Greater occipital and supraorbital nerve blockade for the preventive treatment of migraine: a single-blind,randomized, placebo-controlled study

Objective: Nerve injections have been used for the acute and preventive treatment of migraine in recent decades. Most of these injections focused on greater occipital nerve (GON) blockade. However, few studies were placebo controlled, and only a few of them investigated GON and supraorbital nerve (SON) blockade together. This study aimed to evaluate the efficacy of GON and SON blockade with local anesthetics for the preventive treatment of migraine without aura.

Methods: Eighty-seven patients diagnosed with migraine without aura were included in the study. Patients were divided randomly. One group was injected with 1% lidocaine, the other group was injected with 0.9% saline. GON and SON injections were done bilaterally. The injections were repeated weekly for 3 weeks. Patients were followed up for 2 months to assess clinical response.

Results: Seventy-one patients completed the study. After 2 months, the number of headache days decreased significantly from 12.8?±?10.9 to 5.3?±?7.4, and VAS decreased from 8.3?±?1.0 to 5.5?±?1.9 in the blockade group. The number of headache days decreased from 12.4?±?10.3 to 7.5?±?7.2 and VAS decreased from 8.2?±?1.1 to 7.4?±?1.3 in the placebo group. Response was seen in 65.1% of the patients in the blockade group (65.4% for episodic migraine, 64.7% for chronic migraine) and 28.6% of the patients in the placebo group. The difference was significant.

Conclusions: The results suggest that GON and SON blockade with lidocaine was more effective than the placebo in the prophylactic treatment of both episodic and chronic migraine.     

Effectiveness and safety of micronized purified flavonoid fraction for the treatment of concomitant varicose veins of the pelvis and lower extremities

Objective: Concomitant varicose veins of the pelvis (VVP) and lower extremities (VVLE) frequently coexist. This study evaluated the effectiveness and safety of micronized purified flavonoid fraction (MPFF) in the treatment of patients with both conditions.

Methods: Female outpatients with concomitant VVP and VVLE received MPFF 1000?mg once daily for 2 months (Group 1), or 1000?mg twice daily for 1 month followed by 1000?mg once daily for 1 month (Group 2), based on pelvic pain intensity. Change in pain intensity during treatment was evaluated on a 10?cm visual analog scale. All patients underwent transvaginal and transabdominal duplex ultrasound scanning, radionuclide phlebography of the lower extremities, and emission computer tomography of the pelvic veins at inclusion and end of treatment.

Results: In Group 1 (N?=?35), MPFF was associated with a twofold reduction in pain syndrome severity (pelvic, perineal and lower leg pain) in all patients after 1 month, and a reduction in chronic pelvic pain (CPP) from 3.4?±?1.2 to 0.83?±?0.18?cm at 2 months. Leg pain significantly decreased from 2.8?±?0.6 at baseline to 0.94?±?0.11 after 2 months. In Group 2 (N?=?30), MPFF decreased CPP severity from 6.3?±?0.8 to 1.2?±?0.12, perineal pain from 3.6?±?0.9 to 0.88?±?0.22 and leg pain from 4.6?±?0.5 to 0.9?±?0.1. Radionuclide phlebography confirmed the clinical improvement in both treatment groups, with a substantial increase in linear blood flow velocity in the internal iliac veins (～10% in Group 1 and 35% in Group 2) and a reduction in mean transit times of the radiopharmaceutical. MPFF also reduced blood stasis in the pelvic venous plexuses. Gastralgias were reported in two patients but resolved rapidly and did not lead to treatment withdrawal.

Conclusion: Phlebotropic treatment with MPFF is an effective and safe method of conservative therapy in patients with concomitant VVP and VVLE.     

Comparison of a twice daily injection of insulin aspart 50 with insulin aspart 30 in patients with poorly controlled type 2 diabetes

Objective: To compare the efficacy and safety of a twice daily injection of insulin aspart (BIAsp) 30 and BIAsp50 in patients with type 2 diabetes mellitus (T2DM) poorly controlled with oral hypoglycemic agents (OHAs).

Methods: In this 12 week prospective, randomized, parallel trial, a total of 80 T2DM patients, 59?±?10?years old with a disease duration of 9.3?±?6.6?years and HbA1c >7% despite large doses of metformin and sulfonylurea administration, were randomized to receive BIAsp30 (n?=?40) or BIAsp50 (n?=?40). The primary endpoint was a change in HbA1c at week 12.

