A novel approach to prepare insulin-loaded poly(lactic-co-glycolic acid) microcapsules and the protein stability study

The purpose of this study was to propose a new preparation method to fabricate insulin-loaded poly(lactic-coglycolic acid) (PLGA) microparticles satisfying protein loading, release profiles, burst release, and particularly stability of the encapsulated protein. Insulin-loaded microcapsules were produced by a single phase o/o solvent evaporation method. The characteristics of the microcapsules were determined by various methods: the surface morphology and size of microparticles by atomic force microscopy and scanning electron microscopy, insulin crystalinity and drug-polymer interactions by XRD, DSC, and FTIR, chemical integrity and aggregation of insulin using HPLC and SDS-PAGE, the protein secondary structure by far ultraviolet-circular dichroism (CD), the antigenicity activity of insulin with ELISA techniques. PLGA microparticles showed smooth surfaces with microcapsule. Encapsulation efficiency of 51% and constant insulin release rate with initial insulin burst release of 24% was obtained. Encapsulated and released insulin was in the intact form and it was dispersed in crystalline state in the polymer matrix. Ease of manufacturing under mild preparation conditions, high level of drug entrapment, desirable release pattern with relatively low initial burst effect and an ability to preserve protein structure are the advantages which are offered by the developed protein encapsulation method.     

Particle size design of PLGA microspheres for potential pulmonary drug delivery using response surface methodology

The large surface area, good vascularization, immense capacity for solute exchange and ultra-thinness of the alveolar epithelium are unique features of the lung facilitating systemic drug delivery via pulmonary administration. The efficacy and safety of many new and existing inhaled therapies may be enhanced through advanced controlled-release systems by using polymer particles. Poly (D,L-lactic-co-glycolic acid) (PLGA) is well known by its safety in biomedical preparations which has been approved for human use by the FDA. The optimum aerodynamic particle size distribution for most inhalation aerosols has generally been recognized to be in the range of 1-5 microns. PLGA microspheres, therefore, were prepared by a developed oil-in-oil solvent evaporation method and characterized. A four-factor, three levels Box-Behnken design was used for the optimization procedure with temperature, stirring speed, PLGA and surfactant concentration as independent variables. Particle size and polydispersity of microspheres were considered as dependent variables. PLGA microparticles were prepared successfully in desired size for pulmonary delivery by solvent evaporation method. It was found that the particle size of microspheres could be easily controlled. It was also proved that response surface methodology could efficiently be applied for size characterization and optimization of PLGA microparticles for pulmonary drug delivery.     

赤芝子实体中灵芝酸类成分的研究

自赤芝［Ｇａｎｏｄｅｒｍａｌｕｃｉｄｕｍ（Ｆｒ．）Ｋａｒｓｔ．］子实体的二氯甲烷提取物中分离得到一个新的四环三萜化合物，命名为灵芝酸ＤＭ（ｇａｎｏｄｅｒｉｃａｃｉｄＤＭ，Ｉ）。根据光谱（ＵＶ，ＩＲ，１ＨＮＭＲ，１３ＣＮＭＲ，ＭＳ２ＤＮＭＲ）分析，确定其结构为Ｉ式。同时还分离得到二个已知的灵芝酸类化合物，即灵芝酸Ａ（ｇａｎｏｄｅｒｉｃａｃｉｄＡ，Ｉ）和灵芝酸Ｃ（ｇａｎｏｄｅｒｉｃａｃｉｄＣ，ＩＩ）。     

Factors predictive of response and survival in patients with metastatic colorectal cancer in Taiwan

