38.8%, respectively. CBR at 24 weeks was 65.5% (95% CI 47.3% to 80.1%). In 18 patients with measurable lesions, ORR was 55.6% (95% CI 33.7% to 75.4%). ESR1-mutant circulating tumor DNA (ctDNA) allele fraction decreased from baseline to week 4 in 21/26 (80.8%) patients. CONCLUSIONS: Lasofoxifene plus abemaciclib had an acceptable safety profile, was well tolerated, and exhibited meaningful antitumor activity in women with ESR1-mutated, ER+/HER2- mBC after disease progression on prior CDK4/6i. Observed decreases in ESR1-mutant ctDNA with lasofoxifene concordant with clinical response suggest target engagement. If the ELAINE 2 findings are confirmed in the initiated, phase III, ELAINE 3 trial, these data could be practice-changing and help address a critical unmet need.

Lasofoxifene versus fulvestrant for ER+/HER2- metastatic breast cancer with an ESR1 mutation: results from the randomized, phase II ELAINE 1 trial.

BACKGROUND: Acquired estrogen receptor alpha (ER/ESR1) mutations commonly cause endocrine resistance in ER+ metastatic breast cancer (mBC). Lasofoxifene, a novel selective ER modulator, stabilizes an antagonist conformation of wild-type and ESR1-mutated ER-ligand binding domains, and has antitumor activity in ESR1-mutated xenografts. PATIENTS AND METHODS: In this open-label, randomized, phase II, multicenter, ELAINE 1 study (NCT03781063), we randomized women with ESR1-mutated, ER+/human epidermal growth factor receptor 2 negative (HER2-) mBC that had progressed on an aromatase inhibitor (AI) plus a cyclin-dependent kinase 4/6 inhibitor (CDK4/6i) to oral lasofoxifene 5 mg daily or IM fulvestrant 500 mg (days 1, 15, and 29, and then every 4 weeks) until disease progression/toxicity. The primary endpoint was progression-free survival (PFS); secondary endpoints were safety/tolerability. RESULTS: A total of 103 patients received lasofoxifene (n = 52) or fulvestrant (n = 51). The most current efficacy analysis showed that lasofoxifene did not significantly prolong median PFS compared with fulvestrant: 24.2 weeks (∼5.6 months) versus 16.2 weeks (∼3.7 months; P = 0.138); hazard ratio 0.699 (95% confidence interval 0.434-1.125). However, PFS and other clinical endpoints numerically favored lasofoxifene: clinical benefit rate (36.5% versus 21.6%; P = 0.117), objective response rate [13.2% (including a complete response in one lasofoxifene-treated patient) versus 2.9%; P = 0.124], and 6-month (53.4% versus 37.9%) and 12-month (30.7% versus 14.1%) PFS rates. Most common treatment-emergent adverse events with lasofoxifene were nausea, fatigue, arthralgia, and hot flushes. One death occurred in the fulvestrant arm. Circulating tumor DNA ESR1 mutant allele fraction (MAF) decreased from baseline to week 8 in 82.9% of evaluable lasofoxifene-treated versus 61.5% of fulvestrant-treated patients. CONCLUSIONS: Lasofoxifene demonstrated encouraging antitumor activity versus fulvestrant and was well tolerated in patients with ESR1-mutated, endocrine-resistant mBC following progression on AI plus CDK4/6i. Consistent with target engagement, lasofoxifene reduced ESR1 MAF, and to a greater extent than fulvestrant. Lasofoxifene may be a promising targeted treatment for patients with ESR1-mutated mBC and warrants further investigation.

Rivastigmine in Parkinson's Disease Dementia with Orthostatic Hypotension.

