Matricryptins and matrikines: biologically active fragments of the extracellular matrix

Numerous extracellular proteins and glycosaminoglycans (GAGs) undergo limited enzymatic cleavage resulting in the release of fragments exerting biological activities, which are usually different from those of the full‐length molecules. In this review, we define matrikines and matricryptins, which are bioactive fragments released from the extracellular matrix proteins, proteoglycans and GAGs and report their major biological activities. These fragments regulate a number of physiopathological processes including angiogenesis, cancer, fibrosis, inflammation, neurodegenerative diseases and wound healing. The challenges to translate these fragments from molecules biologically active in vitro and in experimental models to potential drugs are discussed in the last part of the review.     

Prediction of the length of the femur from its fragments in a Sri Lankan population

ABSTRACT

In many forensic circumstances, the femur is fragmented due to decomposition and mutilation, thus necessitating development and application of regression equations to estimate the length of the femur from its fragments. In this study, regression equations were derived for the prediction of the femoral length, utilizing 11 standard femoral measurements (maximum length of femur, epicondylar breadth, vertical head diameter, transverse head diameter, vertical neck diameter, transverse neck diameter, neck length, subtrochanteric anterior–posterior diameter, subtrochanteric transverse diameter, mid-shaft anterior–posterior diameter and mid-shaft transverse diameter) obtained from 86 femora belonging to a contemporary Sri Lankan population. Univariate regression equations with moderate to high correlations (0.226 to 0.714) were formulated for the estimation of the maximum femoral length from the measurements of fragments of the femur. Of the single variables, the transverse neck diameter emerged as the best predictor (r = 0.714, SEE = 18.6596) of the maximum length of the femur. The multiple regression equations using a combination of variables yielded higher correlations (0.774–0.857) with lower error estimates (14.18–16.96). The regression equations presented in this study can be used to estimate the length of the femur of unknown, mutilated and dismembered human skeletal remains with considerable accuracy.     

DIABETOGENIC ACTION OF HUMAN GROWTH HORMONE Synthesis and Activity of C-terminal Fragments

Three peptides corresponding to the C-terminal region of human growth hormone have been synthesized by the solid-phase method: HGH-(177–191), HGH-(178–191) and HGH-(179–191). The diabetogenic activities of these synthetic peptides are reported. The data indicate that extension of HGH-(179–191) at its NH₂-terminus is required for in vivo activity. The reduced and S-carbamido-methylated form of HGH-(177–191) was also active, indicating that the disulphide bond is possibly not a prerequisite for biological activity.     

Vipera ammodytes bites treated with antivenom ViperaTAb: a case series with pharmacokinetic evaluation

Context: In clinical practice it is difficult to differentiate between V. berus and V. ammodytes venomous bites. In the past this was not a concern, but due to the current shortage in Viperfav? and European viper venom antiserum availability, V. a. ammodytes venomous bites have recently been treated with ViperaTAb^®, which is a pharmaceutical formulation containing a monospecific ovine Fab fragments against the venom of V. berus.

Objective: To evaluate ViperaTAb^® in V. a. ammodytes envenomations.

Materials and methods: This is a prospective case series of three consecutive patients envenomed by V. a. ammodytes snakebite treated with ViperaTAb^®. V. ammodytes venom, neurotoxic ammodytoxins, and Fab fragment levels were determined in serum samples and a pharmacokinetic analysis of the antivenom Fab fragments was carried out.

Results: Three patients bitten by V. a. ammodytes with extensive local swelling, neurological symptoms and recurrent thrombocytopenia were treated with ViperaTAb^®. V. ammodytes venom was detected in serum of all three patients. Ammodytoxins were detected in the serum of only the most severely envenomed patient who developed neurological symptoms. In the presented moderate cases, a dose of 8?mL of ViperaTAb^® reduced swelling and improved systemic effects, such as thrombocytopenia. However, this dose of ViperaTAb^® was not effective in the most severely envenomed patient with the highest serum values of V. ammodytes venom. In this case ViperaTAb^® did not stop local swelling and it had no effect on neurological signs. ViperaTAb^®’s systemic clearance, distribution and elimination half-lives were 4.3–13.4?mL/h/kg, 1.2–3.2?h and 14.1–55.4?h, respectively.

Conclusions: In patients envenomed by V. a. ammodytes venom, ViperaTAb^® reduces moderate swelling and temporarily improves systemic effects, except neurological symptoms. ViperaTAb^® application induces a decrement of V. ammodytes venom level in the blood, but did not affect serum concentration of neurotoxic ammodytoxins in the one patient with measurable concentrations.     

不育男性精子-透明质酸结合率与精液参数的相关性分析

目的分析不育男性精子-透明质酸结合率与精液参数的相关性。方法选取2017年10月至2018年12月广东省中医院生殖医学科诊治的162例男性不育患者作为研究对象。对这162例男性不育患者的精液参数进行回顾性分析,采用Spearman相关性分析分别分析精液常规参数、精子功能及精浆指标与精子-HA结合率间的相关性。结果精子顶体酶活性与精子-HA结合率呈正相关(r=0.253,P=0.017);精子核蛋白不成熟度与精子-HA结合率呈负相关(r=-0.601,P=0.007);精浆锌浓度与精子-HA结合率呈正相关性(r=0.351,P=0.049)。而精液常规参数、MAR阳性率、精子DNA碎片率、精浆α-葡糖苷酶、柠檬酸、果糖、弹性蛋白酶浓度与精子-HA结合率均无明显相关性,差异均无统计学意义(均P>0.05)。结论精子-HA结合率是反映精子成熟度及受精能力的重要指标,与其他精子不成熟度及功能指标有一定的相关性。     

