Bioequivalence study of two formulations of itraconazole 100 mg capsules in healthy volunteers under fed conditions: a randomized,three-period,reference-replicated,crossover study

Background: The aim of the study was to evaluate the bioequivalence of two itraconazole 100 mg capsule formulations.

Research design and methods: The single-center, open-label, randomized, three-period, three-sequence, reference-replicated, cross-over study included 38 healthy subjects under fed conditions. In each study period (separated by a 14-day washout), a single oral dose of the test (T) or reference (R) product was administered. Blood samples were collected at pre-dose and up to 72.0 h after administration. The calculated pharmacokinetic parameters, based on the plasma concentrations of itraconazole and hydroxy itraconazole, were AUC_0-72h, AUC_0-∝, C_max, T_max, T_1/2 and K_el.

Results: The 90% CI for the test/reference geometric means ratio for the parent compound, itraconazole, was in the range from 85.29% to 116.07% for AUC_0-72h. Since the coefficient of variation (CV) for the reference product was 44.95% for C_max, the 90% CI for this parameter for itraconazole was 93.49–133.78%, which was within the proposed limits of the EMA for bioequivalence of 72.15–138.59%. During the study, 4 subjects encountered a total of 14 mild adverse events.

Conclusions: The use of the reference-scaling approach with 3-period design (TRR, RTR, and RRT) was an efficient way to demonstrate that two commercially available oral itraconazole formulations met the predetermined bioequivalence criteria.     

The influence of noradrenergic blockade on vasospasm and the quantity of cerebral dopamine ß-hydroxylase following subarachnoid haemorrhage in rabbits

BACKGROUND: The aim of this study of experimental subarachnoid haemorrhage (SAH) and exclusion of the sympathetic nervous system (SNS) in rabbits was to find out if changes in the central noradrenergic areas of the hypothalamus and brain stem could be ascertained, in parallel with measurement of the intensity of chronic cerebral vasospasm in the basilar arteries. METHODS: Histologic specimens were prepared by perfusion fixation on day 8 after the SAH. The spastic effect of experimentally induced SAH in New Zealand rabbits was investigated: firstly, using our previously developed method for measuring the corrugation coefficient (CC) of the vessel intima on precisely defined locations of the basilar artery (BA) with the aid of computer image analysis; and secondly, by immunohistochemical assessment of the concentration and localization of dopamine beta-hydroxylase (DBH), using anti-DBH, at precisely defined sites of the hypothalamus and brain stem of the same rabbit. RESULTS: The intima of the BA, assessed by CC, was significantly less corrugated and had significantly less DBH in group A (the control group without SAH and without additional interventions; mean CC = 1.192, P = 0.004; median DBH = 0.50, P = 0.044), in group C (SAH and alpha-blocker phenoxybenzamine; mean CC = 1.142, P = 0.000; median DBH = 0.75, P = 0.001), and in group D (SAH and cervical gangliectomy; mean CC = 1.210, P = 0.003; median DBH = 0.50, P = 0.002) compared with group B (rabbits with SAH and without medication). Group B showed a significantly more intensive accumulation of DBH (median DBH = 1.15) and, according to the CC (mean CC = 1.369), more intensive corrugation of the intima of BA than all other groups. The correlation between CC and DBH for all the rabbits (groups A, B, C and D together) was significantly positive (Spearman Rho = 0.470; p = 0.010). CONCLUSIONS: The results of this study demonstrated: firstly, an intensive excitatory influence of SAH on the quantity of DBH in central noradrenergic areas in the hypothalamus and brain stem; secondly, a very effective influence of peripheral and systemic sympathetic exclusion on lowering the quantity of central sympathetic DBH; thirdly, that the changes in the BA of individual rabbits occur simultaneously with corresponding changes in DBH-containing neurons, thus suggesting the likelihood of SNS involvement in the pathogenesis of post-SAH vasospasm in rabbits.     