Results: The changes in HbA1c from baseline were ?2.5%?±?1.0% in the BIAsp50 group and ?2.5%?±?1.2% in the BIAsp30 group (p?=?.897). No difference was observed in the rate of HbA1c target achievement (<7.0%) between BIAsp50 (42.5%) and BIAsp30 (32.5%) (p?=?.495). The change in fasting plasma glucose (FPG) in the BIAsp50 group was lower than that in the BIAsp30 group (p?<?.001), while the change in two-hour postprandial blood glucose (2hPBG) was higher and blood glucose excursion was lower in the BIAsp50 group than that in the BIAsp30 group (p?<?.001, p?<?.001). A significant improvement in HbA1c was observed with BIAsp50 in subgroups with baseline blood glucose excursion >7.8?mmol/L or 2hPBG >17.6?mmol/L compared with BIAsp30. There were no differences in hypoglycemia or body weight between groups.

Conclusions: Compared with BIAsp30, BIAsp50 showed greater efficacy in patients with baseline BG excursion >7.8?mmol/L or 2hPBG >17.6?mmol/L as well as good safety for hypoglycemia.

Clinical trial registration: ChiCTR-IIR-16008958.     

Clinical efficacy of once-daily micronized purified flavonoid fraction 1000 mg tablet in patients with symptomatic chronic venous disease

Aim: To investigate the clinical efficacy of micronized purified flavonoid fraction (MPFF) 1000?mg given as a single 1000?mg tablet once daily in patients suffering from chronic venous disease (CVD) vs MPFF 500?mg twice daily.

Methods: In an international, randomized, double-blind, parallel-group study, patients classified C0s to C4 according to Clinical Etiological Anatomic Pathophysiologic [CEAP] classification and with leg pain graded as superior to 4?cm on a 10-cm visual analog scale (VAS), were treated for 8 weeks with either MPFF 1000?mg once daily or MPFF 500?mg twice daily. The present post-hoc analysis focuses on the effect of treatment over time in patients randomized to the MPFF 1000?mg group. Leg pain was assessed at each follow-up visit by VAS. VAS scores over time were compared between each visit using paired Student t-tests.

Results: In total, 87 patients out of 174 were randomized to the MPFF 1000?mg group. Mean age?±?SD was 49.1?±?12.2 years, most of the patients were female (81.6%), the main CEAP classes of the most affected leg were C1 (20.7%), C2 (39.1%), C3 (33.33%), and the mean duration of CVD was 14.6?±?10.9 years. Patients with previous CVD treatment represent 27.6% of the patients. A MPFF 1000?mg tablet once daily was associated with a significant and continuous reduction in leg pain throughout the treatment period: –1.54?cm (±1.45) from baseline to week 2 (p?<?.01), –1.11?cm (±1.06) from week 2 to week 4 (p?<?.01), –1.57?cm (±1.05) from week 4 to week 8 (p?<?.01).

Conclusions: The new MPFF 1000?mg dose regimen in once daily tablets was associated with a rapid and continuous reduction in leg pain throughout the 8-week treatment period.     

Atorvastatin potentiates the effect of selenomethionine on thyroid autoimmunity in euthyroid women with Hashimoto’s thyroiditis

Objective: In many studies, selenium supplementation decreased serum titers of thyroid antibodies. The aim of the study was to investigate whether statin therapy determines selenium action on thyroid autoimmunity.

Methods: This prospective case-control study enrolled 42 euthyroid women with Hashimoto’s thyroiditis and normal vitamin D status, 20 of whom had been treated with atorvastatin (40?mg daily) for at least 6 months. All patients received selenomethionine (200 µg daily) for 6 months. Plasma levels of lipids, serum titers of thyroid peroxidase (TPOAb) and thyroglobulin (TgAb) antibodies, as well as serum levels of thyrotropin, free thyroid hormones, and 25-hydroxyvitamin D were determined at the beginning and at the end of the study.

