Neratinib and Ado-Trastuzumab-Emtansine for Pre-treated and Untreated HER2-positive Breast Cancer Brain Metastases: Translational Breast Cancer Research Consortium Trial 022.

PURPOSE: Treatment options for HER2-positive breast cancer brain metastases (BCBM) remain limited. We previously reported central nervous system (CNS) activity for neratinib and neratinib-capecitabine. Preclinical data suggest that neratinib may overcome resistance to ado-trastuzumab-emtansine (T-DM1) when given in combination. In TBCRC 022's cohort 4, we examined the efficacy of neratinib plus T-DM1 in patients with HER2-positive BCBM. PATIENTS AND METHODS: In this multicenter, phase II study, patients with measurable HER2-positive BCBM received neratinib 160 mg daily plus T-DM1 3.6 mg/kg intravenously every 21 days in three parallel-enrolling cohorts (cohort 4A-previously untreated BCBM, cohorts 4B and 4C- BCBM progressing after local CNS-directed therapy without [4B] and with [4C] prior exposure to T-DM1). Cycle 1 diarrheal prophylaxis was required. The primary endpoint was the Response Assessment in Neuro-Oncology-Brain Metastases (RANO-BM) by cohort. Overall survival (OS) and toxicity were also assessed. RESULTS: Between 2018-2021, 6, 17, and 21 patients enrolled to cohorts 4A, 4B, and 4C. Enrollment was stopped prematurely for slow accrual. The CNS objective response rate in cohorts 4A, 4B, and 4C was 33.3% (95% confidence interval [CI]: 4.3-77.7%), 35.3% (95% CI: 14.2-61.7%), and 28.6% (95% CI: 11.3-52.2%), respectively; 38.1-50% experienced stable disease for ≥6 months or response. Diarrhea was the most common grade 3 toxicity (22.7%). Median OS was 30.2 months (cohort 4A; 95% CI: 21.9, not reached [NR]), 23.3 months (cohort 4B; 95% CI: 17.6, NR), and 20.9 months (cohort 4C; 95% CI: 14.9, NR). CONCLUSION: We observed Intracranial activity for neratinib plus T-DM1, including those with prior T-DM1 exposure, suggesting synergistic effects with neratinib. Our data provide additional evidence for neratinib-based combinations in patients with HER2-positive BCBM, even those who are heavily pre-treated.

Long-term quality of life in patients with vestibular schwannoma managed with microsurgery.

OBJECTIVE: Little is known about the long term (greater than 10 years) quality of life in patients with vestibular schwannoma. This study aimed to evaluate long-term outcomes in patients with vestibular schwannoma. METHOD: A retrospective cohort study was performed across 2 academic institutions, with patients followed at least 10 years after vestibular schwannoma surgery (2000 to 2007). Telephone interviews were used to assess quality of life using the Glasgow Benefit Inventory and short form 12 item (version 2) health survey. RESULTS: A total of 99 out of 110 patients were included. Increasing age and symptom burden were associated with poorer quality of life (p = 0.01 and 0.02, respectively). The presence of imbalance, headache and facial nerve dysfunction were all associated with poorer quality of life scores (p = 0.01, 0.04 and 0.02, respectively). CONCLUSION: Identifying and managing post-operative symptoms may improve quality of life in vestibular schwannoma patients and can guide clinical decision making.

A pilot study of cabergoline for the treatment of metastatic breast cancer.