Induction of osteoclast progenitors in inflammatory conditions: key to bone destruction in arthritis

The inflammatory milieu favors recruitment and activation of osteoclasts, and leads to bone destruction as a serious complication associated with arthritis and with other inflammatory processes. The frequency and activity of osteoclast progenitors (OCPs) correspond to arthritis severity, and may be used to monitor disease progression and bone resorption, indicating the need for detailed characterization of the discrete OCP subpopulations. Collectively, current studies suggest that the most potent murine bone marrow OCP population can be identified among lymphoid negative population within the immature myeloid lineage cells, as B220^?CD3^?CD11b^–/loCD115⁺CD117⁺CX3CR1⁺ and possibly also Ter119^?CD11c^?CD135^loLy6C⁺RANK^?. In peripheral blood the OCP population bears the monocytoid phenotype B220^?CD3^?NK1.1^?CD11b⁺Ly6C^hiCD115⁺CX3CR1⁺, presumably expressing RANK in committed OCPs. Much less is known about human OCPs and their regulation in arthritis, but the circulating OCP subset is, most probably, comprised among the lymphoid negative population (CD3^?CD19^?CD56^?), within immature monocyte subset (CD11b⁺CD14⁺CD16^?), expressing receptors for M-CSF and RANKL (CD115⁺RANK⁺). Our preliminary data confirmed positive association between the proportion of peripheral blood OCPs, defined as CD3^?CD19^?CD56^?CD11b⁺CD14⁺, and the disease activity score (DAS28) in the follow-up samples from patients with psoriatic arthritis receiving anti-TNF therapy. In addition, we reviewed cytokines and chemokines which, directly or indirectly, activate OCPs and enhance their differentiation potential, thus mediating osteoresorption. Control of the activity and migratory behaviour of OCPs as well as the identification of crucial bone/joint chemotactic mediators represent promising therapeutic targets in arthritis.     

An early shoulder repositioning program in birth-related brachial plexus injury: a pilot study of the Sup-ER protocol

Background

Birth-related brachial plexus injury (BRBPI) occurs in 1.2/1,000 births in British Columbia. Even in children with “good” recovery, external rotation (ER) and supination (Sup) are often weaker, and permanent skeletal imbalance ensues. A preventive early infant shoulder passive repositioning program was created using primarily a novel custom splint holding the affected arm in full ER and Sup: the Sup-ER splint. The details of the splint and the shoulder repositioning program evolved with experience over several years. This study reviews the first 4 years.

Methods

A retrospective review of BCCH patients managed with the Sup-ER protocol from 2008 to 2011 compared their recovery scores to matched historical controls selected from our database by two independent reviewers.

Results

The protocol was initiated in 18 children during the study period. Six were excluded due to the following: insufficient data points, non-compliance, late splint initiation, and loss to follow-up. Of the 12 matches, the Sup-ER group final score at 2 years was better than controls by 1.18 active movement scale (AMS) points (p = 0.036) in Sup and 0.96 AMS points in ER (but not statistically significant (p = 0.13)). Unexpectedly, but importantly, during the study period, zero subjects were assessed to have the active functional criteria to indicate brachial plexus reconstruction, where previously we operated on 13 %.

Conclusions

Early application of passive shoulder repositioning into Sup and ER may improve outcomes in function of the arm in infants with BRBPI. A North American multi-site randomized control trial has been approved and has started recruitment.     

