Ovariectomy Enhances Acetylcholinesterase Activity But Does Not Alter Ganglioside Content in Cerebral Cortex of Female Adult Rats

In the present work we investigated the effect of ovariectomy on acetylcholinesterase (AChE) activity and ganglioside content in cerebral cortex of female rats. We also studied the activity of butyrylcholinesterase (BuChE) in serum of these animals. Adult Wistar rats were divided into three groups: (1) naive females (control), (2) sham-operated females and (3) castrated females (ovariectomy). Thirty days after ovariectomy, rats were sacrificed by decapitation without anaesthesia. Blood was collected and the serum used for BuChE determination. Cerebral cortex was homogenized to determine AChE activity and extracted with chlorophorm:methanol for ganglioside evaluation. Results showed that rats subjected to ovariectomy presented a significant increase of AChE activity, but did not change the content and the profile of gangliosides in cerebral cortex when compared to sham or naive rats. BuChE activity was decreased in serum of rats ovariectomized. Our findings suggest that the alteration in the activity of brain AChE, as well as serum BuChE activity caused by ovariectomy may contribute to the impaired cognition and/or other neurological dysfunction found in post-menopausal women.     

Tacrine(10)-hupyridone,a dual-binding acetylcholinesterase inhibitor,potently attenuates scopolamine-induced impairments of cognition in mice

Tacrine(10)-hupyridone (A10E) was designed as a dual-binding acetylcholinesterase (AChE) inhibitor from the modification of tacrine and a fragment of huperzine A. We have found that A10E effectively inhibited AChE in a mixed competitive manner, with an IC₅₀ of 26.4 nM, which is more potent than those of tacrine and huperzine A. Most importantly, we have shown, for the first time that A10E attenuated scopolamine-induced cognitive impairments without affecting motor function in mice. A10E effectively attenuated impairments of learning and memory to a similar extent as donepezil, an inhibitor of AChE used for treating Alzheimer’s disease (AD). In addition, A10E significantly decreased AChE activity in the brain of mice, suggesting that A10E might cross the brain blood-barrier. Taken together, our results demonstrated that A10E, a designed dual-binding AChE inhibitor, could effectively reverse cognitive impairments, indicating that A10E might provide therapeutic efficacy for AD treatment.     

Rivastigmine in the treatment of Alzheimer’s disease: an update

Alzheimer’s disease is the most common form of dementia in industrialized countries. In the European Union, about 54% of dementia cases are believed to be due to Alzheimer’s disease. The condition is an age-related neurodegenerative disorder characterized by multiple cognitive deficiencies, including loss of memory, judgment, and comprehension. These manifestations are accompanied by behavioral and mood disturbances. Although no cure has yet been discovered for Alzheimer’s disease, symptomatic therapies are now widely available and offer significant relief to patients and benefits to caregivers in terms of reduced care burden. At the start of the 21st century, health technology assessments recommended three agents for the symptomatic treatment of mild to moderate Alzheimer disease: rivastigmine, donepezil, and galantamine. Rivastigmine (Exelon^®, Novartis Basel—Switzerland) is a slowly reversible inhibitor of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE), while donepezil (Aricept^®, Pfizer, New York, USA) and galantamine (Reminyl^®, Janssen, New Jersey,USA) show no functional inhibition of BuChE, and are considered AChE-selective, rapidly-reversible inhibitors. The efficacy of all three agents has been evaluated in large, double-blind, placebo-controlled clinical trials of up to 6 months’ duration. Rivastigmine treatment in mild to moderate Alzheimer’s disease improves cognition, activities of daily living, and global function.     

