Clinicopathological response to neoadjuvant therapies and pathological complete response as a biomarker of survival in human epidermal growth factor receptor-2 enriched breast cancer - A retrospective cohort study.

BACKGROUND: Human epidermal growth factor receptor-2 (HER2) is overexpressed in 20-25% of breast cancers. Complete eradication of disease following neoadjuvant therapies and chemotherapy has been referred to as pathological complete response (pCR). AIMS: To determine clinicopathological predictors of pCR to neoadjuvant therapies and to evaluate pCR as a surrogate to enhanced survival. METHODS: Consecutive female patients with HER2 positive (HER+) breast cancer managed surgically in a single institution between 2005 and 2015 were included. Descriptive statistics and binary logistic regression were used to determine predictors of pCR. Appraisal of pCR as a predictor of survival was performed using Kaplan-Meier curves and Cox regression analysis. RESULTS: 451 patients were included with a mean age of 56.6 ± 13.4 years (range 23-95). Disease-free (DFS) and overall survival (OS) was 82.3% (371/451) and 82.6% (376/451) respectively with a median follow-up of 108.0 months (range 3-184.0). 118 were treated in the neoadjuvant setting (26.2%): tumour size <50 mm (Odds Ratio (OR): 12.156, P = 0.023) and progesterone receptor negativity (OR: 2.762, P = 0.008) independently predicted breast pCR, while ductal carcinoma (OR: 3.203, P = 0.030) and grade 3 disease (OR: 2.788, P = 0.018) predicted axillary pCR. Both breast and axillary pCR predicted enhanced DFS (Hazard Ratio (HR): 0.470 & HR: 0.449) and OS (HR: 0.383 & HR: 0.307). Axillary pCR independently predicted improved OS (HR: 0.326). CONCLUSION: pCR is sensitive biomarker and surrogate to survival outcomes in HER2+ breast cancer. Patients likely to achieve pCR may be predicted from traditional clinicopathological characteristics and molecular parameters.

Orchestrating the antioxidant defenses in the epididymis.

BACKGROUND: During the post-testicular maturation that occurs in the epididymis, spermatozoa need to face biochemical and morphological changes that may make them vulnerable to oxidative damage. During spermatogenesis and the epididymal maturation, the spermatozoon acquires antioxidant enzymes needed to face possible increases of reactive oxygen species (ROS) produced by its own aerobic metabolism but also due to ROS produced in high quantities by abnormal spermatozoa. OBJECTIVES: Provide an up-to-date review of the enzymatic antioxidant system in the epididymis. MATERIAL AND METHODS: A thorough literature review was performed for papers concerning the players of the antioxidant defenses in the epididymis. RESULTS: The antioxidant system in the epididymis is composed by superoxide dismutases, catalase, glutathione peroxidases, peroxiredoxins, glutathione-S-transferases, thioredoxins and thioredoxin reductase. They work together to maintain low levels of ROS during the epididymal maturation. Knockout models revealed that the absence of one of the enzyme impact sperm quality affecting a variety of proteins involved in motility, the ability to fertilize oocyte, and promotes oxidative damage to the sperm DNA. DISCUSSION AND CONCLUSIONS: These findings suggest that each enzyme is playing a specific role, and in most of the cases, no compensatory mechanisms are put in place when one enzyme is absent. This review highlights the different antioxidant enzymes in the epididymis and their role during maturation of the spermatozoon.

Characterization of sperm chromatin quality in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy.

BACKGROUND: Although the incidences of testicular cancer and Hodgkin's lymphoma have increased in young men over the past decade, combination chemotherapy has improved survival. As fertility is of importance to these patients, characterization of sperm chromatin structure is needed. We assessed sperm chromatin in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy, in comparison with control community and idiopathic infertile volunteers. METHODS: DNA damage was assessed with the sperm chromatin structure assay (SCSA), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and comet assays; reactive thiols (SH) and DNA compaction were determined with the monobromobimane (mBBr) and chromomycin A3 (CMA3) assays, respectively. RESULTS: Both testicular cancer (37%) and Hodgkin's lymphoma (81%) patients had normospermic samples with increased DNA damage, compared with controls. Cancer patients also had higher reactive thiols and CMA3 staining, indicating low DNA compaction. CONCLUSIONS: Sperm DNA integrity and compaction were affected in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy. Although SCSA, TUNEL and comet assays all detected DNA damage, the latter was optimal for use in cancer patients. A combination of the comet assay with tests that evaluate sperm DNA compaction, such as flow cytometry-based CMA3 and mBBr assays, is a reliable strategy to characterize sperm chromatin quality in cancer patients at the time of sperm banking.

