Intraarterial delivery of adenovirus vectors and liposome-DNA complexes to experimental brain neoplasms

This study investigated the intraarterial delivery of genetically engineered replication-deficient adenovirus vectors (AVs) and cationic liposome-plasmid DNA complexes (lipoDNA) to experimental brain tumors. Adenovirus or lipoDNA was injected into the internal carotid artery (ICA) of F344 rats harboring intracerebral 9L gliosarcomas, using bradykinin (BK) to selectively permeabilize the blood-tumor barrier (BTB). Brain and internal organs of the animals were collected 48 hr after vector injection and stained for expression of the marker gene product, beta-galactosidase (beta-Gal). Intracarotid delivery of AV to 9L rat gliosarcoma without BTB disruption resulted in transgene expression in 3-10% of tumor cells distributed throughout the tumor. Virus-mediated expression of beta-gal gene products in this tumor model was particularly high in small foci (< or = 0.5 mm), which had invaded the normal brain tissue surrounding the main tumor mass. In these foci more than 50% of tumor cells were transduced. BK infusion increased the amount of transgene-expressing cells in larger tumor foci to 15-30%. In the brain parenchyma only a few endothelial cells expressed beta-gal owing to AV-mediated gene transfer. Intracarotid delivery of lipoDNA bearing a cytoplasmic expression cassette rendered more than 30% of the tumor cells positive for the marker gene without BTB disruption. The pattern of distribution was in general homogeneous throughout the tumor. BK infusion was able to increase further the number of transduced tumor cells to more than 50%. Although lipoDNA-mediated gene transfer showed increased efficacy as compared with AV-mediated gene transfer, it had less specificity since a larger number of endothelial and glial cells also expressed the transgene. AV and lipoDNA injections, in the absence and presence of BK, also resulted in transduction of peripheral organs. AV showed its known predilection for liver and lung. In the case of lipoDNA, parenchymal organs such as liver, lung, testes, lymphatic nodes, and especially spleen, were transduced. These findings indicate that intracarotid application of AV and lipoDNA vectors can effectively transduce tumor cells in the brain, and that BTB modulation by BK infusion can further increase the number of transgene-expressing tumor cells.     

Does Tamm-Horsfall mucoprotein inhibit or promote calcium oxalate crystallization in human urine?

Using two different experimental techniques, Tamm-Horsfall mucoprotein (THM) has been reported both to inhibit and to promote calcium oxalate (CaOx) crystallization in ultrafiltered human urine. In this study, these two techniques were used to compare the effects of THM on CaOx crystallization in the same ultrafiltered urine samples.

Urine was collected from 10 healthy men and ultrafiltered (10000 Da). Each sample was divided and to one half was added sufficient human THM to give a final concentration of 35 mg/L. CaOx crystallization was induced in the samples by addition of an oxalate load and by evaporation.

Using the evaporation technique THM significantly increased the deposition of CaOx determined as ¹⁴C-oxalate, from 9772 cpm to 43652 cpm (P < 0.01). Using the oxalate load method THM had no effect on the metastable limits of the urine with respect to CaOx, and significantly increased the volume of particulate material deposited from 26000 to 39995 μm³/μl - an increase of 54%. This increase was reduced to 21% when values were corrected for the volume of THM particles recorded in control samples to which no oxalate load was added. Using ¹⁴C-oxalate, it was shown that this increase in volume could not be attributed to an enhanced deposition of crystalline CaOx, but was probably the result of an increased polymerization of THM in the presence of CaOx crystals. Despite this, the average size of the particles precipitated in the presence of THM (6.5 μm) was significantly (P < 0.01) less than that observed in the absence of THM (12.1 μm).

It was concluded that the effect of THM on CaOx crystallization in urine depends upon the methodology used to assess it and that promotion would only be expected in vivo in cases of extreme dehydration. Under usual physiological conditions THM would be expected to inhibit CaOx crystal aggregation and to have little effect, if any, on the amount of crystalline material deposited.     

