Therapeutic potential of phosphodiesterase type 4 inhibition in chronic autoimmune demyelinating disease

It was recently demonstrated that selective phosphodiesterase type 4 (PDE4) inhibition suppresses the clinical manifestations of acute experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), and inhibits the production of tumor necrosis factor-α (TNF-α), a pathogenetically central cytokine. Since the most common presentation of MS in humans is a relapsing–remitting course, we investigated the therapeutic potential of PDE4 inhibition in the relapsing–remitting EAE model of the SJL mouse. Administration of rolipram, the prototypic PDE4 inhibitor, reduced the clinical signs of EAE during both the initial episode of disease and subsequent relapses. In parallel, there was marked reduction of demyelination and also less inflammation throughout the central nervous system (CNS) of rolipram-treated animals. Gene expression of proinflammatory cytokines in the CNS was reduced in most of the rolipram-treated animals. Additional experiments demonstrated that PDE4 inhibition acted principally by inhibiting the secretion of Th1 cytokines, however, the encephalitogenic potential of myelin basic protein-specific T cells was not impaired. Our findings suggest that PDE4 inhibitors are a promising cytokine-directed therapy in chronic demyelinating disease.     

Temporal dependence of ectopeptidase expression in alveolar epithelial cell culture: implications for study of peptide absorption

There is little data available regarding the extent of peptide metabolism encountered following inhalation to the lung. We have studied the activity of five ectopeptidases in primary rat alveolar epithelial cells, A549 cells and pulmonary macrophages. Peptidase activity was assayed in the plasma membrane fractions (PMF) of primary type II alveolar epithelial cells (ATII cells) after 2 days in culture and after 7 days in culture when they had formed monolayers of type I-like cells (ATI-like cells). Dipeptidyl peptidase IV (DPP) activity fell from 36.65 mU/mg protein to 16.29 mU/mg protein between day 2 and day 7 in culture, aminopeptidase N (AMN) activity increased from 16.16 mU/mg protein to 23.99 mU/mg protein, angiotensin-converting enzyme (ACE) activity was lost (4.29 mU/mg protein at day 2, not detected at day 7), and carboxypeptidase M (CPM) activity was acquired (not detected at day 2, 5.20 mU/mg protein at day 7). The profile of exopeptidase expression in A549 cells was similar to that of primary rat alveolar cells at day 7 in culture (DPP 24.24 mU/mg protein, AMN 47.74 mU/mg protein, CPM 4.28 mU/mg protein, ACE not detected). Macrophages expressed high levels of aminopeptidases (DPP 46.85 mU/mg protein, AMN 28.28 mU/mg protein) but carboxypeptidase activity was not detected. Low neutral endopeptidase 24.11 (NEP) activity was found in all cell types studied (0.96–2.41 mU/mg protein). The qualitative and quantitative changes in the peptidase activity of primary cultured rat alveolar cells between day 2 and day 7 in culture has implications for the use of alveolar cell monolayers as drug absorption models to investigate peptide absorption from the lung. Ectopeptidase activity in cultured alveolar cells can be used to infer the peptide-metabolising capacity of the surface of the alveolar epithelium. The aminopeptidase activity in particular, if representative of enzyme activity in vivo, would offer a significant metabolic barrier to systemic delivery of peptides via the lung.     

Targeting neutrophil collagenase/matrix metalloproteinase-8 and gelatinase B/matrix metalloproteinase-9 with a peptidomimetic inhibitor protects against endotoxin shock

Gram-negative sepsis, bacterial meningitis and endotoxin shock are life-threatening disorders, associated with the rapid release of neutrophil enzymes. Neutrophil collagenase/matrix metalloproteinase-8 (MMP-8) and gelatinase B/matrix metalloproteinase-9 (MMP-9) are contained in granules, are quickly exocytosed upon granulocyte activation and efficiently cleave intact and denatured collagens, respectively. Genetic ablation of gelatinase B protects against endotoxin-induced mortality. Therefore, we designed and synthesized a peptidomimetic gelatinase B inhibitor Regasepin1, and compared the selectivity for the collagenases MMP-1, MMP-8 and MMP-13. Regasepin1 was found to inhibit, almost to the same degree, the neutrophil enzymes MMP-8 and MMP-9 and the monocytic tumor necrosis factor-alpha (TNF-alpha) converting enzyme (TACE/ADAM-17) in vitro. With the use of mass spectrometry analysis, the plasma half-life of inhibitor levels was determined after an intraperitoneal bolus injection in mice. Plasma peak levels of the inhibitor were reached at 50 min after intraperitoneal injection and the subsequent half-life in the circulation exceeded 40 min. Regasepin1 protected mice against lethal endotoxinemia by intraperitoneal and intravenous injection routes. This proves the principle that early neutrophil MMP inhibition followed by TACE blockade may become a treatment strategy of gram-negative sepsis, endotoxinemia and other life-threatening inflammatory reactions.     