The relationship between oestrogen and executive functioning in ALS females with emerging Frontotemporal Lobar Degeneration (FTLD) supports a neuroendocrine model of FTLD attenuation.

OBJECTIVE: The prevalence of ALS cognitive or behavioural impairment (ci or bi) consistent with Frontotemporal Degeneration (FTLD) approachs 50%, while ∼5-10% progress to dementia. Our goal was to explore ci and bi differencs between bulbar and limb onset, as well as the neuroprotective potential of oestrogen in emerging FTLD. METHODS: We applied Mann Whitney U to evaluate differences in cognitive and behavioural profiles between site of onset in 78 female and 83 male non-demented ALS participants classified by current consensus criteria with ci. For females, we also examined differences by oestrogen level. FINDINGS: Between group analyses found significantly worse Letter Fluency (LF) for bulbar onset, and worse Category Fluency (CF) for bulbar females. Significantly worse performance was found for low oestrogen females for LF and Similarities, with significantly worse LF for low oestrogen bulbar onset. No significant differences were found for behavioural subgroups, while moderate-severe range traits were higher in occurrence for bulbar and low oestrogen bulbar onset. CONCLUSIONS: Findings support our previously published mesocortical pathway associated "bottom-up" model of FTLD emergence in ALSbi, extending it with a hierarchal hypothesis involving ascending cerebellar pathways in ALSci and ALSbi, further suggesting a role for oestrogen in mitigating female FTLD progression.

Acrosome reaction in bovine spermatozoa: role of reactive oxygen species and lactate dehydrogenase C4.

After capacitation, mammalian spermatozoa accomplish the acrosome reaction (AR), a well-controlled exocytosis process crucial to fertilize mature oocytes that involves several protein kinases such as protein kinase A (PKA), C (PKC), and tyrosine kinase (PTK). Reactive oxygen species (ROS) are involved in both bovine sperm capacitation and AR. Lactate dehydrogenase C4 (LDH-C4) was associated with bovine and mouse sperm capacitation. Our aims were to study the participation of LDH-C4 to contribute with the status redox required for AR and the role of ROS in the regulation of PKA, PKC, and PTK involved in the exocytotic event. Sodium oxamate, an inhibitor of LDH-C4, prevented the AR induced by lysophosphatidylcholine (LPC) or NADH. Hydrogen peroxide promoted and superoxide dismutase (scavenger of superoxide), catalase (scavenger of hydrogen peroxide), diphenyleneiodinum, diphenyliodonium, cibacron blue, and lapachol (inhibitors of NADPH oxidase) prevented the AR, suggesting that ROS and a sperm oxidase are involved in the AR induced by these compounds. Inhibitors of PKA, PKC, and PTK also prevented the AR induced by LPC or NADH, suggesting the involvement of these kinases in the process. These results suggest that LDH-C4 may participate in the regulation of the redox status required to achieve the AR in bovine spermatozoa and that ROS are key elements in the regulation of protein kinases associated with the AR process.

Phase I and pharmacologic study of paclitaxel administered weekly in patients with relapsed ovarian cancer.