绿藻硫酸多糖SHW及其低分子量片段的结构及抗凝活性研究*

目的 对绿藻多糖SHW及其低分子量片段的结构及抗凝活性进行研究。方法 通过热水提取、强阴离子交换色谱和凝胶渗透色谱,从绿藻硬毛藻中提取分离得到硫酸多糖SHW;通过可控酸解方法制备SHW低分子量片段;采用高效凝胶渗透色谱、高效液相色谱、红外光谱及甲基化方法对多糖结构进行表征;通过活化部分凝血活酶时间、凝血酶时间、凝血酶原时间研究多糖SHW及其低分子量片段体外抗凝血活性。结果 多糖SHW及其低分子量片段主要由阿拉伯糖组成,含有少量半乳糖和氨基葡萄糖;糖链中阿拉伯糖主要以→4)-Arap-(1→和→3,4)Arap-(1→的形式存在,半乳糖以→4)-Galp-(1→存在形式,硫酸根主要位于→4)-Arap-(1→的C-3位。SHW及其低分子量片段A1～A6的分子量分别为492 kDa、200 kDa、170kDa、100kDa、26kDa、14kDa 和10 kDa。SHW 具有较高的抗凝血活性,且其抗凝活性随着其分子量的减少而降低。结论 海藻多糖SHW是一种具有抗凝活性的新颖硫酸多糖,其抗凝活性与分子量密切相关。     

Anti-neuropilin 1 antibody Fab′ fragment conjugated liposomal docetaxel for active targeting of tumours

Abstract

Neuropilin-1, a transmembrane receptor entailed in wide range of human tumour cell lines and diverse neoplasms, mediates the effects of VEGF and Semaphorins during the processes of cellular proliferation, survival and migration. In view of this, we had developed and evaluated in vitro and in vivo efficacy of anti-neuropilin-1 immunoliposomes against neuropilin-1 receptor expressing tumours. The PEGylated liposomes loaded with docetaxel were prepared using thin film hydration method. Functionalised PEGylated liposomes were prepared using post-insertion technique. Anti-neuropilin-1 immunoliposomes were prepared by covalently conjugating Fab′ fragments of neuropilin-1 antibody to functionalised PEGylated liposomes via thioether linkage. In vivo evaluation of Taxotere and liposomal formulations was performed using intradermal tumour model to demonstrate anti-angiogenic and tumour regression ability. The modified Fab′ fragments and immunoliposomes were found to be immunoreactive against A549 cells. Further, docetaxel loaded PEGylated liposomes and PEGylated immunoliposomes demonstrated higher in vitro cytotoxicity than Taxotere formulation at the same drug concentration and exposure time. The live imaging showed distinctive cellular uptake of functional immunoliposomes. Further, significant decrease in micro-blood vessel density and tumour volumes was observed using bio-engineered liposomes. The results clearly highlight the need to seek neuropilin-1 as one of the prime targets in developing an anti-angiogenic therapy.     

Study of different routes of immunization using outer membrane vesicles of Neisseria meningitidis B and comparison of two adjuvants

Outer membrane vesicles (OMVs) of Neisseria meningitidis contain important antigens to trigger an immune response against meningococci and have been studied as vaccines compounds. The immune response to a vaccine may be affected by its constitution and route of administration. Therefore, Swiss mice were immunized by different routes with OMVs of N. meningitidis B with dimethyl dioctadecyl ammonium bromide in bilayer fragments (DDA-BF) or aluminum hydroxide (AH) as adjuvants. The adjuvants and different routes were compared regarding the immune responses by ELISA, western blot, delayed type hypersensitivity (DTH) and histopathologic analysis. The antigenic preparation generated humoral and cellular immune responses. In quantitative analyzes, in general, AH was superior to DDA-BF. However, analysis such as IgG avidity index, bactericidal activity and immunoblot, revealed no important differences regarding the adjuvant or route of immunization. Regarding the parameters tested, it was not possible to define a superiority between the adjuvants and routes of immunization proposed by this study.     

小腿开放性骨折游离骨块回植术后口服骨伤复元汤的疗效研究

目的 探讨小腿开放性骨折游离骨块回植术后口服骨伤复元汤的临床疗效.方法 72例接受游离骨块回植术的小腿开放性骨折患者,以治疗方法不同分为常规组和实验组,各36例.常规组予以常规治疗,实验组在常规治疗基础上口服骨伤复元汤治疗.对比两组患者术后不同时间段患肢疼痛程度、骨愈合时间及术后骨痂生长情况.结果 术后第2、4天,两组...     

丙型肝炎病毒（HCV）高变区合成肽的抗原性及动物免疫观察

根据ＨＣＶ蛋白的可亲性、柔韧性和亲水性合成了覆盖ＨＣＶ蛋白高变区（Ｅ／ＮＳ１）的１３个多肽片段，应用建立的抗－ＨＣＶＥＬＩＳＡ方法检测ＨＣＶ感染者血清标本，发现ＨＣＶ高变区存在两个抗原决定簇位点，分别位于ＨＣＶ的蛋白氨基酸位点３８１～４２１、４８４～４８９处。应用ＨＣＶ高变区多肽片段ＳＰ９、ＳＰ（１０）和ＳＰ２以及核衣壳区（Ｃ）多肽ＳＰ（１４）免疫家兔得到抗－ＳＰ多克隆抗体。