Supplementation with vitamins E plus C or soy isoflavones in ovariectomized rats: effect on the activities of Na<Superscript>+</Superscript>, K<Superscript>+</Superscript>-ATPase and cholinesterases

Since a previous study demonstrated that ovariectomized rats present an activation of Na⁺, K⁺-ATPase and acetylcholinesterase (AChE) activities, in the present study we investigated the influence of vitamins E plus C or soy isoflavones on the effects elicited by ovariectomy on the activities of these enzyme in hippocampus of ovariectomized rats. We also determined the effect of the same compounds on the reduction of serum butyrylcholinesterase (BuChE) activity caused by ovariectomy. Female adult Wistar rats were assigned to one of the following groups: sham (submitted to surgery without removal of the ovaries) and ovariectomized. Seven days after surgery, animals were treated for 30 days with a single daily intraperitoneous injection of vitamins E (40 mg/kg) plus C (100 mg/kg) or saline (control). In another set of experiments, the rats were fed for 30 days on a special diet with soy protein or a standard diet with casein (control). Rats were sacrificed after treatments and the hippocampus was dissected and serum was separated. Data demonstrate that vitamins E plus C reversed the activation of Na⁺, K⁺-ATPase and AChE in hippocampus of ovariectomized rats. Conversely, soy protein supplementation reversed the increase of AChE activity, but not of Na⁺, K⁺-ATPase activity, caused by ovariectomized group. Neither treatment was able to reverse the reduction of serum BuChE activity. Furthermore, treatments with vitamins E plus C or soy were unable to reverse the decrease in estradiol levels caused by ovariectomy. Our findings show that the treatment with vitamins E plus C significantly reversed the effect of ovariectomy on hippocampal Na⁺, K⁺-ATPase and AChE activities. However, a soy diet that was rich in isoflavones was able to reverse just the increase of AChE. Neither treatment altered the reduction in serum BuChE activity. Taken together, these vitamins and soy may have a protective role against the possible brain dysfunction observed in some menopause women. Vitamins E plus C and soy isoflavones may be a good alternative as a novel therapeutic strategy.     

Homocysteine Inhibits Butyrylcholinesterase Activity in Rat Serum

In the present work we investigated the in vitro effects of homocysteine (Hcy) and methionine (Met), metabolites accumulated in homocystinuria, on butyrylcholinesterase (BuChE) activity in rat serum. We also studied the kinetics of the inhibition of BuChE activity caused by Hcy. For determination of BuChE we used serum of 60-day-old Wistar rats, which was incubated in the absence (control) or presence of Hcy (0.01–0.5 mM) or Met (0.2–2.0 mM). The kinetics of the interaction of Hcy and BuChE was determined using the Lineweaver–Burk double reciprocal plot. Results showed that serum BuChE activity was not altered by Met, but it was significantly inhibited (37%) by 500 M Hcy, a concentration similar to those found in blood of homocystinuric patients. The apparent K _m values, in the absence and presence of 500 M of Hcy, were 0.034 and 0.142 mM, respectively, and V _max of BuChE for acetylcholine (ACh) as substrate was 1.25 mol ACSCh/h/mg of protein. The K _i value obtained was 120 M, and the inhibition was of the competitive type, suggesting a common binding site for Hcy and ACh. It is proposed that inhibition of cholinesterase activity may be one of the mechanisms involved in the neurological dysfunction observed in homocystinuria.     

Retreatment of patients with chronic hepatitis C,subtype 1b and cirrhosis,who failed previous direct‐acting antiviral therapy including first‐ and second‐generation NS5A inhibitors with ombitasvir/paritaprevir/ritonavir,dasabuvir + sofosbuvir + ribavirin

The use of direct‐acting antiviral agents (DAAs) in patients with chronic HCV genotype 1 infection results in sustained virologic response (SVR) rates of 95%‐97%, but 3%‐5% of patients experience virologic failure. We observed 17 patients infected with HCV subtype 1b who failed previous treatment with DAA, including 13 subjects (76.5%) with liver cirrhosis. Twelve subjects (70.6%) previously received NS5A inhibitors of the first generation (ledipasvir or daclatasvir) and five subjects (29.4%) – the second generation (velpatasvir). All patients were retreated with a combination of ombitasvir/paritaprevir/ritonavir and dasabuvir (3D) with sofosbuvir (SOF) and ribavirin (RBV). We compared SVR₁₂ rates depending on fibrosis stage, presence of just single or double NS5A mutations (L31M/V/I and/or Y93H), and on the generation of previously used NS5A inhibitor. Observed SVR₁₂ rates were as follows: 94.1% (16/17 patients) overall; 100% in patients without cirrhosis (n = 4) vs 92.3% in those with cirrhosis (n = 13); 100% with single L31M/V/I or Y93H mutation (n = 7) vs 88.9% with double mutations (n = 9); 100% in patients who previously failed first generation (n = 12) vs 80.0% in those failed second‐generation NS5A inhibitors (n = 5). Retreatment with 3D + 0SOF + RBV was highly effective and safe in patients with chronic HCV GT1b infection who failed previous use of NS5A inhibitors. Fibrosis stage, baseline presence of NS5A RAS mutations and the generation of previously used NS5A inhibitors may impact the probability of achieving SVR₁₂, but statistical significance was not demonstrated in our small retrospective cohort. Further studies in a larger population are needed to confirm or not the predictive value of these baseline factors.     