Potential of Using Wood Biomass Ash in Low-Strength Composites

Reducing greenhouse gas emissions and dependence on fossil fuels is the cornerstone of all European climate and energy strategies. Consequently, renewable energy sources are becoming more competitive with fossil fuels. The largest source of bioenergy in the European Union is biomass-fired power plants. Therefore, the European coal phase-out strategy led to an increased use of wood biomass as a sustainable fuel, generating large amounts of wood biomass ash (WBA). In the research studies reported so far, WBA has been mainly used in cementitious composites. However, given the similarities between the chemical composition of WBA and hydraulic lime (HL), this research focused on its potential classification as a building lime. Overall, three different sources of fly WBA were considered for the preparation of binders as mixtures of WBA and coal fly ash (CFA) in different ratios. The contribution of each binder mixture on the paste and mortar properties was analyzed based on the chemical composition, setting time, volume stability, and contribution to the mortar strength (compressive and flexural). In general, it can be concluded that the studied binders can meet the criteria of EN 459-1. However, special attention should be paid to the volume deformations and the setting time.     

Palladium(II) Complexes with N‐Heteroaromatic Bidentate Hydrazone Ligands: The Effect of the Chelate Ring Size and Lipophilicity on in vitro Cytotoxic Activity

Novel Pd(II) complex with N‐heteroaromatic Schiff base ligand, derived from 8‐quinolinecarboxaldehyde (q8a) and ethyl hydrazinoacetate (haOEt), was synthesized and characterized by analytical and spectroscopy methods. The structure of novel complex, as well as structures of its quinoline and pyridine analogues, was optimized by density functional theory calculations, and theoretical data show good agreement with experimental results. A cytotoxic action of the complexes was evaluated on cultures of human promyelocytic leukemia (HL‐60), human glioma (U251), rat glioma (C6), and mouse fibrosarcoma (L929) cell lines. Among investigated compounds, only complexes with quinoline‐based ligands reduce the cell numbers in a dose‐dependent manner in investigated cell lines. The observed cytotoxic effect of two isomeric quinoline‐based complexes is predominantly mediated through the induction of apoptotic cell death in HL‐60 cell line. The cytotoxicity of most efficient novel Pd(II) complex is comparable to the activity of cisplatin, in all cell lines investigated.     

Markers of inflammation and microvascular complications in type 1 diabetes

Aims

Long-term hyperglycemia, characteristic for type 1 diabetes, causes enhanced oxidative stress, chronic inflammation and endothelial dysfunction which are the key events in the development of vascular complications. On the other hand, some data shows that existence of chronic degenerative complications may cause increased inflammatory marker levels in diabetic patients, mainly as a repercussion of malfunctioned endothelial repair process. Our study aims to determinate a degree of inflammation in type 1 diabetes patients and to investigate its relation to development of the chronic microvascular complications.

Methods

General information, anthropometric, glucose metabolism, lipid and lipoprotein parameters, levels of C reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were analyzed and screening tests for detection of the chronic microvascular complications were conducted in 76 type 1 diabetes patients.

Results

Forty six patients had at least one of the complications. They were older and had longer duration of diabetes (p=0.015; p<0.0001) and higher values of total (p=0.021), LDL-cholesterol (p=0.048) and triglycerides (p=0.002). Levels of CRP (p=0.004) and TNF-α (p=0.048) were higher in patients with chronic microvascular complications in regard to patients without diagnosed microangiopathy.

Conclusion

Low grade chronic inflammation is characteristic for type 1 diabetes patients with developed chronic microvascular complications.     

Effects of high fat diet,ovariectomy, and physical activity on leptin receptor expression in rat brain and white fat tissue

Aim

To evaluate in a rat animal model whether ovariectomy, high fat diet (HFD), and physical activity in the form of running affect leptin receptor (Ob-R) distribution in the brain and white fat tissue compared to sham (Sh) surgery, standard diet (StD), and sedentary conditions.

Methods

The study included 48 female laboratory Wistar rats (4 weeks old). Following eight weeks of feeding with standard or HFD, rats were subjected to either OVX or Sh surgery. After surgery, all animals continued StD or HFD for the next 10 weeks. During these 10 weeks, ovariectomy and Sh groups were subjected to physical activity or sedentary conditions. Free-floating immunohistochemistry and Western blot methods were carried out to detect Ob-R in the brain and adipose tissue.