PURPOSE: The prolactin (PRL) receptor is over-expressed in breast cancer, and pre-clinical data indicate that it contributes to breast oncogenesis. Cabergoline is a potent dopamine receptor agonist of D2 receptors and has a direct inhibitory effect on pituitary PRL secretion. METHODS: A phase II study of cabergoline in patients with metastatic breast cancer was conducted. The primary end point of the study was to determine the clinical benefit rate (CBR) at 2 months. Eligible patients had tumors of any receptor status with no limit of prior lines of therapy. Measurable and unmeasurable diseases were allowed. Cabergoline 1 mg orally, twice weekly (1 cycle = 4 weeks) was given until disease progression or unacceptable toxicity. PRL receptor immunohistochemical staining was performed on available baseline tumor tissue; serial serum PRL levels were assessed. RESULTS: Twenty women were enrolled; 18 were evaluable for CBR. Tumor receptor status was distributed as follows: HR-any/HER2+ 2(10%), HR+/HER2- 18 (90%). The CBR was 33% (6/18), median progression free survival was 1.8 months, and median overall survival was 10.4 months. Two patients experienced disease control for over 12 months. Most common treatment-related adverse events were nausea (30%), fatigue (25%), and elevation in alkaline phosphatase (15%). Nine patients had baseline tissue for analysis; there was no association between baseline tumor PRL receptor expression and clinical benefit (p = 0.24). Change in serum PRL level and response were not correlated after 2 months of treatment (p = 0.64). CONCLUSION: Cabergoline was well tolerated, and while the ORR was low, a small subset of patients experienced extended disease control.

Developmental therapeutics for patients with breast cancer and central nervous system metastasis: current landscape and future perspectives.

Breast cancer is the second-leading cause of metastatic disease in the central nervous system (CNS). Recent advances in the biological understanding of breast cancer have facilitated an unprecedented increase of survival in a subset of patients presenting with metastatic breast cancer. Patients with HER2 positive (HER2+) or triple negative breast cancer are at highest risk of developing CNS metastasis, and typically experience a poor prognosis despite treatment with local and systemic therapies. Among the obstacles ahead in the realm of developmental therapeutics for breast cancer CNS metastasis is the improvement of our knowledge on its biological nuances and on the interaction of the blood­brain barrier with new compounds. This article reviews recent discoveries related to the underlying biology of breast cancer brain metastases, clinical progress to date and suggests rational approaches for investigational therapies.

Obtaining from grape pomace an enzymatic extract with anti-inflammatory properties.

Grape pomace, a winemaking industry by-product, is a rich source of bioactive dietary compounds. Using proteases we have developed an enzymatic process for obtaining a water-soluble extract (GP-EE) that contains biomolecules such as peptides, carbohydrates, lipids and polyphenols in soluble form. Of especial interest is its high polyphenol content (12%), of which 77% are flavonoids and 33% are phenolic acids. The present study evaluates in vitro the potential anti-inflammatory effect of GP-EE by monitoring the expression of inflammatory molecules on N13 microglia cells stimulated with lipopolysaccharide (LPS). GP-EE decreases the mRNA levels of the inflammatory molecules studied. The molecules under study were as follows: inducible nitric oxide synthase (iNOS), tumor necrosis factor- α (TNF-α), interleukin-1ß (IL-1ß), the ionized calcium binding adaptor molecule-1(Iba-1) and the Toll like receptor-4 (TLR-4), as well as the iNOS protein level in LPS-stimulated microglia. Our findings suggest that, as a result of its ability to regulate excessive microglial activation, GP-EE possesses anti-inflammatory properties. Therefore, acting as a chemopreventive agent, it may be of therapeutic interest in neurodegenerative diseases involving neuroinflammation. We can, therefore, propose GP-EE as a useful natural extract and one that would be beneficial to apply in the field of functional foods.

Antiproliferative and immunoactivatory ability of an enzymatic extract from rice bran.

The validation of natural products as source of functional foods or nutraceuticals has become an important issue in current health research. Thus, the present work has tested on MOLT-4 cells (human T cell acute lymphoblastic leukemic) the antiproliferative effect of a water-soluble enzymatic extract from rice bran (EERB). Present work shows that EERB induces cellular death in MOLT-4 cells in a dose-dependent way (0-10mg/mL) but not in non-tumoral lymphocytes. Flow cytometric analysis of MOLT-4 cells treated with EERB showed the presence of death cells by apoptosis rather than necrosis. Additionally, EERB also exerts an immunoactivatory effect on N13 microglia cells, by inducing TNF-alpha (tumour necrosis factor-α) expression, which plays a key role in the innate immune response to infection. Accordingly, we can propose EERB as a useful natural standardized extract with antiproliferative and immunoactivatory ability that would be beneficial to apply in the functional food field.

Protection against free radicals (UVB irradiation) of a water-soluble enzymatic extract from rice bran. Study using human keratinocyte monolayer and reconstructed human epidermis.