Protease inhibitors and indoleamines selectively inhibit cholinesterases in the histopathologic structures of Alzheimer disease.

Neurofibrillary tangles and amyloid plaques express acetylcholinesterase and butyrylcholinesterase activity in Alzheimer disease. We previously reported that traditional acetylcholinesterase inhibitors such as BW284C51, tacrine, and physostigmine were more potent inhibitors of the acetylcholinesterase in normal axons and cell bodies than of the acetylcholinesterase in plaques and tangles. We now report that the reverse pattern is seen with indoleamines (such as serotonin and its precursor 5-hydroxytryptophan), carboxypeptidase inhibitor, and the nonspecific protease inhibitor bacitracin. These substances are more potent inhibitors of the cholinesterases in plaques and tangles than of those in normal axons and cell bodies. These results show that the enzymatic properties of plaque and tangle-associated cholinesterases diverge from those of normal axons and cell bodies. The selective susceptibility to bacitracin and carboxypeptidase inhibitor indicates that the catalytic sites of plaque and tangle-bound cholinesterases are more closely associated with peptidase or protease-like properties than the catalytic sites of cholinesterases in normal axons and cell bodies. This shift in enzymatic affinity may lead to the abnormal protein processing that is thought to play a major role in the pathogenesis of Alzheimer disease. The availability of pharmacological and dietary means for altering brain indoleamines raises therapeutic possibilities for inhibiting the abnormal cholinesterase activity associated with Alzheimer disease.     

Evaluation of the Relationship Between Adenosine Deaminase,Myeloperoxidase, Cholinesterase,Preeclampsia Severity,and Neonatal Outcomes

We aimed to evaluate whole blood adenosine deaminase (ADA), myeloperoxidase (MPO), butyrylcholinesterase (BChE), and acetylcholinesterase (AChE) activities and to investigate whether there was a correlation between these enzymes and severity of preeclampsia and neonatal outcomes. Sixty-one pregnant women with mild (n = 31) or severe (n = 30) preeclampsia and 50 healthy controls were included in this study. Whole blood adenosine deaminase, myeloperoxidase, butyrylcholinesterase, and acetylcholinesterase activities were measured. Adenosine deaminase and myeloperoxidase activities were significantly higher in both mild and severe preeclamptic women than they were in the controls. There was also a significant difference between the severe and the mild preeclamptic groups with respect to these enzyme activities. Although BChE activity was lower in the severe preeclamptic women than it was in the healthy controls (P < .05), AChE activity was similar in all groups (P > .05). We noted an inverse correlation between ADA activity and birth weight (r = ?0.337) (P < .05) and between MPO activity and Apgar scores at 1 and 5 minutes (r = ?0.438 and r = ?0.475, respectively, P < .01). We concluded that elevated ADA and MPO but not AChE activities may correlate with disease severity and neonatal outcomes in preeclamptic women. Further studies are needed to elucidate the exact roles of ADA and MPO in the pathophysiology of preeclampsia.     

Organophosphate increases the sensitivity of human exocrine pancreas to acetylcholine.