Monoclonal antibodies specific for kappa chain, lambda chain, and IgG1 of human gammaglobulin

Hybrid cell lines secreting antibodies specific for human gammaglobulin (HGG) were prepared by cell fusion and cloning techniques. The monoclonal antibodies were tested for their antibody reacts with a different antigenic determinant of HGG. One reacts with isolated kappa (kappa) light chains, one with isolated lambda (lambda) light chains, and one with the Fc fragment of IgG1 molecules. The reactivity patterns of two additional monoclonal antibodies are more complex. One reacts with a determinant present on the Fc of all IgG subclasses and the other binds to a determinant on the Fab of IgG molecules. The two monoclonal antibodies reactive with light chains also bind to surface components of human B cells. The murine immunoglobulin (Ig) class of each clone product was identified.     

Allergic reactions to streptokinase consistent with anaphylactic or antigen-antibody complex-mediated damage

Streptokinase used as a thrombolytic agent may produce immunologic drug reactions. We report a second case of IgE-mediated anaphylaxis and a late reaction to streptokinase. This late reaction was consistent with an IgG-mediated antigen-antibody disease with transient renal involvement, fever, and cutaneous lesions. The second case responded to prednisone therapy.     

Pre-hospital delays and intravenous thrombolysis in urban and rural areas

Kozera G, Chwojnicki K, Gójska‐Grymaj?o A, G?secki D, Schminke U, Nyka WM. Pre‐hospital delays and intravenous thrombolysis in urban and rural areas.
Acta Neurol Scand: 2012: 126: 171–177.
© 2011 John Wiley & Sons A/S. Introduction– It is crucial to understand the reasons behind pre‐ and in‐hospital delays to improve nationwide access to effective treatment for acute stroke. Aims– To evaluate the pre‐ and in‐hospital delays and to compare the intravenous (IV) thrombolysis rates in the urban and rural areas of the Province of Pomerania, Poland. Materials & methods– We evaluated the medical records of 2134 patients treated in the stroke units (SUs) and consecutively reported to the Pomeranian Stroke Register from June 2006–December 2007. Results– The time of ischaemic stroke onset was known in 488 (59%) of the 834 urban patients and in 744 (70%) of the 1063 rural patients (P < 0.001). The proportion of patients who called the emergency medical services with a delay of >45 min was similar in both locations: urban, 314/488 (64.3%) vs rural, 490/744 (65.8%). Although the proportion of patients who reached the emergency room within 3 h was higher in the rural areas (29.0% vs 24.3%; P = 0.02), only 4.2% of these patients received IV thrombolysis compared with 23.1% in the urban areas (P < 0.001). The proportion of patients who did not seek any kind of professional medical help prior to admission was lower in the rural areas (29/744 (3.9%) vs urban 50/488 (10.2%)) (P < 0.001). Conclusions– Pre‐hospital delays reduced the number of patients eligible for IV thrombolysis in both rural and urban areas. The low proportion of patients treated with IV thrombolysis in rural SUs may be attributed to ineffective in‐hospital procedures.     