The antioxidant capacity of a water-soluble enzymatic extract from rice bran (EERB) has been tested in two cell models: keratinocyte monolayers and human reconstructed epidermis. Cells were incubated in culture medium in presence of different amounts of EERB and were UVB irradiated. Cell population assessment (MTT assay) and MDA (malonaldehyde) production were evaluated. The EERB did not induce cytotoxic effect for concentrations inferior or equal to 100 microg/mL. Human keratinocyte monolayers were protected of irradiation preventing 33% the lipid peroxidation process at concentration of 10 microg/ml of EEBR. In reconstructed human epidermis, 100 microg/mL decreased lipid peroxidation process by 44% (p<0.01) with regards to irradiated negative control. This effect was comparable to that of vitamin E at 600 microg/mL. Our data indicate that EERB is potentially able to efficiently counteract UVB-induced response. The EERB, diluted at 10% with water has very good skin compatibility. This product showed a sun protection factor of 4.8+/-0.3. Thus we can propose EERB as a useful natural standardized extract in skin photoprotection with promising applications in the field of dermatology.

Manejo endoscopico de las fístulas de líquido céfalo raquídeo del piso craneal anterior / Endoscopic treatment of cerebrospinal fluid fistulae of skull base

La fístula de líquido céfalo raquídeo (LCR) implica un quiebre de todas las barreras entre el espacio subaracnoideo y el tracto aéreo digestivo superior. Es una condición seria y fatal. La meningitis bacteriana es la mayor causa de morbilidad y mortalidad asociada a esta condición. Existen múltiples clasificaciones que tratan de sistematizar este problema, algunas de ellas son de escaso valor para el clínico. Existe una evolución en los métodos diagnósticos y terapéuticos en esta patología. Se presenta el marco teórico de la patología, evolución de la técnica quirúrgica, conceptos actuales en la reparación endoscópica de las fístulas de LCR y la experiencia quirúrgica desarrollada por los autores en 24 pacientes tratados en el Hospital Regional de Concepción durante el periodo abril 2001 a agosto 2008.

Cerebrospinal Fluid (CSF) fistulae involves a breakdown of all barriers that separates de subarachnoid space from the upper aero digestive tract. It is a serious and sometimes fatal condition. Meningitis is the mayor cause of morbidity anc mortality. There are many classifications trying to deal witt this issue, some of them with mean less clinical value. There is an evolution in diagnostic and therapeutic methods in this pathology We present the theoretical frame of this pathology, the evolution of the surgical technique, the current concepts in the endoscopic repair of CSF fistulae and the surgical experience developed by the authors in 24 patients treateo at the Hospital regional de Concepcion between April 2001 and August 2008.

Macrophage inducible nitric oxide synthase gene expression is blocked by a benzothiophene derivative with anti-HIV properties.

Nitric oxide (NO) has been shown to mediate multiple physiological and toxicological functions. The inducible nitric oxide synthase (iNOS) is responsible for the high output generation of NO by macrophages following their stimulation by cytokines or bacterial antigens. The inhibition of TNF alpha-stimulated HIV expression and the anti-inflammatory property of PD144795, a new benzothiophene derivative, have been recently described. We have now analyzed whether some of these properties could be mediated by an effect of PD144795 on NO-dependent inflammatory events. We show that PD144795 suppresses the lipopolysaccharide-elicited production of nitrite (NO(-)(2)) by primary peritoneal mouse macrophages and by a macrophage-derived cell line, RAW 264.7. This effect was dependent on the dose and timing of addition of PD144795 to the cells. Suppression of NO(-)(2) production was associated with a decrease in the amount of iNOS protein, iNOS enzyme activity and mRNA expression. The effect of PD144795 was partially abolished by coincubation of the cells with LPS and IFN gamma. However, the inhibitory effect of PD144795 was not abrogated by the simultaneous addition of LPS and TNF alpha, which indirectly suggests that the effect of PD144795 was not due to the inhibition of TNF alpha synthesis. Additionally, PD144795 did not block NF-kappa B nuclear translocation induced by LPS. Inhibition of iNOS gene expression represents a novel mechanism of PD144795 action that underlines the anti-inflammatory effects of this immunosuppressive drug.