Human pancreas contains two cholinesterase isoenzymes: acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). In the present study, binding potency of two organophosphates for human cholinesterases were compared by the Ellman method. Echothiophate was found to have much greater potency than iso-OMPA for both cholinesterases. Using Karnovsky histochemical stains on human pancreatic tissue, the same results were confirmed. Dose-response studies with acetylcholine were done on viable pancreas fragments from nine human donors, without pancreatic disease (group I). Cold-preservation time was less than 30 h. Pancreas was minced into fragments, after the technique of Scheele and Palade, placed in Eagle's medium, and gassed with O2. Amylase release was measured by the Phadebas Method and corrected for basal release. There was a dose-dependent response to acetylcholine at 1 and 2 h, with a shift in peak amylase release to the left, when fragments were preincubated in 10(-4) M echothiophate. This indicated a 100-fold increase in sensitivity to acetylcholine. In three patients with chronic pancreatitis (Group II), there were variable patterns of response of amylase release to acetylcholine, and higher basal outputs. In Group III, prolonged storage conditions of over 40 h were tested for 4 pancreas donor tissues. There was no response to acetylcholine. These studies show that for up to 30 h cold storage, fragments of pancreas from human organ donors respond to acetylcholine in dose-dependent manner. An organophosphate, echothiophate (10(-4) M) which inhibits both cholinesterases, increases pancreatic sensitivity to acetylcholine, and these results are similar to findings from canine pancreas fragments, which also showed increased sensitivity.     

Predictive score for hepatocellular carcinoma after hepatitis B e antigen loss in patients treated with entecavir or tenofovir

The risk of developing hepatocellular carcinoma (HCC) after hepatitis B e antigen seroclearance (ESC) remains unclear. We established and validated a new risk prediction model for HCC development after ESC in patients with chronic hepatitis B (CHB) receiving antiviral therapy (AVT). Between 2006 and 2016, 769 patients (training cohort) and 1,061 patients (validation cohort) with CHB who experienced ESC during AVT using entecavir (ETV) or tenofovir disoproxil fumarate (TDF) were recruited. In the multivariate analysis, male sex (hazard ratio [HR] = 2.092; 95% confidence interval [CI] = 1.152‐3.800), cirrhosis (HR = 5.141; 95% CI = 2.367‐11.167) and fibrosis‐4 index (FIB‐4) of >3.25 (HR = 2.070; 95% CI = 1.184‐3.620) were the independent risk factors for HCC development (all P < .05). Accordingly, a novel HCC‐ESC_AVT model was developed (1x[sex: male = 1, female = 0] + 3x(cirrhosis = 1, noncirrhosis = 0) + 1x(FIB‐4: >3.25 = 1, ≤3.25 = 0). The cumulative risk for HCC development was significantly different among the risk groups based on the HCC‐ESC_AVT category (0‐1, 2‐4 and 5 for the low‐, intermediate‐ and high‐risk groups, respectively) (overall P < .001, log‐rank test). The area under the receiver operating characteristic curve (AUC) for predicting HCC development 3, 5 and 10 years after ESC was 0.791, 0.771 and 0.790, respectively (all P < .05). The predictive value of the HCC‐ESC_AVT model was similar in the validation cohort (AUC = 0.802, 0.774 and 0.776 at 3, 5 and 10 years, respectively; all P < .05). Hence, we have developed and validated a new HCC‐ESC_AVT model for HCC development, which includes male sex, cirrhosis and FIB‐4 of >3.25 as constituent variables.     

Risk of dipeptidyl peptidase‐4 (DPP‐4) inhibitors on site‐specific cancer: A systematic review and meta‐analysis

The long‐term impact of dipeptidyl peptidase‐4 (DPP‐4) inhibition is unknown, and there are concerns about the influence of DPP‐4 inhibition on carcinogenesis of the pancreas and thyroid. As DPP‐4 is a rather unselective enzyme present in many tissues, we focused on all specific cancer types. PubMed and EMBASE were searched between January 2005 and April 2017 to identify studies comparing DPP‐4 inhibitors with either placebo or active drugs on cancer risk. Studies were included if they reported on at least one specific cancer outcome and had a follow‐up of at least 1 year after start of drug use. Methodological quality of the studies was assessed by the Cochrane Collaboration's tool and the Newcastle‐Ottawa Scale. Twenty‐five studies met the inclusion criteria (12 randomized controlled trials and 13 observational studies). Sample sizes of the DPP‐4 inhibitor groups ranged from 29 to 8212 patients for randomized controlled trials and from 2422 to 71 137 patients for observational studies. Mean age ranged from 51 to 76 years, and mean follow‐up was 1.5 years. None of the pooled (sensitivity) analyses, except the observational studies studying breast cancer (hazard ratio [95% CI]: 0.76 [0.60‐0.96]), showed evidence for an association between DPP‐4 inhibitors and site‐specific cancer. Also for pancreatic and thyroid cancer, no statistically significant risk was found. Based on the current literature, it is not possible to conclude whether DPP‐4 inhibitors were associated with an increased risk of site‐specific cancer. Future studies should address the methodological limitations and follow patients for a longer period to determine the long‐term cancer risk of DPP‐4 inhibitors.     