OBJECTIVE: The purpose of this study was to evaluate if the cognitive benefit of rivastigmine is affected by the presence of orthostatic hypotension (OH) in patients with Parkinson's disease dementia (PDD). METHODS: We conducted a post hoc analysis on 1,047 patients with PDD from 2 randomized controlled trials comparing rivastigmine versus placebo at week 24 (n = 501) and rivastigmine patch versus capsule at week 76 (n = 546). A drop ≥ 20 mm Hg in systolic blood pressure (SBP) or ≥ 10 in diastolic blood pressure (DBP) upon standing classified subjects as OH positive (OH+); otherwise, OH negative (OH-). The primary end point was the Alzheimer's Disease Assessment Scale - Cognitive subscale (ADAS-Cog) at week 24 and the Mattis Dementia Rating Scale (MDRS) at week 76, using intention-to-treat with retrieved dropout at week 24 and observed cases at week 76, consistent with the original analyses. RESULTS: Overall safety was comparable between OH+ (n = 288, 27.5%) and OH- (n = 730, 69.7%), except for higher frequency of syncope (9.2%) in the OH+ placebo arm. The placebo-adjusted effect of rivastigmine on ADAS-Cog at week 24 was 5.6 ± 1.2 for OH+ and 1.9 ± 0.9 in OH- (p = 0.0165). Among subjects with OH, the MDRS change from baseline at week 76 was higher for rivastigmine capsules versus patch (10.6 ± 2.9 vs -1.5 ± 3.0, p = 0.031). The overall prevalence of OH was lower for rivastigmine than placebo at week 24 (28.3% vs 44.6%, p = 0.0476). INTERPRETATION: The cognitive benefit from rivastigmine is larger in patients with PDD with OH, possibly mediated by a direct antihypotensive effect. ANN NEUROL 2021;89:91-98.

Differential expression of interferon-induced genes and other tissue-based biomarkers in acute graft-versus-host disease vs. lupus erythematosus in skin.

BACKGROUND: In both acute graft-versus-host disease (GVHD) and lupus erythematosus (LE), the patient's own tissues are subjected to immunological assault via complex mechanisms influenced by interferon (IFN) and other cytokines. Although not typically confused clinically, these entities have overlapping histopathological findings in the skin. AIM: To assess whether GVHD can be differentiated from LE using molecular methods on skin specimens. METHODS: We developed a quantitative reverse transcription PCR assay based on previously identified tissue-based biomarkers of cutaneous GVHD, and compared gene expression in GVHD with that in LE. RESULTS: Both entities showed robust expression of IFN-induced genes and of genes encoding proteins involved in antigen presentation, cell signalling and tissue repair. Levels of gene expression differed significantly in GVHD compared with LE, particularly those of IFN-induced genes such as MX1, OAS3, TAP1 and STAT3 (P < 0.01). Three logistic regression models could differentiate the two entities with a high degree of certainty (receiver operating characteristic area under the curve of 1.0). CONCLUSION: The study demonstrates the feasibility of distinguishing between microscopically similar inflammatory dermatoses using tissue-based molecular techniques.

Short communication: Annatto in Cheddar cheese-derived whey protein concentrate is primarily associated with milk fat globule membrane.

The yellow color of Cheddar cheese whey arises from a residual amount of annatto that partitions into the whey during Cheddar cheese manufacture. Bleaching of the color using hydrogen peroxide or benzoyl peroxide is often a prerequisite to produce an acceptable neutral-colored whey protein concentrate and isolate. However, the use of these strong oxidizing agents often generates off-flavors as a result of lipid oxidation and results in loss of nutritive value due to protein oxidation. The objective of this study was to determine the extent of partitioning of annatto between protein, milk fat globule membrane (MFGM), and aqueous (serum) phases of cheese whey so that a simple method can be developed to remove annatto from cheese whey. The MFGM was separated from Cheddar cheese whey using a recently developed novel method. Quantitative analysis of the distribution of annatto in the fat-free whey protein isolate (WPI), the MFGM fractions, and the serum phase revealed that annatto was not bound to the protein fraction but was mostly distributed between the serum phase and the MFGM fraction. The results showed that a colorless WPI or whey protein concentrate could be produced from Cheddar cheese whey by separation of MFGM from the whey, followed by diafiltration. This approach will negate the need for using bleaching agents.

Experience of clomiphene use in a district general hospital.

Clomiphene citrate (CC) has been widely used for ovulation induction since the 1960s, due to its ease of administration and minimal side-effects. We conducted a retrospective study of 565 women who attended the infertility clinic at a district general hospital (Victoria Hospital, Kirkcaldy) between January 2004 and October 2008. The database was analysed for age, pregnancy rates, type of infertility and pregnancy outcomes. The conception rate with clomiphene citrate was 25%. The drug was well tolerated in 97.5% of women. The miscarriage rate with CC was 4.3%. We believe that these data give useful insights on CC therapy to gynaecologists involved in infertility clinics in a district general hospital setting.

Incompatibility of mixing of proteins in adsorbed binary protein films at the air-water interface.