Age-related changes in membrane lipid composition, fluidity and respiratory burst in rat peritoneal neutrophils.

The O2*(-) production has been studied in rat peritoneal neutrophils of different age (3, 12 and 24 months), in order to analyse whether the neutrophil respiratory burst is modified with increasing age. To stimulate NADPH oxidase, the enzyme responsible for the respiratory burst, two stimuli that act in different way have been used: phorbol myristate acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (N-FMLP). Production of O2*(-) decreased with age in neutrophils stimulated with N-FMLP (about 40%), but not in the stimulated with PMA. No difference in NADPH oxidase activity was found with age. The NADPH is supplied to the respiratory burst mainly by the pentose phosphate shunt. A progressive and significant decrease in the two most important enzymes of this route, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase, was detected as a function of age; in spite of this reduction, the NADPH produced by cells from old animals seems not limiting for the O2*(-) production. The N-FMLP-induced decrease in the O2*(-) production may be related to the age-dependent increase in the membrane fluidity observed. A decline in the cholesterol/phospholipid ratio and a rise in the total polyunsaturated fatty acids content were found, that correlated well with the increase in the membrane fluidity. The decrease (50%) of phosphatidylinositols in the 24-month-old animals may be also related to the age-impairment in the respiratory burst found after stimulation with N-FMLP. These studies suggest that the age-related alterations in neutrophil may result in diminished neutrophil function and increased susceptibility to infection in the ageing.

Liver lipid composition and antioxidant enzyme activities of spontaneously hypertensive rats after ingestion of dietary fats (fish, olive and high-oleic sunflower oils).

Hypertension is associated with greater than normal lipoperoxidation and an imbalance in antioxidant status, suggesting that oxidative stress is important in the pathogenesis of this disease. Although many studies have examined the effect of antioxidants in the diet on hypertensión and other disorders, less attention has been given to the evaluation of the role of specific dietary lipids in modulating endogenous antioxidant enzyme status. Previously, we have described that liver antioxidant enzyme activities may be modulated by consumption of different oils in normotensive rats. The purpose of the present study was to examine the effects of feeding different lipidic diets (olive oil, OO, high-oleic-acid sunflower oil, HOSO, and fish oil, FO) on liver antioxidant enzyme activities of spontaneously hypertensive rats (SHR). Plasma and liver lipid composition was also studied. Total triacylglycerol concentration increases in plasma and liver of animals fed on the HOSO and OO diets and decreases in those fed on the FO diet, relative to rats fed the control diet. The animals fed on the oil-enriched diet show similar hepatic cholesterol and phospholipid contents, which are higher than the control group. Consumption of the FO diet results in a decrease in the total cholesterol and phospholipid concentration in plasma, compared with the high-oleic-acid diets. In liver, the FO group show higher levels of polyunsaturated fatty acids (PUFA) of the (n - 3) series, in relation to the animals fed on the diets enriched in oleic acid. Livers of FO-fed rats, compared with those of OO- and HOSO-fed rats showed: (i) significantly higher activities of catalase, glutathione peroxidase and Cu/Zn superoxide dismutase; (ii) no differences in the NADPH-cytochrome c reductase activity. The HOSO diet had a similar effect on liver antioxidant enzyme activities as the OO diet. In conclusion, it appears that changes in the liver fatty acid composition due mainly to n - 3 lipids may enhance the efficiency of the antioxidant defence system and may yield a benefit in the hypertension status. The two monounsaturated fatty acids oils studied (OO and HOSO), with the same high content of oleic acid, but different content of natural antioxidants, had similar effects on the antioxidant enzyme activities studied.

Effects of two highly monounsaturated oils on lipid composition and enzyme activities in rat jejunum.