Kinetics of the interaction between anti‐FVIII antibodies and FVIII from therapeutic concentrates,with and without von Willebrand factor,assessed by surface plasmon resonance

The presence of VWF in plasma‐derived FVIII (pdFVIII/VWF) products has been pointed out as a key difference with recombinant FVIII (rFVIII) products with regard to immunogenicity. A Surface Plasmon Resonance (SPR) study was designed to characterize in detail the interaction between anti‐FVIII (IgGs) from a severe haemophilia A patient, and FVIII from concentrates of different sources. Full‐length rFVIII (preincubated or not with purified VWF), B domain‐deleted (BDD)‐rFVIII and pdFVIII/VWF were analysed. To ensure reproducible conditions for accurate determination of kinetic constants, a capture‐based assay format was developed using protein G surfaces for specific and reversible coupling of endogenous anti‐FVIII antibodies. Concentration ranges (nm ) of FVIII products tested were 9–0.03 (rFVIII) and 6–0.024 (pdFVIII/VWF). The association with antibodies was monitored for 3–5 min, whereas dissociation of the complex was followed for 5–20–240 min. A strong interaction of rFVIII and BDD‐rFVIII with patient's IgG was detected with the K _D values in the low picomolar range (5.9 ± 3.0 and 12.7 ± 6.9 pm , respectively) and very slow dissociation rates, while pdFVIII/VWF showed only marginal binding signals. The VWF complexed rFVIII displayed reduced binding signals compared with uncomplexed rFVIII, but the K _D was still in the picomolar range (4.1 ± 1.9 pm ) indicating insufficient complex formation. rFVIII, alone or bound to exogenously added VWF, showed high affinity for anti‐FVIII IgGs from a severe haemophilia A patient whereas pdFVIII/VWF did not. These results are in agreement with those studies that point towards rFVIII concentrates to be more immunogenic than pdFVIII concentrates.     

Teaching Geriatrics Using an Innovative,Individual‐Centered Educational Game: Students and Educators Win. A Proof‐of‐Concept Study

Given the increasingly aging population, nearly every doctor will encounter elderly adults who present with multiple complex comorbidities that can challenge even experienced physicians. This may explain why many medical students do not have a positive attitude toward elderly adults and find the complexity of their problems overwhelming. It was hypothesized that a recently developed medical school geriatrics course, based on the game GeriatriX and designed specifically to address the complexities associated with decision‐making in geriatrics, can have a positive effect on attitudes toward geriatrics and on perceived knowledge of geriatrics. The effects of this game‐based course were evaluated as a proof of concept. The assessment was based on the Aging Semantic Differential (ASD) and a validated self‐perceived knowledge scale of geriatric topics. The usability of (and satisfaction with) GeriatriX was also assessed using a 5‐point Likert scale. After completion of the course, the ASD changed significantly in the geriatrics course group (n = 29; P = .02) but not in a control group that took a neuroscience course (n = 24; P = .30). Moreover, the geriatrics course group had a significant increase in self‐perceived knowledge for 12 of the 18 topics (P = .002), whereas in the control group self‐perceived knowledge increased significantly for one topic only (sensory impairment) (P = .04). Finally, the geriatrics students reported enjoying GeriatriX. This proof‐of‐concept study clearly supports the hypothesis that a 4‐week course using a modern educational approach such as GeriatriX can improve students' self‐perceived knowledge of geriatrics and their attitudes toward elderly adults.     