5-Fluorouracil in combination with leucovorin has been shown to be active in therapeutic trials of metastatic colorectal carcinoma. In this study, we administered these drugs to 72 patients with metastatic colorectal carcinoma. Thirty-six of them without previous exposure to 5-fluorouracil were treated with weekly bolus injections of 5-fluorouracil (425 mg/m2) and leucovorin (25 mg/m2) supplemented with oral levamisole. Another 36 patients with or without prior 5-fluorouracil treatment received 5-fluorouracil 3,000 mg/m2 and leucovorin 300 mg/m2 in a 48-hour continuous infusion every two weeks. Clinical efficacy and toxicity were assessed by WHO criteria. Variables were tested for relations to response and survival by univariate and multivariate analysis. The response rate was 19.4% in weekly bolus arm and 13.9% in biweekly high-dose infusion arm (P = 0.527). Median survivals in the two arms were 18.4 months (weekly) and 21 months (biweekly) respectively (P = 0.708). Gastrointestinal side effects including nausea, vomiting, diarrhea and mucositia were the major toxicities of these regimens. By multivariate analysis, the only factor to influence response rate was the site of metastases (P = 0.009). The only factor to affect survival was performance status of the patient (P = 0.0001). We concluded that the two 5-fluorouracil based regimens are well-tolerated and shown to have a response rate comparable with previous reports of similar regimens in patients with metastatic colorectal cancer. Only liver metastases seemed to have a better response to therapy. Performance status is the most important prognostic factor in patients with metastatic colorectal cancer.      

Toxic cardiogenic shock in a patient receiving weekly 24-h infusion of high-dose 5-fluorouracil and leucovorin

A 54-year-old man was treated with weekly 24-h infusion of high-dose 5-fluorouracil (2600 mg/m2) and leucovorin (100 mg/m2) for metastatic colon cancer. At first, he tolerated the treatment well and no significant toxicity was identified. After a total of eight courses of treatment, a stable disease was observed, but mild shortness of breath was found on occasion. The patient had no previous history of cardiac disease and the heart performance assessed by left ventricular ejection fraction before treatment was normal. Unfortunately, acute pulmonary edema with lethal cardiogenic shock occurred during the ninth course of treatment, in spite of intensive medical treatment. The chest X-ray showed extreme cardiomegaly. Repeated assessment of his heart function by echocardiogram and ventricular ejection fraction revealed a very poor cardiac performance. Toxic cardiogenic shock during weekly 24-h infusion of high-dose 5-fluorouracil and leucovorin is extremely rare. To the best of our knowledge, no case has been reported in the English literature. We report a case and the relevant literature about the incidence, clinical picture and possible pathophysiology on 5-fluorouracil-related cardioxicity is reviewed.      

Pharmacokinetic study of niosome-loaded insulin in diabetic rats

Background and the purpose of the study

Encapsulation of human insulin in lipid vesicular systems such as niosomes was sought as a route to protect this protein against proteolytic enzymes and to improve its oral bioavailability. The purpose of this study was to assess the effect of insulin encapsulation in niosomes on oral bioavailability in diabetic rats.

Methods

Recombinant human insulin was entrapped in multilamellar niosomes composed of polyoxyethylene alkyl ether surfactants (Brij 52 and Brij 92) or sorbitan monostearate (Span 60) and cholesterol. The amount of insulin released in simulated intestinal fluid (SIF) and simulated gastric fluid (SGF) were measured at 37°C. The protection of entrapped insulin against pepsin, α-chymotrypsin and trypsin were evaluated in comparison with free insulin solution. Diabetes was induced by IP injection of streptozotocin (65 mg/kg) in male wistar rats and effects of orally administered niosomes and subcutaneously injected insulin on hypoglycemia and elevation of insulin levels in serum were compared.

Results and conclusion

The extent and rate of insulin release from Brij 92 and Span 60 vesicles were lower than that of Brij 52 niosomes (P<0.05). Vesicles protected insulin in comparison with free insulin solution against proteolytic enzymes (P<0.05) significantly. Animals treated with oral niosome-encapsulated insulin (100 IU/kg) showed decreased levels of blood glucose and elevated serum insulin, which in the case of Brij 92 niosomes, hypoglycemic effect was significant (P<0.05).Niosomes were also stable in solubilizing bile salt solutions and could effectively prolong the release of insulin in both SGF and SIF. Results of this study showed that niosomes may be utilized as oral carriers of insulin; however, to increase bioavailability of insulin, further studies on the protease inhibitor co-encapsulation in niosomal formulations might be helpful.     