多西他赛联合顺铂和替吉奥胶囊治疗宫颈癌的临床疗效观察

目的 研究多西他赛联合顺铂和替吉奥胶囊治疗宫颈癌的临床疗效和安全性。方法 将2011年1月-2013年12月就诊于重庆市大足区人民医院的宫颈癌患者120例随机分为联合治疗、多西他赛、替吉奥胶囊组,每组40例,3组患者均在1个疗程的前3天每天静脉滴注注射用顺铂30 mg/m²,联合治疗组在此基础上在第1天静脉滴注多西他赛注射液,60 mg/m²,同时口服替吉奥胶囊60 mg/m²,2次/d,连服2周。多西他赛组静脉滴注多西他赛注射液,用法用量同联合治疗组。替吉奥胶囊组口服替吉奥胶囊,用法用量同治疗组。21 d为一个疗程,3组患者共治疗2个疗程。比较3组患者治疗后的临床疗效、肿瘤直径和不良反应发生情况。结果 治疗后3组患者的肿瘤直径均显著减小,同组治疗前后差异有统计学意义（P<0.05、0.01）;治疗后,多西他赛组和替吉奥胶囊组患者肿瘤直径较联合治疗组大,差异有统计学意义（P<0.05）。联合治疗组、多西他赛组和替吉奥胶囊组的总有效率分别为95.0%、77.5%、80.0%,后两组的总有效率均显著低于联合治疗组,差异有统计学意义（P<0.05）。3组患者均发生化疗的毒副反应,发生率为100%,其中主要包括恶心、呕吐、腹泻、脱发、肾功能损伤、骨髓抑制。3组患者不良反应发生率差异无统计学意义。结论 多西他赛联合顺铂和替吉奥胶囊治疗宫颈癌具有良好的临床疗效,可显著减小肿瘤直径,且不良反应可控。     

反相高效液相色谱法测定贞芪扶正颗粒中黄芪甲苷的含量

[目的]反相高效液相色谱法测定贞芪扶正颗粒中黄芪甲苷的含量。[方法]采用反相高效液相色谱法，流动相为乙腈-水(30：70)，检测波长为205nm。[结果]线性方程为Y=751565．09X-41668．30，r=0．9998，黄芪甲苷在0．53μg～2．65μg范围内线性关系良好，平均回收率为98．17％，RSD％为1．61％(n=6)。[结论]建立的检测方法简便、准确。     

Effects of trypsin inhibitor on the remant pancreas after 85 percent pancreatectomy in rats

Major resection of the pancreas leads to disorders of the endocrine and exocrine pancreas. The effect of a trypsin inhibitor on the remnant pancreas was studied in rats after 85 percent pancreatectomy. Impairments of the glucose elimination rate and the integrated insulin response after 85 percent pancreatectomy were improved by means of oral administration of a synthetic trypsin inhibitor for 4 and 12 weeks. The pancreatic insulin content in the animals treated with trypsin inhibitor for 13 weeks increased to about 1.3 times than that obtained in control animals. The exocrine pancreatic function in 85 percent pancreatectomized rats treated with trypsin inhibitor for 4 weeks and 12 weeks showed substantial improvement as shown by the test with N-benzoyl-L-tyrosil-p-aminobenzoic acid (BT-PABA). The pancreatic amylase, lipase, and protein contents in the animals treated with trypsin inhibitor were increased to 1.9, 1.7 and 2.1-fold, respectively, as compared to control animals for 13 weeks. Histologic examination showed a decrease in abnormal islets of Langerhans, and a tendency toward hypertrophy of the acinar cells. These results suggest that oral administration of a trypsin inhibitor to rats is effective in improving pancreatic endocrine and exocrine functions after 85 percent pancreatectomy.     

NK1受体阻滞剂对电针内关穴抗大鼠AMI损伤效应的影响

目的：探讨NK1受体阻滞剂对电针大鼠内关穴抗AMI损伤效应的自主神经及中枢SP、NOS的影响。方法：静注NK1受体阻滞剂观察心交感神经、迷走神经放电频率变化;观察治疗前后下丘脑室旁核、延髓迷走神经复合区及脊髓外侧角SP、NOS灰度值的变化。结果：阻滞剂组与阻滞剂加电针组对AMI大鼠迷走神经和交感神经放电频率无显著性差异;阻滞剂组、阻滞剂加电针组与电针内关组对AMI大鼠迷走神经和交感神经放电频率有显著性差异（△P〈0.05）;阻滞剂组与阻滞剂加电针组SP在PVN、延髓迷走神经复合区及脊髓外侧角的阳性表达降低,NOS表达降低。结论：静注NK1受体阻滞剂阻断了电针内关对AMI大鼠迷走神经和交感神经放电的良性调节,阻断了SP在PVN、延髓迷走神经复合区及脊髓外侧角的阳性表达,引起NOS表达降低,SP在电针内关穴对AMI保护效应中起到关键作用。