PURPOSE: Paclitaxel has shown significant activity in advanced ovarian cancer. In vitro studies with paclitaxel have suggested that fractionated brief infusion schedules may be more effective than the standard 24-hour infusion. We commenced a phase I evaluation of escalating-dose paclitaxel (40, 50, 60, 80, 100 mg/m2) administered weekly as a 1-hour infusion in patients with recurrent ovarian cancer. All patients had received prior paclitaxel and cisplatin therapy. All patients received standard premedication. PATIENTS AND METHODS: Eighteen patients are assessable on this phase I study. The mean age was 54 years (range, 48 to 74). The median number of prior chemotherapy regimens was three (range, two to five). The mean paclitaxel-free interval was 10.1 months (range, 1 to 24). RESULTS: A total of 194 cycles of therapy were administered, with a mean of 10 (range, one to 12) per patient. No mucositis or grade III neuropathy was seen. Alopecia occurred in one out of 18 assessable patients. The mean neutrophil nadir was 4.0 x 10(9)/L. At the top dose level (100 mg/m2) delivered, dose-intensity was 90.75% of that planned and greater than two fold the standard dose-intensity. Partial responses were seen in four of 13 assessable patients (30%). Two patients with progression of disease on standard three-week paclitaxel schedules switched to a weekly schedule with demonstrated response. Increasing paclitaxel dose correlated with measured area under the curve (AUC) (R2 = .614). Dose-limiting toxicity was reached at 100 mg/m2 with two of three patients experiencing a treatment delay, thus defining a maximum-tolerated dose of 80 mg/m2 in this group of heavily pretreated patients on this weekly schedule. CONCLUSION: (1) Paclitaxel administered as a 1-hour infusion is well tolerated; (2) this schedule of administration does not result in cumulative myelosuppression; and (3) this schedule of administration results in dose-intensive paclitaxel delivery with a favorable toxicity profile.

Phase II study of "dose-dense" high-dose chemotherapy treatment with peripheral-blood progenitor-cell support as primary treatment for patients with advanced ovarian cancer.

PURPOSE: We performed a pilot phase II study to evaluate the potential for delivery of rapidly sequenced high-dose chemotherapy treatments rescued with autologous peripheral-blood progenitor cells (PBP) in patients with previously untreated, advanced ovarian cancer. PATIENTS AND METHODS: A single cycle of mobilization was used, primed with cyclophosphamide (CPA)/paclitaxel (Txl) and filgrastim (granulocyte colony-stimulating factor [G-CSF]), followed by three cycles of high-dose carboplatin (CBDCA)/Txl and one cycle of high-dose melphalan (MEL), each rescued by PBP. We then analyzed the outcome for a total of 56 consecutive patients treated with high-dose chemotherapy as part of this program. RESULTS: In the phase II pilot, 21 patients were enrolled. There were no treatment-related deaths through 98 high-dose treatments, although 34 treatments were complicated by hospitalization, primarily for neutropenic fever. Seventy-six percent of patients experienced grade 3 to 4 gastrointestinal toxicity and 62% experienced grade 2 to 3 neuropathy. Five of 15 (33%) patients who underwent second-look surgery attained a pathologic complete response. In the overall analysis, 56 patients were reviewed. Forty-four patients were assessable for response by second-look surgery or clinical progression. Fifteen of 44 patients achieved a pathologic complete response (34%). The pathologic complete response rate in optimal-disease patients was 12 of 22 (55%), while only three of 22 (13%) suboptimal stage III and IV patients achieved a pathologic complete response. CONCLUSION: The Gynecologic Oncology Group has initiated a pilot phase II trial of this approach in patients with optimally debulked stage III ovarian cancer. There is no evidence to support the use of this or other aggressive regimens outside of a clinical trial.

Effect of anxiolytics and antidepressants on extinction and negative contrast.

Anxiolytics, particularly the benzodiazepines and barbiturates tend to retard, but not prevent, extinction, promote recovery from negative contrast, and elevate S- responding in discrimination training. Anxiolytics, administered during acquisition, tend to eliminate the partial reinforcement extinction effect, but this result is substantially influenced by parametric considerations. Behaviors that are energized in extinction may have a different pharmacological profile than behaviors that decline. Conclusions regarding the effects of antidepressants must be more tentative but, in general, acutely administered antidepressants are relatively ineffective in all of these paradigms. However, antidepressants may enhance the efficiency of responding on DRL schedules whereas anxiolytics tend to disrupt such behavior.

Crystals of an ATPase fragment of bovine clathrin uncoating ATPase.

A 44,000 Mr amino-terminal, clathrin-independent ATPase fragment of the bovine clathrin uncoating ATPase has been crystallized in a form suitable for X-ray diffraction studies. The crystals are orthorhombic, space group P2(1)2(1)2(1), a = 145.3 A, b = 65.0 A, c = 46.9 A, with one protein molecule per asymmetric unit (1 A = 0.1 nm).

Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways.

The effect of nitric oxide (NO*) on the capacitation rates of cryopreserved bull spermatozoa and the participation of protein kinases in the capacitation process were evaluated. A pool of spermatozoa from four bulls were incubated in TALP medium in the presence of heparin (10 IU/ml) or sodium nitroprusside (SNP, 0.05-100 microM), a NO* donor. The participation of NO* was confirmed by the use of scavengers, i.e. methylene blue (50,100 microM) and hemoglobin (20-40 microg/ml). The role of nitric oxide synthase in heparin-induced capacitation was evaluated using enzyme inhibitors Nomega-nitro-L-arginine methyl ester (L-NAME) and Nomega-nitro-L-arginine (L-NA) in concentrations ranging from 1 to 500 microM. The effects of protein kinase A (PKA), protein kinase C (PKC) and protein tyrosine kinase (PTK), on NO*-induced capacitation were evaluated by incubation with specific inhibitors of these enzymes (H-89, 50 microM; bisindolylmaleimide I, 0.1 microM and genistein, 3 microM). The role of hydrogen peroxide or superoxide anion in NO*-induced capacitation was evaluated by incubation with catalase (20-100 microg/ml) or superoxide dismutase (SOD, 0.05-0.5 mg/ml), respectively. Capacitation percentages were determined by the fluorescence technique with chlortetracycline (CTC). SNP concentrations employed had no effect on progressive motility or sperm viability. Capacitation values of the 0.05 microM SNP treatment (31 +/- 5.15%) were similar to those of heparin treated samples (33 +/- 4.27%). Inhibitors of nitric oxide synthase (NOS) diminished capacitation percentages in a dose-dependent manner as did the addition of NO*- scavengers (P <0.05). The presence of PKA, PKC and PTK inhibitors likewise decreased capacitation percentages (6.25 +/- 0.71, 12.75 +/- 1.41, 9.00 +/- 1.41%, respectively). The presence of catalase or SOD in the incubation medium had no effect on capacitation percentages. These results indicate that NO* may be generated by a sperm NOS during heparin-induced capacitation and that exogenous NO* acts as a capacitation inducer and involves the participation of PKA, PKC and PTK as part of the intracellular mechanisms that lead to capacitation in cryopreserved bull spermatozoa.

Dose escalation of paclitaxel with high-dose carboplatin using peripheral blood progenitor cell support in patients with advanced ovarian cancer.

A phase I study of escalating doses of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) given in combination with high-dose carboplatin was conducted to identify the antitumor efficacy and maximum tolerated dose of paclitaxel in patients who had received sequential cycles of paclitaxel/cyclophosphamide as prior treatment for ovarian carcinoma. Eighteen patients with advanced ovarian cancer were treated in this study. Induction therapy consisted of two cycles of cyclophosphamide 3.0 g/m2 plus high-dose paclitaxel 300 mg/m2 plus filgrastim and leukapheresis to harvest peripheral blood progenitor cells, followed by four courses of rapidly cycled high-dose carboplatin with planned dose escalation of paclitaxel (150, 200, 250, and 300 mg/m2) rescued with peripheral blood progenitor cells. The study was amended after accrual of 11 patients, and the remaining seven patients received a single cycle of induction therapy with paclitaxel/cyclophosphamide, followed by four courses of rapidly cycled high-dose carboplatin with planned dose escalation of paclitaxel through levels 200 and 250 mg/m2. All 18 patients have completed therapy. Of the 15 who are evaluable for response, the pathologic complete response was 33% (five of 15 patients). The administration of escalating doses of paclitaxel in combination with high-dose carboplatin following sequential cycles of paclitaxel/cyclophosphamide induction resulted in significant nonhematopoietic toxicity. Induction with a single cycle of paclitaxel/cyclophosphamide resulted in excellent progenitor cell mobilization, and significantly ameliorated the toxicity of this approach. The response rates thus far obtained are promising and warrant further evaluation.

Photodynamic action of aluminium phthalocyanine tetrasulphonate (A1PcS4) on smooth muscle: effects of thiols and a cyclic GMP analogue.