Vacuum-assisted closure as a surgical assistant in life-threatening necrotizing fasciitis in children

Necrotizing fasciitis is a severe soft tissue infection that can involve skin, subcutaneous fat, fascia and muscle. It can result in devastating sequelae including tissue necrosis, sepsis, toxic shock syndrome, cardiopulmonary collapse and death. To control rapidly spreading necrosis, early diagnosis and aggressive surgical treatment with extensive radical debridement of the affected areas is necessary, as well as systemic administration of broad-spectrum antimicrobials and, very often, intensive care support.The subatmospheric negative pressure dressing has been previously used in acute and complex wounds management. The concept of using vacuum-assisted closure dressing as another management component is presented in the current article.     

Inhibitory activity of xanthine oxidase by fractions Crateva adansonii

ObjectiveTo study the inhibitory effect of various extracts from Crateva adansonii (C. adansonii) used traditionally against several inflammatory diseases such as rheumatism, arthritis, and gout, was investigated on purified bovine milk xanthine oxidase (XO) activity.MethodsXanthine oxidase inhibitory activity was assayed spectrophotometrically and the degree of enzyme inhibition was determined by measuring the increase in absorbance at 295 nm associated with uric acid formation. Enzyme kinetics was carried out using Lineweaver-Burk plots using xanthine as the substrate.ResultsAmong the fractions tested, the chloroform fraction exhibited highest potency (IC₅₀ 20.2±1.6 μg/mL) followed by the petroleum ether (IC₅₀ 30.1±2.2 μg/mL), ethyl acetate (IC₅₀ 43.9±1.4 μg/mL) and residual (IC₅₀ 98.0±3.3 μg/mL) fractions. The IC₅₀ value of allopurinol used, as the standard was 5.7±0.3 μg/mL.ConclusionsEnzyme inhibition mechanism indicated that the mode of inhibition was of a mixed type. Our findings suggest that the therapeutic use of these plants may be due to the observed Xanthine oxidase inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.     

Electrical Performance and Automatic Capture Characteristics of a 3.5-mm2 Passive Fixation Lead during 1-Year Follow-Up

Background: Bipolar low polarization electrodes are recommended for a regular AutoCapture™ (St. Jude Medical, Inc., Sylmar, CA, USA) function in order to effectively detect the evoked response (ER) signal. The objective of this national multicenter registry was to evaluate the electrical performance and the AutoCapture™ characteristics of the bipolar ventricular pacing lead IsoFlex S, model 1636T or 1646T (St. Jude Medical), in combination with single- and dual-chamber pacemakers.
Methods: Ventricular pacing and sensing thresholds, lead impedance, ER amplitude, and polarization signals were measured at discharge and routine follow-up visits after 1, 3, 6, 9, and 12 months. AutoCapture™ activation was recommended based on the results of the ER sensitivity test.
Results: Of the 252 patients initially included, 109 (43%) have completed the follow-up. The mean ventricular pacing threshold was 0.43 ± 0.19 V at discharge and 0.68 ± 0.32 V at 12 months postimplant. The values for the ventricular sensing threshold were between 9.51 ± 4.12 and 9.99 ± 4.09 mV at discharge and at the 12-month follow-up. The unipolar lead impedance decreased from 533 ± 94 to 476 ± 73 ohms during the follow-up. The mean ER amplitude was 16.47 ± 6.70 mV at discharge and 17.42 ± 7.43 mV after 12 months, and the corresponding mean polarization signals were 0.59 ± 1.00 and 0.74 ± 1.24 mV, respectively. AutoCapture™ activation was recommended in at least 95% of the patients investigated over the 12-month follow-up.
Conclusion: The bipolar ventricular pacing lead IsoFlex S 1636/1646T shows a good electrical performance and is mostly compatible with the AutoCapture™ algorithm.