The effects of two monounsaturated fatty acid (MUFA) oils, olive oil (OO) and high-oleic sunflower oil (HOSO), with high content in oleic acid but differing in their non-fatty acid fraction, on brush-border membrane (BBM) lipid composition and fluidity and on mucosal enzyme activities of rat jejunum were studied. Animals were given semipurified diet with linoleic acid to prevent essential fatty acid deficiency (control group) or semipurified diet containing 10% of either OO or HOSO for 12 weeks. There was a significant decrease in the content of jejunal BBM phospholipids together with an increase in the level of free cholesterol in both oil-fed rats, when compared to control group. Although the increase in the BBM free cholesterol level was not statistically significant in HOSO-fed rats, a significant decrease in the phospholipid/free cholesterol ratio was found in both OO and HOSO-fed animals compared to control group. Rat jejunal BBM had a high level of free fatty acids which was increased in BBM isolated from OO and HOSO-fed animals. There was no statistical significant difference in the phospholipid distribution between the control and the OO group. However, HOSO-fed animals showed the lowest level of phosphatidylethanolamine together with the highest phosphatidylcholine content and the phosphatidylcholine/sphingomyelin ratio. The fatty acid pattern of jejunal BBM lipids was modified according to the major fatty acids in the oils. There was a decrease in both stearic acid (18:0) and linoleic acid (18:2 n-6), together with an increase in oleic acid (18:1 n-9) in jenunal BBM isolated from both oil experimental groups. All these results were accompanied by a significant increase in the BBM fluidity (as assessed by steady-state fluorescence polarization of diphenylhexatriene) isolated from oil-fed rat, when compared to control group. OO and HOSO-fed animals had the lowest activities of sucrase and maltase, while alkaline phosphatase activity only was decreased in HOSO-fed animals. The specific activity of maltase was not modified in any experimental rats. In summary, both MUFA oils induced similar effects on jejunal BBM lipid composition, fluidity, sucrase, maltase and lactase activities. Furthermore, HOSO intake resulted in a lowest alkaline phosphatase activity which was accompanied by changes in individual phospholipid composition. All these results suggest that effects of MUFA oils on jejunal BBM lipid composition and hydrolase activities are most likely due to the presence of high content of oleic acid rather than other components contained in the non-fatty acid of olive oil.

Effects of dietary fats (fish, olive and high-oleic-acid sunflower oils) on lipid composition and antioxidant enzymes in rat liver.

The effects of two oleic-acid-rich diets (containing olive oil, OO, and high-oleic-acid sunflower oil, HOSO) on plasma and liver lipid composition detoxification enzyme activities, were compared with those of a fish-oil (FO) diet and a control diet. Compared with the control diet, plasma and hepatic total triacylglycerol concentrations were increased in the animals fed on the HOSO and OO diets and decreased in those fed on the FO diet. The animals fed on FO showed the highest level of cholesterol in the liver and had lower plasma cholesterol concentrations when compared with those fed on the two oleic-acid-rich diets. In comparison with the animals fed on the diets enriched in oleic acid, the FO group showed higher hepatic levels of polyunsaturated fatty acids of the n-3 series and lower levels of fatty acids of the n-6 series. Livers of FO-fed rats, compared with those of OO- and HOSO-fed rats showed: (1) significantly higher activities of catalase (EC 1.11.1.6) glutathione peroxidase (EC 1.11.1.9) and Cu/Zn superoxide dismutase (EC 1.15.1.1); (2) no differences in the NADPH-cytochrome c reductase (EC 1.6.99.3) activity. The HOSO diet had a similar effect on liver antioxidant enzyme activities as the OO diet. In conclusion, it appears that changes in the liver fatty acid composition due mainly to n-3 lipids may enhance the efficiency of the antioxidant defence system. The two monounsaturated fatty acids oils studied (OO and HOSO), with the same high content of oleic acid but different contents of natural antioxidants, had similar effects on the antioxidant enzyme activities measured.

Changes in superoxide dismutase activity in liver and lung of old rats.

Superoxide dismutase activity was measured in liver and lung from 3 and 24 month-old rats. Both total SOD and Mn-SOD activity decreased significantly in the liver of old rats. Recent results from our laboratory have indicated that during aging, the activity of Cu/Zn-SOD decreases in rat liver and that there is an accumulation of altered protein. It was also shown that the old Cu/Zn-SOD had one histidine fewer than the young one. In the present study, the immunoprecipitation experiments showed that the amount of immunoprecipitable Mn-SOD from liver of old rats was greater than from young ones, but when amino acid residues were measured in purified young and old Mn-SOD from liver, no change was observed. In lung, no significant age-related differences in total SOD, Cu/Zn-SOD and Mn-SOD activity were found, nor was there accumulation of altered protein during aging.