Results: At baseline, there were no differences between both treatment arms in plasma lipids, titers of thyroid antibodies, serum levels of thyrotropin, free thyroid hormones, and 25-hydroxyvitamin D. Selenometionine decreased titers of TPOAb (from 843?±?228 to 562?±?189?U/mL) and TgAb (from 795?±?286 to 501?±?216?U/mL) in atorvastatin-treated women, as well as titers of TPOAb (from 892?±?247 to 705?±?205?U/mL) and TgAb (from 810?±?301 to 645?±?224?U/mL) in statin-naive women. The changes in antibody titers were more pronounced in women receiving atorvastatin (between-group difference: 94 [32–156] [TPOAb]; 129 [52–206] [TgAb]). Treatment-induced changes in TPOAb and TgAb correlated positively with baseline thyroid antibody titers. Circulating levels of lipids, free thyroxine, free triiodothyronine, and 25-hydroxyvitamin D remained at similar levels throughout the study.

Conclusions: The obtained results indicate that the decrease in titers of thyroid antibodies was potentiated by atorvastatin use.     

High one-year mortality following hospitalization for severe hypoglycemia among patients with diabetes mellitus: findings of a retrospective cohort study at an acute tertiary care hospital in Singapore

Objectives: Little is known about the 1-year short-term mortality rate following hospital admissions with severe hypoglycemia. This study aimed to determine the factors associated with increased 1-year mortality rate following hospitalization in diabetes patients admitted with severe hypoglycemia to the Singapore General Hospital.

Methods: Clinical, biochemical, and 1-year mortality data from diabetes patients who were admitted with severe hypoglycemia in the year 2014 were extracted from institutional medical records. Patients who passed away during the episode of admissions with severe hypoglycemia were excluded from the analysis. The clinical and biochemical factors between patients who survived and those who did not survive within 1 year following admission were compared using logistic regression analysis.

Results: Three hundred and four patients (181 female and 123 male) were admitted with severe hypoglycemia in 2014, and the mean capillary blood glucose on admission was 2.3?±?0.7?mmol/L. Sixty-three (20.7%) patients died within 1-year post-discharge from the hospital. Compared with patients who survived 1-year post-discharge from the hospital, non-survivors were older (69.3?±?11.0 vs 75.5?±?11.2 years, p?<?.001), had longer lengths of stay (LOS) (5.0?±?7.4 vs 9.0?±?12.8 days, p?=?.02), and had a higher Charlson Comorbidity Index (CCI) (4.1?±?1.9 vs 5.9?±?2.4, p?<?.001). Factors associated with increased 1-year mortality risk were age (odds ratio [OR]?=?1.06; 95% confidence interval [CI]?=?1.03–1.09, p?<?.01), LOS in hospital (OR?=?1.01; 95% CI?=?1.01–1.08, p?<?.01), and CCI (OR?=?1.51; 95% CI?=?1.31–1.75, p?<?.01), respectively.

Conclusions: Older diabetes patients with more comorbidities and longer LOS were at increased risk of dying within a year of discharge after hospitalization with severe hypoglycemia. Admission with severe hypoglycemia has important prognostic implications. Healthcare professionals should address hypoglycemia and other health issues during the hospital admissions.     

Interaction of deoxyschizandrin and schizandrin B with liver uptake transporters OATP1B1 and OATP1B3

1. Deoxyschizandrin and schizandrin B have diverse pharmacological effects, including hepatoprotective activity. We aim to study their hepatic uptake and their effects on the hepatic uptake of other clinical drugs mediated by OATP1B1 and OATP1B3.

2. Deoxyschizandrin exhibited a high affinity for OATP1B1 with K_m of 17.61?±?0.43?μM but a low affinity for OATP1B3. Similarly, schizandrin B also showed a strong affinity for OATP1B1 with K_m of 18.45?±?1.23?μM but a weak affinity for OATP1B3.

3. Atorvastatin and rifampicin could inhibit the uptake of deoxyschizandrin and schizandrin B mediated by OATP1B1.

4. Intriguingly, both deoxyschizandrin and schizandrin B significantly promoted the uptake of atorvastatin (with EC₅₀ of 50.58?±?8.08 and 24.70?±?5.82 µM, respectively) and rosuvastatin (with EC₅₀ of 13.46?±?2.70 and 8.99?±?4.73 µM, respectively) mediated by OATP1B1. Deoxyschizandrin could markedly promote the uptake of fluvastatin but inhibit the uptake of sodium taurocholate (TCNa) mediated by OATP1B1.