In Vitro Homocysteine Inhibits Platelet Na+, K+-ATPase and Serum Butyrylcholinesterase Activities of Young Rats

Homocystinuria is an inborn error of sulphur amino acid metabolism, resulting in accumulation of tissue homocysteine. This disease is characterized predominantly by vascular and nervous system dysfunction. In the present study we investigated the in vitro effects of homocysteine, the main metabolite accumulated in homocystinuria, on platelet Na+,K+-ATPase and serum butyrylcholinesterase (BuChE) activities of young rats. Platelet and serum of 29-day-old Wistar rats were incubated in the absence (control) or presence of homocysteine (0.01-0.5 mM). Results showed that Na+,K+-ATPase and BuChE activities were significantly inhibited by homocysteine. It is proposed that inhibition of Na+,K+-ATPase and BuChE activities might be one useful peripheral marker for the neurotoxic effects of homocysteine.     

CD14 expression on Kupffer cells during the course of carbon tetrachloride‐mediated liver injury

OBJECTIVE: To investigate the expression of CD14 on Kupffer cells during the course of carbon tetrachloride (CCl₄)‐mediated liver injury and its role in the activation of Kupffer cells. METHODS: Rats were administered CCl₄ twice weekly for up to 8 weeks. Kupffer cells were isolated from normal and CCl₄‐treated rats by the combined ‘collagenase‐pronase’ perfusion method, discontinuous density gradient centrifugation. On the day after isolation, the cells were incubated with RPMI‐1640 containing varying doses of lipopolysaccharide (LPS) for 6 h. Supernatants were then collected for measuring the concentration of tumor necrosis factor‐alpha (TNF‐α) by enzyme‐linked immunosorbent assay (ELISA). The expression of CD14 mRNA on Kupffer cells were determined by RT‐PCR. The plasma concentrations of endotoxin were determined by chromogenic substrate Limulus amebocyte lysate assay. RESULTS: Basic TNF‐α production of Kupffer cells isolated from CCl₄‐treated rats at 4 and 6 weeks was significantly higher than that of normal (P < 0.05). Following LPS stimulation the production of TNF‐α was markedly increased in Kupffer cells from the 2‐, 4‐ and 6‐week treatment groups (P < 0.05). Moreover, LPS‐induced TNF‐α production was dose‐dependent. CD14 mRNA expression on Kupffer cells isolated from CCl₄‐treated rats was elevated following 2 weeks of CCl₄ administration and the maximum elevation occurred at 6 weeks. Gene expression was decreased in Kupffer cells after 8 weeks of CCl₄ treatment. CCl₄ administration elicited extensive changes in liver morphology, including steatosis, inflammation and necrosis. The plasma concentrations of endotoxin of CCl₄ ‐ treated rats were increased during the time of liver injury. CONCLUSION: Up‐regulation of CD14 expression in Kupffer cells during CCl₄‐mediated chronic liver injury indicates cell activation and that they are more sensitive to LPS stimulation. Kupffer cells are critical effector cells in the early stage of liver injury.     

Comparative analysis of pulmonary hypertension in patients treated with imatinib,nilotinib and dasatinib

Pulmonary hypertension (PH) is a rare, but life‐threatening, adverse event in patients treated with tyrosine kinase inhibitors (TKIs), such as dasatinib, but has not been fully evaluated in patients treated with imatinib or nilotinib. We used echocardiography to noninvasively assess the incidence of PH in 105 patients with chronic myeloid leukaemia (CML) treated with imatinib (n  = 37), nilotinib (n  = 30) or dasatinib (n  = 38). The mean triscupid regurgitation peak gradient (TRPG), which reflects pulmonary arterial pressure, was 22·7 mmHg in the imatinib group, 23·1 mmHg in the nilotinib group and 23·4 mmHg for dasatinib group. These values were not significantly different, but higher than those (19·0 mmHg) in newly diagnosed CML patients. A TRPG > 31 mmHg, marking possible PH onset, was detected in 9 of 105 patients: one (2·7%) treated with imatinib, three (10·0%) with nilotinib and five (13·2%) with dasatinib. Only three patients complained of dyspnoea, whereas the other six were asymptomatic. In addition, there was a tendency toward correlation of TRPG value and age or TKI treatment duration. These results suggested that treatment with not only dasatinib, but also imatinib and nilotinib, can be associated with subclinical PH. Noninvasive echocardiography is useful for screening, especially in older patients with long‐term TKI treatment.     