Competitive adsorption of proteins from several binary protein solutions to the air-water interface has been studied. With a few exceptions, the equilibrium composition of the saturated monolayer of mixed protein films at various bulk concentration ratios did not follow a Langmuir-type competitive adsorption model. The deviation from ideal behavior results from incompatibility of mixing of proteins in the film at the air-water interface. This immiscibility alters the ratio of the binding affinity of the proteins in a protein 1/protein 2/water ternary film compared to that in a protein 1/water and protein 2/water binary film. A method to determine the extent of incompatibility between two proteins in a mixed protein film has been developed. It is shown that the incompatibility index derived for 19 protein 1/water and protein 2/water systems studied show a linear relationship with the absolute difference between Flory-Huggins protein-solvent interaction parameters, that is, /chi(1s) - chi(2)/, of the constituent proteins. On the basis of the evidence, it is theorized that, because of incompatibility, proteins in a mixed protein film at interfaces may undergo two-dimensional phase separation.

Lateral phase separation in adsorbed binary protein films at the air-water interface.

Lateral phase separation in two-dimensional mixed films of soy 11S/beta-casein, acidic subunits of soy 11 (AS11S)/beta-casein, and alpha-lactalbumin/beta-casein adsorbed at the air-water interface has been studied using an epifluorescence microscopy method. No distinct lateral phase separation was observed in the mixed protein films when they were examined after 24 h of adsorption from the bulk phase. However, when the soy 11S/beta-casein and AS11S/beta-casein films were aged at the air-water interface for 96 h, phase-separated regions of the constituent proteins were evident, indicating that the phase separation process was kinetically limited by a viscosity barrier against lateral diffusion. In these films, beta-casein always formed the continuous phase and the other globular protein the dispersed phase. The morphology of the dispersed patches was affected by the protein composition in the film. In contrast with soy 11S/beta-casein and AS11S/beta-casein films, no lateral phase separation was observed in the alpha-lactalbumin/beta-casein film at both low and high concentration ratios in the film. The results of these studies proved that proteins in adsorbed binary films exhibit limited miscibility, and the deviation of competitive adsorption behavior of proteins at the air-water interface from that predicted by the ideal Langmuir model (Razumovsky, L.; Damodaran, S. J. Agric. Food Chem. 2001, 49, 3080-3086) is in fact due to thermodynamic incompatibility of mixing of the proteins in the binary film. It is hypothesized that phase separation in adsorbed mixed protein films at the air-water and possibly oil-water interfaces of foams and emulsions might be a source of instability in these dispersed systems.

Incompatibility and Phase Separation in a Bovine Serum Albumin/beta-Casein/Water Ternary Film at the Air-Water Interface.

Thermodynamic incompatibility and two-dimensional phase separation in a bovine serum albumin (BSA)/beta-casein/water ternary film at the air-water interface has been studied using an epifluorescence microscopy technique. The incompatibility between BSA and beta-casein at the air-water interface was deduced from deviation of the experimental equilibrium composition of the proteins in the mixed saturated monolayer film from that predicted by the Langmuir-type competitive adsorption model at various bulk concentration ratios. Fluorescence microscopy of the mixed monolayer film showed distinct phase-separated BSA-rich and beta-casein-rich regions coexisting with inhomogeneous mixed regions. BSA always tended to be the dispersed phase and beta-casein the continuous phase. It is suggested that because the free energy at the "interfaces" between the phase-separated regions is generally higher than at other regions of the film, they may act as zones of instability in protein-stabilized foams and possibly emulsions. Copyright 2000 Academic Press.

Emulsifying properties of acidic subunits of soy 11S globulin.

The emulsifying properties of the acidic subunits (AS11S) isolated from soy glycinin (11S) have been studied. The isolated AS11S existed in solution mainly as a dimer species. Circular dichroic analysis indicated only a slight increase in aperiodic structure and no significant difference in beta-sheet structure when compared with those of soy 11S. At similar experimental conditions, the emulsifying properties of AS11S were superior to those of soy 11S and heat-denatured 11S. Emulsions prepared with 1% AS11S remained very stable without any visible oil separation for more than a month under gentle agitating conditions, whereas those prepared with 1% 11S collapsed and separated into phases within 2-3 days. The AS11S-stabilized emulsions were very stable below 0.15 M ionic strength. Studies on the rate of adsorption and surface tension reduction at the air-water interface showed that AS11S was significantly more surface active than soy 11S. It is proposed that, because the mass fraction of acidic subunits in soy 11S is approximately 60% and it is relatively easy to separate the acidic subunits from soy 11S, it may be industrially feasible to develop an economical process to isolate functional acidic subunits for use in emulsion-based food products.