5. The promotion on hepatic uptake of statins mediated by OATP1B1 might lead to enhanced efficacy of cholesterol lowering and reduced risk of myopathy for hyperlipidemia patients when given statins together with deoxyschizandrin or schizandrin B.     

In vivo study of mPEG–PCL as a nanocarriers for anti-inflammatory drug delivery of simvastatin

Purpose: In this study, methoxy poly (ethylene glycol)-poly (ε-caprolactone) (mPEG–PCL) di-block copolymers were synthesized. The purpose of this work is to investigate the in vivo anti-inflammatory effects of simvastatin-loaded micelles.

Methods: The structure of synthesized copolymers was characterized by using HNMR, FTIR, and GPC techniques. Simvastatin was encapsulated in micelles through a single-step nano-precipitation method, leading to the formation of simvastatin-loaded mPEG–PCL (simvastatin-mPEG–PCL) micelles. In this study, the anti-inflammatory effects of simvastatin/mPEG–PCL micelles versus indomethacin were investigated in acute inflammation-induced rats. The paw edema thickness was measured 1, 2, 3, and 4?h after injection of formulation. The inhibition of edema in various groups were calculated and reported by percentages.

Results: The results showed that the zeta potential of micelles was about ?14.9?±?0.47?mV and the average size was in range of 66.10?±?0.34?nm. Simvastatin was encapsulated in mPEG–PCL micelles with a loading capacity of 9.63?±?0.87% and an encapsulation efficiency of 64.20?±?0.79%. Simvastatin and simvastatin-mPEG–PCL micelles showed significant anti-inflammatory activity in the present study.

Conclusions: This study revealed that simvastatin and simvastatin/mPEG–PCL micelles both have anti-inflammatory effects and suggested that statins have potential anti-inflammatory activity along with their lipid lowering properties.     

QAPSA: post-marketing surveillance of capsaicin 8% patch for long-term use in patients with peripheral neuropathic pain in France

Objective: To describe the characteristics of patients treated with the capsaicin 8% patch, prescribing conditions, long-term effects of repeat treatment on pain intensity and perception, quality of life, and impact on concomitant medication.

Methods: A national, longitudinal, prospective, non-interventional, post-marketing open study was conducted in 50 French pain centers. Adult volunteer non-diabetic patients with peripheral neuropathic pain receiving capsaicin 8% patch treatment were consecutively enrolled. Treatment could be repeated over a 12-month period, with 6?months’ follow-up after last application.

Results: A total of 684 patients (age: 53.0?±?14.9?years, mean?±?standard deviation; post-traumatic/surgical peripheral neuropathic pain: 76.3%; pain intensity: 6.2?±?1.7; pain duration: 3.0?years, median) were treated with 1 to 5 patches at 3/4?month intervals; 70.3% were naive to capsaicin 8% patch treatment at inclusion. Six months after last application, treatment was considered as successful for 21.8% (95% confidence interval: 17.5%–26.7%) of patients by a stringent criterion combining improvement according to the patient’s global impression of change (PGIC) and at least 30% improvement on a numerical pain rating scale (NPRS). Clinically relevant improvement in health-related quality of life was observed at end-of-study. No unexpected safety concerns were observed with capsaicin 8% patch repeat treatment.

Conclusions: The data of this post-marketing study meets the request by the French authorities for additional data on conditions of use in everyday practice. They confirmed the tolerance and long-term effect of capsaicin 8% patch in patients with peripheral neuropathic pain in real-world conditions.     

Stress-related upper gastrointestinal bleeding in adult neurocritical care patients: a Chinese multicenter,retrospective study

Objective: China has limited data on stress-related gastrointestinal ulcers in patients admitted for neurosurgical care. This study evaluated the incidence of upper gastrointestinal bleeding (UGIB) and use of stress ulcer prophylaxis (SUP) in Chinese neurocritical care patients (Glasgow Coma Scale [GCS] score ≤10).