1. The smooth muscle system of the guinea-pig taenia caeci has been used in vitro to characterize the photodynamic action of aluminium phthalocyanine tetrasulphonate (A1PcS4) in the presence or absence of the thiol reductants L-cysteine (Cys), N-acetyl-L-cysteine (NAC), DL-dithiothreitol (DTT) or reduced glutathione (GSH). 2. In all photodynamic experiments the muscle was exposed to A1PcS4 (10(-5) M) for 30 min, followed by a 30 min washout period before photon irradiation at 32,000 lux (lambda > 570 nm) for 30 min. Photodynamic contractions were measured relative to the contractile response to carbachol (5 x 10(-5) M) and relaxation responses were determined in muscle precontracted with either carbachol 5 x 10(-5) M or KCl 23.5 mM. 3. Photon-activation of A1PcS4-sensitized smooth muscle evoked a triphasic response: an initial transient contraction and subsequent relaxation followed by a secondary sustained contraction. Cys 10 mM, NAC 10 mM and DTT 5 mM had no effect on the initial photodynamic contraction but significantly decreased the magnitude of the sustained contraction from mean values of 98% to 18%, 95% to 72% and 93% to 6% of the standard carbachol contraction (5 x 10(-5) M), respectively; GSH 10 mM was without significant effect on either the initial or sustained contraction. 4. In the absence of extracellular calcium the A1PcS4-sensitized smooth muscle did not respond to photon activation but re-introduction of calcium after cessation of illumination produced a sustained contraction which was markedly inhibited by Cys 10 mM. 5. In precontracted AlPcS4-treated muscle preparations photon activation produced a triphasic relaxation response, i.e. a rapid relaxation followed by a transient contraction and a secondary more sustained relaxation. The sustained phase of photodynamic relaxation was potentiated significantly by Cys 10 mM,NAC 10 mM, DTT 5 mM and GSH 10 mM, the relaxation being approximately doubled in magnitude from mean values of 34% to 68%, 30% to 73%, 34% to 68%, and 48% to 77%, respectively, relative to the standard carbachol (5 x l0-5 M) response.6. The cyclic GMP analogue, 8-(4-chlorophenylthio)-guanosine-3':5'-cyclic monophosphate (8-PCPTcGMP)(2 x 10-4 M) alone caused a triphasic relaxation response similar to that produced by photon activation of an AIPcS4-sensitized precontracted preparation in the presence of thiol reductants. The pattern of 8-PCPT-cGMP-induced relaxation was similar in muscle precontracted with carbachol 5 x 10-5M or KCI 23.5 mM.7. It is concluded that the rapid generation of reactive intermediates by photon-activation of boundAlPcS4 leads to membrane permeabilization, calcium entry and muscle contraction. These effects may be opposed by a direct stimulatory action of singlet oxygen on guanylate cyclase which is enhanced by the action of thiol reagents and mimicked by the cyclic GMP analogue, 8-PCPT-cGMP.

Effects of intrahippocampal administration of colchicine on incentive contrast and on radial maze performance.

The behavior of rats given intradentate injections of the neurotoxin colchicine was examined in three experimental settings. In Experiment 1, colchicine-treated, artificial cerebrospinal fluid (CSF)-treated, and untreated animals did not differ in the intake of 32% and 4% sucrose solutions, nor did they differ in degree of successive negative contrast when the 32% solution was changed to 4% sucrose. In Experiment 2, the colchicine-treated and CSF-treated animals did not differ in degree of suppression in the intake of a 0.15% saccharin solution when it preceded 32% sucrose in once-daily pairings (anticipatory contrast), nor did they differ in reversal performance when saccharin-sucrose and saccharin-saccharin pairings were reversed. In Experiment 3, the colchicine-treated animals were substantially impaired in radial-arm maze performance compared with CSF-treated controls. These results suggest that a completely functioning hippocampus is not necessary for the memory of reward quality, the comparison of rewards, the suppression of behavior when reward is decreased, the formation of associations between two levels of reward, and the reversal of this association, as long as these processes are reflected in consummatory behavior. The data are interpreted in terms of differences between instrumental behavior and sensory memory and/or consummatory behavior--an interpretation that is not incompatible with a deficiency in working memory in the animals with lesions.