Effect of transglutaminase-catalyzed polymerization of beta-casein on its emulsifying properties.

beta-Casein was polymerized to various extents by the transglutaminase-catalyzed cross-linking reaction. Under the reaction conditions used, dimer, trimer, and tetramer of beta-casein were produced as predominant species at short reaction times (20-90 min) and large polymers at longer reaction times (3-24 h). Examination of the emulsifying properties of these cross-linked beta-casein polymers showed that although the emulsifying activity index decreased, the storage stability of the emulsions increased with increasing degree of polymerization. Experiments with mixtures of native and polymerized beta-casein also showed that increasing the fraction of polymerized beta-casein in the mixture increased the emulsion stability. This enhancement of emulsion stability might be due either to an enhancement in steric stabilization, attributable to the branched nature of the trasgutaminase-catalyzed polymers, or to stronger cohesive interactions between polymerized beta-casein molecules than between monomeric beta-casein molecules in the film.

A New Methodology for Studying Protein Adsorption at Oil-Water Interfaces.

A new methodology has been developed for studying the adsorption behavior of proteins at oil-water interfaces. This technique employs the radiotracer method for monitoring adsorption of 14C-labeled proteins at the oil-water interface. The uniqueness of the new method lies in the formation of a 1000 Å thick triglyceride oil film on the water surface. beta-casein was used to generate a standard curve for relating interfacial radioactivity (µCi/m2) to cpm at the oil-water interface. Adsorption isotherm of beta-casein was determined in the bulk protein concentration range 1.5 x 10(-5)-3.8 x 10(-3)% by weight of solution. The saturated monolayer coverage was found to be about 7.3 mg/m2. This value was quite different from other values reported in the literature. Adsorption studies with another protein, lysozyme, at the oil-water interface also revealed a high surface concentration of 3.0 mg/m2. The most significant difference between the adsorption of beta-casein at the oil-water and air-water interfaces was the lack of an induction period for the development of interfacial pressure in the former. This difference may be attributed to the attractive dispersion interaction between protein molecules and the oil phase. Copyright 1998 Academic Press.

Stability of protease Q against autolysis and in sodium dodecyl sulfate and urea solutions.

Protease Q, a recently discovered subtilisin-like protease, exhibited unusual stability in 10% sodium dodecyl sulfate (SDS) and 8 M urea solutions. In 2% SDS it exhibited higher activity than the control, and in 10% SDS it retained 50% of its original activity when azocoll was used as substrate. Kinetic studies showed that the decrease in the activity of protease Q in SDS solutions followed a first order kinetics with a half-life of 446, 278, 78, and 24 h in 1%, 2%, 5%, and 10% SDS, respectively, at 25 degrees C. Protease Q was also stable against autolysis. In 20 mM Tris-HCl buffer (pH 8.3) containing 1.0 mM CaCl2, protease Q had a half life of 2822 h and 725 h at room temperature and at 37 degrees C, respectively. The enzyme was able to completely hydrolyze beta-lactoglobulin, beta-casein, and keratin in 8-10 M urea.

Trichotillomania: symptom or syndrome? A need for revision.

A 40-year-old religious Hindu male presented with his fourth episode of mania along with repetitive plucking of hair. This unusual behaviour was explained by him as a way to 'attain divinity by discarding the impure demoniac hair'. He had not shown this symptom in the previous episodes. With treatment of the mood disorder the hair plucking also disappeared. Hair removal has an important religious-cultural meaning in eastern India. Possible implication of the symptom is discussed. A case is made for reporting hair plucking as a symptom rather than a syndrome and it may at times represent a culture-specific symptom.

Isolation and characterization of titin T1 from bovine cardiac muscle.

Methods were developed for isolation of bovine cardiac titin in the apparently full-sized T1 form. The pure protein could be obtained either in the denatured state after chromatography of myofibril extracts on Sephacryl S-1000 or in the native state using hydroxylapatite chromatography. Titin-specific chicken polyclonal antibodies against denatured titin stained the I band regions and weakly stained the M line area of cardiac myofibrils but primarily stained the A-I junction positions in skeletal muscle myofibrils. Bovine cardiac titin contained 176 +/- 5 mumol of sulfhydryl groups per gram in the presence of 6 M guanidine hydrochloride or 0.1% SDS; only 45% of these groups were reactive under non-denaturing condition. Circular dichroism measurements indicated that titin contained approximately 53% beta-sheet and 47% random coil at pH 7.0. These results are consistent with the idea that titin is primarily composed of immunoglobulin C2 and fibronectin type III like motifs throughout its length, including the flexible I band region.