Methods: This multicenter, retrospective study was performed from January 2015 to July 2015. Medical records of 1468 patients hospitalized during 2014 were reviewed. An estimated UGIB incidence rate of 4.4% was considered for precision of 1.3% for estimation of UGIB. The primary endpoint was evaluation of overall incidence of any overt UGIB in ≤14 days after cerebral lesion. Secondary endpoints included incidence of UGIB with or and without clinically significant complications, time to UGIB, associated risk factors and SUP used.

Results: We analyzed 1416 patients (mean age: 53.7?±?14.00 years; males: 62.4%) with cerebral lesions. Overall incidence rate of UGIB ≤14 days was 12.9% (95% CI: 11.2%–14.7%), 0.76% with and 12.1% without significant clinical complications. Average time and duration of bleeding were 2.9?±?3.37 days and 4.2?±?8.4 days, respectively. The most significant risk factors for UGIB were mechanical ventilation for >48?hours (p?<?.0001), UGIB history (p?=?.0026) and use of anticoagulants (p?<?.0001). Acid-suppression drugs were administered for SUP in 79.0% of the patients, whereas 40.5% received hemostatic drugs.

Conclusions: The rate of UGIB incidence was higher than the estimated rate in neurocritical care patients in China, suggesting the need for better management and treatment for stress-related mucosal disease in China. History of UGIB, mechanical ventilation and/or anticoagulants significantly affected UGIB.

ClinicalTrials registry number: NCT02316990.     

Effects of quercetin on the pharmacokinetics of losartan and its metabolite EXP3174 in rats

1. This study investigates the influence of quercetin on the pharmacokinetics of losartan and its metabolite EXP3174 in rats.

2. The pharmacokinetic profiles of losartan and EXP3174 of orally administered losartan (10?mg/kg) with or without pretreatment with quercetin (20?mg/kg/day for 7 days) were investigated. Additionally, Caco-2 cell transwell model and rat liver microsome incubation experiments were also conducted to investigate its potential mechanism.

3. The results showed that when the rats were pretreated with quercetin, the C_max (2.16?±?0.40 vs. 1.33?±?0.21?mg/L) and the AUC_(0–t) (13.89?±?1.22 vs. 7.34?±?0.75?mg·h/L) of losartan increased significantly (p?<?.05), and while the C_max (0.76?±?0.09 vs. 1.14?±?0.18?mg/L) of EXP3174 decreased significantly compared to the control (p?<?.05). The t_1/2 of losartan was prolonged from 3.27?±?0.45?h to 4.74?±?0.51?h (p?<?.05). The results also indicated that quercetin could increase losartan absorption rate by inhibiting the activity of P-gp and decrease its metabolic stability by inhibiting the activity of CYP450 enzyme.

4. These results indicated that the herb–drug interaction between quercetin and losartan might occur when they are co-administered in rats, quercetin could increase the systemic exposure of losartan and decrease the plasma concentration of EXP3174, possibly by inhibiting the activity of P-gp or CYP450 enzyme.     

Optimisation of the microencapsulation of lavender oil by spray drying

Background: Lavender oil consists of around 100 components and is susceptible to volatilisation and degradation reactions.

Aim: Microencapsulate lavender oil by spray drying using a biocompatible polymeric blend of gum acacia and maltodextrin to protect the oil components. Effect of total polymer content, oil loading, gum acacia, and maltodextrin proportions on the size, yield, loading, and encapsulation efficiency of the microparticles was investigated.

Methods: Morphology and oil localisation within microparticles were assessed by confocal laser scanning electron microscope. Structural preservation and compatibility were assessed using Fourier transform infra-red spectroscopy, differential scanning calorimetry, and gas chromatography–mass spectrometry.

Results: Lavender microparticles of size 12.42?±?1.79?µm prepared at 30 w/w% polymer concentration, 16.67 w/w% oil loading, and 25w/w% gum acacia showed maximum oil protection at high loading (12?mg w/w%), and encapsulation efficiency (77.89 w/w%).

Conclusion: Lavender oil was successfully microencapsulated into stable microparticles by spray drying using gum acacia/maltodextrin polymeric blend.     

Impact of lixisenatide dose range on clinical outcomes with fixed-ratio combination iGlarLixi in patients with type 2 diabetes

Objective: To evaluate the lixisenatide dose range delivered by the iGlarLixi SoloSTAR pen (5–20 µg), alone or in fixed-ratio combination with insulin glargine (iGlar; iGlarLixi).

