Analysis of odontoblast gene expression using a novel approach,laser capture microdissection

Studying the mechanisms of molecular interactions in developing tissues demands sensitive molecular biological in vivo and in vitro techniques. Laser capture microdissection (LCM) allows for the isolation of mRNA in histological sections even from single cells, thus enabling the identification of in vivo gene expression products in closely circumscribed tissue areas. The aims of this study were to assess the optimal fixation, processing, and staining conditions to retrieve RNA from microdissected odontoblasts. Fluorometric assays and RT-PCR analysis of alpha 1(I) collagen, dentin sialophosphoprotein (Dspp), and osteocalcin (OC) confirmed that the total RNA isolated from day 0 and day 3 captured odontoblasts was sufficient in quantity and quality. Our results indicate that individual odontoblasts obtained by LCM are morphologically intact and chemically unaltered, allowing accurate molecular and biochemical analyses.     

Prognostic factors affecting survival in multiple myeloma

70 patients suffering from multiple myeloma were observed by authors in the last 15 years and three months. In the meantime fifty-two out of them have died, and 18 patients are under permanent care. 43 IgG, 17 IgA, 6 Bence-Jones, 2 IgD types were diagnosed according to the paraprotein distribution, one patient proved to be nonsecretory, and an other one to osteosclerotic form as well. The median survival time was 27 months in the group of deceased patients. In the group followed-up 50.8 months survival time was observed up to the closing of the study. Several prognostic factors were investigated. According to the classification by Durie and Salmon the survival time was 60 months in the patients with stage I, 33 months in stage II., and 9 months in stage III respectively. The prognosis is much poorer in patients into the "B" category: the survival time was 14 months. Classified in the basis of the type of the myeloma-cell, the cases with well matured cells have had the best prognosis with survival time of 46 months, while the most unfavourable prognosis was observed in patients with blast-cell type, with a median survival time of 10 months. The greatest number of patients suffered from multiple myeloma of IgG paraprotein type, in this group the serum IgA level was found to be significantly decreased in the patients died due to inevitable infections. The survival was injured significantly by the occurrence of concomitant severe diseases, to.(ABSTRACT TRUNCATED AT 250 WORDS)     

Animal models for the optokinetic system of the human]

BACKGROUND: We compare the neuronal and behavioural consequences of abnormal visual experience during early infancy for the optokinetic system of cat and monkey with the neuroopthalmological results in man. MATERIAL AND METHODS: Optokinetic eye movements were recorded with the search coil method and electrooculography. In addition, the response properties of single neurons in the visual cortex and pretectum of anesthetized and paralyzed cats and monkeys were determined in electrophysiological experiments. RESULTS: Our data show that monocular deprivation and strabismus lead to an increase of asymmetry of monocular optokinetic nystagmus and a decrease in gain of slow phase optokinetic eye movements during monocular stimulation as well as to a significant decrease of binocular convergence in the oculomotor system of both cat and monkey. Although these changes are basically similar in both species they are not identical. These differential effects may be explained by structural differences. CONCLUSIONS: Assuming a similar neuronal substrate for the optokinetic system in all primates the monkey represents a more suitable animal model for the human visual system than the cat.     

Die lokalisierte Form einer sympathischen Reflexdystrophie

Introduction

The reflex sympathetic dystrophy (RSD) syndrome usually shows a distally generalized distribution pattern of symptoms. Here we report a case with a distally localized form of RSD.

Patient and methods

In a 53-year-old woman, following a local lesion in the nail bed of the left thumb, a neuroma developed at the side of the lesion during the next half year. She was finally operated upon. Following that intervention, a complex and painful clinical syndrome occurred that for the most part affected only the thumb. A clinical neurological examination was carried out, including distal suprasystolic compression of the affected extremity after bandaging it (the so-called ischemia test). For diagnostic and therapeutic reasons, afterwards a conventional blockade of the ipsilateral stellate ganglion was applied.

Results

The clinical investigation showed a triad of autonomic (swelling, side difference of skin temperature, hyperhydrosis), motor (reduced movement ability, tremor) and sensory disturbances (spontaneous pain, allodynia), which nearly exclusively affected the entire left thumb. The spontaneous pain showed an orthostatic component (the pain being diminished or exaggerated when the extremity was elevated or lowered, respectively) and was suppressed by the ischemia test. Following the sympathetic block, all symptoms disappeared within one day (follow-up period: 5 months).

Conclusion

In contrast to the common clinical picture of RSD, with a distally generalized distribution of symptoms, the present case showed a so-called localized form of RSD, its triad affecting only the thumb with the lesion. Typically, the pain showed an orthostatic component and was suppressed by the ischemia test. The sympathetic block was immediately successful, proving the occurrence of this form of RSD for the first time. In similar clinical cases, this form of RSD should be considered as a differential diagnosis.     