Results

StD-ovariectomy-sedentary group had a greater number of Ob-R positive neurons in lateral hypothalamic nuclei than StD-Sh-sedentary group. There was no difference in Ob-R positive neurons in arcuatus nuclei between all groups. Ob-R distribution in the barrel cortex was higher in HFD group than in StD group. Ob-R presence in perirenal and subcutaneous fat was decreased in StD-ovariectomy group.

Conclusion

HFD and ovariectomy increased Ob-R distribution in lateral hypothalamic nuclei, but there was no effect on arcuatus nuclei. Our results are first to suggest that HFD, ovariectomy, and physical activity affect Ob-R distribution in the barrel cortex, which might be correlated with the role of Ob-R in election of food in rats.Obesity is one of the leading health issues worldwide, associated with an increased risk of morbidity and mortality (1). In 1997, the World Health Organization (WHO) formally recognized obesity as a global epidemic (2). Increase in body fat stores and obesity is caused by an imbalance between energy intake and energy expenditure (3,4). Since childhood obesity is a predictor of an increased death rate, the “obesity epidemic” may reverse the current declining rate of death from cardiovascular diseases (5). Factors that contribute to obesity can be environmental (6), social (7), behavioral (8), psychological (9), and genetic (10,11).Women generally have more body fat than men (12). Nevertheless similar odds ratios were recorded in women and men for the association of abdominal obesity with acute myocardial infarction (13). Weight gain is common after menopause, indicating an association between hormones and fat stores (14). A large scale observational study found that both the body mass index and the level of physical activity were independent predictors of mortality and that a higher level of physical activity did not eliminate the risk associated with adiposity. At the same time, women who were both lean and physically active had the lowest mortality (15). In animal studies menopause can be induced by ovariectomy (OVX) (16).Obesity can also be called a disorder of appetite and it is controlled by complex homeostatic mechanisms involving the hypothalamus and brainstem (17). Many gut peptides like cholecystokinin, ghrelin, glucagon-like peptide-1 (GLP-1), and -2 and peptide YY (PYY) act on the brain to control eating behavior (18). There are two different system for controlling feeding behavior: short-term and long-term (19). Short-term regulation involves neural signals from the GI tract and its hormones, like insulin, glucagon, and ghrelin (20). A hormone that functions mainly within long-term regulation is leptin (16 kD), a hormonal product of the obesity (ob) gen, primarily secreted by adipocytes (21) and released in the brain. It generates a feeling of satisfaction and acts like an appetite-suppressing agent. Circulating leptin levels are lower in ovariectomized rats (22).Food intake is regulated via neural circuits located in the hypothalamus (23). Leptin acts via its leptin or Ob receptors (Ob-R) and is primarily expressed in hypothalamic neurons (19) especially in arcuate, ventromedial, and dorsomedial nuclei (24). Leptin is transported across the blood-brain barrier (BBB) by a saturable transporter (25). Ob-R is also detected in nonhypothalamic areas in the mice and in human brain neocortex, cerebellum, entorinal cortex, amygdale, and rostral medulla (26). Adipocytes, endothelial cells, and macrophages also have leptin receptor at its surface, which suggests autocrine and paracrine action for leptin in human adipose tissue (27). Association between the expression of Ob-R in target tissues and physiological and hormonal controlled processes is still unclear. Leptin receptors mRNA is found in each of the major components of the CNS “feeding” circuitry – the brainstem, hypothalamus, and is distributed reward centers (Allan brain) (28). Therefore, the aim of the current study was to evaluate whether HFD affects Ob-R distribution compared with StD specifically in the barrel field and piriform cortex compared to standard feeding centers in the hypothalamus. We supposed that the combination of OVX and HFD is interesting for further research on selected brain regions, which might be alleviated by physical activity. We also supposed that changes in Ob-R level in white fat tissue would correlate with the changes in brain regions.