Methods: Data from three clinical studies were analyzed to assess lixisenatide efficacy and safety: a phase 2a trial assessing gastric emptying effects (ACT6011); a phase 2b dose-ranging trial (DRI6012); and a randomized controlled phase 3 trial comparing iGlarLixi with its components of iGlar and lixisenatide (LixiLan-O). Efficacy metrics included glycated hemoglobin A_1c (A1C), post-prandial glucose (PPG) values following a standardized breakfast, fasting plasma glucose (FPG), and weight change. Occurrence of gastrointestinal adverse events was also assessed.

Results: ACT6011: lixisenatide doses from 5–20?μg once daily (QD) suppressed PPG; maximal reductions in mean PPG area under the curve were achieved with doses ≥12.5 µg QD, but doses as low as 5?μg achieved 44% of maximal reduction. DRI6012: lixisenatide doses 5–20?μg QD resulted in significant, dose-dependent decreases in A1C, percentage of patients achieving A1C <7.0%, and 2-h PPG levels; doses of 20?μg achieved complete suppression of PPG. LixiLan-O: iGlarLixi decreased 2-h PPG across the entire dose range. Lixisenatide dose was unrelated to reductions in FPG with iGlarLixi. Similar reductions in A1C were seen with iGlarLixi across all lixisenatide doses.

Conclusions: This analysis demonstrates the clinical benefit of lixisenatide alone or in the formulation of iGlarLixi over the entire dose range of lixisenatide contained in iGlarLixi (5–20 µg), supporting the selection of the lixisenatide dose range delivered by the iGlarLixi SoloSTAR pen.     

The effect of complexation with cyclodextrins on the antioxidant and antimicrobial activity of ellagic acid

Purpose: The aim of the paper was to develop the simple procedures for preparation of inclusion complexes of ellagic acid (EA) with cyclodextrins (CDs) and to investigate their antioxidant and antimicrobial activity.

Methods: The structural characterization was carried out using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and nuclear magnetic resonance (NMR) methods. The phase solubility technique was used to investigate the interactions between ‘host’ and ‘guest’ molecules and to estimate the molar ratio between them. The antioxidant and antimicrobial activity of EA and inclusion complexes were determined.

Results: The apparent stability constants were found to be 117?dm³ mol^?1 for the complex with β-CD and 161?dm³ mol^?1 for the complex with (2-hydroxypropyl)-β-cyclodextrin (HP-β-CD). The results of phase-solubility studies showed that EA formed the inclusion complexes with CDs in the molar ratio of 1:1. The calculated half-maximal inhibitory concentration was 41.18?μg cm^?3 for butyl hydroxy toluene, 1.96?μg cm^?3 for EA, 0.88?μg cm^?3 for inclusion complex with HP-β-CD, and 1.27?μg cm^?3 for inclusion complex with β-CD.

Conclusion: The stability constants indicated the rapid release of EA from the inclusion complexes in the aqueous medium at 25?°C. The antioxidant activity of EA was increased, while the antimicrobial activity was preserved after complexation with CDs.     

Characterisation of peppermint (Mentha piperita L.) essential oil encapsulates

Aim: The aim was to choose the optimal encapsulation system and to incorporate encapsulates loaded with essential oil into the ice cream as a model food product.

Methods: Ca-alginate beads were produced by electrostatic extrusion process. Gelatine/alginate coacervates were processed with coacervation. Carnauba wax microparticles were produced using melt dispersion process. Morphological properties, chemical, and thermal stabilities of encapsulates were tested by SEM, FTIR spectral, and thermogravimetric analysis.

Results: Alginate provided sufficient emulsion stability over 1?h. Ca-alginate showed higher encapsulation efficiency (EE) (98.4?±?4.3%) compared to carnauba wax (94.2?±?7.8%) and gelatine/alginate coacervates (13.2?±?1.2%). The presence of essential oil in all three types of encapsulates confirmed with FTIR. The encapsulation process ensured controlled release and thermal stability of the oil.

Conclusions: Ca-alginate matrix as the most suitable for peppermint essential oil encapsulation. The sensory analysis showed that ice cream incorporating encapsulates is a promising system for the consumption of health beneficial peppermint essential oil.     