Ventral root avulsion versus transection at the cervical 7 level of the cat spinal cord

The results of unilateral ventral root avulsions at the cervical 7 (C7) level were compared using light microscopy with unilateral transactions of the ventral root C7 at the surface of the cat spinal cord after survival times ranging from one to 90 days. Sections were stained for Nissl, acetylcholinesterase, neurofilaments, glial fibrillary acidic protein, and B-50/GA P43. A distinct difference in neuronal reaction was found between the effects of avulsion and transection of ventral root C7. After avulsion, changes in the Nissl-and acetylcholinesterase-stained sections and the sections stained with antibodies against neurofilaments remained present up to 90 days, although the number of neurons in the ventral horn on the avulsed side had significantly decreased at 60 and 90 days. This was in contrast to the transection where the changes in the stainings tended to return to the normal situation between 30 and 60 days and showed the normal pattern 90 days after transection. No decrease in the number of neurons was found. Both lesions were situated in the transitional region of the nerve roots. After avulsion axons break in the white matter of the CNS, while the axons were transected in the PNS compartment of the CNS-PNS transitional region. After avulsion, not only were the axons disconnected, as in transection, but the connections of recurrent motor axon collaterals probably were damaged. This may explain the different reaction after avulsion and transection.     

Depth perception and cortical physiology in normal and innate microstrabismic cats.

Evidence is presented that innate microstrabismus and abnormal cortical visual receptive-field properties can occur also in cats without any apparent involvement of the Siamese or albino genetic abnormalities in their visual system. A possible cause for microstrabismus in these cats may be sought in an abnormally large horizontal distance between blind spot and area centralis indicated by a temporal displacement of the most central receptive fields on both retinae. Depth perception was found to be impaired in cats with innate microstrabismus. Behavioral measurements using a Y-maze revealed in four such cats that the performance in recognizing the nearer of two random-dot patterns did not improve when they were allowed to use both eyes instead of only one. The ability of microstrabismic cats to perceive depth under binocular viewing conditions only corresponded to the monocular performance of five normal cats. Electrophysiological recordings were performed in the visual cortex (areas 17 and 18) of four awake cats, two normal, and two innate microstrabismic animals. Ocular dominance and orientation tuning of single neurons in area 17 and 18 were analyzed quantitatively. The percentage of neurons in area 17 and 18 which could be activated through either eye was significantly reduced to 49.7% in the microstrabismic animals when compared to the normal cats (74.8%). "True binocular cells," which can only be activated by simultaneous stimulation of both eyes, were significantly less frequent (1.6%) in microstrabismic cats than in normal animals (10.4%). However, subthreshold binocular interactions were identical in both groups of animals. In the strabismic animals, long-term binocular stimulation of monocular neurons did not give a clear indication of alternating use of one or the other eye. The range of stimulus orientations leading to discharge rates above 50% of the maximal response, i.e. the half-width of the orientation tuning curves, was the same in the two groups of cats. However, orientation sensitivity, i.e. the alternation in discharge rate per degree change in stimulus orientation, was higher in cortical cells of normal cats than in those of microstrabismic cats. In normal and microstrabismic cats, no clear sign of an "oblique effect," i.e. the preference of cortical neurons for vertical and horizontal orientations compared to oblique orientations, could be found neither in the incidence of cells with horizontal or vertical preferred orientation nor in the sharpness of orientation tuning and sensitivity of these neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Cat Distal Forelimb Joints and Locomotion: An X-ray Study

The complex construction of the joint apparatus of the cat distal forelimb, which allows the paw three degrees of freedom, poses special requirements on the neural signals controlling the paw position. To understand the electromyography (emg) signals of the distal forelimb muscles during locomotion, it is necessary to know the kinematics of the forelimb joints in detail. As no such information is available, we used the pulsed X-ray technique in trained cats during treadmill locomotion to analyse the angular excursions of the wrist, the metacarpophalangeal (MCP) and the proximal interphalangeal (PIP) joints. X-ray illuminations were done in either the parasagittal or the frontal plane. At the beginning of the stance phase the wrist (WR) and the MCP joints extended slowly, and the PIP joints flexed. Whereas the WR and the PIP joints maintained a constant angular position of approximately 200 degrees and 60 degrees, respectively, throughout the stance phase, extension continued in the MCP joints from 240 degrees at touch-down to 300 degrees at the end of the stance phase. Slightly before lift-off (100 ms) the WR and the MCP joints flexed rapidly. This flexion changed approximately 150 ms after lift-off into a slow extension. The PIP joints extended rapidly at the beginning and at the end of the swing phase, during the interposed period of the swing phase they displayed a slow flexion. Rotatory movements of the forelimb in the radioulnar joints were present during the swing and stance phases. During the swing phase the limb first supinated (starting 100 ms after lift-off); pronation occurred immediately before ground contact. During the stance phase the supination angle was kept constant until 100 ms before lift-off, when a short pronation was found. The paw was kept in an ulnar deviated position throughout the complete step cycle. Ulnar deviation decreased at the end of the swing and stance phases. The results of this study increase our understanding of how the body weight is transmitted on to the ground. They suggest four main functions for the skeletomotor apparatus and the underlying neural commands to secure the forward movement of the animal during the stance phase: (i) preparation and stabilization of a force-transmitting platform; (ii) stabilization of the wrist and the carpal/metacarpal joints; (iii) stabilization of the supination angle; (iv) antigravity control of the extension in the MCP.