Influence of the age and sex on respiratory burst of human monocytes.

The respiratory burst reaction has been studied in monocytes from men and women of different age. Phorbol myristate acetate (PMA) was used to stimulate NADPH oxidase. Superoxide anion production was found to be dependent on age and sex (it decreased 45% in men and 70% in women during aging).

Nitric oxide and superoxide anion production decrease with age in resident and activated rat peritoneal macrophages.

Superoxide anion and nitric oxide production have been studied in resident and activated peritoneal macrophages of 3-, 12-, and 24-month-old rats. Some key enzymes involved in the metabolism of glucose were also studied in relation to aging. Production of O2 and NO was reduced in all cases in middle-aged (12 months) and old (24 months) animals. Malic enzyme and citrate synthase activity shows a progressive reduction with age. Hexoquinase, pyruvate quinase, and lactate dehydrogenase activities decrease sharply from 3 to 12 months with no significant change between 12 and 24 months. Taken as a whole, the results of enzyme activity suggest that aging may reduce the capacity for glucose utilization in macrophages.

Respiratory burst is decreased by human hyperlipemic serum in rat peritoneal macrophages.

The effect of hyperlipemic human serum on superoxide anion (O2-) production by rat peritoneal macrophages was investigated. Phorbol myristate acetate (PMA)-stimulated O2- production was inhibited when cells were preincubated with hyperlipemic human serum. This inhibition was specifically carried out by a lipid fraction and was dependent on both cholesterol and triglyceride serum levels. This inhibitory effect was not exerted by a direct effect on NADPH-oxidase activity, nor by a putative superoxide dismutase activity present in the serum. With human neutrophils, we observed a decreased mobility of the cytosolic factor p47-phox to the membrane during the activation process, caused by hyperlipemic serum. We did not find any effect of hyperlipemic serum on NO2- production by cultured rat macrophages. These results suggest that a pathological increase of circulating plasma lipids may be associated with an impaired inflamatory capacity of macrophages.

Decrease in free-radical production with age in rat peritoneal macrophages.

The respiratory-burst reaction has been studied in rat peritoneal macrophages of different ages (3, 12 and 24 months) using phorbol 12-myristate 13-acetate (PMA) to stimulate NADPH oxidase. Production of O2-. and H2O2 decreased with age (about 50 and 75% respectively); however, no difference in NADPH oxidase activity was found. NO. production was also reduced with age (40%). Furthermore, a progressive and significant decrease in the pentose phosphate flux was detected as a function of age in control and PMA-stimulated macrophages. The NADPH/NADP+ ratio decreased with age in control and PMA-stimulated macrophages. Glucose uptake was lower in middle-aged (12 months) and old (24 months) animals but no differences were found between these groups.

Inhibitory effect of cyclosporin A and FK506 on nitric oxide production by cultured macrophages. Evidence of a direct effect on nitric oxide synthase activity.

Casein-elicited peritoneal macrophages from mice were cultured either alone or with interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS), and the effect of cyclosporin A (CsA) and FK506 on NO2- production (due technical difficulties NO2- was taken as the index for NO) was analysed. We observed an inhibitory effect of CsA and FK506 on NO2- production. The IC50 for NO2- production by casein-elicited macrophages was 0.1 microgram/ml for CsA and 0.3 microgram/ml FK506. The effect of both drugs was dose-dependent and was more clear in non-stimulated macrophages. The presence of IFN-gamma and LPS in the culture increased NO2- production by casein-elicited macrophages and partially eliminated the inhibition exerted by CsA and FK506. Both drugs acted directly on the nitric oxide synthase (NOS), since CsA and FK506 reduced by 35% and by 17%, respectively, NOS activity in the crude cytosolic fraction. However, CsA and FK506 did not alter 14CO2 production from [1-14C]glucose, suggesting that the pentose monophosphate pathway activity was not modified. These data add new insight into the interpretation of the immunosuppressive properties of both drugs.