A randomized,double-blinded,placebo-controlled,parallel trial of vitamin D3 supplementation in adult patients with migraine

Background: Vitamin D levels have been linked to certain pain states, including migraine. This study investigated whether vitamin D supplementation would be beneficial for adult patients with migraine (ClinicalTrials.gov Identifier: NCT01695460).

Methods: A randomized, double-blinded, placebo-controlled parallel trial was conducted in migraine patients (36 women and 12 men, 18–65 years of age). A 4-week baseline period was conducted before randomization to 24 weeks of treatment. Participants were assigned to receive D3-Vitamin (n?=?24, 18 women and 6 men, 100?μg/day D3-Vitamin) or placebo (n?=?24,?18 women and 6 men). Migraine attacks and related symptoms were assessed by self-reported diaries. The response rate (i.e. experiencing a 50% or greater reduction in migraine frequency from baseline to week 24), change in migraine severity, and number of migraine days were recorded. Changes in migraine-related symptoms, HIT-6^TM scores, and pain sensitivity tests (pressure pain threshold and temporal summation) were also evaluated. Serum levels of both 25 (OH)D and 1,25 (OH)₂D were assessed from baseline to week 24.

Results: The number of headache days changed from 6.14?±?3.60 in the treatment group and 5.72?±?4.52 in the placebo group at baseline to 3.28?±?3.24 and 4.93?±?3.24 by the end of the trial, respectively. Migraine patients on D3-Vitamin demonstrated a significant decrease (p?<?.001) in migraine frequency from baseline to week 24 compared with placebo. However, migraine severity, pressure pain thresholds, or temporal summation did not show a significant change. 25(OH)D levels increased significantly for the D3-Vitamin group during the first 12 weeks of treatment. There was no significant change in 1,25(OH)₂D. No side-effects were reported or noted.

Conclusions: D3-Vitamin was superior to placebo in reducing migraine days in migraine patients. Larger studies are required to confirm that vitamin D₃ might be one of the prophylactic options for adult patients with migraine.     

Bufalin induces apoptosis via mitochondrial ROS-mediated caspase-3 activation in HCT-116 and SW620 human colon cancer cells

Objective: Bufalin has been reported to kill various types of cancer including human colorectal cancer. Our previous study demonstrated that bufalin induced cell death via autophagy in HT-29 and Caco-2 colon cancer cells, but the action of bufalin remains unclear. This study was conducted to investigate the role of bufalin in other colon cancer HCT-116 and SW620 cells as well as its potential mechanism.

Methods: The effect of bufalin in HCT-116 and SW620 colon cancer cells was detected by assessing cell viability and cell death. Apoptotic cells were analyzed by Western blot and trypan blue dye exclusion assay. Mitochondrial ROS production was analyzed by flow cytometry after DCFDA and DHR-123 staining. The potential mechanism was investigated via pharmacological inhibitors.

Results: Bufalin had high potency against HCT-116 and SW620 cells with IC50 values of 12.823?±?1.792?nM and 26.303?±?2.498?nM in HCT-116 and SW620 cells, respectively. Bufalin decreased cell viability, increased cell death as well as caspase-3 downstream target (cleaved PARP) accumulation, and these actions were significantly blocked by pan-caspase inhibitor zVAD-FMK. Mechanistically, ROS production, but neither the NAD(P)H oxidase, AMPK, ERK nor p38, is responsible for bufalin-induced apoptotic cell death. Moreover, bufalin-induced ROS generation is derived from mitochondria.

Conclusion: Bufalin significantly induces apoptosis in HCT-116 and SW620 colon cancer cells via mitochondrial ROS-mediated caspase-3 activation. We believe that our novel findings will greatly alter our current understanding on the anti-cancer mechanism of bufalin in colon cancer cells and will pave the way for further exploiting the clinical application.     

Comparisons of acute inflammatory responses of nose-only inhalation and intratracheal instillation of ammonia in rats

Objective: To establish a rat model with respiratory and pulmonary responses caused by inhalation exposure to non-lethal concentrations of ammonia (NH₃) that can be used for evaluation of new medical countermeasure strategies for NH₃-induced acute lung injury (ALI). This is of great value since no specific antidotes of NH₃-induced injuries exist and medical management relies on supportive and symptomatically relieving efforts.