Influence of ethanol on contrast in consummatory behavior.

Negative contrast that occurs when rats are shifted from 32% to 4% sucrose was reduced by IP injections of ethanol (1.0 g/kg) on postshift day 2, but not on postshift day 1. Smaller doses (0.25 and 0.5 g/kg) were ineffective, while larger doses (1.5 and 2 g/kg) produced sedation. A dose of 0.75 g/kg had effects similar to the 1.0 g/kg dose when administered on post-shift day 2. These results parallel those obtained with chlordiazepoxide and differ somewhat from amobarbital treatment.

Amobarbital sodium reduces successive gustatory contrast.

Amobarbital sodium (17.5 mg/kg) produced equivalent reductions in negative contrast when injected for the first time on either day 1 or 2 following a shift from 32% to 4% sucrose. These results differed from those obtained in earlier studies with chlordiazepoxide.

The effects of morphine in the consummatory contrast paradigm.

Rats shifted from 32% to 4% sucrose show a negative contrast effect, licking significantly less than animals that receive only 4% sucrose. The effects of morphine sulfate (0.5, 1.0, 2.0, 4.0, 8.0, and 16.0 mg/kg) on negative contrast were investigated in four experiments. Contrast was reduced on both the 1st and 2nd postshift day by the 4.0 and 8.0 mg/kg doses, but the effects were less robust than those seen with the benzodiazepines. The effects of morphine on contrast were dissociable from simple increases in sucrose consumption. Naloxone (0.25, 0.5, and 1.0 mg/kg) had no effect on contrast or sucrose intake. However, the contrast-reducing effect of morphine (4.0 mg/kg) was blocked by pretreatment with naloxone (0.50 mg/kg). The results are discussed in terms of other anxiolytic screening paradigms that have obtained "partial anxiolytic effects" using morphine.

Chlordiazepoxide and ethanol additively reduce gustatory negative contrast.

Negative contrast that occurs when rats are shifted from 32% to 4% sucrose has been shown to be reduced by chlordiazepoxide (CDP) and ethanol (ETOH). In a previous experiment, doses of 0.75 and 1.0 g/kg ETOH substantially reduced contrast while doses of 0.25 and 0.5 g/kg ETOH were much less effective. In this study, doses of 6 and 8 mg/kg CDP were shown to attenuate the negative contrast effect while smaller doses (2 and 4 mg/kg) influenced contrast to a lesser degree. Evidence for an additive effect of CDP and ETOH on contrast reduction was obtained when 4 mg/kg CDP and 0.5 g/kg ETOH were administered together.

Conditions under which chlordiazepoxide influences gustatory contrast.

Rats shifted from 32% sucrose to 4% sucrose consumed less 4% than animals without prior experience with 32% sucrose. The influence of chlordiazepoxide (CDP) on this successive negative contrast obtained in sucrose ingestion was investigated in four experiments. The results indicated that (1) rats injected with CDP during both preshift experience with 32% sucrose and post-shift experience with 4% sucrose showed an essentially unchanged contrast effect compared with saline-injected rats, (2) CDP injection for the first time on post-shift day 2 eliminated contrast but post-shift day 1 injections had little effect, (3) animals injected with CDP throughout preshift and switched to saline coincident with the sucrose shift showed a contrast effect at least as great as control animals, and (4) injections of CDP tended to elevate lick rate regardless of other conditions. These results indicate a disinhibitory effect of CDP and possible neophobia operating on the first post-shift day.

The effect of minor tranquilizers on psychomotor performance.

The effect of a standard daily regimen of chlordiazepoxide, prazepam (a new benzodiazepine tranquilizer), and placebo were examined in a three way double-blind comparison for a sample of normal volunteers. The criteria include a limited spectrum of psychomotor functions. A learning effect was conspicuous for all drugs on all criteria during the day-long sequence of ten trials. For the most part the differences between the drug groups were insignificant, but there were distinctive modifications in the performance of the chlordiazepoxide group. The clearest effect of the tranquilizer medications was found in the time estimation tests. Paradoxically, the drugs correct a naturally occurring perceptual error.