High level production of soluble single chain antibodies in small-scale Escherichia coli cultures

We have investigated the effect of growth and induction conditions on the production of soluble single-chain Fv antibody fragments in Escherichia coli under the control of wt lac promoter. The scFv was directed into the periplasmic space by a pelB leader sequence. Addition of sucrose to the medium gave a 15–25-fold increase in the yield of soluble scFv-phOx (3.0 mg/l) for bacterial shake-tube cultures and an increase of 80–150-fold (16.5 mg/l) for shake-flask cultures. Using flask culture in the presence of 0.4 M sucrose, a significant amount of scFv was released into the medium. We found that the scFv could be made to accumulate in the periplasm or be secreted into the medium by simply changing the incubation conditions and the concentration of the inducer. The ratio between soluble antibody fragments and insoluble scFv aggregates proved to be dependent on the strength of the promoter. Lowering the incubation temperature below 20°C had no effect on the yield of soluble antibody fragments in the periplasm, but they were no longer secreted into the medium. An example of high level production in shake-flask cultures and one-step purification by immobilized metal affinity chromatography (IMAC) is described for a soluble scFv specific for the T cell surface antigen CD3. The biological activity of the purified anti-CD3 scFv was demonstrated by flow cytometry. This method should be especially useful for the functional screening of a large number of clones in small-scale cultures.     

No correlation between number of MRI-evident lesions in cerebrum and the soleus stretch reflex in multiple sclerosis patients

The aim of the study was to investigate if the stretch reflex of the soleus muscle was useful in quantifying upper motor neuron lesions. The soleus stretch reflex was recorded in 10 healthy subjects and 20 patients with active relapsing-remitting multiple sclerosis and correlated to the number of MRI lesions in cerebrum and clinical scores (expanded disability status scale and regional functional scoring system). The short latency stretch reflex was elicited by rotating the left ankle joint 4 degrees with a rise time in the interval of 40-640 ms. The amplitude of the stretch was larger in multiple sclerosis patients being 88.5 microV in patients and 12.8 microV in controls, P = 0.007. The sensitivity of the stretch reflex expressed as the slope of the best linear fit was increased in MS patients to 2.6 microVs/degree compared with 0.6 microVs/degree (0.1-2.2) in controls, P = 0.009. There was no correlation between amplitude of the stretch reflex and number of MRI lesions (r = -0.03). In conclusion, the soleus stretch reflex might be useful to quantify spasticity but is not useful in detecting dysfunction of upper motor neurons in MS.     

The Reactivity of Psychosis Rating Form (RPRF): background,development and psychometrics

The ICD-10 and DSM-IV classifications have both given low priority to “reactivity” to acute stress as a classificatory principle for functional psychoses. In Scandinavia, reactivity is still considered an important factor in the development of such psychoses. Reactivity is a complex concept, and its various components are historically examined. The Reactivity of Psychosis Rating Form (RPRF) was developed in order to operationalize reactivity. Seven of the 10 elements of RPRF can be rated reliably. Factor analysis of the RPRF yields three factors: stressor, onset and change, that also show high interrater reliability. Our results indicate that RPRF has both construct and discriminant validity. Further studies with the RPRF may elucidate the true status of reactivity in functional psychoses.     

Studying Weak Central Coherence at Low Levels: Children with Autism do not Succumb to Visual Illusions. A Research Note

While anecdotal reports of abnormal perceptual experiences in autism abound, there have been to date no experimental studies showing fundamental perceptual peculiarities. The present paper reports results from a first study of low-level visual integration in autism. Twenty-five subjects with autism, 21 normal 7- and 8-year-olds, and 26 children with learning difficulties were asked to make simple judgements about six well-known visual illusions. Two conditions were used, in an attempt to explore group differences; standard two-dimensional black and white line drawings, and the same figures augmented with raised coloured lines. The subjects with autism were less likely to succumb to the two-dimensional illusions than were the other groups, and were less aided by the three-dimensional disembedded' condition. These striking results are discussed with reference to the 'central coherence' account of autism.     

Organization of visual corticostriatal projections in the cat,with observations on visual projections to claustrum and amygdala

Electrophysiological mapping criteria were employed to identify visual areas 20a, 20b, 21a, 21b, PMLS, AMLS, ALLS, PLLS, DLS, VLS, and PS in the cat, and to guide placement of tracer deposits. Anterograde tracer methods were used to study the corticostriatal projections of these extrastriate visual areas. The experiments demonstrate that all 11 extrastriate areas send projections to two distinct regions within the striatum, an extensive longitudinal zone within the caudate nucleus, and a more compact region within the posterolateral putamen. Cortical visual projections to the putamen terminate in relatively compact sheets or slabs, and appear to overlap extensively, while those to the caudate nucleus are irregularly patchy and more widely dispersed. Retrograde tracer deposits into the visual recipient zone of the caudate nucleus reveal substantial convergence of other cortical inputs to this same domain. Aspects of visuotopic organization are preserved in the visual projections to both the putamen and the caudate nucleus, but unequivocal retinotopic organization could not be inferred from the available material. Ten of the eleven extrastriate visual area also project topographically onto the visual zone of the claustrum. Area PS does not appear to contribute to the corticoclaustral projections. Five of the extrastriate visual areas (ALLS, PLLS, DLS, VLS, PS) also send sparse projections to the amygdaloid complex. c 1993 Wiley-Liss, Inc.     