脑出血大鼠脑组织膜型基质金属蛋白酶、基质金属蛋白酶2和基质金属蛋白酶9表达及益气活血中药的干预效应

目的:观察益气活血中药对脑出血大鼠脑组织中膜型基质金属蛋白酶、基质金属蛋白酶2和基质金属蛋白酶9表达量的影响,从脑出血损伤区微血管系统重建的角度,探讨益气活血中药治疗脑出血的作用机制。方法:实验于2006-03/10在中南大学湘雅医院中西医结合研究所实验室完成。实验材料:补阳还五汤全方(黄芪,当归,赤芍,红花,川芎,地龙,桃仁按20∶3∶3∶3∶2∶3∶3的比例);补阳还五汤益气成分(黄芪按上述比例);补阳还五汤活血成分(当归,赤芍,地龙,川芎,桃仁,红花按3∶3∶3∶2∶3∶3的比例)。用蒸馏水两次水煎,分别浓缩为1.54,0.81和0.73g/mL。实验分组:155只SD大鼠随机分为正常对照组、假手术组、模型组、益气活血组、益气组、活血组。正常对照组5只大鼠,其余每组30只,再随机分为术后灌胃2,4,7,14,21,28d6个观察时间点,各个时间点5只大鼠。实验干预:造模:采用立体定位技术将胶原酶Ⅶ注入大鼠大脑右苍白球制成脑出血大鼠模型。假手术组大鼠仅注入2μL生理盐水,其余手术过程相同。给药:正常对照组:普通饲养,自由饮水;假手术组和模型组术后予蒸馏水灌胃2次/d,2mL/次;益气活血组、益气组、活血组分别给予补阳还五汤全方、补阳还五汤益气成分、补阳还五汤活血成分30.80,16.20,14.60g/(kg·d)(按体表面积计算为临床70kg成人剂量的3倍)灌胃,2次/d,2mL/次。各组大鼠分别于灌胃2,4,7,14,21,28d麻醉下取脑,制备切片;正常组动物于28d处死。实验评估:免疫组织化学染色方法检测各组灌胃不同时间脑组织基质金属蛋白酶2、基质金属蛋白酶9和膜型基质金属蛋白酶的阳性微血管数。结果:155只大鼠均进入结果分析。①正常组、假手术组皮质偶见膜型基质金属蛋白酶、基质金属蛋白酶2和基质金属蛋白酶9表达。②模型组膜型基质金属蛋白酶、基质金属蛋白酶2呈双峰表达,4d为最高峰,至14～21d再有小高峰出现。③益气活血组给药4d时,膜型基质金属蛋白酶、基质金属蛋白酶2为表达低谷,低于模型组(P<0.01)、益气组和活血组(P<0.05);在中后期,益气活血组膜型基质金属蛋白酶表达高峰为7～14d,较模型组提前出现,21d后与模型组比,各治疗组膜型基质金属蛋白酶已表达极少(P<0.01);益气活血组基质金属蛋白酶2在7d后一直呈高水平维持,高于其他各组(P<0.05),28d后开始逐渐下降。④模型组基质金属蛋白酶9在造模后4d达最高峰(P<0.01),两周后几乎无表达。益气活血组基质金属蛋白酶9高峰推迟至7d出现(P<0.05),之后逐渐下降。结论:益气活血中药可通过调节脑出血后膜型基质金属蛋白酶、基质金属蛋白酶2和基质金属蛋白酶9表达,改善出血后脑组织的微环境,有利于微血管系统重建,促进组织修复。     

MELANOMA OF THE FACE: THE SAFETY OF NARROW EXCISION MARGINS

Recent studies have shown that narrower excision margins may be safe, but the optimal or minimum margin for melanoma is unknown. Wide margins of excision are possible on the trunk and limbs, but functional and cosmetic constraints often limit the extent of excision on the face. A collaborative study from two continents (Cape Town, South Africa and Northwood, England) investigated the outcome of different excision margins of 106 patients with stage I melanoma of the face. The margin of excision was measured from the records of the pathological specimen. Thirty patients had margins of less than 1 cm, 64 had margins of between 1 and 2 cm, and 12 had margins greater than 2 cm. Primary apposition or flap closure was possible in 85 patients. Seven patients developed local recurrences and these were not influenced by the excision margin. This study supports the contention that the primary treatment of cutaneous melanoma on the face should be histologically confirmed complete excision, and that this can be achieved with margins of excision less than 1 cm. Local recurrence is not related to the margin of excision or to tumour thickness.     

AN ELECTROMYOGRAPHIC ANALYSIS OF THE SHOULDER COMPLEX MUSCULATURE WHILE PERFORMING EXERCISES USING THE BODYBLADE? CLASSIC AND BODYBLADE? PRO

Background

In spite of the bodyblade (BB®) being used in clinical settings during shoulder and trunk rehabilitation and training for 24 years, there are only five known scientific papers that have described muscle recruitment patterns using the BB®. Moreover, there are no known studies that have examined muscle activity differences between males and females (who both use the bodyblade in the clinic) or between different BB® devices.

Hypothesis/Purpose

The primary purposes of this investigation were to compare glenohumeral and scapular muscle activity between the Bodyblade® Pro (BB®P) and Bodyblade® Classic (BB®C) devices while performing a variety of exercises, as well as to compare muscle activity between males and females. It was hypothesized that glenohumeral and scapular muscle activity would be significantly greater in females compared to males, significantly greater while performing exercises with the BB®P compared to the BB®C, significantly different among various BB® exercises, and greater with two hand use compared to one hand use for the same exercise.

Study Design

Controlled laboratory study using a repeated-measures, counterbalanced design.