Effect of high‐altitude trekking on blood pressure and on asymmetric dimethylarginine and isoprostane production: Results from a Mount Ararat expedition

The study aimed at exploring the mechanisms behind blood pressure and heart rate changes upon acute altitude exposure utilizing urinary excretion of biochemical factors involved in cardiovascular regulation. The study was conducted on 12 lowlander native male mountain climbers, living at sea level, exposed to altitudes ranging from 1800 to 5147 m above sea level over 4 days, during their ascent to Mount Ararat (Turkey). Blood pressure (measured by oscillometric method), heart rate, and blood oxygen saturation (SpO₂) were recorded at rest (on awakening before food intake), in hypoxic conditions at 4200 m and at sea level before and after the altitude expedition. In the same study conditions (ie before‐during‐after the expedition), first‐voided urinary samples were collected and assayed for 8‐iso‐prostaglandin F_2α (8‐iso‐PGF_2α) and asymmetric dimethylarginine (ADMA) determination. Heart rate, and systolic and diastolic blood pressures were higher (P < .05) at high altitude than at the sea level. Furthermore, both urinary 8‐iso‐PGF_2α and ADMA were significantly elevated (P < .01) at high altitude and returned to normal levels soon after returning to sea level. A 4‐day exposure to high‐altitude hypoxia induced a temporary increase in blood pressure and heart rate, confirming previous findings. Blood pressure increase at high altitude was associated with significantly enhanced production of biochemical mediators such as 8‐iso‐PGF2α, catecholamines, and ADMA, although we could not demonstrate a direct link between these parallel significant changes probably due to the forcefully limited sample size of our study, carried out in challenging environmental conditions at very high altitude.     

Undetectable HCV‐RNA at treatment‐week 8 results in high‐sustained virological response in HCV G1 treatment‐experienced patients with advanced liver disease: the International Italian/Spanish Boceprevir/Peginterferon/Ribavirin Name Patients Program

In many countries, first‐generation protease inhibitors (PIs)/peginterferon/ribavirin (P/R) still represent the only treatment option for HCV‐infected patients. Subjects with advanced disease and previous failure to P/R urgently need therapy, but they are under‐represented in clinical trials. All treatment‐experienced F3/4 Metavir patients who received boceprevir (BOC)+P/R in the Italian–Spanish Name Patient Program have been included in this study. Multivariate logistic regression analysis (MLR) was used to identify baseline and on‐treatment predictors of SVR and adverse events (AEs). Four hundred and sixteen patients, mean age 57.7 (range 25–78 years), 70% males, 69.5% (289/416) F4, 14% (41/289) Child–Pugh class A6, 24% (70/289) with varices and 42% (173/416) prior null responders to P/R, were analysed. Overall, SVR rate (all 381 patients who received one dose of BOC) was 49%, (58% in F3, 45% in F4, 61% in relapsers, 51% in partial, 38% in null responders, and 72% in subjects with undetectable HCV‐RNA at treatment‐week (TW)8. Among patients with TW8 HCV‐RNA ≥ 1000 IU/L, SVR was 8% (negative predictive value = 92%). Death occurred in 3 (0.8%) patients, while decompensation and infections were observed in 2.9% and 11%, respectively. At MLR, SVR predictors were TW4 HCV‐RNA ≥ 1log₁₀‐decline from baseline, undetectable TW8 HCV‐RNA, prior relapse, albumin levels ≥3.5 g/dL and platelet counts ≥100 000/μL. Metavir F4, Child‐Pugh A6, albumin, platelets, age and female gender were associated with serious and haematological AEs. Among treatment‐experienced patients with advanced liver disease eligible for IFN‐based therapy, TW8 HCV‐RNA characterised the subset with either high or poor likelihood of achieving SVR. Using TW8 HCV‐RNA as a futility rule, BOC/P/R appears to have a favourable benefit–risk profile.