Phase II, randomized trial of preoperative epirubicin-paclitaxel +/- gefitinib with biomarker evaluation in operable breast cancer

PURPOSE: To evaluate the in vivo effect of adding gefitinib to preoperative chemotherapy on the EGFR-dependent p42/44 MAPK in operable breast cancer (BC) patients. Secondary aims: to evaluate EGFR, (p)-EGFR, Ki67, apoptotic index (TUNEL test) and VEGFR2 expression from baseline to surgery, percentage of pathologic complete response (pCR), and toxicity. PATIENTS AND METHODS: 90 patients with stage II-IIIA BC have been randomized to receive epirubicin 90 mg/sqm and paclitaxel 175 mg/sqm on day 1 plus: gefitinib 250 mg daily from day 5 to 16 (Arm A, intermittent), gefitinib 250 mg daily from day 1 to 21 (Arm B, continuous), or placebo (Arm C). Treatment plan: 4 courses every 3 weeks, followed by surgery. RESULTS: After preoperative therapy, 86/90 patients underwent surgery; 46 patients (51%) received breast conservative surgery. A pCR was observed in 4 patients. No significant differences in the expression of p42/44 MAPK, EGFR, (p)-EGFR, VEGFR2, proliferation index and apoptosis were observed comparing the combined Arms A + B vs C, and comparing Arm A vs B. Hematologic toxicities were not significantly different comparing Arms A + B vs Arm C, and comparing Arm A vs B. Significantly higher skin and mucosal toxicities were observed when comparing the two gefitinib Arms (A + B) vs Arm C (32% vs 9.6%, P = 0.018; 57% vs 29%, P = 0.009 respectively), while no significant differences were observed comparing Arm A vs B. CONCLUSION: Adding gefitinib to chemotherapy did not result in different effects on the EGFR-dependent pathway, proliferation, apoptosis and VEGFR2 expression as compared to placebo, while enhancing skin and mucosal toxicity. The two schedules of gefitinib (intermittent vs continuous) did not result in different biologic effects.     

Increased density of M1 receptors in the hippocampus of juvenile rats chronically deprived of NGF

Binding studies were used to assess the changes in affinity and/or number of M1 muscarinic receptors in hippocampi from juvenile rats chronically deprived of NGF. NGF deprivation was obtained by implanting into right ventricle at postnatal day 2 (P2) hybrydoma cells secreting high levels of monoclonal antibodies against NGF (αD11). Parenteral myeloma cells (P3U) were used as controls. Competition experiments were used to characterise the [³H]-PNZ binding sites in membrane preparations of hippocampi from rats sacrificed at P15. [³H]-PNZ bound M1 receptors both in P3U and αD11 group as shown by displacing potency order of antagonists: TLZ=4-DAMP>PNZ>p-F-HHSiD>MTC. The deprivation of NGF for two weeks significantly increased the number of M1 receptors without changing the K_i values of antagonists with exception of methoctramine which showed an increase in affinity in αD11 group. Similar changes in binding parameters were already observed after the first week of anti-NGF treatment. In contrast, a treatment for a week with implant at postnatal day 15 failed to produce any changes in M1 binding parameters. These results provide further physiological evidence for developmentally regulated modulatory role of NGF in the cholinergic function in the hippocampus.     

A novel compound heterozygous mutation in PYGM gene associated with McArdle’s disease

McArdle’s disease is an autosomal recessive glycogenosis due to mutation in the myophosphorylase gene (PYGM) resulting in a pure myopathy. The clinical onset typically occurs in childhood with cramps, myalgia, and intolerance to physical exercise, although late onset forms are also reported. We describe a case of a 17-year-old male complaining of cramps and myalgia following brief and intense exercise. The patient reported marked improvement in muscle fatigability few minutes after starting aerobic exercise. When he was a child, he had experienced few episodes of vomiting, nausea, and black colored urine following physical activity. Laboratory testings revealed high creatine kinase serum levels. Genetic testings for metabolic myopathies demonstrated a compound heterozygous for two PYGM mutations (p.R570Q and p.K754Nfs*49) allowing the diagnosis of McArdle’s disease. To date, 183 mutations in the PYGM gene are listed in Human Gene Mutation Database Professional 2021.2, but this novel compound heterozygosis has never been reported before.Key words: McArdle’s disease, glycogenosis, PYGM, hyperCKemia, second wind phenomenon     

Liver resection for hepatocellular carcinoma on cirrhosis: univariate and multivariate analysis of risk factors for intrahepatic recurrence