Visual scoring and laser Doppler perfusion imaging of skin irritancy induced by different nicotine patches

Background/aims: Local skin reactions are the most common reason for discontinuation of transdermal nicotine replacement therapy in smoking cessation programs. The aims of the present study were (1) to quantify the intensity of skin reactions induced by different types of nicotine patches and (2) to compare the clinical evaluation of skin erythema using visual scores with independently performed quantitative estimates of skin perfusion.
Methods: Thirty-three subjects were included in the study, each receiving 2 different types of nicotine patches (Nicotinell and Nicorette) and 1 type of placebo patch (Nicorette), placed ventrally on the upper arms according to a randomized protocol. Patches were removed after 24 h (Nicotinell) and 16 h (Nicorette), respectively, according to recommended application times. Visual scoring and laser Doppler perfusion imaging were performed 45 min after removal of patches, in a randomized order.
Results: Nicotinell patches induced the highest cumulative clinical score for skin irritancy. All 3 investigated patches gave rise to a slight but significant skin perfusion increase and individual visual scores and perfusion data correlated.
Conclusion The degree of skin irritancy and underlying perfusion increase induced by 1 daily maintenance dose of transdermal nicotine via a patch is low, but differs between patch types.     

Functional mri of human brain activation combining high spatial and temporal resolution by a cine flash technique

Functional mapping of human brain activation has been accomplished at high spatial and temporal resolution (voxel size 4.9 μl, temporal increment 100 ms). The approach was based on oxygenation-sensitive long-echo time FLASH MRI sequences synchronized to multiply repeated cycles of visual stimulation in a CINE acquisition mode. This high temporal resolution revealed that stimulus-related signal intensity changes in human visual cortex display an initial latency followed by increases extending over several seconds. Furthermore, the temporal characteristics of the complete CINE MRI signal time course depended on the absolute and relative durations of activation and control periods and, for example, caused an apparent absence of a poststimulation “undershoot” phenomenon. Complementing hyperoxygenation due to rapid hemodynamic adjustments, these results suggest signal intensity modulation by enhanced oxygen consumption and concomitant deoxygenation during prolonged and/or repetitive stimulation.     

左旋多巴对弱视眼视诱发电位影响的研究

目的:探讨左旋多巴治疗弱视的效果。方法:正常眼和弱视眼服用单次剂量左旋多巴前后进行图形视诱发电位(pattem visual evoked poten-tial,PVEP)检测。结果:正常眼服药后中空间频率PVEP的N_1P_1振幅和高空间频率PVEP的P_1N_2振幅增大,弱视眼服药后低空间频率PVEP的N_1波潜伏期和中空间频率PVEP的P_1波潜伏期缩短。结论:左旋多巴可改善弱视眼的视功能,可作为弱视的一种新的治疗方法。眼科学报1997;13:182—185。     

New view of the organization of the pulvinar nucleus in Tupaia as revealed by tectopulvinar and pulvinar-cortical projections

The projections of the superficial layers of the superior colliculus to the pulvinar nucleus in Tupaia were reexamined by injecting WGA-HRP into the tectum. The main result was finding two different patterns of terminations in the pulvinar nucleus: a zone remote from the lateral geniculate nucleus, which occupies the dorsomedial and caudal poles of the pulvinar nucleus, was almost entirely filled with terminals in every case irrespective of the location of the injection site; and a second division of the pulvinar nucleus, adjacent to the lateral geniculate nucleus, contained irregular patches--much more densely populated--and the distribution of patches varied from case to case. We call the first projection "diffuse" and the patchy projection "specific." Next we injected several divisions of the extrastriate visual cortex to find the cortical target of each pathway. The diffuse path terminates in the ventral temporal area (Tv). The specific path terminates in the dorsal temporal area (Td) and area 18. We speculated about the significance of the two pathways: the specific path may be responsible for the preservation of vision after removal of the striate cortex; the diffuse path may have an important place in the evolution of the visual areas of the temporal and occipital lobe. We argued that the target of the diffuse path is in a position to relate limbic and visual impulses and relay the product of such integration to the other visual areas, striate as well as extrastriate cortex.