Methods

Twenty young adults, 10 males and 10 females, performed seven BB® exercises using the BB®C and BB®P, which are: 1) BB®1 - one hand, up and down motion, arm at side; 2) BB®2 - one hand, front to back motion, shoulder flexed 90 °; 3) BB®3 - one hand, up and down motion, shoulder abducted 90 °; 4) BB®4 - one hand, side to side motion, shoulder and elbow flexed 45 °; 5) BB®5 - two hands, side to side motion, shoulders and elbows flexed 45 °; 6) BB®6 - two hands, up and down motion, shoulders flexed 90 °; and 7) BB®7 - two hands, front to back motion, shoulders flexed 90 °. EMG data were collected from anterior and posterior deltoids, sternal pectoralis major, latissimus dorsi, infraspinatus, upper and lower trapezius, and serratus anterior during 10 sec of continuous motion for each exercise, and then normalized using maximum voluntary isometric contractions (MVIC). A two-factor repeated measures Analysis of Variance (p < 0.05) was employed to assess differences in EMG activity between BB® devices (BB®C and BB®P) and genders.

Results

As hypothesized, for numerous exercises and muscles glenohumeral and scapular EMG activity was significantly greater in females compared to males and was significantly greater in the BB®P compared to BB®C. There were generally no significant interactions between BB® devices and gender. Overall glenohumeral and scapular muscle activity was significantly greater in BB®3 and BB®6 compared to the remaining exercises, but generally not significantly different between using one hand and using two hands.

Conclusions

It may be appropriate to employ BB® exercises during shoulder rehabilitation earlier for males compared to females and earlier for the BB®C compared to the BB®P given less overall muscle activation in males and BB®C compared to in females and BB®P. There was generally no difference in muscle activity between performing the BB® with one-hand or two-hands. Differences in muscle activity between exercises generally was the similar regardless if the BB®C or the BB®P was employed.

Level of Evidence

Level 2     

The red cell mass-arterial oxygen relationship in normal man: Application to patients with chronic obstructive airway disease

The normal relationship between red cell mass measured, with (51)chromium-labeled red cells, and arterial oxygen saturation (Sa(O2)) over the range from 97.3 to 83.4% was examined by studying 73 normal men residing at sea level and altitudes of 1600 and 3100 m. A simple, linear relationship between Sa(O2) and red cell mass was found over the entire range (r = - 0.7524, P < 0.001). In contrast, a correlation between red cell mass and arterial O(2) tension was found only over the lower half of the range of O(2) tensions where Sa(O2) was also decreased (r = - 0.7731, P < 0.005). This suggested that O(2) saturation rather than tension is the more important determinant of the erythropoietic response to chronic hypoxia. If this response is regulated by tissue O(2) tension, then it will be influenced by O(2) transport, which, in turn, is a function of blood flow and arterial O(2) content, and hence Sa(O2). In nine patients with chronic obstructive airway disease the relationship between red cell mass and Sa(O2) was also determined and was found to be steeper than in the normal subjects (P < 0.05).     

N-Acetylcysteine Rinse for Thick Secretion and Mucositis of Head and Neck Chemoradiotherapy (Alliance MC13C2): A Double-Blind Randomized Clinical Trial

ObjectiveTo determine whether N-acetylcysteine rinse was safe and could improve thickened secretions and dry mouth during and after radiotherapy.Patients and MethodsWe designed a prospective pilot double-blind, placebo-controlled randomized clinical trial (Alliance MC13C2). Adult patients (age ≥18 years) were enrolled if they underwent chemoradiotherapy (≥60 Gy). Patients initiated testing rinse within 3 days of starting radiotherapy. With swish-and-spit, they received 10% N-acetylcysteine (2500 mg daily) or placebo rinse solution 5 times daily during radiotherapy and 2 weeks postradiotherapy. The primary aim was to evaluate N-acetylcysteine in improvement of saliva viscosity with the Groningen Radiotherapy-Induced Xerostomia questionnaire. Secondary aims included evaluating xerostomia improvement by the same questionnaire and with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire–Head and Neck-35 Questions survey and adverse-event profiles. The type I error rate was 20%.ResultsThirty-two patients undergoing chemoradiotherapy were enrolled. Baseline characteristics were balanced for placebo (n=17) and N-acetylcysteine (n=15). N-acetylcysteine was better for improving sticky saliva (area under curve, P=.12). Scores of multiple secondary end points favored N-acetylcysteine, including sticky saliva daytime (P=.04), daytime and total xerostomia (both P=.02), pain (P=.18), and trouble with social eating (P=.15). Repeated measures models confirmed the findings. Taste was a major dissatisifer for N-acetylcysteine rinse; however, both testing rinses were safe and well tolerated overall.ConclusionOur pilot data showed that N-acetylcysteine rinse was safe and provided strong evidence of potential efficacy for improving thickened saliva and xerostomia by patient-reported outcome. A confirmatory phase 3 trial is required.Trial Registrationclinicaltrials.gov Identifier: NCT02123511