OBJECTIVE: To evaluate prognostic factors that could affect disease-free survival and recurrence after liver resection for hepatocellular carcinoma (HCC) on cirrhosis. SUMMARY BACKGROUND DATA: Tumor recurrence is the main cause of poor survival after liver resection for HCC on cirrhosis. METHODS: Two hundred twenty-four liver resections for HCC on cirrhosis were retrospectively reviewed. Univariate and multivariate analyses were performed on several clinicopathologic variables to analyze factors affecting long-term outcome and intrahepatic recurrence. The relation between preoperative aminotransferase level and recurrence rate was evaluated in the overall group, and separately in HCV-positive and in HBsAg-positive patients. Median follow-up was 35.6 months. RESULTS: The 1-, 3-, and 5-year overall survival rates were 83%, 62.8%, and 42.5%, respectively. The 1-, 3-, and 5-year disease-free survival rates were 70.3%, 43%, and 27.4%, respectively. The 1-, 3-, and 5-year recurrence rates were 20.8%, 38.6%, and 54.4% respectively. Tumor recurrence appeared in 93 patients (41.5%) and was the main cause of death in 51 patients (56%). Number of nodules, tumor capsule, microvascular portal vein thrombosis, and preoperative serum aspartate aminotransferase (AST) level significantly affected disease-free survival and recurrence rates. On multivariate analysis, single nodules and preoperative AST level less than twice normal (2N) were related to a better 5-year disease-free survival and lower tumor recurrence. In particular, among HCV-positive patients the recurrence rate was strongly affected by the preoperative AST level. CONCLUSIONS: Child A patients with single nodules are the best candidates for liver resection. Tumor recurrence is strictly linked to the status of the underlying liver disease, and a preoperative AST level equal to 2N seems to be a sensitive cutoff among patients with different risks of recurrence. HCV-positive patients with AST levels above 2N have the highest risk for intrahepatic recurrence and should be monitored carefully or offered alternative treatments.     

Interrelationship among thigh intermuscular adipose tissue,cross-sectional area,muscle strength,and functional mobility in older subjects

The aim of this cross-sectional study was to investigate the association between lower limb strength, muscle mass and composition, and balance ability in elders.Thirthy-four older participants (Age: 65.6 ± 4.73 years; male = 10 and female = 24) were assessed for muscle strength (maximal isometric strength of knee extensors and one repetition maximum by leg press, the one repetition maximum [1RM]), balance and gait capacity (Mini-BESTest), body composition by whole-body dual energy x-ray absorptiometry (obtaining Appendicular Skeletal Muscle Mass Index, ASMMI), and magnetic resonance imaging of thigh to evaluate Intermuscular Adipose Tissue (IMAT) and muscle Cross Sectional Area (CSA).Positive correlations between 1RM and ASMMI (r_s = 0.64, P < .0001) and thigh CSA (r_s = 0.52, P = .0017), but not with thigh IMAT, were found. In addition, significant correlations between knee extensors strength and ASMMI (r_s = 0.48, P = .004) and thigh CSA (r_s = 0.49, P = .0033) and IMAT (r_s = -0.35, P = .043) were observed, whereas no significant correlations between the Mini-BESTest with ASMMI, thigh CSA, and IMAT were observed.Lower limb strength positively correlated with appendicular muscle mass. Further, the maximal isometric strength of knee extensors negatively correlated with thigh IMAT in elderly patients, whereas the dynamic balance ability did not correlate with any of the morphological variables of the muscle (i.e., ASMMI, CSA, and IMAT). A reduced muscle size and strength could affect movement and reduce physical function in older patients. Improving the composition and size of muscle in elder subjects could reduce frailty and risk of falls.     

Fragrant bioethanol: A valued bioproduct from orange juice and essential oil extraction

We identify the composition of a less known, yet important bioproduct obtained in relatively high amount during orange fruit processing for juice and essential oil extraction comprised of bioethanol dissolving valued terpenes. Green cosmetic, perfume and biosolvent applications are anticipated.     

Hippocampal hypoglycaemia-activated K+ channels: single-channel analysis of glucose and voltage dependence

The effect of glucose on kinetics and the voltage-dependent characteristics of glucose-sensitive channels in hippocampal neurons were examined with the cell-attached mode of the patch-clamp technique. Recordings of a 100-pS K⁺ channel in the presence or absence of glucose demonstrate that the increase in channel open state probability (P _o) induced by glucose deprivation (40- to 400-times the control in high-glucose medium) was largely due to a decrease in the global amount of time spent by the channel in a long-lived closed state. TheP _o value of the same 100-pS channel was also found to increase (by approx. 80-times) following a depolarization of 40 mV from rest, the main factor responsible for this being a dramatic shortening of the long closed-times on depolarization. Another glucose-sensitive channel of smaller conductance (approx. 10 pS) showed a similar dependence ofP _o on glucose, but different dependence on voltage, with long openings at the same hyperpolarized potentials where the 100-pS channel was almost always closed. Our results indicate that the action of glucose on the kinetics of hippocampal channels closely resembles that of ATP-sensitive channels in pancreaticβ-cells. Furthermore, they indicate that the two types of glucose-sensitive channels found in hippocampal neurons, differing not only in their single-channel conductance but also in the dependence on voltage, could play different roles in the responses of these cells to modified energetic supply.