Methods: Female Sprague-Dawley rats (8–9?weeks old, 213g?±?2g) were exposed to NH₃ using two different exposure regimens; nose-only inhalation or intratracheal instillation. The experiment was terminated 5?h, 24?h, 14 and 28?days post-exposure.

Results: Nose-only inhalation of NH₃ (9000–15 000?ppm) resulted in increased salivation and labored breathing directly post-exposure. Exposure did not increase inflammatory cells in bronchoalveolar lavage fluid but exposure to 12 000?ppm NH₃ during 15?min reduced body weight and induced coagulation abnormalities by increasing serum fibrinogen levels. All animals were relatively recovered by 24?h. Intratracheal instillation of NH₃ (1%) caused early symptoms of ALI including airway hyperresponsiveness, neutrophilic lung inflammation and altered levels of coagulation factors (increased fibrinogen and PAI-1) and early biomarkers of ALI (IL-18, MMP-9, TGFβ) which was followed by increased deposition of newly produced collagen 14?days later. Histopathology analysis at 5?h revealed epithelial desquamation and that most lesions were healed after 14?days.

Conclusions: This study demonstrates that intratracheal instillation can reproduce several early hallmarks of ALI. Our findings therefore support that the intratracheal instillation exposure regimen can be used for new medical countermeasure strategies for NH₃-induced ALI.     

Montelukast nasal spray: formulation development and in vitro evaluation

Objectives: The aim of this study was to evaluate potential development of montelukast sodium (MTS) as a nasal spray.

Materials and methods: The formulations were prepared using hydroxypropyl cellulose (HPC) and carbomer 940 (C940). The prepared formulations were evaluated for clarity, pH, hydrodynamic size, zeta potential, viscosity, contact angle, surface tension, droplet size distribution, muco-adhesiveness, drug release, and stability. The suitable formulations were also assessed for their effects on nasal epithelial cells.

Results and discussion: At room temperature (25?°C), the formulation containing 0.01% w/v C940 exhibited suitable viscosity and rheological properties. Spray droplets were in ranges of 10–40?µm, which are suitable for nasal administration. The works of adhesion of the formulations containing HPC or C940 were in the range of 0.8–4.0 and 0.2–27 µJ, respectively. In addition, the formulation containing 0.01% w/v C940 and the 0.5–15?µg/50?µL concentrations of MTS showed no signs of cytotoxicity and did not open the tight junction of nasal epithelial cells.

Conclusion: The formulated MTS nasal spray is ideal for nasal administration for the treatment of allergic rhinitis. The formulation containing 0.01% w/v C940 had no toxicity nor alteration on the tight junction of nasal epithelial cells.     

Non-clinical pharmacokinetic profiles of rovatirelin,an orally available thyrotropin-releasing hormone analogue

1. The non-clinical pharmacokinetic profiles of rovatirelin, a novel thyrotropin-releasing hormone (TRH) analogue, were investigated in vivo and in vitro.

2. Rovatirelin orally administered to rats and dogs was rapidly absorbed and bioavailability was estimated to be 7.3 and 41.3%, respectively. The extent of plasma protein binding of rovatirelin in rats, dogs, and humans was low in all species (～15%). The permeability of rovatirelin from blood to brain (permeability-surface area) ranged from 1.04?±?0.14 to 1.29?±?0.28?μL/min/g in rats, and rovatirelin was stable in rat plasma and brain homogenates.

3. The metabolite pattern was qualitatively similar in vitro and in vivo. In animals, rovatirelin aminopentanoic acid (rovatirelin-acid), rovatirelin aminopentanone (rovatirelin-ketone), rovatirelin pyrrolidine (4S)-hydroxy (rovatirelin-OH), (thiazoylalanyl)methylpyrrolidine (TAMP), 3-(4-thiazoyl)-l-alanine (TA), and unknown metabolites were observed. In human hepatocytes, TAMP was mainly formed and no unique human metabolite was observed.

4. The radioactivity from administered [¹⁴C]rovatirelin was predominantly excreted in faeces in rats and dogs, and almost all radioactivity was recovered 168?h after administration. Absorption, brain penetration, and stability of rovatirelin in the brain were greater than for taltirelin.

5. Thus, orally administered rovatirelin is a potentially improved treatment for spinocerebellar degeneration compared with taltirelin.